TW201209402A - Apparatus for loading oligonucleotide spotting devices and spotting oligonucleotide probes - Google Patents

Abstract

An apparatus for loading oligonucleotide spotting devices and spotting oligonucleotide probes, the apparatus having a plurality of oligonucleotide vials, each with a droplet dispenser, a mounting surface for detachably mounting an oligonucleotide spotting device, a chuck for detachably mounting the oligonucleotide spotting device adjacent the mounting surface, and, a control processor for operative control of the oligonucleotide vials, the oligonucleotide spotting device when mounted in the chuck and movement of the mounting surface relative to the oligonucleotide vials, and the oligonucleotide spotting device, wherein, the control processor is configured to activate the droplet dispensers, and move the oligonucleotide spotting device into registration with the oligonucleotide vials.

Description

201209402 VI. Description of the Invention: TECHNICAL FIELD OF THE INVENTION The present invention relates to a diagnostic apparatus using microsystem technology (MST). In particular, the present invention relates to the treatment and analysis of microfluidics and biochemicals for molecular diagnostics. [Prior Art]

Molecular diagnostics have been used to provide an early indication of disease detection before the onset of symptoms. Molecular diagnostic tests are used to detect: • genetic disorders • acquired diseases • infectious diseases • genes associated with health-related genetic factors due to high accuracy and rapid processing time, molecular diagnostic tests can reduce the incidence of ineffective health care, Improve patient outcomes, improve disease management, and individualized patient care. Many techniques for molecular diagnostics are based on the detection and identification of specific nucleic acids (both deoxyribonucleic acid (DNA) and ribonucleic acid (RNA)) extracted and amplified from biological samples such as blood or saliva. The complementary nature of the nucleobase allows the short sequence (oligonucleotide) of the synthesized DNA to bind (hybridize) to a particular nucleic acid sequence for use in nucleic acid assays. If hybridization occurs, the complementary sequence is present in the sample. This makes it possible, for example, to predict the disease that an individual will get in the future, to determine the type and pathogenicity of the infectious pathogen, or to determine the individual's response to the drug. 201209402 Nucleic Acid-Based Molecular Diagnostic Tests There are four nucleic acid-based tests. Independent steps: 1. Sample preparation 2. Nucleic acid extraction 3. Nucleic acid amplification (optional) 4- Detection

Many sample types, such as blood, urine, sputum, and tissue samples, are used for genetic analysis. Diagnostic tests determine the type of sample required' because not all samples represent disease progression. These samples have various components' but usually only one of them is of interest. For example, in the blood, high concentrations of red blood cells can inhibit the detection of pathogenic microorganisms. Therefore, purification and/or concentration steps are typically required at the beginning of the nucleic acid assay.

Blood is one of the most frequently requested sample types. It has three main components: white blood cells, red blood cells, and platelets. Platelets accelerate coagulation and maintain activity in vitro. In order to inhibit coagulation, the sample is mixed with a reagent such as ethylenediaminetetraacetic acid (EDTA) prior to purification and concentration. Red blood cells are typically removed from the sample to concentrate the target cells. In the human body, red blood cells account for about 99% of cellular material, but they do not carry DNA because they do not have a nucleus. In addition, red blood cells contain components such as heme that may interfere with downstream nucleic acid amplification procedures (described below). Red blood cell removal can be accomplished by dissolving the solution differentially red blood cells, leaving the entire remaining cellular material, which can then be separated from the sample by centrifugation. This provides a concentrated target cell from which the nucleic acid is extracted. The exact procedure used to extract nucleic acids depends on the sample and the diagnosis to be performed -6- 201209402 analysis. For example, the protocol used to extract viral RNA is quite different from the protocol used to extract the genome DN A. However, extracting nucleic acids from a target cell typically involves a cell lysis step and subsequent nucleic acid purification. The cell lysis step ruptures the cell and nuclear membrane, releasing the genetic material. This is often done using a lysing detergent such as sodium lauryl sulfate, which also denatures the large amount of protein present in the cell. .

The nucleic acid is then purified by an alcohol precipitation step, which is usually carried out using ice ethanol or isopropanol, or via a solid phase purification step, which is usually carried out in a column of cerium oxide substrate, resin or high. The concentration is carried out on paramagnetic beads in the presence of a chaotropic salt, followed by washing of the nucleic acid, followed by elution with a buffer of low ionic strength. A selective step prior to precipitation of the nucleic acid is the addition of a protease which digests the protein to further purify the sample. Other lysis methods include mechanical lysis via ultrasonic vibration and heating of the sample to 94 °C to disrupt thermal lysis of the cell membrane. Target DNA or RNA can be present in the extracted material in very small amounts, especially if the target is from a pathogenic source. Nucleic acid amplification provides the ability to selectively amplify (i.e., replicate) a particular target in a low concentration to a detectable amount. The most commonly used nucleic acid amplification technique is the polymerase chain reaction (PCR). PCR is well known in the art, and a complete description of this type of reaction is provided in E. van Pelt-Verkuil et al. Principles and Technical Aspects of PCR Amplification, Springer, 2008. PCR is a useful technique for amplifying the 201209402 target DNA sequence in the context of complex DNA. If RNA is to be amplified (by PCR), it is first necessary to transcribe RNA into cDNA (complementary DNA) using an enzyme called reverse transcriptase. Subsequently, the obtained cDNA was amplified by PCR. PCR is an exponential method and can be continued as long as the conditions for maintaining the reaction are acceptable. The components of the PCR reaction are:

1. A short single-stranded DNA having a nucleotide complementary to a region of about 1 to 30 complementary to the Btt-flanking sequence.

2. DNA polymerase-synthesis of DNA thermostable enzymes 3. Deoxyribonucleoside triphosphates (dNTPs) - provide nucleotides that are incorporated into newly synthesized DNA strands. 4. Buffer - an ideal chemistry for DNA synthesis surroundings.

PCR typically involves placing these reactants in a vial containing about the extracted nucleic acid (about 10 to 50 microliters). The tube is placed in a thermal cycler; this reactor is an instrument that allows the reaction to take unequal time at a range of different temperatures. The standard procedures for each thermal cycle involve the variable phase, the annealed phase, and the extended phase. The extension phase is sometimes referred to as the primer extension phase. In addition to these three-step procedures, a two-step thermal protocol can also be used in which the annealing and extension phases are combined. The denatured phase generally involves raising the reaction temperature to 90 to 95 ° C to denature the DNA strand; in the annealed phase, the temperature is lowered to 50 to 60 ° C to bond the primer; then in the extended phase, the temperature is raised to The optimal DNA polymerase activity temperature is 60 to 72 ° C for extension of the primer. This process is repeated about 20 to 40 times, with the end result of generating millions of copies of the target sequence between the primers. Variants of many standard PCR protocols for molecular diagnostics have been developed, -8 - 201209402 which include, for example, multiple primer set PCR, linker-primed PCR, direct PCR, tandem PCR, real-time PCR And reverse transcriptase PCR.

Multiple primer set PCR uses multiple primer sets in a single PCR mix to generate different sizes of amplicons specific for different DNA sequences. Additional information can be obtained from a single experiment by one target (targeting) multiple genes (in other ways, several trials are required). The optimization of multi-initiator PCR is more difficult because it requires the selection of primers with approximate binding temperatures and amplicon with approximate length and base composition to ensure equal amplification efficiency of each amplicon. Linker-primed PCR, also known as ligation adaptor (PCR), is a non-target-specific primer that can extend the nucleic acid of virtually all DNA sequences in a complex DNA mixture. Addition method. This method first digests the target DNA population with a suitable restriction endonuclease (enzyme). Then, using a ligase, a double-stranded oligonucleotide linker with a suitable overhang (also known as a junction) Binding to the end of the target DNA fragment. Next, nucleic acid amplification is performed using an oligonucleotide primer specific for the linker sequence. Thus, all DNA-derived fragments adjacent to the linker oligonucleotide can be Amplification Direct PCR describes a system that performs PCR directly on a sample without any (or minimal) nucleic acid extraction. Many components present in unpurified biological samples, such as protohemoglobin in blood, have long been considered It will inhibit the PCR reaction. Therefore, it is customary to enhance the purification of the target nucleic acid before preparing the PCR reaction mixture. However, the chemical properties and the appropriate variation of the sample concentration -9-201209402 degrees are utilized. It is possible to perform minimal DNA purification or PCR by direct PCR. The PCR chemistry for direct PCR involves the introduction of buffer strength, the use of reactive and processivity polymerases and potential polymerization. An enzyme inhibitor chelated additive.

Repeated sequence PCR utilizes two separate nucleic acid amplification cycles to increase the probability of amplifying the correct amplicon. One type of repeat PCR is nested PCR in which two pairs of PCR primers are used to amplify a single locus in a different nucleic acid amplification cycle. The first pair of primers hybridize to a nucleic acid sequence located in a region other than the target nucleic acid sequence. A second pair of primers (nested primers) is used for the second amplification, the pair of primers being incorporated within the first PCR product to produce a second PCR product comprising the target nucleic acid, and the second product is short. The rationale used in this strategy is that if the wrong locus is amplified due to a mistake during the first nucleic acid amplification, the probability that the wrong locus is re-amplified by the second pair of primers is very low, thus ensuring specificity.

Real-time PCR or quantitative PCR is used to measure the amount of PCR product in real time. The initial amount of nucleic acid in the sample can be determined by using a probe containing a fluorophore or a fluorescent dye and a set of standards in the reaction. This is especially useful for molecular diagnostics where the choice of treatment may vary depending on the amount of pathogen in the sample. Reverse transcriptase PCR (RT-PCR) is used to amplify DNA from RNA. The reverse transcriptase is an enzyme that reverse transcribes RNA into complementary DNA (cDNA). This cDNA is then amplified by PCR. RT-PCR is widely used in expression pro filing to determine the expression of a gene or to identify sequences of an RNA transcript, including transcription initiation and termination sites. It is also used to amplify -10- 201209402 RNA viruses, such as human immunodeficiency virus or hepatitis C virus. The isothermal amplification is another form of nucleic acid amplification that does not rely on the thermal denaturation of the target DNA during the amplification reaction, thus eliminating the need for sophisticated instruments. Therefore, the thermostatic nucleic acid amplification method can be carried out in a field place or simply in an environment other than the laboratory. Some thermostatic nucleic acid amplification methods have been described, including Strand Displacement Amplification, Transcription Mediated Amplification,

Nucleic Acid Sequence Based Amplification, Recombinase Polymerase Amplification, Rolling Circle Amplification, Ramification Amplification, Helicase Activity Dependence Helicase-Dependent Isothermal DNA Amplification and Loop-Mediated Isothermal Amplification Constant-temperature nucleic acid amplification does not rely on continuous heating to denature template DNA to produce a single molecule that acts as a template for continued amplification. It is a method of producing a single-stranded molecule by other methods at a constant temperature, such as enzymatic cleavage of a DNA molecule by specific restriction of an internal nuclease, or separation of a DNA double strand by an enzyme. Strand-substituted amplification (SDA) relies on the ability of specific restriction enzymes to cleave unmodified strands of hemi-m〇dified DNA, and polymerase extensions that lack 5'-3' exonuclease activity and replace downstream stocks Ability. Exponential nucleic acid amplification is then achieved by coupling sense and antisense, wherein the strand from the sense reaction replaces the template for the antisense reaction. The nicking enzyme used in this reaction does not cleave the DNA in a conventional manner, but instead produces an incision in one of the -11 - 201209402 DNA strands, such as N. Alwl, Ν·BstNBl, and Mlyl. SDA is improved by using a combination of a thermostable restriction enzyme (Aval) and a thermostable outer polymerization enzyme (B st polymerase). This combination has been shown to increase the amplification efficiency of the reaction from 1-8 fold amplification to 101 fold amplification, so it is possible to utilize this technique to amplify unique single copy molecules.

Transcription-mediated amplification (TMA) and nucleic acid sequence-based amplification (N A SB A) RNA polymerase is used to replicate RNA sequences rather than corresponding genomic DNA. This technique uses two primers and two or three enzymes, RNA polymerase, reverse transcriptase, and selective RNase® (if reverse transcriptase does not have RNase activity). One of the primers contains a promoter sequence of RNA polymerase. In the first step of nucleic acid amplification, the primer hybridizes to the target ribosomal RNA (rRNA) at a defined site. The reverse transcriptase then extends from the promoter 3's end to generate a DNA copy of the target rRNA. The RNA in the formed RNA:DNA dimer is decomposed by the RNase activity (if any) of the reverse transcriptase or additional RNase. In the next step, the second primer binds to the DNA copy. The new DNA strand is synthesized by reverse transcriptase from the end of this primer to produce a double-stranded DNA molecule. RNA polymerase recognizes the promoter sequence in the DNA template and initiates transcription. Each newly synthesized RNA expander is re-entered as a template for a new replication cycle. In recombinase polymerase amplification (RPA), isothermal amplification of a specific DNA fragment is achieved by binding an oppositely directed oligonucleotide primer to the template DNA and then extending the primers by a DNA polymerase. . Denaturation of the double-stranded DNA (dsDNA) template does not require heating. Instead, RPA uses a recombinase-primer complex to scan dsDNA to facilitate stock exchange at the homologous -12-201209402 locus. The resulting structure is stabilized by the interaction of a single strand of DNA binding protein with the substituted template strand, thereby preventing the primer from exiting via branch migration. Recombinase unwinding allows the strand-substituted DNA polymerase (such as a large fragment of B. subtilis p〇i i (Bsu)) to be accessible to the 3' end of the oligonucleotide, followed by primer extension. Exponential nucleic acid amplification is accomplished by repeated cycles of this process.

The helicase-dependent amplification (HDA) mimics an in vivo system in which DNA helicase is used to generate a single-strand template for primer hybridization, followed by extension of the primer by DNA polymerase. In the first step of the HD A reaction, the de-enzyme is passed along the target DNA to interrupt the hydrogen bond between the two strands, which in turn binds to the single-stranded binding protein. The primers are bonded by exposing the single-strand target region by helicase. The DNA polymerase then uses the free deoxyribonucleoside triphosphate (dNTP) to extend the 3' end of each primer to create two DNA replication strands. The two dsDNA replication strands each enter the next HDA cycle, resulting in amplification of the exponential nucleic acid of the target sequence. Other DNA-based thermostating techniques include rolling circle amplification (RCA), in which a DNA polymerase extends the primer around a circular DNA template to produce a long DNA product consisting of many repeating copies of the loop. Prior to the end of the reaction, the polymerase produced tens of thousands of copies of the circular template and the copies of the strands were tethered to the original target DNA. This approach allows for spatial dissociation of the target and rapid nucleic acid amplification of the signal. A copy of 1 to 12 templates can be generated in one hour. A variant of the branched-type amplification system RCA that uses a closed circular probe (C-probe) or a padlock probe and a DNA polymerase with high processivity to exponentially expand under constant temperature conditions - 13- 201209402 The C-probe.

Circular thermostat amplification (LAMP) provides high selectivity using DNA polymerase and a set of four specially designed primers that recognize a total of six different sequences on the target DN A. The primers within the sense and antisense strand sequences containing the target DNA initiate LAMP. The strands initiated by the exogenous primers then replace the DNA synthesis to release a single strand of DNA. The single-stranded DNA can serve as a template for DNA synthesis initiated by a second primer and an external primer, and the second primer and the foreign primer are hybridized to the other end of the target, and the DNA is synthesized to produce stem-loop DNA. structure. In the subsequent LAMP cycle, an internal primer hybridizes to the loop on the product and initiates the replacement DNA synthesis, producing the original stem-loop DNA and the new stem-loop DNA with twice as long stems. The cycle reaction continued to accumulate 1 〇 9 copies of the target within one hour. The final product is a stem-loop DNA having a plurality of inverted repeats of the target and a cauliflower-like structure, wherein the plurality of loops in the cauliflower-like structure are formed by adhering mutually overlapping target repeats in the same strand. .

After completion of nucleic acid amplification, the amplification product must be analyzed to determine whether or not the expected amplicon (amplification amount of the target nucleic acid) is produced. The method of analyzing the product can be carried out by simply measuring the size of the amplicon by gel electrophoresis to hybridization with DN A to identify the nucleotide composition of the amplicon. Gel electrophoresis is the easiest way to check whether a nucleic acid amplification method produces the desired amplicon. Gel electrophoresis utilizes an electric field applied to a gel matrix to separate DNA fragments. Negatively charged DNA fragments will move in the matrix at different rates, depending on the fragment size. After electrophoresis is completed, the fragments in the gel are stained for visualization. Ethyl bromide is a commonly used dyeing agent, and its -14-201209402 exhibits fluorescence under ultraviolet light. The size of the fragments is determined by comparison with DNA ladders containing DNA fragments of known size and migrating side by side with the amplicons on the gel. Since the oligonucleotide primer binds to a specific site adjacent to the target DNA, the size of the amplification product can be predicted and detected as a band of known size on the gel. In order to determine the correctness of the amplicon, or if several amplicons are generated, DNA hybridization with the amplicon is usually used.

needle. DNA hybridization refers to the formation of double stranded DNA by complementary base pairing. DNA hybridization of about 20 nucleotides in length is required for DNA hybridization to clearly identify a particular amplification product. If the probe has a sequence complementary to the amplicon (target) DNA sequence, hybridization will occur at the appropriate temperature, pH and ion concentration. If hybridization occurs, the gene or DNA sequence of interest is present in the original sample. Optical detection systems are most commonly used to detect hybridization methods. One of the amplicon and probe is labeled with a fluorescent agent or an electrochemiluminescent agent to emit light. These methods differ in that they cause the photo-generating group to produce an excited state, but both can be used to covalently label nucleotide strands. In the case of electrochemiluminescence (ECL), the light system is generated by current stimulating luminescent group molecules or complexes. In the case of fluorescence, it is irradiated with excitation light to cause light to be emitted. The fluorescent system is detected by a light source and a detection unit that provides excitation light of a wavelength absorbed by the fluorescent molecule. The detection unit includes a light sensor (such as a photomultiplier tube or a charge coupled device (CCD) array) that detects the emitted signal and prevents the excitation light from being included in the output of the light sensor -15-201209402 (such as wavelength selection) filter). The fluorescent molecules emit Stokes shifted light in response to the excitation light, which is collected by the detection unit. The Stokes shift is the difference in frequency or wavelength between the emitted light and the absorbed excitation light.

The ECL emission system is detected using a light sensor that is sensitive to the emission wavelength of the ECL species used. For example, transition metal-ligand complexes emit light of a visible wavelength, so conventional photodiodes and CCDs can be used as light sensors. An advantage of ECL is that if the ambient light is shielded, the ECL emits light that is the only light in the detection system, thus increasing sensitivity.

Microarrays allow hundreds of thousands of DN A hybridization experiments to be performed simultaneously. Microarrays are powerful molecular diagnostic tools that screen thousands of genetic diseases or detect the presence of numerous infectious pathogens in a single experiment. The microarray consists of a number of different DNA probes that are fixed at the point of acceptance. The target DN A (amplicon) is first labeled with fluorescent or luminescent molecules (either during nucleic acid amplification or after nucleic acid amplification), followed by application of the target DNA to the probe microarray. The microarray is cultured in a temperature controlled, humidified environment for hours or days to cause hybridization between the probe and the amplicon. After incubation, the microarray must be washed through a series of buffers to remove unbound strands. After cleaning, the microarray surface is dried with a gas stream (usually nitrogen). The rigor of hybridization and cleaning is critical. Insufficient stringency may result in highly non-specific binding. Excessive rigor may result in inability to properly combine, resulting in reduced sensitivity. Hybridization is identified by detecting the light emitted by the labeled amplicon of the hybridizing probe with the complementary probe. -16- 201209402 Fluorescence from microarrays is detected by a microarray scanner, which is usually a computer-controlled inverted scanning fluorescent conjugated focal microscope. This microscope usually uses laser-excited fluorescent dye and light sensing. The transmitter (such as a photomultiplier tube or CCD) detects the transmitted signal. Fluorescent molecules emit Stokes bits (as described above), which are collected by the detection unit.

The emitted fluorescent light must be collected, separated from the unabsorbed excitation wavelength and transmitted to the detector. In a microarray scanner, a conjugate focal configuration of a conjugate focal hole aperture mounted on the image side is typically used to eliminate non-focusing information. This device allows only the light of the focused portion to be detected. Light from above and below the focal plane of the target cannot enter the detector, thus increasing the signal to noise ratio. The detected fluorescent photons are converted into electrical energy by a detector and then converted into a digital signal. This digital signal is translated into a number that represents the intensity of the fluorescence from a given pixel. Each feature of the array consists of one or more of these pixels. The final result of the scan is an image of the surface of the array. Since the exact sequence and position of each probe on the microarray is known, the target sequence to which it is hybridized can be simultaneously identified and analyzed. More information on fluorescent probes can be found at: http://www.premierbiosoft.com/tech_notes/FRET_probe.html and http://www.invitrogen.com/site/us/en/home/References/Molecular- Probes-The-

Handbook/Technical-Notes-and-Product-Highlights/Fluorescence-Resonance-

Energy-Transfer-FRET.html POINT-OF-CARE Molecular Diagnostics' Although molecular diagnostic tests offer many benefits, the growth of such tests in the -17-201209402 bed laboratory is still slower than expected, not yet The mainstream of laboratory medical testing. This is primarily due to the higher complexity and cost of nucleic acid detection compared to assays that do not involve nucleic acid methods. The extensive use of molecular diagnostic testing in clinical settings is closely related to the development of instrumentation, which must significantly reduce costs, provide rapid (automatic) analysis from initial (sample processing) to final (resulting), and does not require The operation of substantial human intervention.

Point-of-care technology provides care in the physician's office, on the hospital bed side, or even in a consumer-focused home environment. This technology offers many advantages including: • Quick results for immediate treatment and improved care quality • Very small amount Sample testing to obtain laboratory numbers • Reduce clinical workload • Reduce laboratory workload and reduce office productivity by reducing administrative work

• Improve cost per patient by reducing hospital stays, outpatients can be diagnosed at the time of initial diagnosis, and reduce sample handling, storage, and delivery • Help with clinical management decisions such as infection control and antibiotic use in laboratory wafers (LOC) Molecular Diagnostics Molecular diagnostic systems based on microfluidic technology provide devices that automate and accelerate molecular diagnostic analysis. The shorter detection time is primarily due to the fact that the required sample volume is less active, automated, and low-cost built-in cascaded diagnostic method steps within the microfluidic device. The volume of the scale of the nanoliter and microliter is also -18- 201209402

Reduce reagent consumption and cost. Laboratory wafer (LOC) devices are a common form of microfluidic device. The LOC device has an MST structure within the MST layer for integrating fluid processing onto a single supporting substrate (usually helium). The unit cost of each LOC device is very low by manufacturing the VLSI (very large integrated circuit) lithography technology of the semiconductor industry. However, controlling the flow of fluid through the LOC device, adding reagents, controlling reaction conditions, and the like requires a large external hydroelectric engineering device. The connection of the LOC devices to these external devices greatly limits the molecular diagnostic use of the LOC devices in a laboratory environment. The cost of external instruments and their operational complexity precludes LOC-based molecular diagnostics as an option in a point-of-care environment. In view of this, there is a need for a molecular diagnostic system based on a LOC device for use in point-of-care. SUMMARY OF THE INVENTION Various aspects of the invention are now described by the following numbered paragraphs. GLE00 1.1. This aspect of the invention provides a microfluidic device for analyzing a sample fluid, the microfluidic device comprising: a sample inlet for receiving the sample; a microsystem technology (MST) layer, the MST layer A functional portion for processing and analyzing the sample; and a CMOS circuit having digital memory for storing data and operational information for operating and controlling the functional portions during the sample processing and analysis. GLE00 1.2 Preferably, the microfluidic device also has a plurality of reagents -19-201209402 reservoirs containing reagents for processing the sample, wherein the data stored in the digital memory is identified with respect to the reagent. GLE001.3 Preferably, the data stored in the digitizer is used for the unique identifier of the microfluidic device. GLE001.4 Preferably, the sample is a biological material-containing organism

A sample, one of the functional portions, is used to amplify a polymerase chain reaction (PCR) portion of the nucleic acid sequence in the sample, and the operational information stored in the digital memory is related to the time and length of the thermal cycle.

GLE001.5 Preferably, the functional portions include a culture portion located upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir having a heater to maintain the sample and the restriction enzyme The culture temperature of the mixture during the digestion of the nucleic acid sequence by the restriction enzyme. GLE001.6 Preferably, the microfluidic device also has a temperature sensor, wherein the CMOS circuit uses the output of the temperature sensor to feedback control the culture portion"

GLE001.7 Preferably, the microfluidic device also has a probe array for hybridizing with the target nucleic acid sequence in the amplicon from the PCR portion. GLE001.8 is preferably stored in the digital memory. The data includes probe identification data to identify probes at various locations within the array of probes. GLE00 1.9 Preferably, each probe is configured to form a probe-target hybrid with a complementary nucleic acid sequence (included in the amplicon). The various probe-target hybrid systems are configured to In response to the input, a photon is emitted -20-201209402, and the CMOS circuit has a photosensor to sense photons emitted by the probe-target hybrid. GLE001.10 Preferably, the data stored in the digital memory comprises hybridization data generated from the light sensor output. GLE00 1.il Preferably, the microfluidic device also has an array of hybrid chambers for interposing probes such that the probes within each hybridization chamber are configured to hybridize to one of the target nucleic acid sequences.

GLE001.12 Preferably, the light sensor is associated with a photodiode array of the hybridization chamber. GLE001.13 Preferably, the CMOS circuit has a pad and the CMOS circuit is configured to transmit hybridization data to an external device. GLE001.14 Preferably, the sample is drawn from a patient and the CMOS circuit is configured to download patient data via the pad and store the patient data in the digital device. GLE001.15 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the CMOS circuit. GLE001.1 6 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary of the liquid A drive stream is used to boil the liquid to release the meniscus from the meniscus anchor to recover the capillary drive flow. GLE001.17 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent the edge of the hole. -21 - 201209402 GLE001.18 Preferably, the microfluidic device also has a support substrate and an upper cover, wherein the CMOS circuit is interposed between the support substrate and the MST layer, the upper cover covers the MST layer and defines the reagents Reservoir. GLE001.19 Preferably, the reagent reservoirs each have a surface tension valve configured to define a meniscus anchor, the meniscus anchor used to form a meniscus to retain the reagent therein until contact When the sample stream is removed to remove the meniscus, the reagent is combined with the sample.

GLE00 1 .20 Preferably, the PCR portion is configured to complete the thermal cycling of the sample in 30 seconds. The microfluidic device with integrated, mass-produced, inexpensive, and energy-intensive integrated input memory Fluid and treat it. The digital memory system is used to store data and control information required by the device and modules of the device during operation. Integrating digital memory into the device provides a microfluidic system that is easy to manufacture, mass-produced, easy to use, and inexpensive.

GLE002.1 This aspect of the invention provides a test module for analyzing a sample fluid, the test module comprising: a barn for receiving the sample; a functional portion for processing and analyzing the sample And digital memory for storing data and job information to manipulate and control these functions during sample processing and analysis. GLE002.2 Preferably, the test module also has a plurality of reagent reservoirs. The reagent reservoirs contain reagents for processing the sample, wherein the data stored in the digits is identified by the reagents. -22- 201209402 GLE002.3 Preferably, the data stored in the digital memory is used for the unique identifier of the test module. GLE002.4 Preferably, the sample is a biological sample containing a genetic material, and one of the functional portions is a polymerase chain reaction (PC R) portion for amplifying a nucleic acid sequence in the sample, and is stored in The job information of the digital memory is about the time and length of the thermal cycle.

GLE002.5 Preferably, the functional units are located in the PCR

The culture portion upstream of the portion and one of the reagent reservoirs is a restriction enzyme reservoir having a heater to maintain a culture temperature of the mixture of the sample and the restriction enzyme during the restriction enzyme digestion of the nucleic acid sequence. GLE002.6 Preferably, the test module also has a CMOS circuit and a temperature sensor, wherein the CMOS circuit has digital memory and uses the output of the temperature sensor to feedback control the culture. GLE002.7 Preferably, the test module also has a probe array for hybridization with a target nucleic acid sequence in an amplicon from the PCR portion. GLE002.8 Preferably, the data stored in the digital memory includes probe identification data to identify probes at various locations within the array of probes. GLE002.9 Preferably, each probe is configured to form a probe-target hybrid with a complementary target nucleic acid sequence (included in the amplicon), the various probe-target hybrid systems are organized Photons are emitted in response to the input, and the CMOS circuit has a light sensor to sense photons emitted by the probe-target hybrid. GLE002.1 0 Preferably, the data packet stored in the digital memory -23-201209402 includes the hybrid data generated from the output of the light sensor. GLE002.1 1 Preferably, the test module also has an array of hybridization chambers for interposing probes such that the probes within each hybridization chamber are configured to hybridize to one of the target nucleic acid sequences. GLE002.1 2 Preferably, the light sensor is associated with a photodiode array of the hybridization chamber.

GLE002.1 3 Preferably, the CMOS circuit has a data interface for transmitting hybrid data to an external device. Preferably, the sample is drawn from the patient and the CMOS circuit is configured to download patient data via the data interface and to store the patient data in the digital memory. GLE002.1 5 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the control circuit.

GLE002.1 6 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary of the liquid A drive stream is used to boil the liquid to release the meniscus from the meniscus anchor to recover the capillary drive flow. GLE002.1 7 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent the edge of the hole. GLE002.1 8 Preferably, the test module also has a LOC device having a sample inlet, a support substrate, a microsystem technology (MS T) layer in fluid communication with the container, between the support substrate and the MS The CMOS circuit and the upper cover between the T layer-24 and the 201209402, wherein the CMOS circuit has the digital memory, the MST layer has the functional portion, and the upper cover covers the MS T layer and defines the reagent reservoirs. GLE002.1 9 Preferably, the reagent reservoirs each have a surface tension valve configured to define a meniscus anchor, the meniscus anchor used to form a meniscus to retain the reagent therein until The reagent is allowed to bind to the sample while contacting the sample stream to remove the meniscus.

GLE002.20 Preferably, the PCR section is configured to complete the thermal cycling of the sample within 30 seconds. The easy-to-use, mass-produced, and inexpensive LOC device with integrated digital memory accepts input fluid and processes it. The digital memory system is used to store data and control information required by the LOC device and modules of the LOC device during operation. The information stored on the Billion includes the features of the module with this LOC device. Integrating digital memory into the device provides a module that is easy to manufacture, mass-produced, easy to use, and inexpensive. GLE003.1 This aspect of the invention provides a on-wafer laboratory (LOC) device for analyzing a sample fluid, the LOC device comprising: a sample inlet for accepting the sample; a microsystem technology (MST) layer, The MST layer has a functional portion for processing and analyzing the sample; and a CMOS circuit having a digital body for storing epidemiological data and configured to download an epidemiological update updated by an external source data. -25- 201209402 GLE003.2 Preferably, the CMOS circuit has a universal serial bus (u S B) device driver for operating to control a USB connection to the external source. GLE003.3 Preferably, the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample, wherein the data stored in the digital memory is identified with respect to the reagent. GLE003.4 Preferably, the data stored in the digital memory

A unique identifier for the LOC device. GLE003.5 Preferably, the sample is a biological sample containing a genetic material, one of the functional portions being a polymerase chain reaction (PCR) portion for amplifying a nucleic acid sequence in the sample.

GLE003.6 Preferably, the functional portions include a culture portion located upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir having a heater to maintain the sample and the restriction enzyme The enzymatic reaction temperature of the mixture.

GLE003.7 Preferably, the LOC device also has a temperature sensor, wherein the CMOS circuit uses the output of the temperature sensor to feedback control the culture. GLE003.8 Preferably, the LOC device also has a probe array for hybridizing to the target nucleic acid sequence in the amplicon from the PCR portion. GLE003.9 Preferably, each probe is configured to A probe-target hybrid is formed with a complementary target nucleic acid sequence (included in the amplicon), and the various probe-target hybridization systems are configured to react to the input and -26-

201209402, and the CMOS circuit has a light sensor to sense photons emitted by the probe hybrid. GLE003.1 0 Preferably, the LOC device also has a hybrid, the hybrid chamber array for containing probes such that each of the hybridization chambers is configured to hybridize to one of the target nucleic acid sequences. GLE003.il Preferably, the light sensor is registered with the hybrid photodiode array. GLE003.12 Preferably, the CMOS circuit has a pad to connect to the USB connector and to transmit hybridization data to an external device. GLE003.1 3 Preferably, the sample is extracted from the patient and the CMOS circuit is configured to download the patient via the USB connector to store the patient data in the digital memory. GLE003.14 Preferably, the PCR portion has an active valve that retains liquid in the PCR portion during thermal cycling and allows the liquid to flow from the activation signal of the CMOS circuit to the hybridization chamber" GLE003.1 5 Preferably, The active valve is provided with a meniscus and a boiling start valve of the heater, the meniscus anchoring device is configured to block the meniscus to prevent the capillary driving flow of the liquid, and the heater uses the liquid to The meniscus is released at the meniscus anchor to recover the flow. GLE003.1 6 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent the edge of the hole. GLE003.1 7 Preferably, the LOC device also has a support cover, wherein the CMOS circuit is interposed between the support substrate and the 丨

a ten-target intercommunication probe for the chamber, and the data is used in response to the anchor to form between the boiled squeezing layer of the heated holding substrate % MST -27-201209402, The upper cover covers the MST layer and defines the reagent reservoirs. GLE003.1 8 Preferably, the reagent reservoirs each have a surface tension valve for setting a meniscus anchor, the meniscus anchor is used for A meniscus is formed to retain the reagent therein, and the reagent is allowed to bind to the sample until the meniscus is removed by contacting the sample stream. GLE003.1 9 Preferably, the PCR section has a thermal cycle time of less than 4 seconds.

GLE003.20 Preferably, the PCR portion has a thermal cycle time of between 0.45 seconds and 1.5 seconds.

The easy-to-use, mass-produced, and inexpensive LOC device with integrated digital memory accepts input fluid and processes it. The digital memory system is used to store data and control information required by the LOC device and the module of the LOC device during operation. The information stored in this memory includes epidemiological updates available at the time and information for analytical and diagnostic purposes. This information provides modules that do not require the independence of external support. Integrating digital memory into the device provides a module that is easy to manufacture, mass-produced, and easy to use and inexpensive. GLE004.1 This aspect of the invention provides a on-wafer laboratory (LOC) device for analyzing a sample fluid, the LOC device comprising: a sample inlet for receiving the sample; a microsystem technology (MST) layer The MST layer has a functional portion for processing and analyzing the sample: and a CMOS circuit having a digital body for storing genetic data and configured to download genetic data updated by an external source-28 - 201209402 GLE004.2 Preferably, the CMOS circuit has a universal serial bus (USB) device driver to operatively control a USB connector connected to the external source. GLE004.3 Preferably, the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample, wherein the data stored in the digital memory is identified with respect to the reagent.

GLE004.4 Preferably, the data stored in the digital memory is used for the unique identifier of the LOC device. GLE004.5 Preferably, the sample is a biological sample containing a genetic material, and one of the functional portions is a polymerase chain reaction (PCR) portion for amplifying a nucleic acid sequence in the genetic material.

GLE004.6 Preferably, the functional portions include a culture portion located upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir having a heater to maintain the sample and the restriction enzyme The enzymatic reaction temperature of the mixture. GLE004.7 Preferably, the LOC device also has a temperature sensor, wherein the CMOS circuit uses the output of the temperature sensor to feedback control the culture. GLE004.8 Preferably, the LOC device also has a probe array for hybridizing to the target nucleic acid sequence in the amplicon from the PCR portion. GLE004.9 Preferably, each probe is configured to Forming a probe-target hybrid with a complementary nucleic acid sequence (included in the amplicon), -29-201209402 The various probe-target hybridization systems are configured to emit in response to an input' and the CMOS The circuit has a light sensor to sense photons emitted by the probe-target hybrid. GLE004.1 0 Preferably, the LOC device also has an array of hybridization chambers for interposing probes such that the probes within each hybridization chamber are organized to hybridize to one of the target nucleic acid sequences.

GLE004.il Preferably, the light sensor is associated with a photodiode array of the hybridization chamber. GLE004.1 2 Preferably, the CMOS circuit has pads for transmitting hybrid data via the USB connector. GLE004.1 3 Preferably, the sample is drawn from a patient and the CMOS circuit is configured to download patient data via the pad and store the patient data in the digital memory.

GLE004.1 4 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the CMOS circuit. GLE004.1 5 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary of the liquid A drive stream is used to boil the liquid to release the meniscus from the meniscus anchor to recover the capillary drive flow. GLE004.1 6 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent the edge of the hole. GLE004.1 7 Preferably, the LOC device also has a support substrate -30-201209402 and an upper cover, wherein the CMOS circuit is interposed between the support substrate and the MST layer, the upper cover covers the MST layer and defines the reagents Reservoir. GLE004.1 8 Preferably, the reagent reservoirs each have a surface tension valve disposed with a meniscus anchor, the meniscus anchor used to form a meniscus to retain the reagent therein until The reagent is allowed to bind to the sample while contacting the sample stream to remove the meniscus. GLE004.19 Preferably, the PCR portion has a heat of less than 4 seconds.

GLE004.20 Preferably, the PCR portion has a thermal cycle time of between 0.45 seconds and 1.5 seconds. The easy-to-use, mass-produced, and inexpensive LOC device with integrated digital memory accepts input fluid and processes it. The digital memory system is used to store data and control information required by the LOC device and modules of the LOC device during operation. The information stored in this memory includes information on genetic information updates available at the time and information for analysis and diagnostic purposes. This information provides modules that do not require the independence of external support. Integrating digital memory into the device provides a module that is easy to manufacture, mass-produced, and easy to use and inexpensive. GLE005.1 This aspect of the invention provides a wafer-on-lab (LOC) device for analyzing a sample fluid. The LOC device comprises: a sample inlet for accepting the sample; a microsystem technology (MST) layer, The MST layer has a functional portion for processing and analyzing the sample: and a CMOS circuit having a digital memory for storing -31 - 201209402 operation information for controlling the operation of the functional portion during the sample processing and analysis The data is encrypted for secure communication with external devices. GLE 005.2 Preferably, the LOC device also has a multi-reservoir. The reagent reservoirs contain reagents for processing the sample, and the data stored in the digital memory is identified with respect to the reagent. GLE005.3 Preferably, the r stored in the digital memory is used for the unique identifier of the LOC device. GLE005.4 Preferably, the sample contains a sample of a genetic material, and one of the functional portions is used to amplify a polymerase chain reaction (PCR) portion of the sample, and is stored in the digital record Information about the time and length of the thermal cycle. GLE005.5 Preferably, the functional portions include a culture portion located upstream of the f portion and one of the reagent reservoirs is a restriction enzyme. The culture portion has a heater to maintain mixing of the sample and the restriction enzyme to limit enzyme digestion. The culture temperature during the nucleic acid sequence. GLE005.6 Preferably, the LOC device also has a warmer, wherein the CMOS circuit uses the output of the temperature sensor to make the culture. GLE005.7 Preferably, the LOC device also has probes for use with the target nucleic acid sequence GLE005.8 in the amplicon from the PCR portion. Preferably, the digital memory is stored in the digital memory including probe identification. Data to identify the data of each bit in the probe array and the reagents of the biological nucleic acid sequence of the data storage system, and the data of the hybridization of the sensory feedback needle array

-32- 201209402 Needle. GLE005.9 Preferably, each probe is configured to form a probe-target hybrid with a complementary target nucleic acid sequence (included in the amplicon), the various probe-target hybrid systems are organized The light is emitted in response to the input, and the CMOS circuit has a light sensor to sense photons emitted by the probe-target hybrid.

GLE005.10 Preferably, the data stored in the digital memory comprises hybridization data generated from the light sensor output. GLE005.1 1 Preferably, the LOC device also has an array of hybridization chambers for interposing probes such that probes within each hybridization chamber are configured to hybridize to one of the target nucleic acid sequences. GLE005.12 Preferably, the light sensor is associated with a photodiode array of the hybridization chamber. GLE005.1 3 Preferably, the CMOS circuit has a pad and the CMOS circuit is configured to transmit hybridization data to an external device. GLE005.14 Preferably, the sample is drawn from a patient and the CMOS circuit is configured to download patient data via the pad and store the patient data in the digital memory. GLE005.1 5 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the CMOS circuit. GLE005.1 6 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary of the liquid Drive flow, the heater is used to boil -33 - 201209402 The liquid releases the meniscus from the meniscus anchor to recover the capillary drive flow. GLE005.1 7 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent the edge of the hole. GLE005.1 8 Preferably, the LOC device also has a support substrate and an upper cover, wherein the CMOS circuit is interposed between the support substrate and the MST layer, the upper cover covers the MST layer and defines the reagent reservoirs. GLE005.1 9 Preferably, the reagent reservoirs each have a surface tension valve configured to define a meniscus anchor, the meniscus anchor used to form a meniscus to retain the reagent therein until The reagent is allowed to bind to the sample while contacting the sample stream to remove the meniscus. GLE005.20 Preferably, the P R R portion is organized to complete the thermal cycling of the sample in 30 seconds. The easy-to-use, mass-produced and inexpensive LOC device with integrated digital memory accepts diagnostic samples and processes them. The digital system is used to store the data and control information required by the LOC device and the diagnostic module of the LOC device during operation. The memory also preserves information on patient test results. The ability to store patient test results allows the diagnostic module to be tested with only a minimal external power supply, and then analyze the patient test results with a fully functional reader. The preservation of the information on the results of the test will ensure that the information is not illegally used through illegal channels. Integrating digital memory into the device provides a module that is easy to manufacture, mass-produced, easy to use, and inexpensive. GLE006.1 This aspect of the invention provides a test module for analyzing a sample 201209402 fluid, the test module comprising: a container for receiving the sample; a functional portion for processing and analyzing the sample A support substrate: and a CMOS circuit on the support substrate for operating and controlling the functional portion during processing and analysis of the sample.

GLE006.2 Preferably, the test module also has a plurality of reagent reservoirs containing reagents for processing the sample, wherein the CMOS circuit has a digital memory for storing identification data of the reagents

GLE006.3 Preferably, the data stored in the digital memory is used to uniquely identify the test module. GLE006.4 Preferably, the sample is a biological sample containing a genetic material, and one of the functional portions is a polymerase chain reaction (PCR) portion for amplifying a nucleic acid sequence in the sample, and is stored in the sample The φ job information of the digital memory is about the time and length of the thermal cycle.

GLE006.5 Preferably, the functional portions include a culture portion located upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir having a heater to maintain the sample and the restriction enzyme The culture temperature of the mixture during the digestion of the nucleic acid sequence by the restriction enzyme. GLE006.6 Preferably, the test module also has a temperature sensor, wherein the CMOS circuit uses the output of the temperature sensor to feedback control the culture. GLE006.7 Preferably, the test module also has a probe array -35-201209402 for hybridization with a target nucleic acid sequence in an amplicon from the PCR portion. GLE006.8 Preferably, the data stored in the digital memory includes probe identification data to identify probes at various locations within the array of probes. GLE006.9 Preferably, each probe is organized to complement each other

The target nucleic acid sequence (included in the amplicon) forms a probe-target hybrid, the various probe-target hybridization systems are configured to emit in response to the input, and the CMOS circuit has a light sensor Photons emitted by the probe-target hybrid are sensed. GLE006.1 0 Preferably, the data stored in the digital body includes the hybrid data generated from the light sensor output. GLE006.1 1 Preferably, the test module also has an array of hybridization chambers for interposing probes such that the probes within each of the hybridization chambers are configured to hybridize to one of the target nucleic acid sequences. GLE006.1 2 Preferably, the light sensor is registered with the hybrid chamber

Photodiode array. GLE006.1 3 Preferably, the control circuit has a data interface for transmitting hybrid data to an external device. GLE006.1 4 Preferably, the sample is drawn from the patient and the control circuitry is configured to download patient data via the data interface and store the patient data in the digital memory. GLE006.1 5 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the control circuit. -36- 201209402 GLE006.1 6 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater. The meniscus anchor is configured to form a meniscus to prevent The capillary drive of the liquid is used to boil the liquid to release the meniscus from the meniscus anchor to recover the capillary drive flow. GLE006.1 7 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent the edge of the hole.

GLE006.1 8 Preferably, the test module also has a LOC device having the support substrate and the CMOS circuit, and has a sample inlet in fluid communication with the container, and a micro system technology (MST) having the functional portion a layer and an upper cover, wherein the upper cover covers the MST layer and defines the reagent reservoirs. GLE006.1 9 Preferably, the reagent reservoirs each have a surface tension valve configured to define a meniscus anchor, the meniscus anchor used to form a meniscus to retain the reagent therein until The reagent is allowed to bind to the sample while contacting the sample stream to remove the meniscus. GLE006.20 Preferably, the PCR portion is configured to complete the thermal cycling of the sample within 30 seconds. The easy-to-use, mass-produced and inexpensive LOC device accepts and processes the sample and electronically controls all of the LOC devices on the wafer of the LOC device. Integrating the control electronics into the LOC device allows the module to utilize the L〇C device without special external support and provides a module that is easy to manufacture and mass-produces, which is easy to use and inexpensive. GLE007.1 This aspect of the invention provides a on-wafer laboratory-37-201209402 (LOC) device for mounting in a test module for analyzing sample fluid and communicating test results with an external device, the LOC The device includes a support substrate; a sample inlet for receiving the sample; a microsystem technology (MST) layer having a functional portion for processing and analyzing the sample;

The CMOS circuit on the support substrate is used to operate and control a communication interface communicated with the external device in the test module. GLE007.2 Preferably, the CMOS circuit has a universal serial bus (USB) device driver for operating a USB connector that communicates with the external device in the test module. GLE007.3 Preferably, the C Μ Ο S circuit is additionally configured to operate to control the functional portion during processing and analysis of the sample. GLE007.4 Preferably, the LOC device also has a plurality of reagents

A reservoir containing reagents for processing the sample, wherein the CMOS circuit has a digital memory for storing identification data relating to the reagent. GLE007.5 Preferably, the data stored in the digital memory is used for the unique identifier of the LOC device. GLE007.6 Preferably, the sample is a biological sample containing a genetic material, and one of the functional portions is a polymerase chain reaction (PCR) portion for amplifying a nucleic acid sequence in the sample, and is stored in the sample The working information of the digital memory is about the time and length of the thermal cycle. -38- 201209402

GLE007.7 Preferably, the functional portions include a culture portion located upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir having a heater to maintain the sample and the restriction enzyme The culture temperature of the mixture during the digestion of the nucleic acid sequence by the restriction enzyme. GLE007.8 Preferably, the LOC device also has a temperature sensor, wherein the CMOS circuit uses the output of the temperature sensor to feedback control the culture.

GLE007.9 Preferably, the LOC device also has a probe array for hybridizing with the target nucleic acid sequence in the amplicon from the PCR portion. GLE007.1 0 is preferably stored in the digit The data therein includes probe identification data to identify probes at various locations within the probe array. GLE007.il Preferably, each probe is configured to form a probe-target hybrid with a complementary nucleic acid sequence (included in the amplicon), φ the various probe-target hybrid systems are grouped The structure is configured to emit in response to the input, and the CMOS circuit has a light sensor to sense photons emitted by the probe-target hybrid. GLE007.1 2 Preferably, the data stored in the digital memory includes the hybrid data generated from the light sensor output. GLE007.1 3 Preferably, the LOC device also has an array of hybridization chambers for interposing probes such that the probes within each of the hybridization chambers are organized to hybridize to one of the target nucleic acid sequences. GLE007.14 Preferably, the light sensor is a register of the 2012 09402 photodiode array of the hybridization chamber. GLE007.1 5 Preferably, the CMOS circuit is configured to transmit the hybrid data to an external device via the test module communication interface. GLE007.1 6 Preferably, the sample is drawn from a patient and the CMOS circuit is configured to download patient data via the communication interface and store the patient data in the digital memory.

GLE007.1 7 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the control circuit. GLE007.1 8 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus paver is configured to form a meniscus to prevent capillary of the liquid A drive stream is used to boil the liquid to release the meniscus from the meniscus anchor to recover the capillary drive flow.

GLE007.1 9 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent the edge of the hole. GLE007.20 Preferably, the PCR portion is configured to complete the thermal cycling of the sample within 30 seconds. The easy-to-use, mass-produced and inexpensive LOC device accepts and processes the sample and electronically controls the on-chip electronic control of the LOC device to communicate with the host's data and commands. Integrating the electronics into the LOC device allows the module to utilize the LOC device without special external support and provides a module that is easy to manufacture, mass-produced, easy to use, and inexpensive. -40- 201209402 GLE008.1 This aspect of the invention provides a on-wafer laboratory (LOC) device for mounting in a test module for analyzing sample fluid and communicating test results with an external device, the LOC The device comprises a support substrate; the sample is inserted into the □, which is used to receive the sample;

a microsystem technology (MST) layer having a functional portion for processing and analyzing the sample; and a CMOS circuit on the support substrate having a versatile serial bus (USB) device driver for operation Controlling a USB connector that communicates with the external device in the test module. GLE008.2 Preferably, the C Μ 0 S circuit is otherwise configured to operate to control the functional portion during processing and analysis of the sample. GLE008.3 Preferably, the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample, wherein the CMOS circuitry has digital memory for storing identification data relating to the reagents. GLE008.4 Preferably, the data stored in the digital memory is used for the unique identifier of the LOC device. GLE008.5 Preferably, the sample is a biological sample containing a genetic material, and one of the functional portions is a polymerase chain reaction (PCR) portion for amplifying a nucleic acid sequence in the sample, and stored in the sample The working information of the digital memory is about the time and length of the thermal cycle.

GLE008.6 Preferably, the functional portions include a culture portion located upstream of the PCR -41 - 201209402 portion and one of the reagent reservoirs is a restriction enzyme reservoir having a heater to maintain the sample The culture temperature of the mixture with the restriction enzyme during the digestion of the nucleic acid sequence by the restriction enzyme. GLE00 8.7 Preferably, the LOC device also has a temperature sensor, wherein the CMOS circuit uses the output of the temperature sensor to feedback control the culture. GLE00 8.8 Preferably, the LOC device also has a probe array for hybridizing with the target nucleic acid sequence in the amplicon from the PCR portion. GLE00 8.9 Preferably, the data stored in the digital memory includes The probe identifies the data to identify probes at various locations within the array of probes. GLE008.1 0 Preferably, each probe is configured to form a probe-target hybrid with a complementary nucleic acid sequence (included in the amplicon), the various probe-target hybrid systems are grouped The structure is configured to emit in response to the input, and the CMOS circuit has a light sensor to sense photons emitted by the probe-target hybrid. GLE008.il Preferably, the data stored in the digital memory comprises hybridization data generated from the light sensor output. GLE008.1 2 Preferably, the digital memory comprises a random access memory (RAM) and a flash memory, the ram is configured to store the hybrid data and the flash memory system is configured to store a program Data to operate the function and the probe identification data. GLE00 8.1 3 Preferably, the LOC device also has a hybrid array 201209402 column. The hybrid chamber array is adapted to contain probes such that the probes within each hybridization chamber are organized to hybridize to one of the target nucleic acid sequences. GLE008.14 Preferably, the light sensor is associated with a photodiode array of the hybridization chamber. GLE008.1 5 Preferably, the USB device driver is configured to transmit the hybrid data to the external device via the USB connector.

GLE008.1 6 Preferably, the sample is drawn from a patient and the CMOS circuit is configured to download patient data via the USB connector and to store the patient data in the digital memory. GLE008.1 7 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the control circuit. GLE008.1 8 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary of the liquid A drive stream is used to boil the liquid to release the meniscus from the meniscus anchor to recover the capillary drive flow. GLE 008.1 9 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent the edge of the hole. GLE008.20 Preferably, the PCR portion is configured to complete the thermal cycling of the sample within 30 seconds. The easy-to-use, mass-produced and inexpensive LOC device with integrated USB device controller accepts diagnostic samples and processes them. Integrating the USB device controller into the LOC device allows the module to utilize the LOC device without the need for special -43-201209402 external support, and provides a module that is easy to manufacture, easy to use, and inexpensive. GLE009.1 This aspect of the invention provides a wafer-on-lab (LOC) device for fluid separation, the LOC device comprising a support substrate; a sample inlet for receiving the sample: a microsystem technology (MST) layer The MST layer has a functional portion to analyze and analyze the sample: and a CMOS electric CMOS circuit interposed between the support substrate and the MST layer has a controller to control the operation of the functional portion during analysis of the sample. GLE009.2 Preferably, the CMOS circuit has digital bits for storing data and operational information of the functional units controlled by the controller during processing and analysis of the sample. GLE009.3 Preferably, the LOC device also has a multi-reservoir, the reagent reservoir contains reagents for processing the sample, and the data stored in the digital memory is unique to the LOC device, the unique identifier It is related to the identification of the reagent. GLE009.4 Preferably, the sample contains a sample of a genetic material, and one of the functional portions is used to amplify a polymerase chain reaction (PCR) portion of the sample, and is stored in the digital record Information about the time and length of the thermal cycle. GLE009.5 Preferably, the functional parts include a culture portion located upstream of the f portion, and one of the reagent reservoirs is a restriction enzyme, a sample can be used for the treatment, and the treatment and the memory are used. The PCR reservoir in which the reagent stores the identifier of the biological nucleic acid sequence, 44 - 201209402 The culture portion has a heater to maintain the culture temperature of the mixture of the sample and the restriction enzyme during the restriction enzyme digestion of the nucleic acid sequence. GLE009.6 Preferably, the LOC device also has a temperature sensor, wherein the CMOS circuit uses the output of the temperature sensor to feedback control the culture. GLE009.7 Preferably, the LOC device also has a probe array for hybridization with a target nucleic acid sequence in an amplicon from the PCR portion

GLE009.8 Preferably, the data stored in the digital memory includes probe identification data to identify probes at various locations within the array of probes. GLE009.9 Preferably, each probe is configured to form a probe-target hybrid with a complementary target nucleic acid sequence (included in the amplicon), the various probe-target hybrid systems are organized Photons are emitted in response to the input, and the CMOS circuit has a light sensor to sense photons emitted by the probe-target hybrid. GLE009.1 0 Preferably, the data stored in the digital memory comprises hybridization data generated from the light sensor output. Preferably, the LOC device also has an array of hybridization chambers for interposing probes such that the probes within each of the hybridization chambers are organized to hybridize to one of the target nucleic acid sequences. GLE009.1 2 Preferably, the light sensor is a register of the photodiode array of the hybridization chamber. GLE009.1 3 Preferably, the CMOS circuit has pads, and the -45-201209402 C Μ O S circuit is configured to transmit hybridization data to an external device. GLE009.1 4 Preferably the sample is drawn from the patient and the CMOS circuit is configured to download patient data via the pad and store the patient data in the digital memory. GLE009.1 5 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the CMOS circuit.

GLE009.1 6 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary of the liquid A drive stream is used to boil the liquid to release the meniscus from the meniscus anchor to recover the capillary drive flow. GLE009.1 7 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent the edge of the hole.

GLE009.1 8 Preferably, the LOC device also has an upper cover, wherein the upper cover covers the MST layer and defines the reagent reservoirs. GLE009.1 9 Preferably, the reagent reservoirs each have a surface tension valve configured to define a meniscus anchor, the meniscus anchor used to form a meniscus to retain the reagent therein until The reagent is allowed to bind to the sample while contacting the sample stream to remove the meniscus. GLE009.20 Preferably, the PCR portion is configured to complete the thermal cycle of the sample within 30 seconds. The easy-to-use, mass-produced and inexpensive L0C device receives the diagnostic sample and processes it, from the L 0 C Integrated Controller Control for Units -46 - 201209402 All functions of this LOC unit. Integrating the controller into the LOC device allows the module to utilize the LOC device without special external support, and provides a module that is easy to manufacture, mass-produced, easy to use, and inexpensive. GLE010.1 This aspect of the invention provides a wafer-on-lab (LOC) device for analyzing a sample fluid, the LOC device comprising: a support substrate;

a sample inlet for receiving the sample; a microsystem technology (MST) layer having a functional portion for processing and analyzing the sample; and a CMOS circuit interposed between the support substrate and the MST layer, The CMOS circuit has digital memory for storing data and operational information for controlling the functions of the functional components during processing and analysis of the sample. GLE010.2 Preferably, the LOC device also has a plurality of reagent reservoirs containing reagents for processing the sample, wherein the information stored in the digital memory is identified by the reagent. GLE010.3 Preferably, the data stored in the digital memory is used for the unique identifier of the LOC device. GLE010.4 Preferably, the sample is a biological sample containing a genetic material, and one of the functional portions is a polymerase chain reaction (PCR) portion for amplifying a nucleic acid sequence in the sample, and is stored in the sample The information on the operation of the digital body is about the time and length of the thermal cycle.

GLE010.5 Preferably, the functional portions include a culture portion located upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir, -47-201209402, the culture portion has a heater to maintain the sample The culture temperature of the mixture with the restriction enzyme during the digestion of the nucleic acid sequence by the restriction enzyme. GLE 0106 Preferably, the LOC device also has a temperature sensor, wherein the CMOS circuit uses the output of the temperature sensor to feedback control the culture. GLE010.7 Preferably, the LOC device also has a probe array for hybridizing with the target nucleic acid sequence in the amplicon from the PCR portion, GLE010.8 is preferably stored in the digital body The data includes probe identification data to identify probes at various locations within the array of probes. GLE010.9 Preferably, each probe is organized to complement each other

The target nucleic acid sequence (included in the amplicon) forms a probe-target hybrid, the various probe-target hybridization systems are configured to emit photons in response to the input, and the CMOS circuit has a light sensor Photons emitted by the probe-target hybrid are sensed. GLE 010.10 Preferably, the data stored in the digital memory comprises hybridization data generated from the light sensor output. Preferably, the LOC device also has an array of hybridization chambers for interposing probes such that the probes within each of the hybridization chambers are organized to hybridize to one of the target nucleic acid sequences. GLE 010.12 Preferably, the light sensor is associated with a photodiode array of the hybridization chamber. GLE010.13 Preferably, the CMOS circuit has pads, and the -48-201209402 CMOS circuit is configured to transmit hybridization data to an external device. GLE01 0.14 Preferably, the sample is drawn from a patient and the CMOS circuit is configured to download patient data via the pad and store the patient data in the digital memory. GLE 010.15 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the CMOS circuit.

GLE0 10.16 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary flow of the liquid The heater is used to boil the liquid to release the meniscus from the meniscus anchor to restore the capillary drive flow. GLE 010.17 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent the edge of the hole. GLE 010.18 Preferably, the LOC device also has an upper cover, wherein the upper cover covers the MST layer and defines the reagent reservoirs. GLE 010.19 Preferably, the reagent reservoirs each have a surface tension valve configured to define a meniscus anchor, the meniscus anchor used to form a meniscus to retain the reagent therein until contact When the sample stream is removed to remove the meniscus, the reagent is combined with the sample. GLE010.20 Preferably, the PCR portion is configured to complete the thermal cycling of the sample within 30 seconds. The easy-to-use, mass-produced and inexpensive LOC device accepts and processes the sample, and the integrated data of the LOC device is used to store the intermediate data. Integrating the data RAM into the LOC device allows the module to utilize the L Ο C device without special external support, and provides a module that is easy to manufacture, mass-produced, easy to use, and inexpensive. GLE0 1 1 · 1 This aspect of the invention provides a wafer-on-lab (LOC) device for analyzing a sample fluid, the LOC device comprising: a support substrate;

a sample is introduced into the sample for receiving the sample; a microsystem technology (MST) layer having a functional portion for processing and analyzing the sample: and a CMOS circuit interposed between the support substrate and the MST layer The CMOS circuit has flash memory for storing program data to operate the functions during processing and analysis of the sample. GLE011.2 Preferably, the LOC device also has a plurality of reagents

The reservoir, the reagent reservoir containing reagents for processing the sample, wherein the flash memory also stores information relating to the identification of the reagent. GLE011.3 Preferably, the information includes a unique identifier for the LOC device. GLE0 11.4 Preferably, the sample is a biological sample containing a genetic material, and one of the functional portions is used to amplify a polymerase chain reaction (PCR) portion of the nucleic acid sequence in the sample and is stored in the digit The job information of the memory is about the time and length of the thermal cycle.

GLE0 11.5 Preferably, the functional portions include a culture portion located upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir, -50-201209402, the culture portion has a heater to maintain the sample and The culture temperature of the restriction enzyme mixture during the digestion of the nucleic acid sequence by the restriction enzyme. GLE 011.6 Preferably, the LOC device also has a temperature sensor 'where the CMOS circuit uses the output of the temperature sensor to feedback control the culture. GLE011.7 Preferably, the LOC device also has a probe array for hybridizing to the target nucleic acid sequence in the amplicon from the PCR portion. GLE011.8 Preferably, the flash memory storage probe is identified. Data to identify probes at various locations within the probe array. GLE011.9 Preferably, each probe is configured to form a probe-target hybrid with a complementary target nucleic acid sequence (included in the amplicon), the various probe-target hybrid systems are organized Photons are emitted in response to the input, and the CMOS circuit has a light sensor to sense photons emitted by the probe-target hybrid.

GLE011.10 Preferably, the CMOS circuit has a random access memory (RAM) configured to store the hybrid data produced by the output of the light sensor. Preferably, the LOC device also has an array of hybridization chambers for interposing probes such that probes within each of the hybridization chambers are configured to hybridize to one of the target nucleic acid sequences. GLE011.12 Preferably, the light sensor is associated with a photodiode array of the hybridization chamber. GLE011.13 Preferably, the CMOS circuit has pads, and the -51 - 201209402 CMOS circuit is configured to transmit hybridization data to an external device. G L E 0 1 1 . 1 4 Preferably the sample is drawn from a patient and the CMOS circuit is configured to download patient data via the pad and store the patient data in the digital memory. GLE01 1.15 Preferably the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the CMOS circuit.

GLE01 1 .16 preferably 'the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary of the liquid A drive stream is used to boil the liquid to release the meniscus from the meniscus anchor to recover the capillary drive flow. GLE011.17 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent the edge of the hole.

GLE01 1 .18 Preferably, the LOC device also has an upper cover, wherein the upper cover covers the M ST layer and defines the reagent reservoirs & GLE011.19 Preferably, the reagent reservoirs each have a meniscus setting A surface tension valve of an anchor for forming a meniscus to retain the reagent therein until the sample is removed to remove the meniscus, the reagent is with the sample Combine. GLE01 1 .20 Preferably, the P C R portion is organized to complete the thermal cycling of the sample in 30 seconds. The easy-to-use, mass-produced and inexpensive microfluidic device with integrated program and data flash memory accepts input fluid and processes it. -52- 201209402 This flash memory system is used to store the information and programs required for the device and the modules containing the device during the operation. The integration of flash memory into the device provides a microfluidic system that is easy to manufacture, mass-produced, easy to use, and inexpensive. GLE012.1 This aspect of the invention provides a test module for analyzing a sample fluid and communicating epidemiological data to a database, the test set comprising:

a container for receiving the sample; a functional portion for processing and analyzing the sample: a communication interface for communicating with the epidemiological database; and a controller for operating the control Communication interface. GLE012.2 Preferably, the test module also has a universal serial bus (USB) connector, wherein the communication interface is a device driver for operating the USB connector to communicate with an external device. GLE012.3 Preferably, the test module also has digital memory for storing epidemiological data. GLE012.4 Preferably, the test module also has a plurality of reagent reservoirs containing reagents for processing the sample, wherein the data stored in the digital memory includes the reagent identification. GLE012.5 Preferably, the data stored in the digital memory includes a unique identifier for the test module. GLE012.6 Preferably, the sample is a biological sample containing a genetic material, one of the functional portions being a polymerase chain reaction (PCR) portion for amplifying a nucleic acid sequence in the sample. -53- 201209402

GLE012.7 Preferably, the functional portions include a culture portion located upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir having a heater to maintain the sample and the restriction enzyme The enzymatic reaction temperature of the mixture. GLE012.8 Preferably, the test module also has a CMOS circuit and a temperature sensor, wherein the CMOS circuit has digital memory and uses the output of the temperature sensor to feedback control the culture.

GLE012.9 Preferably, the test module also has a probe array for hybridization with a target nucleic acid sequence in an amplicon from the PCR portion. GLE012.10 Preferably, each probe is configured to form a probe-target hybrid with a complementary nucleic acid sequence (included in the amplicon), the various probe-target hybrid systems are organized The photons are emitted in response to the excitation, and the CMOS circuit has a light sensor to sense photons emitted by the probe-target hybrid. GLE0 12.il Preferably, the test module also has a hybrid chamber array

The hybrid chamber array is adapted to contain probes such that the probes within each of the hybridization chambers are configured to hybridize to one of the target nucleic acid sequences. GLE012.12 Preferably, the light sensor is associated with a photodiode array of the hybridization chamber. GLE0 1 2. 1 3 Preferably, the C Μ Ο S circuit is organized to communicate hybridization information with an external device. GLE012.14 Preferably, the sample is drawn from a patient and the CMOS circuit is configured to download and store the patient data in the digital memory. -54- 201209402 GLE01 2.1 5 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the CMOS circuit. GLE012.16 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary driving of the liquid Flow, the heater is used to boil the liquid to release the meniscus from the meniscus anchor to restore capillary

GLE012.17 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent the edge of the hole. GLE012.18 Preferably, the test module also has a LOC device having a sample inlet, a support substrate, a microsystem technology (MST) layer in fluid communication with the container, and between the support substrate and the MST layer. Between the CMOS circuit and the upper cover, wherein the CMOS circuit has the digital memory, the communication interface, the controller, the MST layer has the function φ portion, and the upper cover covers the MST layer and defines the reagent reservoirs . GLE012.19 Preferably, the reagent reservoirs each have a surface tension valve configured to define a meniscus anchor, the meniscus anchor used to form a meniscus to retain the reagent therein until contact When the sample stream is removed to remove the meniscus, the reagent is combined with the sample. GLE012.20 Preferably, the PCR portion has a thermal cycle time of less than 4 seconds. This easy-to-use, mass-produced 'unusual and portable diagnostic test module accepts biochemical samples and processes and analyzes the sample, updating the epidemiological database based on the diagnostic results derived from the sample -55- 201209402. The update of the epidemiological database provides an enhanced scientific basis for functionalizing the diagnostic test module and the ideal high response to epidemiological conditions. The automated update database based on the diagnostic test module will provide a number of quality and economic benefits to the health information system.

GLE013.1 This aspect of the invention provides a test module for analyzing a sample fluid and communicating local data with an epidemiological database, the test module comprising: a container for receiving the sample; a functional portion, It is used to process and analyze the sample: the communication interface, which is used to communicate with the epidemiological database; and the control (4) 1 foot (4) 乍 (4) aging age 1 1 Φ , the controller is organized to correlate local data Epidemiological data transmitted to the communication interface for communication with the epidemiological database. GLE013.2 Preferably, the test module also has a universal serial

A busbar (USB) connector, wherein the communication interface is a USB device driver for operating and controlling the USB connector to communicate with an external device. GLE 013.3 Preferably, the controller is configured to automatically communicate with the epidemiological database without the need for user activation. GLE013.4 Preferably, the test module also has a user interface. The user interface can be used to input data to the controller to communicate with the epidemiological database. GLE013.5 Preferably, the test module also has digital memory for storing epidemiological data. -56 - 201209402 GLE013.6 Preferably, the test module also has a plurality of reagent reservoirs, the reagent reservoirs containing reagents for processing the sample, wherein the data stored in the digital memory includes the reagent identification . GLE013.7 Preferably, the data stored in the digital memory includes a unique identifier for the test module. GLE013.8 Preferably, the sample is a biological material-containing organism

A sample, one of the functional portions, is used to amplify a polymerase chain reaction (PCR) portion of the nucleic acid sequence in the sample. GLE013.9 Preferably, the test module also has a CMOS circuit and a temperature sensor, wherein the CMOS circuit has digital memory and uses the output of the temperature sensor to feedback control the PCR portion. GLE013.10 Preferably, the test module also has a probe array for hybridization with a target nucleic acid sequence in an amplicon from the PCR portion. GLE013.il Preferably, each probe is configured to form a probe-target hybrid with a complementary target nucleic acid sequence (included in the amplicon), the various probe-target hybrid systems are organized The photons are emitted in response to the excitation, and the CMOS circuit has a light sensor to sense photons emitted by the probe-target hybrid. GLE013.12 Preferably, the test module also has an array of hybridization chambers for interposing probes such that the probes within each of the hybridization chambers are configured to hybridize to one of the target nucleic acid sequences. GLE013.13 Preferably, the light sensor is associated with a photodiode array of the hybridization chamber. GLE013.14 Preferably, the CMOS circuit is configured to communicate -57-201209402 hybrid data to an external device. GLE0 1 3.15 Preferably, the sample is drawn from a patient and the C Μ Ο S circuit is configured to download and store the patient data in the digital memory. GLE013.16 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the CMOS circuit. GLE01 3.1 7 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary driving of the liquid Flow, the heater is used to boil the liquid to release the meniscus from the meniscus anchor to restore the capillary drive flow. GLE013.18 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent the edge of the hole. GLE013.19 Preferably, the test module also has a LOC device having a sample inlet, a support substrate, a microsystem technology (MST) layer in fluid communication with the container, between the support substrate and the MS T layer Between the CMOS circuit and the upper cover, wherein the CMOS circuit has the digital memory, the communication interface, the controller, the MST layer has the functional portion, and the upper cover covers the MST layer and defines the reagent reservoir . GLE013.20 Preferably, the reagent reservoirs each have a surface tension valve for setting a meniscus anchor, the meniscus anchor used to form a meniscus to retain the reagent therein until contact When the sample stream is removed to remove the meniscus, the reagent is combined with the sample. -58- 201209402 This easy-to-use, mass-produced, inexpensive and portable diagnostic test module accepts biochemical samples and processes and analyzes the sample, updating epidemiological data based on the diagnostic results from the sample and the test site data. Library.

Updating the epidemiological database with this diagnostic result and location data provides an enhanced scientific basis for functionalizing the diagnostic test module and the ideal high response to epidemiological conditions. The automated update database based on the diagnostic test module will provide a number of quality and economic benefits for the health information system. GLE014.1 This aspect of the invention provides a test module for analyzing a sample fluid and communicating with a medical library, the test module comprising: a container for receiving the sample; a functional portion, which is used Processing and analyzing the sample; a communication interface for communicating with the medical database; and a controller for operating and controlling the communication interface. GLE014.2 Preferably, the test module of claim 1 further comprises a universal serial bus (USB) connector, wherein the communication interface is a USB device driver for operating and controlling the USB connector to External device communication. GLE014.3 Preferably, the test module of claim 2 includes a digital memory, wherein the medical database stores an electronic health record (EHR), an electronic medical record (EMR), and a personal health record (PHR). And the digital memory system is configured to store data related to EHR, EMR, and PHR. 201209402 GLE01 4.4 Preferably, the test module according to claim 3 of the patent application further comprises a plurality of reagent reservoirs, the reagent reservoirs containing reagents for processing the samples, wherein the data stored in the digital memory This reagent identification is included. GLE014.5 Preferably, the data stored in the digital memory includes a unique identifier for the test module. GLE01 4.6 Preferably, the sample contains cells of different sizes

The biological sample, one of the functional portions, is a polymerase chain reaction (PCR) portion for amplifying a nucleic acid sequence in the sample. GLE0 14.7 Preferably, the test module according to claim 6 further comprises a CMOS circuit and a temperature sensor, wherein the CMOS circuit has a digital body and uses the output of the temperature sensor for feedback control. This PCR section. GLE014.8 Preferably, one of the functional units is a dialysis department

The dialysis section is configured to separate a sample of the cells larger than the predetermined valve to a portion of the sample that is treated separately from the remaining sample containing only cells smaller than the predetermined valve. GLE014.9 Preferably, one of the functional portions is a lysis unit that is configured to release a nucleic acid sequence within a cell that is less than a predetermined valve 歹1J ^ GLE0 14.10. Preferably, The test module of claim 9 of the patent application further comprises a probe array for hybridization with the target nucleic acid sequence in the amplicon from the PCR portion. GLE014.il Preferably, each probe is configured to form a probe-target hybrid with the nucleic acid sequence of the complementary -60-201209402 (included in the amplicon). The system is configured to emit photons in response to excitation, and the CMOS circuit has a light sensor to sense photons emitted by the probe-target hybrid. GLE01 4.1 2 Preferably, the test module of claim 11 further comprises a hybridization chamber array for containing probes to organize the probes within each of the hybridization chambers to One of the target nucleic acid sequences

Hybrid. GLE014.13 Preferably, the light sensor is configured to register a photodiode array of the hybridization chamber. GLE014.14 Preferably, the C Μ Ο S circuit is organized to communicate hybridization data to an external device. GLE014.15 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the CMOS circuit.

GLE014.16 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary driving of the liquid Flow, the heater is used to boil the liquid to release the meniscus from the meniscus anchor to restore the capillary drive flow. GLE014.17 Preferably, the meniscus anchor is a hole having a ring shape and located adjacent to the edge of the hole. GLE014.18 Preferably, the test module of claim 5 is Also included is a LOC device comprising a sample inlet, a support substrate, a microsystem technology (M ST) layer, a CMOS circuit and an upper cover interposed between the support substrate and the MST layer, wherein the LOC device is in communication with the container fluid 201209402, wherein The CMOS circuit has the digital memory, the communication interface, the controller, the MST layer has the functional portion, and the upper cover covers the MST layer and defines the reagent reservoirs.

GLE014.19 Preferably, the reagent reservoirs each have a surface tension valve configured to define a meniscus anchor, the meniscus anchor used to form a meniscus to retain the reagent therein until contact When the sample stream is removed to remove the meniscus, the reagent is combined with the sample. GLE014.20 Preferably, the PCR portion has a thermal cycle time of less than 4 seconds. The easy-to-use, mass-produced, inexpensive, and portable diagnostic test module accepts biochemical samples and processes and analyzes the sample, and updates the patient's database based on the diagnostic results derived from the sample.

Updating the patient's database with the diagnosis and location information to provide improved patient health care services, automatically maintaining the medical record of the patient, and improving the scientific basis for functionalizing the diagnostic test module and epidemiological status The ideal is highly reactive. The automated update database based on the diagnostic test module will provide a number of quality and economic benefits to the health information system. GRE00 1.1 This aspect of the invention provides a microfluidic test module for analyzing a sample fluid and communicating test results with a mobile phone, the microfluidic test module comprising: a housing having a container for receiving a sample; a functional portion, It is used to process and analyze the sample; -62- 201209402 communication interface for communicating with the mobile phone: and a controller for operating and controlling the communication interface. GRE001.2 Preferably, the communication interface is a universal serial bus (USB) device driver operative to control a USB connector that communicates with an external device in the test module. GRE001.3 Preferably, the USB device driver is configured to draw power from the mobile phone to supply power to the controller and the functional portion.

GRE 00 1.4 Preferably, the controller is otherwise configured to operate to control the functional portion during processing and analysis of the sample. GRE00 1.5. Preferably, the controller is configured to download data via the mobile phone. GRE 01.6 Preferably, the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing identification data for the reagents. GRE001.7 Preferably, the data stored in the digital memory is used for the unique identifier of the microfluidic test module. GRE001.8 Preferably, the sample is a biological sample containing a genetic material, and one of the functional portions is a nucleic acid amplification portion for amplifying a nucleic acid sequence in the genetic material. GRE001.9 Preferably, the nucleic acid amplification portion is a polymerase chain reaction (PCR) portion and the data stored in the digitizer includes thermal cycle time and number of cycles.

GRE001.10 Preferably, the functional portions include a culture portion located upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir, -63-201209402, the culture portion has a heater to maintain the The culture temperature of the mixture of the sample and the restriction enzyme during the digestion of the nucleic acid sequence by the restriction enzyme. GRE001.11 Preferably, the microfluidic test module also has a probe array for hybridization to a target nucleic acid sequence in an amplicon from the PCR portion.

GRE001.12 Preferably, the data stored in the digital memory includes probe identification data to identify probes at various locations within the array of probes. GRE00 1 · 1 3 Preferably, the microfluidic test module also has a light sensor, wherein each probe is configured to form a probe with a complementary nucleic acid sequence (included in the amplicon). The subject hybrids are configured to emit photons in response to input, and the light sensors are configured to sense photons emitted by the probe-target hybrid.

GRE001.14 Preferably, the data stored in the digital memory comprises hybridization data generated from the light sensor output. GRE001.15 Preferably, the microfluidic test module also has a hybridization chamber array for interposing probes such that the probes within each hybridization chamber are organized to hybridize to one of the target nucleic acid sequences. GRE001.16 Preferably, the light sensor is associated with a photodiode array of the hybridization chamber. GRE001.17 Preferably, the controller is configured to transmit the hash material to the mobile phone. GRE001.18 Preferably, the sample is drawn from a patient, and the controller is configured to download patient data via the mobile phone and store the patient data in the digital memory. GRE001.19 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the controller.

GRE001.20 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary driving of the liquid Flow, the heater is used to boil the liquid to release the meniscus from the meniscus anchor to restore the capillary drive flow. The easy to use, mass produced, inexpensive, lightweight and portable The fluid diagnostic test module has a complete set of reagents required to receive the sample and process and analyze the sample material using the integrated sensor of the module, and provide an electronic result at the output of the sample. The module's communication system is connected to a mobile phone/smartphone. The mobile phone/smart phone provides power, computing, communication and user interface support for the module. Mobile phones/smart phones are widely and inexpensively available, eliminating the need for dedicated, cumbersome and expensive modular support systems. GRE002.1 This aspect of the invention provides a microfluidic test module for analyzing a sample fluid and communicating test results with a laptop, the microfluidic test module comprising: a §& a housing; a functional unit for processing and analyzing the sample; a communication interface for communicating with the laptop; and a -65-201209402 controller for controlling the communication interface. GRE002.2 Preferably, the communication interface is a universal serial bus (USB) device driver operative to control a USB connector that communicates with an external device in the test module. GRE002.3 Preferably, the USB device driver is configured to draw power from the laptop to supply power to the controller and the functional unit

GRE 002.4 Preferably, the controller is otherwise configured to operate to control the functional portion during processing and analysis of the sample. GRE002.5 Preferably, the controller is configured to download data via the laptop. GRE002.6 Preferably, the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing identification data for the reagents. GRE002.7 Preferably, the data stored in the digital memory

A unique identifier for the microfluidic test module. GRE002.8 Preferably, the sample is a biological sample containing a genetic material, and one of the functional portions is a nucleic acid amplification portion for amplifying a nucleic acid sequence in the genetic material. GRE002.9 Preferably, the nucleic acid amplification portion is a polymerase chain reaction (PCR) portion and the data stored in the digital memory includes thermal cycle time and number of cycles.

GRE002.1 0 Preferably, the functional portions include a culture portion located upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir, -66-201209402, the culture portion has a heater to maintain the The culture temperature of the mixture of the sample and the restriction enzyme during the digestion of the nucleic acid sequence by the restriction enzyme. GRE 002.1 1 Preferably the microfluidic test module also has a probe array for hybridization to a target nucleic acid sequence in an amplicon from the PCR portion. GRE 002.12 Preferably, the data stored in the digital memory includes probe identification data to identify probes at various locations within the array of probes. GRE002.1 3 Preferably, the microfluidic test module also has a light sensor, wherein each probe is configured to form a probe-target with a complementary nucleic acid sequence (included in the amplicon) Hybrids, the various probe-target hybridization systems are configured to emit photons in response to input, and the light sensor is configured to sense photons emitted by the probe-target hybrid. GRE002.1 4 Preferably, the data packet stored in the digital memory includes the hybrid data generated by the light sensor output. GRE002.1 5 Preferably, the microfluidic test module also has an array of hybridization chambers for interposing probes such that probes within each hybridization chamber are configured to hybridize to one of the target nucleic acid sequences . GRE002.1 6 Preferably, the light sensor is associated with a photodiode array of the hybridization chamber. GRE 002.1 7 Preferably, the controller is configured to transmit the hash data to the laptop. GRE002. 1 8 Preferably, the sample is taken from a patient, and the controller is configured to download patient data via the laptop and store the patient data in the digital memory. body. GRE 002.1 9 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the controller.

GRE002.20 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary driving of the liquid Flow, the heater is used to boil the liquid to release the meniscus from the meniscus anchor to recover the capillary drive flow.

The easy-to-use, mass-produced, inexpensive, lightweight, and portable microfluidic diagnostic test module has a complete library of reagents required to receive samples and process and analyze the sample material using the integrated sensor of the module. And provide electronic results at the output of the other. The communication field of the module is interfaced with a laptop/notebook, which provides power, calculation, communication and user interface support for the module. The laptop/notebook system is widely and inexpensively available, eliminating the need for a dedicated, cumbersome and expensive modular support system. GRE003.1 This aspect of the invention provides a microfluidic test module for analyzing a sample fluid and communicating test results with a dedicated reader specifically constructed for the microfluidic test module, the microfluidic test module comprising: a housing having a sample receiving container: a functional portion for processing and analyzing the sample; a communication interface for communicating with the dedicated reader; and -68-201209402 controller for operating control Communication interface. GRE003.2 Preferably, the communication interface is a universal serial bus (USB) device driver operative to control a USB connector that communicates with an external device in the test module. GRE003.3 Preferably, the USB device driver is configured to draw power from the dedicated reader to supply power to the controller and the functional unit

GRE003.4 Preferably, the controller is otherwise configured to operate to control the functional portion during processing and analysis of the sample. GRE003.5 Preferably, the controller is configured to download data via the dedicated reader. GRE003.6 Preferably, the microfluidic test module also has a plurality of reagent reservoirs, the reagent reservoirs containing A reagent for processing the sample and a digital body for storing the identification data of the reagent. GRE003.7 Preferably, the data stored in the digital memory is φ for the unique identifier of the microfluidic test module. GRE003.8 Preferably, the sample is a biological sample containing a genetic material, and one of the functional portions is a nucleic acid amplification portion for amplifying a nucleic acid sequence in the genetic material. GRE003.9 Preferably, the nucleic acid amplification part is a polymerase chain reaction (PCR) part and the data stored in the digital body includes thermal cycle time and number of cycles.

GRE003.1 0 Preferably, the functional portions include a culture portion located upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir, -69-201209402, the culture portion has a heater to maintain The mixture of the sample and the restriction enzyme is cultured during the digestion of the nucleic acid sequence by the restriction enzyme. GRE003.1 1 Preferably, the microfluidic test module also has a probe array for hybridization with a target nucleic acid sequence in an amplicon from the PCR portion.

GRE003.1 2 Preferably, the data stored in the digital memory includes probe identification data to identify probes at various locations within the array of probes. GRE003.1 3 Preferably, the microfluidic test module also has a light sensor, wherein each probe is configured to form a probe-target with a complementary nucleic acid sequence (included in the amplicon) Hybrids, the various probe-target hybridization systems are configured to emit photons in response to input, and the light sensor is configured to sense photons emitted by the probe-target hybrid.

GRE003.1 4 Preferably, the data stored in the digital body includes the hybrid data generated from the light sensor output. GRE003.1 5 Preferably, the microfluidic test module also has an array of hybridization chambers for interposing probes such that probes within each hybridization chamber are configured to hybridize to one of the target nucleic acid sequences . GRE003.1 6 Preferably, the light sensor is a register of the photodiode array of the hybridization chamber. GRE 003.1 7 Preferably, the controller is configured to transmit the hybrid data to the dedicated reader. GRE003.1 8 Preferably, the sample is taken from a patient, and the controller is configured to download patient data and store the patient data in the digital memory via the dedicated reader. . GRE003.1 9 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the controller" GRE003.20 Preferably, the active valve system is provided with a meniscus anchor

And a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary flow of the liquid, the heater being used to boil the liquid to anchor from the meniscus The meniscus is released at the device to restore the capillary drive flow. The easy-to-use, mass-produced, inexpensive, lightweight, and portable microfluidic diagnostic test module has a complete library of reagents required to receive samples and process and analyze the sample material using the integrated sensor of the module. And provide electronic results at the output of the other. The module's communication system is interfaced with a non-expensive and portable dedicated reader that provides power, computing, communication and user interface support for the module. This dedicated reader eliminates the need for cumbersome and expensive modular support systems. GRE004.1 This aspect of the invention provides a microfluidic test module for analyzing a sample fluid and communicating test results with a desktop computer, the microfluidic test module comprising: a housing having a container for receiving a sample; a functional unit for processing and analyzing the sample; a communication interface for communicating with the desktop computer; and a controller for operating and controlling the communication interface. -71 - 201209402 GRE004.2 Preferably, the communication interface is a universal serial bus (USB) device driver for operating a USB connector that communicates with an external device in the test module. GRE004.3 Preferably, the USB device driver is configured to draw power from the desktop computer to supply power to the controller and the function portion GRE004.4. Preferably, the controller is otherwise configured In this way

The function is controlled during the processing and analysis of the product. GRE004.5 Preferably, the controller is configured to download data via the desktop computer. GRE004.6 Preferably, the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing identification data for the reagents. GRE004.7 Preferably, the data stored in the digital memory is a unique identifier for the microfluidic test module.

GRE004.8 Preferably, the sample is a biological sample containing a genetic material, and one of the functional portions is a nucleic acid amplification portion for amplifying a nucleic acid sequence in the genetic material. GRE004.9 Preferably, the nucleic acid amplification part is a polymerase chain reaction (PCR) part and the data stored in the digital body includes thermal cycle time and number of cycles.

GRE004.1 0 Preferably, the functional portions include a culture portion located upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir having a heater to maintain the sample and the restriction enzyme The mixture is subjected to a culture temperature during which the restriction enzyme digests the nucleic acid sequence at -72 to 201209402. GRE004.1 1 Preferably, the microfluidic test module also has a probe array for hybridization to a target nucleic acid sequence in an amplicon from the PCR portion. GRE004.12 Preferably, the data stored in the digital memory includes probe identification data to identify probes at various locations within the array of probes.

GRE004.1 3 Preferably, the microfluidic test module also has a light sensor, wherein each probe is configured to form a probe-target with a complementary nucleic acid sequence (included in the amplicon) Hybrids, the various probe-target hybridization systems are configured to emit photons in response to input, and the light sensor is configured to sense photons emitted by the probe-target hybrid. GRE004.14 Preferably, the data stored in the digital memory comprises hybridization data generated from the light sensor output. GRE004.1 5 Preferably, the microfluidic test module also has an array of hybridization chambers for interposing probes such that probes within each hybridization chamber are configured to hybridize to one of the target nucleic acid sequences . GRE004.16 Preferably, the light sensor is associated with a photodiode array of the hybridization chamber. GRE004.1 7 Preferably, the controller is configured to transmit the hash data to the desktop computer. GRE004.1 8 Preferably, the sample is taken from a patient, and the controller is configured to download patient data via the desktop computer and store the patient data in the digital memory. . GRE004.1 9 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the controller.

GRE004.20 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary driving of the liquid Flow, the heater is used to boil the liquid to release the meniscus from the meniscus anchor to restore the capillary drive flow.

The easy-to-use, mass-produced, inexpensive, lightweight, and portable microfluidic diagnostic test module has a complete library of reagents required to receive samples and process and analyze the sample material using the integrated sensor of the module. And provide electronic results at the output of the other. The module's communication system is interfaced with a desktop computer that provides power, computing, communication and user interface support for the module. Desktop computers are widely available and inexpensive, eliminating the need for dedicated, cumbersome and expensive modular support systems. GRE005.1 This aspect of the invention provides a microfluidic test module for analyzing a sample fluid and communicating test results with an e-book reader, the microfluidic test module comprising: a housing having a container for receiving a sample; The department is for processing and analyzing the sample; the communication interface is for communicating with the e-book reader; and the controller is for operating and controlling the communication interface. GRE005.2 Preferably, the communication interface is a universal serial confluence -74-201209402 row (USB) device driver for operating a USB connector that communicates with the external device in the test module. GRE 00 5.3 Preferably, the USB device driver is configured to draw power from the e-book reader to supply power to the controller and the functional portion. GRE 005.4 Preferably, the controller is otherwise configured to operate to control the functional portion during processing and analysis of the sample.

GRE005.5 Preferably, the controller is configured to download data via the e-book reader. GRE005.6 Preferably, the microfluidic test module also has a plurality of reagent reservoirs, the reagent reservoirs Contains reagents for processing the sample and digital memory for storing identification data for the reagent. GRE005.7 Preferably, the data stored in the digitizer is a unique identifier for the microfluidic test module. GRE005.8 Preferably, the sample is a biological φ sample containing a genetic material, and one of the functional portions is a nucleic acid amplification portion for amplifying a nucleic acid sequence in the genetic material. GRE005.9 Preferably, the nucleic acid amplification part is a polymerase chain reaction (PCR) part and the data stored in the digital memory includes a thermal cycle time and a cycle number of $ GRE005.1 0. Preferably, the functional parts Included in the culture portion upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir having a heater to maintain a mixture of the sample and the restriction enzyme during the restriction enzyme digestion of the nucleic acid sequence Culture temperature. -75- 201209402 GRE005.1 1 Preferably, the microfluidic test module also has a probe array for hybridization with a target nucleic acid sequence in an amplicon from the PCR portion. GRE005.1 2 Preferably, the data stored in the digital memory includes probe identification data to identify probes at various locations within the array of probes.

GRE005.1 3 Preferably, the microfluidic test module also has a light sensor, wherein each probe is configured to form a probe-target with a complementary target nucleic acid sequence (included in the amplicon) Hybrids, the various probe-target hybridization systems are configured to emit photons in response to an input, and the light sensor is configured to sense photons emitted by the probe-target hybrid. GRE005.1 4 Preferably, the data stored in the digital body includes the hybrid data generated from the light sensor output.

GRE005.1 5 Preferably, the microfluidic test module also has an array of hybridization chambers for interposing probes such that probes within each hybridization chamber are organized to hybridize to one of the target nucleic acid sequences . GRE005.1 6 Preferably, the light sensor is a register of the photodiode array of the hybridization chamber. GRE 005.1 7 Preferably, the controller is configured to transmit the hash data to the e-book reader. GRE 005.1 8 Preferably, the sample is drawn from a patient and the controller is configured to download patient data via the e-book reader and store the patient data in the digital memory. -76- 201209402 GRE005.1 9 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to flow to the hybridization chamber in response to an activation signal from the controller . GRE005.20 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary driving of the liquid Flow, the heater is used to boil the liquid to release the meniscus from the meniscus anchor to restore capillary

Drive the stream. The easy-to-use, mass-produced, inexpensive, lightweight, and portable microfluidic diagnostic test module has a complete library of reagents required to receive samples and process and analyze the sample material using the integrated sensor of the module. And provide electronic results at the output of the other. The communication system of the module is interfaced with an e-book reader, and the e-book reader provides power, calculation, communication and user interface support of the module. The e-book reader system can be widely and inexpensively obtained, and is exempt from special use. The cumbersome and expensive module support system needs. GRE007.1 This aspect of the invention provides a microfluidic test module for analyzing a sample fluid and communicating test results with a tablet computer, the microfluidic test module comprising: a housing having a container for receiving a sample; a functional portion, It is used to process and analyze the sample; a communication interface for communicating with the tablet; and a controller for operating and controlling the communication interface. GRE007.2 Preferably, the communication interface is a universal serial bus (USB) device driver for operating and controlling a USB connector in the test module that communicates with the external device -77-201209402. GRE007.3 Preferably, the USB device driver is configured to draw power from the tablet to supply power to the controller and the functional portion. GRE 07.4 Preferably, the controller is otherwise configured to operate to control the functional portion during processing and analysis of the sample. GRE007.5 Preferably, the controller is configured to download data via the tablet computer.

GRE007.6 Preferably, the microfluidic test module also has a plurality of reagent reservoirs containing reagents for processing the sample and digital memory for storing identification data of the reagents. GRE007. 7 Preferably, the data stored in the digital memory is

I Unique identifier for microfluidic test modules. GRE007.8 Preferably, the sample is a biological sample containing a genetic material, and one of the functional portions is a nucleic acid amplification portion for amplifying a nucleic acid sequence in the genetic material.

GRE007.9 Preferably, the nucleic acid amplification part is a polymerase chain reaction (PCR) part and the data stored in the digital memory includes thermal cycle time and number of cycles.

GRE007.1 0 Preferably, the functional portions include a culture portion located upstream of the PCR portion and one of the reagent reservoirs is a restriction enzyme reservoir having a heater to maintain the sample and the restriction enzyme The mixture is at a culture temperature during which the restriction enzyme digests the nucleic acid sequence. GRE007.1 1 Preferably, the microfluidic test module also has a probe array for hybridization with the target nucleic acid sequence -78-201209402 in the amplicon from the PCR portion. GRE 007.12 Preferably, the data stored in the digital memory includes probe identification data to identify probes at various locations within the array of probes.

GRE007.1 3 Preferably, the microfluidic test module also has a light sensor, wherein each probe is configured to form a probe-target with a complementary nucleic acid sequence (included in the amplicon) Hybrids, the various probe-target hybridization systems are configured to emit photons in response to input, and the light sensor is configured to sense photons emitted by the probe-target hybrid. GRE007.14 Preferably, the data stored in the digital memory comprises hybridization data generated from the light sensor output. GRE007.1 5 Preferably, the microfluidic test module also has an array of hybridization chambers for interposing probes such that probes within each hybridization chamber are organized to hybridize to one of the target nucleic acid sequences . GRE007.1 6 Preferably, the light sensor is associated with a photodiode array of the hybridization chamber. GRE007.1 7 Preferably, the controller is configured to transmit the hash data to the tablet. GRE007.1 8 Preferably, the sample is drawn from a patient and the controller is configured to download patient data via the tablet and store the patient data in the digital memory. GRE007.1 9 Preferably, the PCR portion has an active valve for retaining liquid in the PCR portion during thermal cycling and allowing the liquid to respond to -79-201209402 from the activation signal of the controller to the hybridization room. GRE007.20 Preferably, the active valve is provided with a meniscus anchor and a boiling start valve of the heater, the meniscus anchor is configured to form a meniscus to prevent capillary driving of the liquid Flow, the heater is used to boil the liquid to release the meniscus from the meniscus anchor to restore the capillary drive flow.

The easy-to-use, mass-produced, inexpensive, lightweight, and portable microfluidic diagnostic test module has a complete library of reagents required to receive samples and process and analyze the sample material using the integrated sensor of the module. And provide electronic results at the output of the other. The module's communication system is interfaced with a tablet computer that provides power, computing, communication and user interface support for the module. Tablet PCs are widely and inexpensively available, eliminating the need for dedicated, cumbersome and expensive modular support systems. GMV001.1 This aspect of the invention provides a reagent microbottle for a reagent dispensing device comprising:

a container for containing a volume of reagent; a digitizer for storing information about the reagent; a droplet generator for droplets ejecting the reagent from the container; and an electrical contact And for connecting to a control processor in the reagent dispensing device to receive a drive pulse for the drop generator and to provide data to the control processor. GMV001.2 Preferably, the container holds between 28 and 2 microliters of reagent. GMV00 1.3 Preferably, the droplet generator is configured to emit droplets of between -50 and 201209402 in volume from 50 picoliters to 150 picoliters. GMV001.4 Preferably, the droplet generator has a piezoelectric actuator GMV001.5. Preferably, the data includes identification data transmitted to the control processor. GMV001.6 Preferably, the identification data distinguishes the unique identification data of the micro-bottle with all other micro-bottles.

GMV001.7 Preferably, the data is encrypted. The reagent microbottle with non-volatile digital memory is used to receive reagents, store reagents, and dispense reagents using a droplet generator under digital control. The droplet generator provides a self-contained, volumetric and positionally accurate reagent dispensing technique for one of the microbottles to simplify the complexity of the automated manufacturing environment in which the microbottle is utilized and to increase the reliability of the automated manufacturing environment utilizing the microbottle. And the cost of an automated manufacturing environment that utilizes the micro-bottle. The digital memory system is used to store the information needed to functionalize the device in an automated manufacturing environment. The digital memory system provides a simple, reliable, safe, safe and inexpensive reagent dispensing method for one of the microbottles in an automated manufacturing environment. GMV002.1 This aspect of the invention provides a reagent microbottle for a reagent dispensing device, the microbottle comprising: a container for containing a volume of reagent; an integrated circuit having a means for storing The digital memory of the identification data of the bottle; and -81 - 201209402 electrical contact' is used to connect the control processor in the reagent dispensing device to provide information to the control processor for identification with the real micro-bottle Comparison of the list of information. GMV002.2 Preferably, the micro vial also has a droplet generator for ejecting reagent droplets from the container. GMV002.3 Preferably, the container holds between 282 microliters and 400 microliters of reagent.

GMV002.4 Preferably, the droplet generator is configured to emit droplets having a volume between 50 picoliters and 150 picoliters. GMV002.5 Preferably, the droplet generator has a piezoelectric actuator GMV002.6. Preferably, the identification data distinguishes the unique identification data of the micro-bottle with all other micro-bottles. GMV002.7 Preferably, the data is encrypted.

The reagent microbottle with the verification integrated circuit is used to accept reagents, store reagents, and dispense reagents using a droplet generator under digital control. The droplet generator provides a self-contained, volumetric and positionally accurate reagent dispensing technique for one of the microbottles to simplify the complexity of the automated manufacturing environment in which the microbottle is utilized and to increase the reliability of the automated manufacturing environment utilizing the microbottle. And reduce the cost of an automated manufacturing environment that utilizes the micro vials. The verification integrated circuit is used to store micro-bottle validation information used during functionalization of the device in an automated manufacturing environment. The verification integrated circuit provides a simple, reliable, safe, secure, and inexpensive reagent dispensing method for one of the micro vials in an automated manufacturing environment. -82- 201209402 GMV003.1 This aspect of the invention provides a reagent micro-bottle for a reagent dispensing device, the micro-bottle comprising: a container for holding a volume of reagent; an integrated circuit having digital memory Body to store reagent information about the characteristics of the reagent; and

An electrical contact is provided for connection to a control processor in the reagent dispensing device to provide data to the control processor for download to a device having the reagent supply. GMV003.2 Preferably, the micro vial also has a droplet generator for ejecting reagent droplets from the container. GMV003.3 Preferably, the container holds between 282 microliters and 400 microliters of reagent. GMV003.4 Preferably, the droplet generator is configured to eject droplets having a volume between 50 picoliters and 150 picoliters. GMV003.5 Preferably, the droplet generator has a piezoelectric actuator GMV003.6. Preferably, the identification data distinguishes the unique identification data of the micro-bottle with all other micro-bottles. GMV003.7 Preferably, the data is encrypted. The reagent microbottle with non-volatile digital memory is used to receive reagents, store reagents, and dispense reagents using a droplet generator under digital control. The droplet generator provides a self-contained and volumetric and accurate reagent dispensing technique for one part of the micro-bottle to simplify the complexity of the automated manufacturing environment utilizing the micro-bottle and to increase the automated manufacturing of the micro-bottle using the micro-bottle-83 - 201209402 Reliability of the environment and the cost of reducing the automated manufacturing environment in which the micro vials are utilized

The digital memory system is used to store the information needed to functionalize the device in an automated manufacturing environment. The information stored in the memory includes the reagent specification data written into the body by the department of the automated manufacturing environment. This information is read from the memory and is utilized as needed by other parts of the automated manufacturing environment. The digital memory system provides a simple, reliable, safe, secure, and inexpensive reagent dispensing method for one of the micro vials in an automated manufacturing environment. GMV004.1 This aspect of the invention provides an oligonucleotide microbottle for an oligonucleotide dispensing device, the microbottle comprising: a container for containing a volume of reagent; an integrated circuit having Digital memory to store oligonucleotide information about the identity of the oligonucleotide: and

An electrical contact is provided for connection to a control processor in the reagent dispensing device to provide information to the control processor for download to a device having the oligonucleotide supply. GMV004.2 Preferably, the micro vial also has a droplet generator for ejecting oligonucleotide droplets from the container. GMV004.3 Preferably, the container holds between 282 microliters and 400 microliters of reagent. GMV004.4 Preferably, the droplet generator is configured to emit droplets having a volume between 50 picoliters and 150 picoliters. GMV004.5 Preferably, the droplet generator has a piezoelectric actuator -84 - 201209402 GMV004.6 Preferably, the identification data distinguishes the unique identification data of the micro vial from all other micro vials. GMV004.7 Preferably, the data is encrypted.

The oligonucleotide microbottle with non-volatile digital memory is used to receive reagents, store reagents, and dispense reagents using a droplet generator under digital control. The droplet generator provides a self-contained, volumetric and positionally accurate oligonucleotide dispensing technique for one part of the micro-bottle, simplifies the complexity of the automated manufacturing environment in which the micro-bottle is utilized, and increases the automated manufacturing environment in which the micro-bottle is utilized. Reliability and reduced cost of an automated manufacturing environment that utilizes the micro vials. The digital memory system is used to store the information needed to functionalize the device in an automated manufacturing environment. The information stored in the memory includes the oligonucleotide specifications written into the memory by the department of the automated manufacturing environment. This information is read from the memory and is utilized as required by the department of the automated manufacturing environment. The digital memory system provides a simple, reliable, safe, secure, and inexpensive reagent dispensing method for one of the micro vials in an automated manufacturing environment. GRD00 1.1 This aspect of the invention provides a reagent dispensing device for loading a reagent to a microfluidic device having digital memory for data of reagents loaded in the microfluidic device, the reagent dispensing device comprising: A plurality of reagent bottles each having an integrated circuit and a droplet dispenser having a memory for storing information about the reagents in the bottle - 85 - 201209402 mounting surface for detachable Installing the microfluidic device for movement of the microfluidic device relative to the bottle; and controlling a processor for operating the bottle and the mounting surface; wherein the control processor is configured to activate the The droplet dispenser of the selected bottle moves the bottle to register with the microfluidic device and download data from the integrated circuit to the digital memory of the microfluidic device. GRD 001.2 Preferably, the reagent dispensing device also has a camera to optically feedback the registration between the bottle selected by the control processor and the microfluidic device. GRD00 1.3 Preferably, the bottle is for holding a micro vial of between 282 ml and 400 ml. GRD001.4 Preferably, the integrated circuit of each of the micro vials has a unique identifier for individually identifying each of the micro vials, the unique identifier being transmitted to the control processor and the digital memory of the microfluidic device. GRD001.5 Preferably, each of the micro vials has electrical contacts for receiving a firing pulse for the droplet dispenser and allowing the control processor to interrogate the integrated circuit. GRD 01.6 Preferably, the reagent dispensing device also has a rack in which the micro vials are removably mounted for electromechanical control of the micro vials. GRD001 . 7

A platform on which the orthogonal axes move, the frame extending parallel to one of the orthogonal axes. -86- 201209402 GRD001.8 Preferably, the microfluidic device is a laboratory on wafer (LOC) device. GRD001.9 Preferably the droplet dispenser has a piezoelectric actuator GRD001.10. Preferably, the droplet dispenser is configured to emit droplets having a volume between 50 picoliters and 150 picoliters.

GRD001.il Preferably, the reagent dispensing apparatus also has a facility configured to apply a sealing device to the LOC device to enclose a plurality of reservoirs in which the reagents have been loaded. GRD001.12 Preferably, the LOC has a polymerase chain reaction (PCR) moiety and the reagent list has one or more of the following: water, polymerase, primer, buffer, anticoagulant, deoxyribonucleoside III Phosphoric acid (dNTP), lysing reagent, ligase, linker and restriction enzyme. The reagent dispensing device is part of a cost effective automated mass production environment that is used to dispense reagents contained in the reagent micro vials to the reagent reservoir of the φ microfluidic device. The data provided by the reagent dispensing device includes automated computer controlled dispensing of the reagent to the reagent reservoir of the microfluidic device, inspection of reagent data stored in the memory of the micro vial, and loading to the microfluidic The specification list of the reagents of the device is aligned, and the reagent data is stored to the memory of the microfluidic device. The reagent dispensing device provides automated and volumetric and positionally accurate reagent dispensing techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, increase the security of the automated manufacturing environment, and increase the automated manufacturing environment. Safety and reduce the cost of this automated manufacturing cycle -87- 201209402. The data provided by the reagent dispensing device automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment.

GRD002.1 This aspect of the invention provides a reagent dispensing device for loading a reagent to a microfluidic device, the microfluidic device having digital memory for data of reagents loaded in the microfluidic device, the reagent dispensing device comprising : a plurality of reagent bottles each having an integrated circuit and a droplet dispenser having a memory mounting surface for storing information about reagents in the bottle for detachably mounting the a microfluidic device for moving the microfluidic device relative to the bottle; and a control processor for operating the bottle and the mounting surface;

The control processor is configured to automatically interrogate each of the integrated circuits to collect and store information about the reagents in each of the bottles. GRD002.2 Preferably, the control processor is configured to automatically activate a droplet dispenser of the selected bottle, move the bottle i to register with the microfluidic device, and download information from the integrated circuit to The digital memory of the microfluidic device. GRD002.3 Preferably, the reagent dispensing device also has a camera to optically feedback the registration between the bottle selected by the control processor and the microfluidic device. -88- 201209402 GRD002.4 Preferably, the bottle is for containing a micro vial of between 282 ml and 400 ml. GRD002.5 Preferably, the integrated circuit of each of the micro vials has a unique identifier for individually identifying each of the micro vials, the unique identifier being transmitted to the control processor and the digital device of the microfluidic device . GRD002.6 Preferably, each of the micro vials has electrical contacts for receiving

The start pulse for the drop dispenser is received and the control processor is allowed to interrogate the integrated circuit. GRD002.7 Preferably, the reagent dispensing apparatus also has a shelf in which the micro vials are removably mounted for electromechanical control of the micro vials. GRD002.8 Preferably, the mounting surface is a platform that is configured to move along two orthogonal axes that extend parallel to one of the orthogonal axes. GRD002.9 Preferably, the microfluidic device is a on-wafer laboratory (LOC) device. GRD 002.10 Preferably, the droplet dispenser has a piezoelectric actuator GRD002.il. Preferably, the droplet dispenser is configured to emit droplets having a volume between 50 picoliters and 150 picoliters. GRD002.12 Preferably, the LOC has a polymerase chain reaction (PCR) moiety and the reagent list has one or more of the following: water, polymerase, primer, buffer, anticoagulant, deoxyribonucleoside III Phosphoric acid (dNTP), lysing reagent, ligase, linker and restriction enzyme. The reagent dispensing device is part of a cost effective automated mass production environment -89-201209402, which is used to dispense reagents contained in a reagent microbottle to a reagent reservoir of the microfluidic device. The data provided by the reagent dispensing device includes automated computer controlled dispensing of the reagent to the reagent reservoir of the microfluidic device, inspection of reagent data stored in the memory of the micro vial, and loading to the microfluidic The specification list of reagents of the device is compared, and the reagent data is stored to the memory of the microfluidic device and the computer memory of the reagent dispensing device.

The reagent dispensing device provides automated and volumetric and positionally accurate reagent dispensing techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, increase the security of the automated manufacturing environment, and increase the automated manufacturing environment. Safety and reduce the cost of this automated manufacturing environment. The data provided by the reagent dispensing device is automated to provide automated, safe, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment.

GRD003.1 This aspect of the invention provides a reagent dispensing device for loading reagents to a fixed array of microfluidic devices, each of the microfluidic devices having a digital memory of data relating to reagents loaded in the microfluidic device, The reagent dispensing device comprises: a plurality of reagent bottles, each of the bottles having an integrated circuit and a droplet dispenser, the integrated circuit having a memory mounting surface for storing information about the reagents in the bottle, which is used for Disassemblingly mounting a fixed array of the microfluidic device for movement of the fixed array of microfluidic devices relative to the bottle; -90-201209402 and a control processor for operating the bottle and the mounting surface; A control processor is configured to activate a droplet dispenser of the selected vial, move the vial to register with one or more microfluidic devices within the fixed array, and download data from the integrated circuit to the one Or digital memory of a plurality of microfluidic devices.

GRD003.2 Preferably, the fixed array of microfluidic devices are arrays of on-wafer laboratory (LOC) devices mounted on detachable PCB (printed circuit board) wafers. GRD003.3 Preferably, the reagent dispensing device also has a camera to optically feedback the registration between the bottle selected by the control processor and the LOC device. GRD003.4 Preferably, the bottle is for holding a micro vial of between 282 ml and 400 ml. GRD003.5 Preferably, the integrated circuit of each of the micro vials has a unique identifier for individually identifying each of the micro vials, the unique identifier being transmitted to the control processor and the digital memory of the LOC device. GRD 003.6 Preferably, each of the micro vials has electrical contacts for receiving a firing pulse for the droplet dispenser and allowing the control processor to interrogate the integrated circuit. GRD003.7 Preferably, the reagent dispensing apparatus also has a shelf in which the micro vials are removably mounted for electromechanical control of the micro vials. Preferably, the mounting surface is configured to move along two orthogonal axes, the frame extending parallel to one of the orthogonal axes. GRD 003.9 Preferably, the droplet dispenser has a piezoelectric actuator GRD003.1. Preferably, the droplet dispenser is configured to emit droplets having a volume between 50 picoliters and 150 picoliters.

GRD 003.il Preferably, the reagent dispensing apparatus also has a facility configured to apply a sealing device to the LOC device to enclose a plurality of reservoirs in which the reagents have been loaded. GRD003.1 2 Preferably, the LOC has a polymerase chain reaction (PCR) moiety and the reagent list has one or more of the following: water, polymerase, primer, buffer, anticoagulant, deoxyribonucleoside Triphosphate (dNTP), lysing reagent, ligase, linker and restriction enzyme.

GRD 003.1 3 Preferably, the control processor is configured to automatically query each of the integrated circuits to collect and store information about the reagents in each of the bottles. The reagent dispensing device is part of a cost effective automated mass production environment that is used to dispense reagents contained in a reagent microbottle to a reagent reservoir of the array of microfluidic devices mounted on the PCB wafer. The data provided by the reagent dispensing device includes automated computer controlled dispensing of the reagent to the reagent reservoir of the microfluidic device, inspection of reagent data stored in the memory of the micro vial, and loading to the microfluidic The specification list of reagents of the device is aligned, and the reagent data is stored to the memory of the microfluidic device. -92 - 201209402 This reagent dispensing device provides automated, volumetric and positionally accurate reagent dispensing techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, increase the security of the automated manufacturing environment, and increase the Automate the manufacturing environment and reduce the cost of the automated manufacturing environment. The data provided by the reagent dispensing device is automated to provide automated, safe, secure and inexpensive data monitoring and management in the automated manufacturing environment.

Distributing the reagent to the array of microfluidic devices mounted on the PCB wafer accelerates the loading process and reduces the cost of the loading process, and by mounting the microfluidic device onto the PCB wafer and soldering them Loading the reagent to the microfluidic device enhances the chemical and physical integrity of the reagent. GRD004.1 This aspect of the invention provides a reagent dispensing apparatus for loading reagents to a germanium wafer on which a wafer-on-lab (LOC) device is built, each LOC device having a loading on the LOC device a digital memory of the reagent of φ, the reagent dispensing device comprising: a plurality of reagent bottles each having an integrated circuit and a droplet dispenser, the integrated circuit having a reagent for storing the reagent in the bottle a memory of the data » mounting surface for detachably mounting the wafer for movement of the wafer relative to the bottle; and a control processor for operating the bottle and the mounting surface Wherein the control processor is configured to activate a droplet of the selected vial-93-201209402 adapter, move the vial to register with one or more LOC devices on the germanium wafer, and from the product The body circuit downloads data to the digital memory of the one or more L0C devices. GRD004.2 Preferably, the tantalum wafer is partially cut to prepare subdivided into separate LOC devices. GRD004.3 Preferably, the reagent dispensing device also has a camera

The registration between the bottle selected by the control processor and the LOC device is optically fed back. GRD004.4 Preferably, the bottle is for holding a micro vial of between 2 82 ml and 400 ml. GRD004.5 Preferably, the integrated circuit of each of the micro vials has a unique identifier for individually identifying each of the micro vials, the unique identifier being transmitted to the control processor and the digital memory of the LOC device. GRD004.6 Preferably, each of the micro vials has electrical contacts for receiving

The start pulse for the drop dispenser is received and the control processor is allowed to interrogate the integrated circuit. GRD004.7 Preferably, the reagent dispensing apparatus also has a rack in which the micro vials are removably mounted for electromechanical control of the micro vials. GRD004.8 Preferably, the mounting surface is a platform that is configured to move along two orthogonal axes that extend parallel to one of the orthogonal axes. GRD004.9 Preferably, the droplet dispenser has a piezoelectric actuator GRD004.1. Preferably, the droplet dispenser is configured to emit -94-201209402. The volume is between 50 picoliters and 150 skins. Drop the droplets. GRD 004.il Preferably, the reagent dispensing apparatus also has a facility configured to apply a sealing device to the LOC device to enclose a plurality of reservoirs in which the reagents have been loaded.

GRD004.1 2 Preferably, the LOC has a polymerase chain reaction (PCR) moiety and the reagent list has one or more of the following: water, polymerase, primer, buffer, anticoagulant, deoxyribonucleoside Triphosphate (dNTP), lysing reagent, ligase, linker and restriction enzyme. GRD004.13 Preferably, the control processor is configured to automatically query each of the integrated circuits to collect and store information about the reagents in each of the bottles. The reagent dispensing device is part of a cost effective automated mass production environment that is used to dispense reagents contained in the reagent microbottle to a reagent reservoir of the microfluidic device on a partially deep cut wafer. The data provided by the reagent dispensing device includes automated computer controlled φ dispensing the reagent to the reagent reservoir of the microfluidic device, checking reagent data stored in the memory of the micro vial, and having to be loaded to the micro The specification list of the reagents of the fluid device is aligned, and the reagent data is stored to the memory of the microfluidic device. The reagent dispensing device provides automated and volumetric and positionally accurate reagent dispensing techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, increase the security of the automated manufacturing environment, and increase the automated manufacturing environment. Safety and reduce the cost of this automated manufacturing environment. -95- 201209402 The information provided by this reagent dispensing facility automates the automated, secure, secure and inexpensive data monitoring and management techniques in this automated manufacturing environment. Distributing the reagent to the microfluidic device on the partially deep cut crucible wafer speeds up the loading process and reduces the cost of the loading process.

GPD001.1 This aspect of the invention provides an oligonucleotide spotting device for use in a contactless spot-like oligonucleotide probe to a surface having a target to be recognized in a biological sample a nucleic acid sequence complementary to a nucleic acid sequence, the oligonucleotide spotting device comprising: a support substrate; a reservoir array for accommodating the oligonucleotide probe suspended in the fluid; and an array of emitters, each of the emitters The device is configured to be in fluid communication with a corresponding one of the reservoirs:

The injector is configured to eject droplets containing the oligonucleotide probe from the corresponding reservoir onto the surface. Preferably, each of the injectors has a plurality of nozzles such that the injectors are configured to eject droplets from each nozzle respectively. GPD001.3 Preferably, the injector has a chamber and a plurality of actuations The chamber is for containing fluid from the corresponding reservoir, one of the actuators corresponding to each of the nozzles to cause the actuator to eject droplets of fluid from the chamber via the corresponding nozzle. GPD001.4 Preferably, the actuator is a thermal actuator, each actuator being configured to generate a vapor bubble in the fluid. -96- 201209402 GPD001.5 Preferably, the droplets are less than 1 〇〇 liter. GPD001.6 Preferably, the injector is configured to emit a volume of the emitter that is configured to emit droplets having a volume of less than 25 picoliters. GPD001.7 Preferably, the injector is configured to emit droplets having a volume of less than 6 picoliters. GPD001.8 Preferably, the injector is configured to emit volume

Droplets from 0.1 picoliter to 1.6 picoliters. GPD001.9 Preferably, the actuators in each of the injectors are configured to be individually actuated. GPD 001.10 Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir to the chamber. GPD001.il preferably, the oligonucleotide spotting device also has a CMOS circuit for providing a drive pulse to the actuator, the CMOS circuit having a pad for connecting to a microprocessor controller, the microprocessor control The device controls the relative movement between the nozzle and the surface to be spotted by the oligonucleotide probe. GPD001.12 Preferably, the CMOS circuit has a memory for storing specifications relating to the oligonucleotide probes in the reservoir. GPD001.13 Preferably, the support substrate has a reservoir side and an emitter side opposite the reservoir side, wherein the reservoir array is formed on the reservoir side and the emitter array is formed on the emitter side . GPD00 1 .1 4 Preferably, the CMOS circuit is interposed between the reservoir array and the emitter array. Preferably, the array of emitters is configured to eject droplets containing the oligonucleotide probe onto the surface at a density greater than one droplet per square millimeter. GPD00 1.16 Preferably, the array of emitters is configured to eject droplets containing the oligonucleotide probe onto the surface at a density greater than 8 droplets per square millimeter.

GPD 1.00 17.7 Preferably, the array of emitters is configured to eject droplets containing the oligonucleotide probe onto the surface at a density greater than 60 droplets per square millimeter. GPD00 1 . 1 8 Preferably, the emitter array is configured to emit droplets containing the oligonucleotide probe at a density of from 500 droplets per square millimeter to 1500 droplets per square millimeter to On the surface. GPD00 1.19 Preferably, the array of emitters is configured to eject droplets containing the oligonucleotide probe onto the surface at a rate of more than 100 droplets per second.

GPD00 1.20 Preferably, the emitter array is configured to eject droplets containing the oligonucleotide probe onto the surface at a rate of more than 1,400 droplets per second. This mass-produced and inexpensive oligonucleotide spotting device is used as part of a cost-effective automated mass production environment. The oligonucleotide is loaded into the oligonucleotide reservoir of the device and the device ejects the oligonucleotide onto the surface to be spotted. Automated data provided by the oligonucleotide spotting device includes automated computer controlled distribution of the oligonucleotide to the surface to be spotted to receive the oligonucleoside stored in the oligonucleotide reservoir - 98- 201209402 Acid specification, storage of the oligonucleotide specification in the digits of the digits, and delivery of the oligonucleotide specifications to other departments of the automated manufacturing environment. The oligonucleotide spotting device provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automating the manufacturing environment, increasing the security of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. The device can also be used as an intermediary φ fluid operation machine, which is required for the volume and positional correctness of the transfer of the fluid to the microscopic level and positional correctness. The data provided by the oligonucleotide spotting device automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. GPD003.1 This aspect of the invention provides a spotting device for use in a contactless spot-like oligonucleotide probe to a wafer on-lab (LOC) device having an array of hybrid chambers For accepting the φ oligonucleotide probe 'the probe has a nucleic acid sequence complementary to the target nucleic acid sequence to be detected in the biological sample, and the hybrid chamber array is configured to accommodate the predetermined analysis of the biological sample A complete series of oligonucleotide probes, the spotting device comprising: a reservoir array for accommodating the oligonucleotide probe suspended in a fluid; and an array of emitters, each of the emitter systems Arranging to be in fluid communication with a corresponding one of the reservoirs, respectively, such that the emitter ejects droplets containing the oligonucleotide probe from the corresponding reservoir to one of the hybrid chambers; wherein -99 - 201209402 The reservoir array is organized to contain the complete series of oligonucleotide probes required for the predetermined analysis by the LOC device. GPD003.2 Preferably, the reservoir array has more than 1 000 reservoirs. GPD003.3 Preferably, each of the injectors has a plurality of nozzles such that the injectors are configured to eject droplets from the respective nozzles. GPD003.4 Preferably, the injector has a chamber and a plurality of actuators for containing fluid from the corresponding reservoir, one of the actuators corresponding to each of the nozzles to cause the actuator to be The corresponding nozzle ejects droplets of fluid from the chamber. GPD003.5 Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid. GPD003.6 Preferably, the injector is configured to emit droplets having a volume of less than 1 〇〇 picoliter. GPD003.7 Preferably, the injector is configured to emit droplets having a volume of less than 25 picoliters. GPD003.8 Preferably, the injector is configured to emit droplets having a volume of less than 6 picoliters. GPD003.9 Preferably, the injector is configured to emit droplets having a volume ranging from 0.1 picoliters to 1.6 picoliters. GPD003.1 0 Preferably, the actuators in each of the injectors are configured to be individually actuated. GPD003.il Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir to the chamber. -100- 201209402 GPD003.1 2 Preferably, the spotting device also has a CMOS circuit for providing a drive pulse to the actuator, the CMOS circuit having a pad for connecting to a microprocessor controller, the microprocessor The controller operatively controls the relative movement of the nozzle to the surface to be spotted by the oligonucleotide probe. GPD003.1 3 Preferably, the CMOS circuit has a memory for storing specifications relating to the oligonucleotide probes in the reservoir.

GPD003.1 4 Preferably, the spotting device also has a support substrate having a reservoir side and an emitter side opposite the reservoir side, wherein the reservoir array is formed on the reservoir side and the An array of emitters is formed on the side of the emitter. GPD003.1 5 Preferably, the array of emitters is configured to spot the oligonucleotide probe onto the surface at a density greater than 1 probe spot per square millimeter. GPD003.16 Preferably, the array of emitters is configured to spot the oligonucleotide probe onto the surface of the φ at a density greater than 8 probe spots per square millimeter. GPD003.1 7 Preferably, the array of emitters is configured to spot the oligonucleotide probe onto the surface at a density greater than 60 probe spots per square millimeter. GPD003.1 8 Preferably, the emitter array is configured to spot the oligonucleotide at a density of 500 probes per square millimeter to 15 probes per square millimeter. The needle is on the surface. GPD003.1 9 Preferably, the array of emitters is configured to spot the oligonucleotide probe onto the surface -101 - 201209402 at a rate of more than one probe spot per second. GPD003.20 Preferably, the array of emitters is configured to spot the oligonucleotide probe onto the surface at a rate of more than 1,400 probes per second. This mass-produced and inexpensive oligonucleotide spotting device is used as part of a cost-effective automated mass production environment. The oligonucleotide is loaded into the oligonucleotide reservoir of the device and the device ejects the oligonucleotide into the hybridization chamber of the LOC device to be spotted. Automated data provided by the oligonucleotide spotting device includes automated computer controlled distribution of the oligonucleotide to the hybridization chamber of the LOC device to receive oligonucleotide specifications stored in the oligonucleotide reservoir The oligonucleotide is stored in the digital memory of the other, and the oligonucleotide is delivered for storage in the memory of the LOC device to be spotted. The oligonucleotide spotting device provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automating the manufacturing environment, increasing the security of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. The device can also be used as an intermediate fluid operation machine, which is required for the volume and positional correctness of the transfer of the fluid to the microscopic level and positional correctness. Numerous oligonucleotide reservoirs and emitters available on the oligonucleotide spotting device also provide a single step to spot each LOC device. The data provided by the oligonucleotide spotting device is automated to provide automated, safe, secure and inexpensive data monitoring in this automated manufacturing environment -102-201209402 and management techniques. GPD004.1 This aspect of the invention provides a biochemical deposition apparatus for depositing a biochemical substance on a surface without contact, the biochemical deposition apparatus comprising: a reservoir array for containing a plurality of biochemical substances; and an injection Arrays, each of the injectors being configured to be in fluid communication with a respective one of the reservoirs;

The injector is configured to eject droplets containing the biochemical substance from the corresponding reservoir onto the surface. GPD004.2 Preferably, the biochemical substance in the reservoir array is an oligonucleotide probe having a nucleic acid sequence complementary to a target nucleic acid sequence to be detected in a biological sample, and A surface-on-wafer laboratory (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probe. GPD004.3 Preferably, the hybridization chamber array is configured to accommodate a complete series of oligonucleotide probes required for a predetermined analysis of the biological sample, and the reservoir array is configured to comprise The complete series of oligonucleotide probes required for the intended analysis by the LOC device. GPD004.4 Preferably, the reservoir array has more than 1 000 reservoirs. Preferably, each of the injectors has a plurality of nozzles such that the injectors are configured to eject droplets from the respective nozzles. GPD004.6 Preferably, the injector has a chamber and a plurality of actuators for containing a fluid having a suspension oligonucleotide probe-103-201209402 needle supplied by the corresponding reservoir, the actuator One of the nozzles corresponds to each of the nozzles to cause the actuator to eject droplets of fluid from the chamber via the corresponding nozzles. GPD004.7 Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid. GPD004.8 Preferably, the injector is configured to emit droplets having a volume of less than 100 picoliters. GPD004.9 Preferably, the injector is configured to emit volume

Droplets less than 25 picoliters. GPD004.1 0 Preferably, the injector is configured to emit droplets having a volume of less than 6 picoliters. GPD004.il Preferably, the injector is configured to emit droplets having a volume ranging from 0.1 picoliters to 1.6 picoliters. GPD004.1 2 Preferably, the actuators in each of the injectors are configured to be individually actuated. GPD004.1 3 Preferably, each of the injectors has a plurality of reservoirs

Extend to the entrance passage of the room.

GPD004.1 4 Preferably, the biodeposition device also has a CMOS circuit for providing a drive pulse to the actuator, the CMOS circuit having a pad for connecting to a microprocessor controller, the microprocessor controller operability The relative movement between the nozzle and the surface to be spotted by the oligonucleotide probe is controlled. GPD004.1 5 Preferably, the CMOS circuit has a memory for storing specifications relating to the oligonucleotide probes in the reservoir. GPD004.1 6 Preferably, the biochemical deposition device also has a support substrate -104 - 201209402, the support substrate having a reservoir side and an emitter side opposite the reservoir side, wherein the reservoir array is attached to the reservoir The side of the device is formed and the array of emitters is formed on the side of the emitter. GPD004.1 7 Preferably, the array of emitters is configured to eject droplets containing the biochemical onto the surface at a density greater than 8 droplets per square millimeter.

GPD004.1 8 Preferably, the array of emitters is configured to eject droplets containing the biochemical onto the surface at a density greater than 60 droplets per square millimeter. GPD004.1 9 Preferably, the array of emitters is configured to eject droplets containing the biochemical onto the surface at a density of from 500 droplets per square millimeter to 1,500 droplets per square millimeter. GPD 004.20 Preferably, the array of emitters is configured to eject droplets containing the biochemical onto the surface at a rate of more than 100 droplets per second. This mass-produced and inexpensive biochemical deposition unit is used as part of a cost-effective automated mass production environment. The biochemical material is loaded into the bioreactor of the device and the device is deposited on the surface by depositing the biochemical material from the biochemical reservoir onto the surface to be deposited. The data provided by the biochemical deposition device includes automatic computer controlled distribution of the biochemical substance to the surface to be spotted, receiving biochemical material specifications stored in the biochemical material reservoir, and storing the biochemical material specifications in the Digital memory, and the department that delivers the biochemical specifications to the automated manufacturing environment. -105- 201209402 This biochemical deposition device provides automated, volumetric and positional, fast and high-density biochemical deposition technology to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the automated manufacturing environment. The preservation, the safety of the automated manufacturing environment and the cost of the automated manufacturing environment. The device can also be used as an intermediate fluid operation machine, which is required for the volume and positional correctness of the fluid from the macroscopic level to the volume and positional correctness of the microscopic level. The data provided by the biochemical deposition device provides automation, security, preservation and inexpensive data monitoring and management techniques in the automated manufacturing environment. GPD005.1 This aspect of the invention provides a microsystem technology (MST) device for use on a contactless spot-like oligonucleotide probe to a surface having a target to be recognized in a biological sample a nucleic acid sequence complementary to a nucleic acid sequence, the MST apparatus comprising: a monolithic substrate having a reservoir side and an emitter side opposite the reservoir side; a reservoir array formed on the reservoir side; and an emitter An array formed on the side of the emitter, each of the injectors being configured to be in fluid communication with a respective one of the reservoirs; wherein the injector is configured to be ejected from the corresponding reservoir The droplets of the oligonucleotide probe are onto the surface. GPD 005.2 Preferably, each of the injectors has a plurality of nozzles such that the injectors are configured to eject droplets from the respective nozzles. GPD005.3 Preferably, the injector has a chamber and a plurality of actuators -106 - 201209402 'the chamber is for containing a fluid having a suspension oligonucleotide probe supplied by the corresponding reservoir, the actuator One of the nozzles corresponds to each of the nozzles to cause the actuator to eject droplets of fluid from the chamber via the corresponding nozzles. GPD005.4 Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid. GPD005.5 Preferably, the injector is configured to emit droplets having a volume of less than 1 〇 〇 liter.

GPD005.6 Preferably, the injector is configured to emit droplets having a volume of less than 25 picoliters. GPD005.7 Preferably, the injector is configured to emit droplets having a volume of less than 6 picoliters. GPD005.8 Preferably, the injector is configured to emit droplets having a volume ranging from 0.1 picoliters to 1.6 picoliters. GPD 005.9 Preferably, the actuators in each of the injectors are configured to be individually actuated. GPD005.1 0 Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir to the chamber. GPD005.il preferably, the MST device also has a CMOS circuit for providing a drive pulse to the actuator, the CMOS circuit having a pad for connecting to a microprocessor controller, the microprocessor controller operatively controlling the The relative movement between the nozzle and the surface to be spotted by the oligonucleotide probe GPD005. 1 2 preferably, the CMOS circuit has a body for storage and storage of the oligonucleotide in the reservoir Needle related specifications. -107-201209402 GPD005.13 Preferably, the surface is a laboratory on-wafer (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probe, the hybridization chamber array being organized A complete series of oligonucleotide probes required to accommodate the predetermined analysis of the biological sample, and the reservoir array is configured to contain the complete oligonucleotide probe required for the predetermined analysis by the LOC device Needle series. GPD005.1 4 Preferably, the reservoir array has more than one reservoir. GPD005.1 5 Preferably, the CMOS circuit is interposed between the array of registers and the array of emitters. GPD005.1 6 Preferably, the array of emitters is configured to spot the oligonucleotide onto the surface at a density greater than 8 probe spots per square millimeter. GPDGG5_17 Preferably, the array of emitters is configured to spot the oligonucleotide onto the surface at a density of more than 60 probe spots per square millimeter. GPD005 1 8 Preferably, the array of emitters is configured to spot the oligonucleotide at a density of 5,000 probes per square millimeter to a density of 1,500 probes per square millimeter. To the surface. GPDG() 5_19 Preferably, the array of emitters is organized to spot the oligonucleotide onto the surface at a rate of more than 100 probes per second. GPD005-2〇 Preferably, the array of emitters is configured to spot the oligonucleotide onto the surface at a rate of more than 1,4 probes per second. -108 - 201209402

The oligonucleotide spotting device utilizes microsystem technology (M S Τ) to produce a large amount, inexpensively and is used as part of a cost-effective automated mass production environment. The oligonucleotide is loaded into an oligonucleotide reservoir of the device and the device ejects the oligonucleotide onto the surface to be spotted. Automated data provided by the oligonucleotide spotting device includes automated computer controlled distribution of the oligonucleotide to the surface to be spotted, receiving the oligonucleotides stored in the oligonucleotide reservoir The specification, storage of the oligonucleotide size in the digit memory, and delivery of the oligonucleotide specification to other departments of the automated manufacturing environment. The oligonucleotide spotting device provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automating the manufacturing environment, increasing the security of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. The device can also be used as an intermediate fluid operation machine, which is required for the volume and positional correctness of the transfer of the fluid to the microscopic level and positional correctness. The data provided by the oligonucleotide spotting device automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. GPD006.1 This aspect of the invention provides an oligonucleotide spotting device for use in a contactless spot-like oligonucleotide probe to a surface having a target to be recognized in a biological sample A nucleic acid sequence complementary nucleic acid sequence, the oligonucleotide spotting device comprising: a reservoir array for containing the oligonucleotide suspended in a fluid - 109 - 201209402 probe: an emitter array covering the a reservoir array, each of the injectors being configured to be in fluid communication with a respective one of the reservoirs; wherein the injector is configured to emit the oligonucleotide probe from the corresponding reservoir Droplets onto the surface.

GPD006.2 Preferably, the oligonucleotide spotting device also has a support substrate having a reservoir side and an emitter side opposite the reservoir side, wherein the reservoir array is attached to the reservoir side Formed and the emitter array is formed on the emitter side. GPD006.3 Preferably, each of the injectors has a plurality of nozzles such that the injectors are configured to eject droplets from the respective nozzles. GPD006.4 Preferably, the injector has a chamber and a plurality of actuators for containing fluid from the corresponding reservoir, one of the actuators corresponding to each of the nozzles to cause the actuator to be The corresponding nozzle emits droplets of fluid from the chamber.

GPD006.5 Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid. GPD006.6 Preferably, the injector is configured to emit droplets having a volume of less than 100 picoliters. GPD006.7 Preferably, the injector is configured to emit droplets having a volume of less than 25 picoliters. GPD006.8 Preferably, the injector is configured to emit droplets having a volume of less than 6 picoliters. GPD006.9 Preferably, the injector is configured to emit droplets of volume -110 - 201209402 between o.l and picoliter to 1.6 microliters. GPD 006.10 Preferably, the actuators in each of the injectors are configured to be individually actuated. GPD006.il Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir to the chamber.

GPD006.1 2 Preferably, the oligonucleotide spotting device also has a CMOS circuit for providing a drive pulse to the actuator, the CMOS circuit having a pad for connecting to a microprocessor controller, the microprocessor The controller operatively controls the relative movement of the nozzle to the surface to be spotted by the oligonucleotide probe. GPD006.1 3 Preferably, the CMOS circuit has a memory for storing specifications relating to the oligonucleotide probes in the reservoir. GPD006.14 Preferably, the CMOS circuit is interposed between the array of registers and the array of emitters. GPD006.1 5 Preferably, the surface-on-wafer laboratory (LOC) φ device 'the L〇C device has an array of hybridization chambers for accepting the oligonucleotide probe, the hybridization chamber array is organized A complete series of oligonucleotide probes required to accommodate the predetermined analysis of the biological sample, and the reservoir array is configured to contain the complete oligonucleotide probe required for the predetermined analysis by the LOC device Needle series. GPD006.1 6 Preferably, the reservoir array has more than one reservoir. GPD006.1 7 Preferably the emitter array is configured to spot the probe onto the surface at a density greater than 8 probe spots per square millimeter. -111 - 201209402 GPD006.1 8 Preferably, the emitter array has a density of 60 probe spots per square millimeter. The GPD006.1 9 preferably, the emitter array maintains 500 probes per square millimeter. The probe was spotted to a density per square millimeter to the surface. GPD006.20 Preferably, the CMOS circuit mimics a pulse to cause the array of emitters to spot the probe onto the surface at a rate of more than 100 per second. This mass-produced, inexpensive structure with a layered structure is used as a cost-effective automated mass production. The oligonucleotide is injected onto the surface to be spotted by the oligonucleotide loaded on the device. Automating the data provided by the spotting device includes automating the oligonucleotide to the surface to be spotted, receiving the oligonucleotide size in the reservoir, storing the oligonucleotide memory, and transmitting The oligonucleotide is sized to the automation department. The oligonucleotide spotting device provides an automated, fast and high density oligonucleotide spotting technique to simplify the complexity of the environment, increase the security of the automated manufacturing environment of the automated manufacturing environment, and increase the automation Sex and reduce the cost of this automated manufacturing environment. The device is operated by fluid operation, and the volume of the machine is transferred from the macroscopic level. The structure is higher than the 丨 needle to the surface of the 丨 组 以 以 以 以 ; ; ; ; ; ; ; ; ; ; ; ; ; ; ; a probe spotted nucleotide spotting one of the reservoirs in the environment, and the allocation of the oligonucleotide is controlled by the oligonucleotide in the position of the digital manufacturing environment Accuracy, the automation of manufacturing and the safety of the environment can also be used as an intermediary and positional correctness -112- 201209402 required to transfer fluid to the microscopic volume and position correctness. The data provided by the oligonucleotide spotting device automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. GPD007.1 This aspect of the invention provides an oligonucleotide spotting device for use in a contactless spot-like oligonucleotide probe to a surface having a target to be recognized in a biological sample a nucleic acid sequence complementary nuclear φ acid sequence, the oligonucleotide spotting device comprising: a support substrate; a reservoir array on a side of the support substrate, the reservoir being configured for containing the suspension in a liquid An oligonucleotide probe; an emitter array on the other side of the support substrate; and a plurality of inlet channels in fluid communication between the reservoir and the emitter; wherein the emitter is configured to correspond to The reservoir ejects droplets containing the oligonucleotide φ acid probe onto the surface. GPD007.2 Preferably, each of the injectors is configured to be in fluid communication with a respective one of the reservoirs. GPD 007.3 Preferably, each of the injectors has a plurality of nozzles such that the injectors are configured to eject droplets from the respective nozzles. GPD007.4 Preferably, the injector has a chamber and a plurality of actuators for containing fluid from the corresponding reservoir, one of the actuators corresponding to each of the nozzles to cause the actuator to be The corresponding nozzle emits droplets of fluid from the chamber. - 113 - 201209402 GPD007.5 Preferably, the actuator is a thermal actuator, each actuator being configured to generate a vapor bubble in the fluid. GPD007.6 Preferably, the injector is configured to emit droplets having a volume of less than 1 〇〇 picoliter. GPD007.7 Preferably, the injector is configured to emit droplets having a volume of less than 25 picoliters. GPD007.8 Preferably, the injector is configured to emit droplets having a volume of less than 6 picoliters. GPD007.9 Preferably, the injector is configured to emit droplets having a volume ranging from 0.1 picoliters to 1.6 picoliters. GPD 007.10 Preferably, the actuators in each of the injectors are configured to be individually actuated. - GPD007.il Preferably, each of the injectors is in fluid communication with one of the reservoirs via more than one inlet passage. GPD007.1 2 Preferably, the oligonucleotide spotting device also has a CMOS circuit for providing a drive pulse to the actuator, the CMOS circuit having a pad for connecting to a microprocessor controller, the microprocessor The controller operatively controls the relative movement of the nozzle to the surface to be spotted by the oligonucleotide probe. GPD007.1 3 Preferably, the CMOS circuit has a memory for storing specifications relating to the oligonucleotide probes in the reservoir. GPD 007.14 Preferably, the CMOS circuit is interposed between the array of registers and the array of emitters. GPD007.1 5 Preferably, the array of emitters is configured to spot the probe onto the surface at a density of 8 probe spots per square millimeter above -114 - 201209402. GPD007.1 6 Preferably, the emitter array is configured to spot the probe onto the surface at a density of more than 60 probe spots per square millimeter. Preferably, the emitter is The array was organized to spot the probe onto the surface at a density of 500 probes per square millimeter to 1,500 probe spots per square millimeter.

GPD007.1 8 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 100 probes per second" GPD007. Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 1,400 probes per second. GPD 007.20 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 20,000 probe points per second. This mass-produced and inexpensive oligonucleotide spotting device is used as part of a cost-effective automated mass production environment. It is made of fluid on both sides of the tantalum substrate to increase the degree of integration of the device, reduce the size of the device and minimize the cost of the device. The oligonucleotide is loaded into the oligonucleotide reservoir of the device and the device ejects the oligonucleotide onto the surface to be spotted. Automated data provided by the oligonucleotide spotting device includes automated computer controlled distribution of the oligonucleotide to the surface to be spotted, receiving the oligonucleotides stored in the oligonucleotide reservoir Specification - 115 - 201209402. Store the oligonucleotide in its digital memory and transfer the oligonucleotide specification to other departments of the automated manufacturing environment. The oligonucleotide spotting device provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automating the manufacturing environment, increasing the security of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. The device can also be used as an intermediate fluid operation machine, which is required for the volume and positional correctness of the transfer of the fluid to the microscopic level and positional correctness. The data provided by the oligonucleotide spotting device automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. GPD008.1 This aspect of the invention provides an oligonucleotide spotting device for use in a contactless spot-like probe to a surface having a complementary nucleic acid sequence to be recognized in a biological sample. Nucleic acid sequence, the oligonucleotide spotting device comprises: a support substrate: an array of emitters, each of the emitters having an actuator for emitting droplets of liquid containing the oligonucleotide probe; A drive pulse is provided to each of the droplets to eject an actuator to eject a droplet; and a pad is used to electrically connect the CMOS circuit to an external microprocessor controller to operate the illuminator array. GPD00 8.2 Preferably, the CMOS circuit has a digital memory for storing the identification information - 116 - 201209402 for the external microprocessor controller to identify the device. GPD008.3 Preferably, the oligonucleotide spotting device also has a reservoir array for containing the oligonucleotide probe suspended in a liquid, wherein the support substrate has a reservoir side and a reservoir side formed opposite the emitter side, the reservoir array being formed on the reservoir side, the emitter array being formed on the emitter side, each of the emitters being configured for use with the probes A corresponding one of the reservoirs is in fluid communication.

GPD008.4 Preferably, the digital memory stores specifications of the oligonucleotide probe. GPD008.5 Preferably, each of the injectors has a plurality of actuators and corresponding plurality of nozzles associated with each of the droplet ejection actuators to cause actuation of one of the actuators via The nozzle associated with the actuator emits a droplet. GPD008.6 Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid. GPD008.7 Preferably, the injector is configured to emit droplets having a volume of less than 1 〇 〇 liter. GPD008.8 Preferably, the injector is configured to emit droplets having a volume of less than 25 picoliters. GPD008.9 Preferably, the injector is configured to emit droplets having a volume of less than 6 picoliters. GPD 008. 10 Preferably, the injector is configured to emit droplets having a volume ranging from 0.1 picoliter to 1.6 picoliters. GPD008.il Preferably, the actuator -117-201209402 in one of the injectors is organized to be individually actuated. GPD008.1 2 Preferably, each of the injectors has a chamber and a passage containing the liquid for ejecting from the nozzle, the passage extending from the chamber to a reservoir corresponding to the injector. GPD008.1 3 Preferably, the configurations in each of the injectors are individually actuated. GPD 008.14 Preferably, the CMOS circuit is between the column and the emitter array. GPD008.1 5 Preferably, the surface is a real device on a wafer having an array of hybridization chambers for receiving an acid probe, the array of hybridization chambers being organized to accommodate the integrity required for the biological sample analysis A series of oligonucleotide probes, and the reservoir is configured to contain a series of oligonucleotide probes required for predetermined analysis by the LOC device. GPD008.1 6 Preferably, the reservoir array has more than a reservoir. GPD008.1 7 Preferably, the array of emitters is spotted by the density of 8 probe spots per square millimeter; GPD008.1 8 Preferably, the array of emitters is per group Square millimeters 60 probe spotting density spotting the probe to 〇GPD008.1 9 Preferably, the emitter array is spotted by a group of 500 probes per square millimeter to a density point of 1500 per square millimeter The probe is applied to the surface. Multiple inlets Multiple inlets are passed through the reservoir array (LOC). The predetermined array of oligonucleosides is composed of a complete set of 1 000 structures. Constructed above the surface to have a probe point

-118-201209402 GPD008.20 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe to the surface at a rate of more than 10,000 probes per second. on.

This mass-produced and inexpensive oligonucleotide spotting device is used as part of a cost-effective automated mass production environment. The oligonucleotide is loaded into the oligonucleotide reservoir of the device and the device ejects the oligonucleotide onto the surface to be spotted. Automated data provided by the oligonucleotide spotting device includes automated computer controlled distribution of the oligonucleotide to the surface to be spotted, receiving the oligonucleotides stored in the oligonucleotide reservoir The specification, storage of the oligonucleotide size in the digit memory, and delivery of the oligonucleotide specification to other departments of the automated manufacturing environment. The spotting device performs these functions under the control of an external computer. The oligonucleotide spotting device provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automating the manufacturing environment, increasing the security of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. The device can also be used as an intermediate fluid operation machine, which is required to transfer the fluid from the macroscopic level to the correct volume and positional correctness of the fluid to the microscopic level. The information provided is automated to provide automated, secure, secure and inexpensive data monitoring and management techniques in this automated manufacturing environment. GPD009.1 This aspect of the invention provides an oligonucleotide spotting device for use on a contactless spot-like probe to a surface having -119-201209402 and a target to be identified in a biological sample a nucleic acid sequence complementary to a nucleic acid sequence, the oligonucleotide spotting device comprising: a support substrate; an array of emitters each having an actuator for ejecting a droplet of a liquid containing the oligonucleotide probe; A CMOS circuit for providing a drive pulse to each of the droplets to eject an actuator to eject a droplet: wherein the CMOS circuit has digital memory for storing data associated with the device. GPD009.2 Preferably, the CMOS circuit has a pad for electrical connection and an external microprocessor controller for operating the array of emitters 〇GPD009.3. Preferably, the data includes identification data for external micro The processing controller identifies the device. GPD009.4 Preferably, the oligonucleotide spotting device also has a reservoir array for containing the oligonucleotide probe suspended in a liquid, wherein the support substrate has a reservoir side and a reservoir side formed opposite the emitter side, the reservoir array being formed on the reservoir side, the emitter array being formed on the emitter side, each of the emitters being configured for use with the probes A corresponding one of the reservoirs is in fluid communication. GPD009.5 Preferably, the digital memory stores specifications of the oligonucleotide probe. GPD009.6 Preferably, each of the injectors has a plurality of actuators and corresponding plurality of nozzles associated with each of the droplet ejection actuators to enable one of the -120-201209402 actuators Actuation ejects droplets via the nozzle associated with the actuator. GPD009.7 Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid. GPD009.8 Preferably, the injector is configured to emit droplets having a volume of less than 100 picoliters. GPD009.9 Preferably, the injector is configured to emit volume

Droplets less than 25 picoliters. GPD009.1 0 Preferably, the injector is configured to emit droplets having a volume of less than 6 picoliters. GPD009.il Preferably, the injector is configured to have an ejection volume of from 0.1 picoliter to 1.6. The droplets of the skin rise. GPD009.1 2 Preferably, the actuators in one of the injectors are organized to be individually actuated. GPD009.1 3 Preferably, each of the injectors has a chamber and a plurality of inlet φ channels, the chamber containing the liquid for ejecting from the nozzles. The plurality of inlet channels extending from the chamber to a reservoir corresponding to the injector Device. GPD009.1 4 Preferably, the actuators in each of the injectors are configured to be individually actuated. GPD009.1 5 Preferably, the CMOS circuit is interposed between the array of registers and the array of emitters. GPD009.16 Preferably, the surface is a laboratory on wafer (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probe, the array of hybridization chambers being organized to accommodate the organism Sample Preparation -121 - 201209402 The complete series of oligonucleotide probes required for analysis, and the reservoir array is organized to contain the complete oligonucleotide probe required for the intended analysis by the LOC device Needle series. GPD009.1 7 Preferably, the reservoir array has more than 1 000 reservoirs. GPD009.1 8 Preferably, the array of emitters is configured to spot the probe onto the surface at a density greater than 60 probes per square millimeter. 〇GPD009.1 9 preferably, the emitter The array was organized to spot the probe onto the surface at a density of 500 probes per square millimeter to a density of 1,500 probes per square millimeter. GPD 009.20 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 100 probes per second. This mass-produced and inexpensive oligonucleotide spotting device is used as part of a cost-effective automated mass production environment. The oligonucleotide is loaded into the oligonucleotide reservoir of the device and the device ejects the oligonucleotide onto the surface to be spotted. Automated data provided by the oligonucleotide spotting device includes automated computer controlled distribution of the oligonucleotide to the surface to be spotted, receiving the oligonucleotides stored in the oligonucleotide reservoir The specification, storage of the oligonucleotide size in the digit memory, and delivery of the oligonucleotide specification to other departments of the automated manufacturing environment. The oligonucleotide spotting device provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment and increase the reliability of the automated manufacturing environment. Sexuality, increase the security of the automated manufacturing environment, increase the security of the automated manufacturing environment, and reduce the cost of the automated manufacturing environment. The device can also be used as an intermediate fluid operation machine, which is required for the volume and positional correctness of the transfer of the fluid to the microscopic level and positional correctness. The data provided by the oligonucleotide spotting device is automated to provide automated, safe, secure and inexpensive data monitoring in the automated manufacturing environment.

And management technology. GPD010.1 This aspect of the invention provides a spotting device for use in a contactless spot-like oligonucleotide probe to a surface having a complementary nucleic acid sequence to be recognized in a biological sample. a nucleic acid sequence, the spotting device comprising: a reservoir array for accommodating the oligonucleotide probe suspended in the fluid; an emitter array in fluid communication with the reservoir, each emitter having an actuating φ device a droplet for ejecting a liquid containing the oligonucleotide probe; and a CMOS circuit for supplying a driving pulse to each of the droplets to emit an actuator to emit a droplet; wherein the CMOS circuit has a digital memory . GPD010.2 Preferably, the CMOS circuit has a pad for electrical connection and an external microprocessor controller GPD010.3 for operating the array of emitters. Preferably, the digital memory stores identification data for external use. The microprocessor controller identifies the device. -123- 201209402 GPD01 0.4 Preferably, the oligonucleotide spotting device also has a support substrate having a reservoir side and an emitter side opposite to the reservoir side, the reservoir array being attached to the reservoir Formed on the side of the injector, the array of emitters being formed on the side of the injector, each of the injectors being configured for fluid communication with a respective one of the probe receptacles. GPD010.5 Preferably, the digital memory stores specifications of the oligonucleotide probe. GPD 010.6 Preferably, each of the injectors has a plurality of actuators and corresponding plurality of nozzles associated with each of the droplet ejection actuators to cause actuation of one of the actuators via The nozzle associated with the actuator emits a droplet. GPD010.7 Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid. GPD 010.8 Preferably, the injector is configured to emit droplets having a volume of less than 2.0 picoliters. GPD 010.9 Preferably, the actuators in one of the injectors are organized to be individually actuated. GPD 010.10 Preferably, each of the injectors has a chamber and a plurality of inlet passages, the chamber containing the liquid for exiting the nozzle, the plurality of inlet passages extending from the chamber to a reservoir corresponding to the injector. GPD 010.il Preferably, the actuators in each of the injectors are configured to be individually actuated. GPD010.12 Preferably, the CMOS circuit is interposed between the reservoir array and the emitter array. Preferably, the surface is a laboratory on-wafer (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probe, the hybridization chamber array being organized A complete series of oligonucleotide probes required to accommodate the predetermined analysis of the biological sample, and the reservoir array is configured to contain the complete oligonucleotide probe required for the predetermined analysis by the LOC device Needle series. GPD010.14 Preferably, the reservoir array has more than one 〇〇〇

GPD 010.15 Preferably, the array of emitters is configured to spot the probe onto the surface at a density greater than 8 probe spots per square millimeter. Preferably, the emitter array is configured to spot the probe onto the surface at a density greater than 60 probe spots per square millimeter. 〇GPD010.17 Preferably, the emitter array The probe is spotted onto the surface with a density of 500 probes per square millimeter to a density of 1,500 probe points per square millimeter. GPD0 10.18 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 100 probes per second. GPD 010.19 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 1,400 probes per second. CJPD010.2 0 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe to the surface at a rate of more than 20,000 probe points per second -125 - 201209402 per second. on. This mass-produced and inexpensive oligonucleotide spotting device is used as part of a cost-effective automated mass production environment. The oligonucleotide is loaded into the oligonucleotide reservoir of the device and the device ejects the oligonucleotide onto the surface to be spotted. Automated data provided by the oligonucleotide spotting device includes automated computer controlled distribution of the oligonucleotide to the surface to be spotted, receiving the oligonucleotides stored in the oligonucleotide reservoir The specification, storage of the oligonucleotide size in the digit memory, and delivery of the oligonucleotide specification to other departments of the automated manufacturing environment. The oligonucleotide spotting device provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automating the manufacturing environment, increasing the safety of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. The device can also be used as an intermediary fluid to operate, the machine is transformed from the size and positional correctness of the giant level. Required to transfer fluid to the microscopic volume and positional correctness. The data provided by the oligonucleotide spotting device automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. GPD01 1.1 This aspect of the invention provides a spotting device for spotlessly wafer-on-lab (LOC) devices with oligonucleotide probes mounted on a printed circuit board (PCB) And a fixed array of each of the LOC devices having an array of hybridization chambers for receiving the oligonucleotide probe having the target nucleic acid sequence-126- to be recognized in the biological sample

201209402 A complementary nucleic acid sequence, the spotting device comprising: a monolithic support substrate; a reservoir array on one side of the support substrate, the reservoir comprising an oligonucleotide probe suspended in a liquid to spot the PCB LOC And an emitter array on the other side of the support substrate, each of the arrays being in fluid communication with a corresponding one of the reservoirs, the emitter exiting the corresponding reservoir containing the oligonucleotide probe One of the liquid chambers of the needle. GPD 011.2 Preferably, the reservoir array has a reservoir. GPD 011.3 Preferably, each of the injectors has a plurality of structures configured to eject droplets from the respective nozzles. GPD011.4 Preferably, the injector has a chamber for containing fluid from the corresponding reservoir, the actuation φ corresponding to each nozzle to cause the actuator to pass through the corresponding nozzle chamber Droplets of fluid. GPD 011.5 Preferably, the actuator is a thermal actuator configured to generate vapor bubbles in the fluid. GPD 011.6 Preferably, the injector is configured to have droplets of less than 100 picoliters. GPD 011.7 Preferably, the injector is configured to have droplets of less than 25 picoliters. GPD011.8 Preferably, the injector is configured to dispense all of the actuators above a sufficient amount to cause the droplet to be dispensed to the 1000 nozzles, with one of the actuators being separated therefrom, Each of the actuated injection volume ejection volume ejection volume -127 - 201209402 is less than 6 picoliters of droplets. GPD01 1.9 Preferably, the injector is configured to emit droplets having a volume ranging from 0.1 picoliters to 1.6 picoliters. GPD01 1.10 Preferably, the actuators in each of the injectors are configured to be individually actuated. GPD011.il Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir to the chamber. GPD01 1.12 Preferably, the spotting device also has a CMOS circuit for providing a drive pulse to the actuator, the CMOS circuit having a pad for connecting to a microprocessor controller, the microprocessor controller operatively controlling the The relative movement between the nozzle and the surface to be spotted by the oligonucleotide probe. GPD011.13 Preferably, the CMOS circuit has a memory for storing specifications relating to the oligonucleotide probes in the reservoir. GPD0 11.14 Preferably, the reservoir array is integrally formed on one side of the monolithic support substrate. GPD0 1 1 . 1 5 Preferably, the array of emitters is configured to spot the probe onto the surface at a density greater than 8 probe spots per square millimeter. Preferably, the emitter array is configured to spot the probe onto the surface at a density of more than 60 probes per square millimeter. 〇GPD0 1 1 . The emitter array is configured to spot the probe onto the surface at a density of from 500 probes per square millimeter to 1 500 probe spots per square millimeter. GPD01 1 · 18 Preferably, the CMOS circuit is configured to generate a -128-201209402 drive pulse to cause the emitter array to spot the probe at a rate of more than 100 probes per second. On the surface. GPD01 1.19 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 1,400 probes per second.

GPD01 1.20 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 20,000 probe points per second. This mass-produced and inexpensive oligonucleotide spotting device is used as part of a cost-effective automated mass production environment. The oligonucleotide is loaded into an oligonucleotide reservoir of the device and the device ejects the oligonucleotide into a hybridization chamber of an array of LOC devices mounted on a PCB wafer. Data automation provided by the oligonucleotide spotting device includes automated computer controlled spotting of oligonucleotides to an array of LOC devices mounted on a PCB wafer, receiving oligonucleotides stored in oligonucleotide reservoirs The glucosinolate size, the storage of the oligonucleotide in its digital memory, and the delivery of the oligonucleotide specification to the LOC device or other department of the automated manufacturing environment. The oligonucleotide spotting device provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automating the manufacturing environment, increasing the security of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. The device can also be used as an intermediate fluid operation machine, which is required for the volume and positional correctness of the transfer of the fluid to the microscopic level and positional correctness. -129- 201209402 The data provided by the oligonucleotide spotting device is automated to provide automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. An array of spotted oligonucleotides to the LOC device mounted on the PCB wafer accelerates the loading process and reduces the cost of the loading process, and after mounting the LOC device to the PCB wafer and soldering them, The LOC device enhances the chemical and physical integrity of the oligonucleotide. GPD012.1 This aspect of the invention provides an oligonucleotide spotting device for use in a contactless spot on which a wafer on a wafer-on-lab (LOC) device array is constructed, the LOC device being Organizing to detect a target nucleic acid sequence in a biological sample using the oligonucleotide probe and each LOC device has an array of hybridization chambers for receiving the oligonucleotide probe, the oligonucleotide spotting device comprising : a reservoir array on one side of the support substrate, the reservoir comprising a sufficient amount of an oligonucleotide probe suspended in a liquid to spot all of the LOC devices on the wafer; and covering the reservoir array to The moving array of emitters is fixed such that the emitter ejects droplets containing the oligonucleotide probe from the corresponding reservoir to one of the hybridization chambers. GPD012.2 Preferably, the oligonucleotide spotting device also has a support substrate having a reservoir side and an emitter side opposite the reservoir side, wherein the reservoir array is attached to the reservoir side Formed and the emitter array is formed on the emitter side" GPD012.3. Preferably, each of the injectors has a plurality of nozzles, and the emitters are configured to emit droplets from the respective nozzles at -130-201209402. . GPD 012.4 Preferably, the injector has a chamber and a plurality of actuators for containing fluid from the corresponding reservoir, one of the actuators corresponding to each of the nozzles to cause the actuator to be The corresponding nozzle emits droplets of fluid from the chamber. GPD 012.5 Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid.

GPD 012.6 Preferably, the injector is configured to emit droplets having a volume of less than 100 picoliters. GPD 012.7 Preferably, the injector is configured to emit droplets having a volume of less than 25 picoliters. GPD 012.8 Preferably, the injector is configured to emit droplets having a volume of less than 6 picoliters. GPD 012.9 Preferably, the injector is configured to emit droplets having a volume between 〇.1 picoliter and 1.6 picoliters. GPD012.10 Preferably, the actuator system in each of the injectors Preferably, each of the injectors has a plurality of inlet channels extending from the reservoir to the chamber. GPD012.12 Preferably, the oligonucleotide spotting device also has a CMOS circuit for providing a drive pulse to the actuator, the CMOS circuit having a pad for connecting to a microprocessor controller, the microprocessor control The operability controls the relative movement between the nozzle and the surface to be spotted by the oligonucleotide probe. -131 - 201209402 GPD012.13 Preferably, the CMOS circuit has a memory for storing specifications relating to the oligonucleotide probes in the reservoir. GPD012.14 Preferably, the CMOS circuit is interposed between the array of registers and the array of emitters. GPD012.15 Preferably, the LOC device has an array of hybridization chambers for receiving the oligonucleotide probes, the array of hybridization chambers configured to contain the complete oligonucleoside required for the predetermined analysis of the biological sample A series of acid probes, and the reservoir array is configured to contain the complete series of oligonucleotide probes required for the predetermined analysis by the LOC device. GPD 012.16 Preferably, the reservoir array has more than 1 000 reservoirs. GPD 012.17 Preferably, the array of emitters is configured to spot up to 50,000 probes per square millimeter to each square. A density of 1,500 probes of millimeters was spotted onto the surface. GPD 012.18 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 100 probes per second. GPD0 12.19 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 1,400 probes per second. GPD 012.20 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 20,000 probe points per second. The mass-produced and inexpensive oligonucleotide spotting device is used as a -132-201209402

Cost-effective automation is part of a large manufacturing environment. The oligonucleotide is loaded into the oligonucleotide reservoir of the device and the device ejects the oligonucleotide into the hybridization chamber of the array of LOC devices on the partially deep cut wafer. The data provided by the oligonucleotide spotting device includes automated computer controlled spotting oligonucleotides to an array of LOC devices on partially deep cut wafers, received and stored in an oligonucleotide reservoir. Oligonucleotide size, storage of the oligonucleotide specification in the digits of the digits, and delivery of the oligonucleotide specification to the LOC device or other department of the automated manufacturing environment. The oligonucleotide spotting device provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automating the manufacturing environment, increasing the security of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. The device can also be used as an intermediate fluid operation machine, which is required for the volume and positional correctness of the macroscopic stage φ to transfer the fluid to the microscopic level and positional correctness. The data provided by the oligonucleotide spotting device automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. The array of oligonucleotides to the LOC device on the partially deep cut wafer accelerates the loading process and reduces the cost of the loading process. GPD0 1 3 · 1 This aspect of the invention provides an oligonucleotide spotting device for use in a contactless spot-like probe onto a surface having a nucleic acid sequence that is intended to be recognized in a biological sample Complementary nucleic acid sequence, -133- 201209402 The oligonucleotide spotting device comprises: a monolithic support substrate: an emitter array formed on one side of the support substrate such that the emitter emits the oligonucleotide probe Droplets onto the surface; wherein, when in use, each of the emitters in the array are configured to emit droplets having a volume of less than 100 picoliters.

GPD 013.2 Preferably, each of the injectors in the array is configured to emit droplets having a volume of less than 25 picoliters. GPD 013.3 Preferably, each of the injectors in the array is configured to emit droplets having a volume of less than 6 picoliters. GPD 013.4 Preferably, each of the injectors in the array is configured to emit droplets having a volume between 0.1 picoliter and 1.6 picoliters. GPD013.5 Preferably, the monolithic support substrate has a reservoir side and

An exiting side opposite the reservoir side, the emitter array being formed on the exit side, the reservoir array being formed on the reservoir side, each of the emitters being configured for use with the probes A corresponding one of the reservoirs is in fluid communication. GPD 013.6 Preferably, each of the injectors has a plurality of nozzles such that the injectors are configured to eject droplets from the respective nozzles. GPD013.7 Preferably, the injector has a chamber and a plurality of actuators for containing fluid supplied by the corresponding reservoir, one of the actuators corresponding to each of the nozzles to cause the actuator Droplets of fluid from the chamber are ejected through the corresponding nozzles. GPD 013.8 Preferably, the actuator is a thermal actuator, each actuation -134 - 201209402 being configured to create a vapor bubble in the fluid. GPD 013.9 Preferably, the actuators in one of the injectors are organized to be individually actuated. GPD013.10 Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir to the chamber.

GPD013.il preferably, the oligonucleotide spotting device also has a CMOS circuit for providing a drive pulse to the actuator, the CMOS circuit having a pad for connecting to the control microprocessor, the control microprocessor The operative control of the relative movement between the nozzle and the surface to be spotted by the probe. GPD013.12 Preferably, the CMOS circuit has a memory for storing specifications relating to the probe in the reservoir. . GPD0 1 3 . 1 3 Preferably, the array of emitters is configured to spot the probe onto the surface at a density greater than 8 probe spots per square millimeter. GPD013.14 Preferably, the emitter array is configured to spot the probe onto the surface at a density greater than φ of 60 probes per square millimeter. 较佳GPD013.15 Preferably, the emitter The array was organized to spot the probe onto the surface at a density of 500 probes per square millimeter to 1,500 probe spots per square millimeter. GPD013.16 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 100 probes per second. GPD013.17 Preferably, the CMOS circuit is configured to generate a -135-201209402 drive pulse to cause the emitter array to spot the probe at a rate of more than 1,400 probes per second. On the surface. GPD013.18 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 20,000 probe points per second.

GPD013.19 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the array of emitters to be spotted at 3,000 probes per second to 1, 1 per second. The needle is spotted at a speed to spot the probe onto the surface. GPD013.20 Preferably, the reservoir array is integrally formed on the reservoir side of the single substrate.

This mass-produced and inexpensive oligonucleotide spotting device is used as part of a cost-effective automated mass production environment. The oligonucleotide is loaded into an oligonucleotide reservoir of the device, and the device ejects the oligonucleotide onto the surface to be spotted" by the oligonucleotide spotting device Automating includes automated computer controlled distribution of the oligonucleotide to the surface to be spotted, receiving the oligonucleotide specifications stored in the oligonucleotide reservoir, and storing the oligonucleotide size in the digits of the oligonucleotide Remember the body and transfer the oligonucleotide specifications to other departments of the automated manufacturing environment. The oligonucleotide spotting device provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automating the manufacturing environment, increasing the security of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. The device can also be used as an intermediary -136- 201209402 fluid-operated machine, which is required for the volume and positional correctness of the transfer from the fluid to the microscopic level and positional correctness. In particular, the ability of the device to spot the necessary low volume probes provides low probe cost, thus allowing for this inexpensive detection system. The data provided by the oligonucleotide spotting device automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment.

GPD014.1 This aspect of the invention provides an oligonucleotide spotting device for use in a contactless spot-like probe to a surface having a complementary nucleic acid sequence to be recognized in a biological sample. a nucleic acid sequence, the oligonucleotide spotting device comprising: an array of emitters, each emitter having an actuator for ejecting a droplet of liquid containing the probe; a CMOS circuit for providing a drive pulse to each The actuator emits droplets; wherein, when in use, the array of emitters spot the probe onto the surface at a rate of more than 100 probes per second. GPD 014.2 Preferably, the array of emitters spot the probe onto the surface at a rate of more than 1,400 probes per second. GPD 014.3 Preferably, the array of emitters spot the probe onto the surface at a rate of more than 2 000 probes per second. GPD014.4 Preferably, the emitter array spots the probe to the surface at a rate of between 1,300 probes per second to 1, 〇〇〇, 探针 probe spotting per second. on. - 137 - 201209402 GPD 014.5 Preferably, the CMOS circuit has a pad for connecting an external control microprocessor for operational control of the array of emitters. GPD 014.6 Preferably, the CMOS circuit has digital memory for storing identification data for identification by the external microprocessor controller. GPD014.7 Preferably, the digital memory stores probe type data and probe position data, the probe type data identifies a probe type in the device, and the probe position data identifies each of the probe type storage Location.

GPD014.8 Preferably, the oligonucleotide spotting device also has a support substrate having a reservoir side and an emitter side opposite the reservoir side, the emitter array being formed on the emitter side The reservoir array is formed on the reservoir side, each of the injectors being configured to be in fluid communication with a respective one of the probe reservoirs for injection containing a probe from the corresponding reservoir Droplets of the needle onto the surface. GPD 014.9 Preferably, each of the injectors has a plurality of nozzles such that the injectors are configured to eject droplets from the respective nozzles.

GPD014.10 Preferably, the injector has a chamber and a plurality of actuators for containing fluid supplied by the corresponding reservoir, one of the actuators corresponding to each of the nozzles to cause the actuator Droplets of fluid from the chamber are ejected through the corresponding nozzles. GPD014.il Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid. GPD 014.12 Preferably, the actuators in one of the injectors are organized to be individually actuated. GPD014.13 Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir - 138 - 201209402 to the chamber. GPD014.14 Preferably, the surface is a laboratory on wafer (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probe, the array of hybridization chambers being organized to accommodate the organism The complete series of oligonucleotide probes required for the predetermined analysis of the sample, and the reservoir array is organized to contain the complete series of oligonucleotide probes required for the predetermined analysis by the LOC device.

GPD014.15 Preferably, the reservoir array has more than 1 000 reservoirs. GPD014.16 Preferably, the CMOS circuit is configured to generate drive pulses such that the array of emitters is above 100 per second. The probe spotted the spot to the probe onto the surface. GPD 014.17 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 1,400 probes per second. GPD 014.18 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 20,000 probe points per second. GPD014.19 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the array of emitters to be spotted at a rate of between 3,000,000 probes per second to 1,000,000 probes per second. The probe is onto the surface. GPD014.20 Preferably, the reservoir array is integrally formed on the reservoir side of the single substrate. -139- 201209402 This mass-produced and inexpensive oligonucleotide spotting device is used as part of a cost-effective automated mass production environment. The oligonucleotide is loaded into the oligonucleotide reservoir of the device and the device ejects the oligonucleotide onto the surface to be spotted. Automated data provided by the oligonucleotide spotting device includes automated computer controlled distribution of the oligonucleotide to the surface to be spotted, receiving the oligonucleotides stored in the oligonucleotide reservoir The specification, storage of the oligonucleotide size in the digit memory, and delivery of the oligonucleotide specification to other departments of the automated manufacturing environment. The oligonucleotide spotting device provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automating the manufacturing environment, increasing the security of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. The high spotting rate of the device in turn provides high spot throughput and reduces the overall cost of the product inspection system. The device can also be used as an intermediate fluid operation machine, which is required for the volume and position correctness transfer fluid to the microscopic volume and positional correctness. The data provided by the oligonucleotide spotting device automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. GPD015.1 This aspect of the invention provides a biochemical deposition apparatus for depositing a biochemical substance on a surface without contact, the biochemical deposition apparatus comprising: a support substrate; -140-201209402 'Storage on one side of the substrate An array of containers for containing a plurality of biochemical materials; and an array of emitters on the other side of the support substrate, each of the emitters being configured for fluid communication with the reservoir; When in use, the array of emitters ejects droplets containing the biochemical onto the surface at a rate of more than 100 droplets per second. GPD015.2 Preferably, the emitter array is higher than per second

The rate of 1,400 droplets ejects droplets containing the biochemical onto the surface. GPD 015.3 Preferably, the array of emitters ejects droplets containing the biochemical onto the surface at a rate of more than 20,000 droplets per second. GPD 015.4 Preferably, the array of emitters ejects droplets containing the biochemical onto the surface at a rate of between 300,000 droplets per second to 1,000,000 droplets per second. GPD015.5 Preferably, the biochemical substance in the reservoir array is an oligonucleotide probe having a nucleic acid sequence complementary to the target nucleic acid sequence to be detected in the biological sample, and A surface-on-wafer laboratory (LOC) device having an array of hybridization chambers for receiving the oligonucleotide probe. GPD015.6 Preferably, the hybridization chamber array is configured to accommodate a complete series of oligonucleotide probes required for a predetermined analysis of the biological sample, and the reservoir array is configured to comprise the LOC A complete series of oligonucleotide probes required for the intended analysis of the device. -141 - 201209402 GPD015.7 Preferably, the reservoir array has more than 1 000 reservoirs. GPD 015.8 Preferably, each of the injectors has a plurality of nozzles such that the injectors are configured to eject droplets from the respective nozzles. GPD015.9 Preferably, the injector has a chamber and a plurality of actuators

a chamber for containing a fluid having a suspension oligonucleotide probe supplied by the corresponding reservoir, one of the actuators corresponding to each of the nozzles for causing the actuator to exit the chamber via the corresponding nozzle Droplets of Fluid" GPD 015.10 Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid. GPD015.il Preferably, the injector is configured to emit droplets having a volume of less than 1 〇〇 picoliter. GPD0 15.12 Preferably, the injector is configured to emit droplets having a volume of less than 25 picoliters. GPD015.13 Preferably, the injector is configured to emit volume

Droplets less than 6 picoliters. GPD015.14 Preferably, the injector is configured to emit droplets having a volume between 0.1 picoliter and 1.6 picolitres. GPD0 15.15 Preferably, the actuators in each of the injectors are configured to be individually actuated. GPD 015.16 Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir to the chamber.

GPD015.17 Preferably, the biodeposition device also has a CMOS circuit for providing a drive pulse to the actuator, the CMOS circuit having a -142-201209402 pad for connecting to the microprocessor controller, the microprocessor control The operability controls the relative movement between the nozzle and the surface to be spotted by the oligonucleotide probe. GPD015.18 Preferably, the CMOS circuit has a memory for storing specifications relating to the oligonucleotide probes in the reservoir. GPD015.19 Preferably, the emitter array is configured to emit droplets containing the biochemical substance at a density of more than 8 droplets per square millimeter to

GPD 015.20 Preferably, the array of emitters is configured to eject droplets containing the biochemical onto the surface at a density greater than 60 droplets per square millimeter. This mass-produced and inexpensive biochemical deposition unit is used as part of a cost-effective automated mass production environment. The biochemical material is loaded into the bioreactor of the device, and the device is deposited on the surface to be deposited by depositing the biochemical material from the biochemical reservoir onto the surface to be deposited. The data provided by the biochemical deposition device includes automatic computer controlled distribution of the biochemical substance to the surface to be spotted, receiving biochemical material specifications stored in the biochemical material reservoir, and storing the biochemical material specifications in the A number of billions of bodies, and the delivery of the biochemical specifications to the department of the automated manufacturing environment. The biochemical deposition device provides automated, volumetric and positional, fast and high density biochemical deposition techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the security of the automated manufacturing environment. Increase the safety of this automated manufacturing environment and reduce the cost of this automated manufacturing environment. The high deposition rate of the device in turn provides high deposition flux and reduces overall product cost. The device can also be used as an intermediate fluid operation machine, which is required for the volume and positional correctivity of the volume and position of the macroscopic level to be transferred to the microscopic level. The data provided by the biochemical deposition device provides automation, security, preservation and inexpensive data monitoring and management techniques in the automated manufacturing environment. GPD0 16.1 This aspect of the invention provides an oligonucleotide spotting device for use in a contactless spot-like probe having a nucleic acid complementary to a target nucleic acid sequence to be recognized in a biological sample. a sequence, the oligonucleotide spotting device comprising: an array of emitters, each emitter having an actuator for ejecting a droplet of liquid containing the probe; a CMOS circuit for providing a drive pulse to each of the The actuator is configured to eject droplets: wherein the array of emitters is configured to spot the probe onto the surface at a density greater than 1 probe spot per square millimeter. GPD 016.2 Preferably, the array of emitters is configured to spot the probe onto the surface at a density greater than 8 probe spots per square millimeter. GPD 016.3 preferably 'the emitter array is configured to spot the probe onto the surface at a density greater than 60 probe spots per square millimeter. 〇GPD016.4 preferably' the emitter array The probe was spotted onto the surface with a density of 5,000 probes per square millimeter to 1,500 probe points per square millimeter - 144 - 201209402. GPD 016.5 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 100 probes per second. GPD 016.6 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot the probe onto the surface at a rate of more than 1,400 probes per second.

GPD 016.7 Preferably, the CMOS circuit is configured to generate drive pulses to cause the emitter array to spot the probe onto the surface at a rate of more than 20,000 probe points per second. GPD016.8 Preferably, the CMOS circuit is configured to generate a drive pulse to cause the emitter array to spot at a rate of between 300,000 probes per second to 1,000,000 probe spots per second. The needle is on the surface. GPD 016.9 Preferably, the CMOS circuit has pads for connecting the external φ portion control microprocessor for operational control of the emitter array. GPD016.10 Preferably, the CMOS circuit has digital memory for storing identification data for identification by the external microprocessor controller. Preferably, the digital memory stores probe type data and probe position data, the probe type data identifies a probe type in the device and the probe position data identifies a storage of each probe type Location. GPD016.12 Preferably, the oligonucleotide spotting device also has a support substrate having a reservoir side and an emitter side opposite the reservoir side, the emitter array being formed on the emitter side The reservoir array -145-201209402 is formed on the reservoir side, each of the injectors being configured to be in fluid communication with a respective one of the probe reservoirs for use in the injection containing the corresponding A droplet of the probe of the reservoir onto the surface. GPD016.13 Preferably, each of the injectors has a plurality of nozzles such that the injectors are configured to eject droplets from the respective nozzles. GPD016.14 Preferably, the injector has a chamber and a plurality of actuators for containing fluid supplied by the corresponding reservoir, one of the actuators corresponding to each of the nozzles to cause the actuator Droplets of fluid from the chamber are ejected through the corresponding nozzles. GPD0 16.15 Preferably, the actuator is a thermal actuator, each actuator being configured to generate a vapor bubble in the fluid. GPD 016.16 Preferably, the actuators in one of the injectors are organized to be individually actuated. GPD 016.17 Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir to the chamber. GPD016.18 Preferably, the surface-based wafer-on-lab (LOC) device 'the LOC device has an array of hybridization chambers for accepting the oligonucleotide probe'. The hybrid chamber array is organized to accommodate the organism The complete series of oligonucleotide probes required for the predetermined analysis of the sample, and the reservoir array is organized to contain the complete series of oligonucleotide probes required for the predetermined analysis by the LOC device. GPD016.19 Preferably, the reservoir array has more than one reservoir. GPD016.20 Preferably, the reservoir array is integrally formed on the -146-201209402 reservoir side of the monolithic substrate. The mass-produced and inexpensive oligonucleotide spotting device is used as a cost-effective automation. A part of a large manufacturing environment. The oligonucleotide is loaded into the oligonucleotide reservoir of the device and the device ejects the oligonucleotide onto the surface to be spotted. Automated data provided by the oligonucleotide spotting device includes automated computer controlled distribution of the oligonucleotide to the surface to be spotted to receive an oligonucleoside φ stored in an oligonucleotide reservoir The acid specification, the storage of the oligonucleotide specification in the digits of the human body, and the delivery of the oligonucleotide specification to other departments of the automated manufacturing environment. The oligonucleotide spotting device provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automating the manufacturing environment, increasing the security of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. The device can also be used as an intermediate fluid operation machine, which is required for the volume and positional correctness of the macroscopic stage φ to transfer the fluid to the microscopic level and positional correctness. In particular, the device provides a low end product size with the necessary high density spotting capabilities, thereby allowing the inexpensive inspection system. The data provided by the oligonucleotide spotting device automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. GPD017.1 This aspect of the invention provides a biochemical deposition apparatus for depositing a biochemical substance on a surface without contact, the biochemical deposition apparatus comprising: -147- 201209402 support substrate: a reservoir on one side of the substrate An array configured to contain a plurality of biochemical materials; and an array of emitters on the other side of the support substrate, the emitter being in fluid communication with the reservoir and configured to be above each square The density of one droplet of millimeters ejects droplets containing the biochemical onto the surface. GPD 017.2 Preferably, the array of emitters is configured to eject droplets containing the biochemical onto the surface at a density greater than 8 droplets per square millimeter. GPD 017.3 Preferably, the array of emitters is configured to eject droplets containing the biochemical onto the surface at a density greater than 60 droplets per square millimeter. GPD 017.4 Preferably, the array of emitters is configured to eject droplets containing the biochemical onto the surface at a density of from 500 droplets per square millimeter to 1,500 droplets per square millimeter. GPD 017.5 Preferably, the array of emitters is configured to eject droplets containing the biochemical onto the surface at a rate of more than one droplet per second. GPD 017.6 Preferably, the array of emitters is configured to eject droplets containing the biochemical onto the surface at a rate of more than 1,400 droplets per second. GPD 017.7 Preferably, the array of emitters is configured to eject droplets containing the biochemical onto the surface at a rate of more than 20,000 droplets per second. -148- 201209402 GPD017.8 Preferably, the emitter array is configured to emit droplets containing the biochemical to the surface at a rate of between 300,000 droplets per second to 1,000,000 droplets per second. on. GPD017.9 Preferably, the biochemical system in the reservoir array

An oligonucleotide probe having a nucleic acid sequence complementary to a target nucleic acid sequence to be detected in a biological sample, and the surface is a laboratory on-wafer (LOC) device having a hybridization chamber An array is used to accept the oligonucleotide probe. GPD 017.10 Preferably, the hybridization chamber array is configured to accommodate a complete series of oligonucleotide probes required for predetermined analysis of the biological sample, and the reservoir array is organized to comprise the LOC A complete series of oligonucleotide probes required for the intended analysis of the device. GPD017.il Preferably, the reservoir array has more than one reservoir. GPD 017.12 Preferably, each of the injectors has a plurality of nozzles, and the emitters are configured to emit droplets from the respective nozzles. GPD017.13 Preferably, the injector has a chamber and a plurality of actuators for containing a fluid having a suspension oligonucleotide probe supplied by the corresponding reservoir, one of the actuators corresponding to Each of the nozzles causes the actuator to eject droplets of fluid from the chamber via the corresponding nozzle. GPD 017.14 Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid. GPD 017.15 Preferably, the injector is configured to emit droplets having a volume of less than 100 picoliters. - 149 - 201209402 GPD 017.16 Preferably, the actuators in each of the injectors are configured to be individually actuated. GPD 017.17 Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir to the chamber.

GPD 017.18 Preferably, the biochemical deposition apparatus also has a CMOS circuit for providing a drive pulse to the actuator, the CMOS circuit having a pad for connecting to a microprocessor controller, the microprocessor controller being operatively controlled The relative movement of the nozzle to the surface to be spotted by the oligonucleotide probe. GPD 017.19 Preferably, the CMOS circuit has a memory for storing specifications relating to the oligonucleotide probes in the reservoir. GPD 017.20 Preferably, the injector is configured to emit droplets having a volume ranging from 0.1 picoliters to 1.6 picoliters.

This mass-produced and inexpensive biochemical deposition unit is used as part of a cost-effective automated mass production environment. The biochemical material is loaded into the bioreactor of the device and the device is deposited on the surface by depositing the biochemical material from the biochemical reservoir onto the surface to be deposited. The data provided by the biochemical deposition device includes automatic computer controlled distribution of the biochemical substance to the surface to be spotted, receiving biochemical material specifications stored in the biochemical material reservoir, and storing the biochemical material specifications in the Digital memory, and the department that delivers the biochemical specifications to the automated manufacturing environment. The biochemical deposition device provides automated, volumetric and positional, fast and high-density biochemical deposition technology to simplify the complexity of the automated manufacturing environment -150-201209402, increase the reliability of the automated manufacturing environment, and increase the automated manufacturing environment. The preservation, the safety of the automated manufacturing environment and the cost of the automated manufacturing environment. The device can also be used as an intermediate fluid operation machine, which is required for the volume and positional correctness of the fluid from the macroscopic level to the volume and positional correctness of the microscopic level. In particular, the device provides a low end product size with the ability to deposit biochemical materials at the necessary high density, thus allowing for this inexpensive product.

The data provided by the biochemical deposition device provides automation, security, preservation and inexpensive data monitoring and management techniques in the automated manufacturing environment. GAL001.1 This aspect of the invention provides a robotic system for spotting oligonucleotides comprising: an oligonucleotide spotting device for use in contactless spot-like oligonucleotides onto a surface, The oligonucleotide spotting device has an array of emitters, at least one reservoir in fluid communication with one or more of the injectors, and a CMOS drive circuit each having a droplet ejection actuator for emitting the oligo a droplet of a liquid of a nucleotide to a surface, the CMOS driving circuit for providing a driving pulse for droplet ejection to each of the droplet ejection actuator; and for loading an oligonucleotide to the oligonucleotide A device for spotting a device having a platform and a plurality of oligonucleotide containers mounted for movement relative to the platform, the platform being for detachably mounting a plurality of the oligonucleotide spotting devices, each of the oligo The nucleotide container has a droplet dispenser to eject a droplet of fluid containing the oligonucleotide to a reservoir of the oligonucleotide spotting device. GAL001.2 Preferably, the oligonucleotide spotting device has a pad-151 - 201209402 for electrically connecting the CMOS driver circuit and the device to enable the device to download oligonucleotide data to the CMOS driver circuit Memory inside. GAL001.3 Preferably, the apparatus has a camera to optically align the platform with the drop dispenser. GAL001.4 Preferably, the oligonucleotide spotting device has a support substrate having a reservoir side having an reservoir side and an emitter side opposite the reservoir side, the at least one reservoir being tied to The reservoir side is formed, and the emitter array is formed on the emitter side. GAL001.5 Preferably, the oligonucleotide spotting device has an array of reservoirs on the reservoir side, each of the reservoirs being in fluid communication with two or more of the injectors. GAL001.6 Preferably, the oligonucleotide is configured to be a probe that hybridizes to a target nucleic acid sequence in a biological sample, and the liquid system probe solution is such that the droplet ejection actuator is via the respective The individual nozzles of the injector emit droplets of the probe solution. GAL001.7 Preferably, the oligonucleotide spotting device has at least one common chamber for containing a fluid to be ejected by the plurality of nozzles. GAL001.8 Preferably, each of the common chambers has a plurality of chamber inlets in fluid communication with the reservoir. GAL001.9 Preferably, the oligonucleotide spotting device has a reservoir array and a common chamber array in fluid communication with one of the reservoirs, respectively. GAL 001.10 Preferably, the droplet ejection actuators each have a heater for generating vapor bubbles in the fluid to eject droplets via nozzles corresponding to the droplet ejection actuators. Preferably, the injectors are each configured to emit droplets of the probe solution having a volume of less than 100 picoliters. GAL001.12 Preferably, the injectors are each configured to emit droplets of the probe solution having a volume of less than 25 picoliters. GAL001.13 Preferably, the injectors are each configured to emit droplets of the probe solution having a volume of less than 6 picoliters.

GAL001.14 Preferably, the injectors are each configured to emit droplets of the probe solution having a volume between 0.1 picoliters and 1.6 picoliters. GAL001.15 Preferably, the container is for use in an aliquot of the probe solution, each bottle having a quality assurance wafer containing a memory for storing information identifying the oligonucleotide. GAL001.16 Preferably, the bottles each have a drop dispenser and an electrical contact for receiving an actuation signal to activate the drop dispenser. GAL001.17 Preferably, the droplet dispenser has a piezoelectric actuator GAL001.18. Preferably, the apparatus is configured to transfer data from the quality assurance wafer of the bottle to the oligonucleotide point. The CMOS circuit of the device. GAL001.19 Preferably, the bottle is suspended from a shelf of an oligonucleotide spotting device adjacent one of the platforms, the frame being configured to establish an electrical connection with each of the bottles. The apparatus for loading an oligonucleotide spotting device is used as part of a cost-effective automated mass production environment to dispense oligonucleotides contained in oligonucleotide microbottles to the oligonucleoside Acid spotting device in the oligo-153- 201209402 nucleotide reservoir. The data provided by the apparatus is automated including automated computer controlled distribution of the oligonucleotide to the oligonucleotide reservoir of the oligonucleotide spotting device, and examination of the oligonucleoside stored in the memory of the microbottle. The acid data is aligned with a specification list of oligonucleotides that must be loaded into the oligonucleotide spotting device, and the oligonucleotide data is stored into the memory of the oligonucleotide spotting device. The device for loading an oligonucleotide spotting device provides automated and volumetric and positionally accurate oligonucleotide dispensing techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the automation The preservation of the manufacturing environment, increasing the safety of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. The data provided by the device for loading the oligonucleotide spotting device is automated to provide automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. GPA001.1 This aspect of the invention provides an oligonucleotide spotting robot for spotting an oligonucleotide probe to a microfluidic device having an oligonucleoside loaded on the microfluidic device a digital memory of the acid probe, the oligonucleotide dispensing robot comprising: a reservoir array for accommodating the oligonucleotide probe suspended in the fluid; an emitter array, each of the emitter systems Arranging to be in fluid communication with a respective one of the reservoirs: a mounting surface for removably mounting the microfluidic device for movement of the microfluidic device relative to the injector; and -154 -

201209402 a control processor for operationally controlling the injector and the face; wherein the control processor is configured to activate the injector, the selective emitter to activate registration with the microfluidic device and to download the Digital memory. GPA 001.2 Preferably, the oligonucleotide spotting machine camera optically feeds the registration between the emitter devices selected by the control processor. GPA001.3 Preferably, the reservoir array has a reservoir CMOS CMOS circuit that exceeds the reservoir GPA001.4. Preferably, the oligonucleotide spotting machine is interposed between the reservoir array and the emitter array. The array is configured to drive the array of emitters based on the microprocessor number from the control, wherein the CMOS circuit stores information of the nucleotide probes. GPA001.5 Preferably, the mounting surface is a platform that is configured to move orthogonal axes, and the array of injectors is immediately adjacent to and facing the platform. GPA001.6 Preferably, the microfluidic device is a wafer (LOC) device having an array of hybridization chambers for nucleotide probes having nucleic acids complementary to the acid sequence to be recognized in the biological sample a sequence, the hybridization chamber array is a complete oligonucleotide probe required for predetermined analysis of a tissue biological sample, the reservoir array being configured to include the installation of the LOC device for the installation of the data The person also has a circuit with the micro-flowing 1 000 people, and the control letter about the oligo to attach the laboratory to the nucleus of the oligo to accommodate the system along the platform, and the predetermined score - 155- 201209402 Analysis of the complete range of oligonucleotide probes required. GPA 001.7 Preferably, each of the injectors has a plurality of nozzles such that the injectors are configured to eject droplets from the respective nozzles. GPA001.8 Preferably, the injector has a chamber and a plurality of actuators for containing fluid from the corresponding reservoir, one of the actuators corresponding to each of the nozzles to cause the actuator to be The corresponding nozzle emits droplets of fluid from the chamber.

GPA001.9 Preferably, the actuator is a thermal actuator, each actuated

The I device is configured to generate vapor bubbles in the fluid. GPA001. 10 Preferably, the injector is configured to emit droplets having a volume of less than 100 picoliters. GPA001.il Preferably, the injector is configured to emit droplets having a volume of less than 25 picoliters. GPA 001.12 Preferably, the ejector is configured to emit droplets having a volume of less than 6 picoliters.

GPA 001.13 Preferably, the ejector is configured to eject droplets having a volume ranging from 0.1 picoliter to 1.6 picoliters. GPA 001.14 Preferably, the actuators in each of the injectors are configured to be individually actuated. GPA00 1.15 Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir to the chamber. GPA 001.16 Preferably, the array of emitters is configured to spot the oligonucleotide probe onto the LOC device at a density greater than 1 probe spot per square millimeter. Preferably, the array of emitters is configured to spot the oligonucleotide onto the LOC device at a density greater than 8 probe spots per square millimeter. GPA 001.18 Preferably, the array of emitters is configured to spot the oligonucleotide onto the LOC device at a density of more than 60 probe spots per square millimeter.

GPA001.19 Preferably, the emitter array is configured to spot the oligonucleotide at a density of 500 probes per square millimeter to a density of 15 探针0 probes per square millimeter. On the LOC device. GPA 001.20 Preferably, the array of emitters is configured to spot the oligonucleotide onto the LOC device at a rate of more than 100 probe spots per second. The oligonucleotide spotting robot is used as part of a cost-effective automated mass production environment. The loaded oligonucleotide spotting device is picked up by the robot, which places the oligonucleotide spotting device and commands φ the device to eject the oligonucleotide onto the surface to be spotted. Automating the data provided by the oligonucleotide spotting robot includes automated computer controlled distribution of the oligonucleotide to the surface to be spotted, reading the reservoir stored in the oligonucleotide spotting device Specification of the oligonucleotide in question, checking the oligonucleotide size stored in the reservoir of the oligonucleotide spotting device to compare with the relevant database, storing the oligonucleotide specification in the The memory of the spotted device, and the delivery of the oligonucleotide specification to other departments of the automated manufacturing environment. The oligonucleotide spotting robot provides automated, volumetric and positional precision -157-201209402, fast and high-density oligonucleotide spotting technology to simplify the complexity of the automated manufacturing environment and increase the reliability of the automated manufacturing environment Sexuality increases the security of the automated manufacturing environment, increases the security of the automated manufacturing environment, and reduces the cost of the automated manufacturing environment. The data provided by the oligonucleotide spotting robot automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. GPA003.1 This aspect of the invention provides an oligonucleotide spotting robot for spotting an oligonucleotide probe to an array of on-wafer laboratory (LOC) devices, each LOC device having a loading on the LOC a digital memory of the data of the oligonucleotide probe of the device, the oligonucleotide dispensing robot comprising: a reservoir array for accommodating the oligonucleotide probe suspended in the fluid: an array of emitters, Each of the injectors is configured to be in fluid communication with a respective one of the reservoirs; a mounting surface for detachably mounting the array of LOC devices for output relative to the array of LOC devices a movement of the device; and a control processor for operatively controlling the injector and the mounting surface; wherein the control processor is configured to activate the emitter, move the selected injector to activate A plurality of the LOC devices are registered and download data that is particularly relevant to each of the LOC devices to the digital device of the LOC device. -158- 201209402 GPA003.2 Preferably, the oligonucleotide spotting robot also has a camera to optically feedback the registration between the emitter selected by the control processor and the LOC device. GPA003.3 Preferably, the reservoir array has more than one reservoir. GPA003.4 Preferably, the oligonucleotide spotting robot also has

a CMOS circuit interposed between the reservoir array and the emitter array, the CMOS circuit being configured to drive an array of the emitters according to a control signal from the control microprocessor, wherein the CMOS circuit stores Information on nucleotide probes. GPA003.5 Preferably, the array of LOC devices is mounted to a printed circuit board (PCB), which in turn is detachably mounted to the mounting surface, the mounting surface being configured to move along two orthogonal axes The platform, and the array of injectors is mounted adjacent to the platform and facing the platform. GPA003.6 Preferably, each of the LOC devices has a hybrid array φ column for accepting the oligonucleotide probe having a nucleic acid sequence complementary to the target nucleic acid sequence to be recognized in the biological sample, The hybridization chamber array is configured to accommodate a complete series of oligonucleotide probes required for a predetermined analysis of the biological sample, and the reservoir array is configured to contain the predetermined analysis required by the LOC device Complete range of oligonucleotide probes. GPA 003.7 Preferably, each of the injectors has a plurality of nozzles such that the injectors are configured to eject droplets from the respective nozzles. GPA003.8 Preferably, the injector has a chamber and a plurality of actuators for containing fluid from the corresponding reservoir, one of the actuators corresponding to -159 - 201209402 corresponding to each of the nozzles The actuator emits droplets of fluid from the chamber via the corresponding nozzle. GPD003.9 Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid. GPA003.10 Preferably, the injector is configured to emit droplets having a volume of less than 100 picoliters.

GPA003.il Preferably, the injector is configured to emit droplets having a volume of less than 25 picoliters. GPA003.1 2 Preferably, the injector is configured to emit droplets having a volume of less than 6 picoliters. GPA 003.1 Preferably, the injector is configured to eject droplets having a volume ranging from 0.1 picoliters to 1.6 picoliters. GPA 003.1 4 Preferably, the actuators in each of the injectors are configured to be individually actuated.

GPA 003.1 5 Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir to the chamber. GPA003.1 6 Preferably, the array of emitters is configured to spot the oligonucleotide probe onto the LOC device at a density greater than 1 probe spot per square millimeter. GPA 003.1 Preferably, the array of emitters is configured to spot the oligonucleotide onto the LOC device at a density greater than 8 probe spots per square millimeter. GP A003.1 8 Preferably, the array of emitters is configured to spot the oligonucleotide onto the -160-201209402 LOC device at a density of more than 60 probe spots per square millimeter. GPA003.19 Preferably, the emitter array is configured to spot the oligonucleotide at a density of 500 probes per square millimeter to 15 probes per square millimeter. On the LOC device. GPA003.20 Preferably, the array of emitters is configured to spot the oligonucleotide onto the LOC device at a rate of more than 100 probes per second.

The oligonucleotide spotting robot is used as part of a cost-effective automated mass production environment. The loaded oligonucleotide spotting device is picked up by the robot. The robot places the oligonucleotide spotting device and commands the device to emit the oligonucleotide to the array of LOC devices mounted on the PCB wafer. In the hybridization room. The data provided by the oligonucleotide spotting robot includes automated computer controlled distribution of the oligonucleotide to the hybridization chamber of the array of LOC devices mounted on the PCB wafer, read and stored in the oligo Specification of the oligonucleotide in the reservoir of the gluconate spotting device, checking the oligonucleotide specifications stored in the reservoir of the oligonucleotide spotting device to compare with the relevant database, and storing the oligo Nucleotide specifications are in the memory of the LOC device to be spotted, and the oligonucleotide specifications are delivered to other departments of the automated manufacturing environment. The oligonucleotide spotting robot provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automating the manufacturing environment, increasing the security of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. -161 - 201209402 The data provided by the oligonucleotide spotting robot is automated to provide automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. An array of spotted oligonucleotides to the LOC device mounted on the PCB wafer accelerates the loading process and reduces the cost of the loading process, and after mounting the LOC device to the PCB wafer and soldering them, The LOC device enhances the chemical and physical integrity of the oligonucleotide. GPA004.1 This aspect of the invention provides an oligonucleotide spotting robot for spotting an oligonucleotide probe to a wafer, on which a wafer-on-lab (LOC) device is constructed, Each LOC device has a digital memory of data relating to reagents loaded in the LOC device, the oligonucleotide dispensing robot comprising: a reservoir array for containing the oligonucleotide probe suspended in the fluid; An array of injectors, each of the injectors being configured to be in fluid communication with a respective one of the reservoirs; a mounting surface for detachably mounting the array of LOC devices for array of the LOC devices Movement relative to the injector: and a control processor for operatively controlling the injector and the mounting surface; wherein the control processor is configured to activate the emitter and move the selected injector to The registration with one or more of the LOC devices is activated and data associated with each of the LOC devices is downloaded to the digital memory of the LOC device. Preferably, the oligonucleotide spotting robot also has a camera to optically feedback the registration between the injector selected by the control processor and the LOC device. GPA004.3 Preferably, the reservoir array has more than one reservoir.

GPA004.4 Preferably, the oligonucleotide spotting robot also has a CMOS circuit between the reservoir array and the emitter array. The CMOS circuit is configured to be based on the microprocessor from the control. A control signal drives an array of the emitters, wherein the CMOS circuitry stores information about the oligonucleotide probes. GPA004.5 Preferably, the germanium wafer is partially cut to prepare for cutting into separate LOC devices, and the germanium wafer is detachably mounted to the mounting surface, the mounting surface being structured The platform is moved along two orthogonal axes, and the array of emitters is mounted adjacent to the platform and facing the platform. GPA004.6 Preferably, each of the LOC devices has an array of hybridization chambers for receiving the oligonucleotide probe having a nucleic acid sequence complementary to a target nucleic acid sequence to be recognized in a biological sample, the hybridization chamber The array is configured to accommodate a complete series of oligonucleotide probes required for a predetermined analysis of the biological sample, and the reservoir array is configured to contain the integrity required for the predetermined analysis performed by the LOC device Oligonucleotide Probe Series ◊ GPA004.7 Preferably, each of the injectors has a plurality of nozzles such that the emitters are configured to eject droplets from each nozzle. GPA004.8 Preferably, the injector has a chamber and a plurality of actuators - 163 - 201209402 for containing fluid from the corresponding reservoir, one of the actuators corresponding to each of the nozzles to An actuator emits droplets of fluid from the chamber via the corresponding nozzle. GPA 04.9 Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid. GPA004.1 0 Preferably, the injector is configured to emit droplets having a volume of less than 1 〇〇 picoliter.

GPA004.il Preferably, the injector is configured to emit droplets having a volume of less than 25 picoliters. GPA004.1 2 Preferably, the injector is configured to emit droplets having a volume of less than 6 picoliters. GPA004.1 3 Preferably, the injector is configured to emit droplets having a volume ranging from 0.1 picoliters to 1.6 picoliters. GPA004.1 4 Preferably, the actuators in each of the injectors are configured to be individually actuated.

GPA 004.1 5 Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir to the chamber. GPA004.1 6 Preferably, the emitter array is configured to spot the oligonucleotide probe onto the LOC device at a density greater than 1 probe spot per square millimeter" GPA004.1 7 Preferably, the array of emitters is configured to spot the oligonucleotide onto the LOC device at a density greater than 8 probe spots per square millimeter. GPA004.1 8 Preferably, the emitter array is configured to spot the oligonucleotide onto the LOC device at a density of 60 probe spots per square millimeter above -164 - 201209402" GPA004.19 Preferably, the array of emitters is configured to spot the oligonucleotide onto the LOC device at a density of 500 probes per square millimeter to a density of 1,500 probes per square millimeter. GPA004.20 Preferably, the emitter array is configured to spot the oligonucleotide to the LOC at a rate of more than one probe spot per second.

The oligonucleotide spotting robot is used as part of a cost-effective automated mass production environment. The loaded oligonucleotide spotting device is picked up by the robot, which places the oligonucleotide spotting device and commands the device to emit the oligonucleotide to the LOC device on the partially deep cut wafer The array is in the hybridization chamber. The data provided by the oligonucleotide spotting robot includes automated computer controlled distribution of the oligonucleotide to the hybridization φ chamber of the array of LOC devices on the partially deep cut wafer, read and stored in The specification of the oligonucleotide in the reservoir of the oligonucleotide spotting device, and the specification of the oligonucleotide stored in the reservoir of the oligonucleotide spotting device are compared with the relevant database, The oligonucleotide is stored in the cell of the LOC device to be spotted, and the oligonucleotide is transferred to other sectors of the automated manufacturing environment. The oligonucleotide spotting robot provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automated manufacturing environment security, increased automation of the automated manufacturing environment and reduced costs of the automated manufacturing environment. The data provided by the oligonucleotide spotting robot automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. The array of oligonucleotides to the LOC device on the partially deep cut wafer accelerates the loading process and reduces the cost of the loading process. GPA005.1 This aspect of the invention provides an oligonucleotide spotting robot for spotting an oligonucleotide probe onto a substrate, the oligonucleotide dispensing robot comprising: a reservoir array for accommodating An oligonucleotide probe suspended in a fluid; an array of emitters, each of the emitters being configured to be in fluid communication with a respective one of the reservoirs; a mounting surface for detachably Mounting the substrate for movement of the substrate relative to the emitter; and controlling a processor for operatively controlling the emitter and the mounting surface: wherein the array of emitters is configured to be higher than 1 mm per square millimeter The density of the probe spotted the probe onto the surface. GPA 005.2 Preferably, the array of emitters is configured to spot the probe onto the substrate at a density greater than 8 probe spots per square millimeter. GPA005.3 Preferably, the array of emitters is configured to spot the probe onto the substrate at a density greater than 60 probe spots per square millimeter - 166 - 201209402 GPA 005.4 preferably, The array of emitters was organized to spot the probe onto the substrate at a density of 500 probes per square millimeter to a density of 1,500 probes per square millimeter. GPA005.5 Preferably, the substrate is on-wafer laboratory (LOC)

Means, the LOC device having digital memory for data of an oligonucleotide probe loaded on the LOC device, and the control microprocessor is configured to activate the emitter and move the selected emitter to Activating the registration with the LOC device and downloading the data to the digital memory. GPA005.6 Preferably, the oligonucleotide spotting robot also has a camera to optically feedback the registration between the emitter selected by the control processor and the LOC device. GPA005.7 Preferably, the reservoir array has more than 1000 reservoirs. GPA005.8 Preferably, the oligonucleotide spotting robot also has a CMOS circuit between the reservoir array and the emitter array, the φ CMOS circuit being configured to be based on the microprocessor from the control A control signal drives the array of emitters, wherein the CMOS circuit stores information about the oligonucleotide probe. GPA 005.9 Preferably, the mounting surface is a platform that is configured to move along two orthogonal axes and the array of emitters is mounted adjacent to the platform and facing the platform. GPA005.1 0 Preferably, the LOC device has an array of hybridization chambers for receiving the oligonucleotide probe having a nucleic acid sequence complementary to the target nucleic acid sequence to be recognized in the biological sample, the hybridization chamber Array-167-201209402 is configured to accommodate a complete series of oligonucleotide probes required for predetermined analysis of the biological sample, and the reservoir array is configured to include a predetermined analysis by the LOC device A complete range of oligonucleotide probes is required. GPA 005. il. Preferably, each of the injectors has a plurality of nozzles such that the injectors are configured to eject droplets from the respective nozzles. GPA005.1 2 Preferably, the injector has a chamber and a plurality of actuators for containing fluid from the corresponding reservoir, one of the actuators corresponding to each of the nozzles to cause the actuator Droplets of fluid from the chamber are ejected through the corresponding nozzles. GPA 005.1 Preferably, the actuator is a thermal actuator, each actuator being configured to create a vapor bubble in the fluid. GPA 005.1 Preferably, the injector is configured to eject droplets having a volume of less than 100 picoliters. GPA 005.1 5 Preferably, the injector is configured to emit droplets having a volume of less than 25 picoliters. GPA 005.1 6 Preferably, the injector is configured to emit droplets having a volume of less than 6 picoliters. GPA 005.1 Preferably, the injector is configured to emit droplets having a volume ranging from 0.1 picoliter to 1.6 picoliters. GPA 005.1 8 Preferably, the actuators in each of the injectors are configured to be individually actuated. GPA 005.1 9 Preferably, each of the injectors has a plurality of inlet passages extending from the reservoir to the chamber. GPA 005.20 Preferably, the array of emitters is organized to spot the oligonucleotide onto the substrate at a rate of -168 to 201209402 per 100 probes per second.

The oligonucleotide spotting robot is used as part of a cost-effective automated mass production environment. The loaded oligonucleotide spotting device is picked up by the robot, which places the oligonucleotide spotting device and instructs the device to eject the oligonucleotide into the hybridization chamber of the LOC device to be spotted. Automating the data provided by the oligonucleotide spotting robot includes automated computer controlled distribution of the oligonucleotide to the hybridization chamber of the LOC device to be spotted, reading and storing in the oligonucleotide spotting The specification of the oligonucleotide in the reservoir of the device, the specification of the oligonucleotide stored in the reservoir of the oligonucleotide spotting device, the alignment with the relevant database, and the storage of the oligonucleotide specification In the memory of the LOC device to be spotted, and transmitting the oligonucleotide specification to other departments of the automated manufacturing environment. The oligonucleotide spotting robot provides automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify the complexity of the automated manufacturing environment, increase the reliability of the automated manufacturing environment, and increase the Automating the manufacturing environment, increasing the security of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. In particular, the robot provides the low end LOC device size with the necessary high density spotting capabilities, thus allowing for this inexpensive inspection system. The data provided by the oligonucleotide spotting robot automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment. GSS 00 1.1 This aspect of the invention provides a system for microarray spotting and genetic analysis, the system comprising: -169- 201209402 a container containing a probe for hybridization to different target nucleic acid sequences; In each of the integrated circuits of the container, each of the integrated circuits has a container for storing probe specifications of probes in the container, and a microfluid for supporting a probe array selected from the container. a device, the selected probe corresponding to a desired genetic test analysis, the microfluidic device having a device digital memory; an oligonucleotide spotting robot configured to spot the selected probe to the microfluidic device Forming the probe array, the oligonucleotide spotting robot having a control microprocessor for downloading the specification data to the device digital device; and a device reader for accessing the device digital device The specification data of the memory is used to analyze the hybridization data from the microfluidic device. GSS001.2 Preferably, the microfluidic device has: a support substrate: a microsystem technology (MST) layer for supporting the selected probe array: and a CMOS circuit interposed between the MST layer and the support substrate The CMOS circuit has a light sensor for detecting probe hybridization in the selected probe array. GSS001.3 Preferably, the probe is a fluorescence resonance energy transfer (FRET) probe. GSS001.4 Preferably, the device digital memory also stores device identification data for uniquely identifying the microfluidic device from -170 to 201209402. Preferably, the system also has a test module in which the microfluidic device is mounted, the test module being configured for transferring data between the device digital memory and the reader. GSS00 1.6 Preferably, the test module is coupled to the reader via a universal serial bus (USB) connector.

GSS00 1.7 Preferably, during use, the CMOS circuit is powered by the reader via the USB connector. GSS00 1.8 Preferably, the test module also has an excitation source for generating a fluorescent emission from the FRET probe that has hybridized to any of the target nucleic acid sequences. GSS00 1.9 preferably, during use, extinguishes the excitation light prior to activation of the photosensor, and the CMOS circuit is configured to activate the photosensor for a predetermined period of time after deactivation of the excitation source. GSS001.10 Preferably, the distance between the light sensor and the probe is less than 249 microns. GSS001.il Preferably, the system also has a container for receiving a biological sample containing the nucleic acid sequence of the target, the container being in fluid communication with the inlet of the microfluidic device. GSS001.12 Preferably, the microfluidic device has a fluid flow path from the inlet to the endpoint sensor, the fluid flow path being configured to carry the target nucleic acid sequence into contact with the probe array by capillary action. GSS001.13 Preferably, the microfluidic device has a plurality of reagent reservoirs for processing the different reagents required for the biological sample. -171 - 201209402 GSS001.14 Preferably, each of the container digits stores identification data, the identification data distinguishes the container for the microfluidic device from the other containers, and the control microprocessor is configured The identification data is downloaded to the device digital memory of the microfluidic device to be spotted. GSS 001.15 Preferably, the containers each have a droplet generator for ejecting droplets of the probe suspension onto the microfluidic device. GSS001.16 Preferably, the system also has a mounting surface for mounting the microfluidic device for movement of the microfluidic device relative to the container such that the control microprocessor controls both the container and the mounting portion Activating the droplet generator 〇GSS001.17 that is selected and moved to a container that is registered with the microfluidic device. Preferably, the system also has a camera for optically feeding back the container selected by the control microprocessor and the microfluid Registration between devices. GSS001.18 Preferably, the data stored in the container and the digits of the device are encrypted. GSS 001.1 9 Preferably, the droplet generator is configured to emit droplets having a volume of less than 6 picoliters. GSS 001.20 Preferably, the selected probe array comprises more than one probe in an area of less than 1500 microns by 1500 microns. The system for variable microarray spotting and gene analysis provides an automated, fast, simple, and low-cost collection of easy-to-use and inexpensive application-specific/application-optimized detection systems that use a library of LOC devices, Oligonucleotide spotting device, device for loading oligonucleotide spotting device, -172-201209402 oligonucleotide spotting robot and oligonucleotide probe stored in micro-bottle with digital counting body The library of needles. All processing steps from the probe to the LOC device are automated. GSL001.1 This aspect of the invention provides a system for loading reagents into a microfluidic device for genetic analysis, the system comprising: a container containing a reagent for processing a biological sample in the microfluidic device:

Each of the integrated circuits is fixed to an integrated circuit of each of the containers, each of the integrated circuits having a container digital memory for storing reagent specifications of the reagents in the container; and a microfluidic device for performing the desired gene test analysis, the micro-fluid device The fluid device has a device digital memory; a reagent dispensing device for loading a selection reagent to the microfluidic device, the reagent dispensing device having control for accessing the container digital memory and transmitting the specification data to the device digital memory a microprocessor; and a device reader for accessing reagent specification data of the device digital memory to analyze test analysis results from the microfluidic device. GSL001.2 Preferably, the microfluidic device has: a support substrate; a microsystem technology (MST) layer for supporting an oligonucleotide probe array, the oligonucleotide probe array being used for the biological sample a hybridization of the nucleic acid sequence in the target; and a CMOS circuit between the MST layer and the support substrate. The CMOS circuit has a light sensor for detecting hybridization in the probe array - 173 - 201209402 GSL001.3 Preferably, the probe is a fluorescence resonance energy transfer (FRET) probe. Preferably, the device digital memory also stores device identification data for uniquely identifying the microfluidic device. GSL0 0 1.5 Preferably, the system also has a test module in which the microfluidic device is mounted, the test module being configured for transferring data between the device digital memory and the reader. G S LO 0 1.6 Preferably, the test module is coupled to the reader via a universal serial bus (USB) connector. GSL001.7 Preferably, during use, the CMOS circuit is powered by the reader via the USB connector. GSL001.8 Preferably, the test module also has an excitation source for generating a fluorescent emission from the FRET probe that has hybridized to any of the target nucleic acid sequences. Preferably, GSL 00 1.9 extinguishes the excitation light prior to activation of the light sensor during use, and the CMOS circuit is configured to activate the light sensor for a predetermined period of time after deactivation of the excitation light source. GSL001 .1 0 Preferably, the distance of the light sensor from the probe is less than 249 microns. GSL001.il Preferably, the system also has a container for receiving a biological sample containing the nucleic acid sequence of the target, the container being in fluid communication with the inlet of the microfluidic device. GSL001.12 Preferably, the microfluidic device has a fluid flow path from the inlet to the end point sensor of -174-201209402, the fluid flow path being configured to carry the target nucleic acid sequence by capillary action and the probe Needle array contact. GSL001.13 Preferably, the microfluidic device has a plurality of reagent reservoirs for processing the reagents required for the biological sample.

GSL001.14 Preferably, each of the container digital memory stores identification data 'the identification data distinguishes the container for the microfluidic device from the other container, and the control microprocessor is configured to download the identification data. To the device digital memory of the microfluidic device to be spotted. GSL001.15 Preferably, the containers each have a droplet generator for ejecting the reagent droplets to the reagent reservoir. GSL001.16 Preferably, the system also has a mounting surface for mounting the microfluidic device for movement of the microfluidic device relative to the container such that the control microprocessor controls both the container and the mounting portion Activating the droplet generator GSL001.17 that is selected and moved to a container that is registered with the microfluidic device. Preferably, the system also has a camera for optically feeding back the container selected by the control microprocessor and the microfluidic device Registration between. GSL001.18 Preferably, the data stored in the container digital memory and the device's digital memory is encrypted. GSL001.19 Preferably, the droplet generator is configured to emit droplets having a volume of less than 6 picoliters. GSL001.20 Preferably, the probe array contains more than 1000 probes in an area of less than 1500 microns by 1500 microns. -175- 201209402 This system for reagent loading and genetic analysis of variable LOC devices provides an automated, fast, simple and cost-effective way to aggregate easy-to-use and inexpensive application-specific/application-optimized detection systems using LOC A library of devices, reagent dispensing devices, and libraries of reagents stored in micro vials with digital memory. All processing steps from reagent receipt to LOC device loading are automated.

GCA001.1 This aspect of the invention provides an apparatus for dispensing a reagent and loading an oligonucleotide spotting device, the apparatus comprising: a plurality of reagent bottles each having a droplet dispenser; a plurality of oligonucleotides a glycoside acid bottle, each of which has a droplet dispenser; a mounting surface for detachably mounting the microfluidic device for movement and detachable mounting of the oligonucleotide point relative to the reagent bottle Sample device; and

a control processor for operatively controlling movement of the reagent vial and the oligonucleotide vial and the mounting surface relative to the reagent vial and the oligonucleotide vial; wherein the control processor is configured to initiate any of the A droplet dispenser, moving the microfluidic device to register the reagent bottle and moving the oligonucleotide spotting device to register the oligonucleotide vial. GCA001.2 Preferably, each of the reagent bottles has an integrated circuit for storing reagent specification data, and each of the oligonucleotide bottles has an integrated circuit for storing oligonucleotide specification data, and the control processor is configured The reagent specification data is downloaded to the digital memory in the microfluidic device and the oligonucleotide specification data is downloaded to the digital memory in the oligonucleotide spotting device. -176- 201209402 GCA001 .3 Preferably, the apparatus also has a camera to optically feedback the registration between the bottle selected by the control processor and the microfluidic device. GCA001.4 Preferably, the reagent bottle and the oligonucleotide bottle have a volume of between 282 microliters and 400 microliters. GCA001.5 Preferably, the integrated circuit of each of the micro vials has a unique identifier for individually identifying each of the micro vials, the unique identifier being transmitted to the control processor.

GCA 01.6 Preferably, each of the micro vials has electrical contacts for receiving a firing pulse for the droplet dispenser and allows the control processor to interrogate the integrated circuit. GCA001.7 Preferably, the apparatus also has a shelf, wherein the microcapsules are removably mounted to the shelf for electromechanical control of the micro vials. GCA001.8 Preferably, the mounting surface is a platform that is configured to move along two orthogonal axes that extend parallel to one of the orthogonal axes. GCA001.9 Preferably, the droplet dispenser has a piezoelectric actuator GCA001.10. Preferably, the droplet dispenser is configured to emit droplets having a volume between 50 picoliters and 150 picoliters. GCA001.11 Preferably, the microfluidic device is a LOC device for gene analysis of biological samples, the LOC device having a polymerase chain reaction (PCR) portion, the reagent list having one or more of the following: water, polymerase, primer, Buffers, anticoagulants, deoxyribonucleoside triphosphates (dNTPs), lysing reagents, ligases, linkers and restriction enzymes. GCA001.12 Preferably, the apparatus also has a GCA001.1 3 for applying a film to the 177-201209402 LOC device to cover the reagent reservoir formed on the outer surface. Preferably, the LOC device is constructed. One of the arrays of LOC devices that are configured to mount the wafer to load reagents to all LOCs of the array. GCA001.14 Preferably, the LOC device is a LOC device mounted on a circuit board (PCB) In one of the arrays, the platform is detachably mounted to load reagents to the array. The combined reagent dispensing device and apparatus for loading oligonucleosides are used as part of a cost-effective automated mass system for dispensing reagent reservoirs of microfluidic devices contained in reagent microbottles And the data provided by the oligonucleotide reservoir containing the oligonucleotide microoligonucleotide to the oligonucleotide spotting device, including automated computer control, to the reagent storage of the microfluidic device And dispensing the oligonucleotide to the oligonucleotide reservoir of the acid spotting device, inspecting the reagent data stored in the microbody to be compared with the list of microfluidic devices that must be loaded, and inspecting and storing The data in the memory of the micro vial is compared with the list of oligonucleotides that must be loaded on the oligonucleotide spotting device, the reagent data is stored in the microfluidic device, and the oligonucleotide data is stored in the In the oligonucleotide spotting device, the combined 5 agent dispensing device and the device for loading the oligonucleoside are provided with an automated and volumetric and positionally accurate reagent and application. The 矽 wafer is detachable. The reagent is placed in the measuring flask in an acid-spotting environment in the printed electrostructure. The memory acid spot-like oligonucleotide-178-201209402 distribution technique in the memory of the oligodeoxynucleotide in the memory of the oligonucleoside bottle is provided to simplify the automated manufacturing The complexity of the environment, increasing the reliability of the automated manufacturing environment, increasing the security of the automated manufacturing environment, increasing the safety of the automated manufacturing environment, and reducing the cost of the automated manufacturing environment. The data provided by the combined reagent dispensing device and the equipment used to load the oligonucleotide spotting device automatically provides automated, secure, secure and inexpensive data monitoring and management techniques in the automated manufacturing environment.

GCA002.1 This aspect of the invention provides an apparatus for dispensing reagents, loading oligonucleotide spotting devices, and spotting oligonucleotide probes, the apparatus comprising: a plurality of reagent bottles, each of which has a droplet dispenser; a plurality of oligonucleotide bottles each having a droplet dispenser; a mounting surface for detachably mounting the microfluidic device for movement of the microfluidic device relative to the reagent bottle Removably mounting an oligonucleotide spotting device; a fixture for detachably mounting the oligonucleotide spotting device adjacent to the mounting surface; and a control processor for operating the reagent and the oligo a glycoside bottle, an oligonucleotide spotting device mounted on the jig, and movement of the mounting surface relative to the reagent and the oligonucleotide bottle, and the oligonucleotide spotting device; wherein the control processor is The tissue is configured to activate any of the droplet dispensers, move the microfluidic device to register the reagent bottle, and move the oligonucleotide spotting device to register the oligonucleotide vial. -179- 201209402 GCA002.2 Preferably the control processor is configured to operate an oligonucleotide spotting device on the fixture to spot the oligonucleotide probe onto the mounting surface Microfluidic device. GCA002.3 preferably 'each of the reagent bottles has an integrated circuit for storing reagent specification data, each of the oligonucleotide bottles has an integrated circuit for storing oligonucleotide specification data, and the control processor is configured The reagent specification data is downloaded to the digital memory in the microfluidic device and the oligonucleotide specification data is downloaded to the digital memory in the oligonucleotide spotting device. GCA002.4 Preferably, the apparatus also has a camera to optically feedback the registration between the bottle selected by the control processor and the microfluidic device. GCA002.5 Preferably, the reagent bottle and the oligonucleotide bottle have a volume of between 282 microliters and 400 microliters. GCA002.6 Preferably, the integrated circuit of each of the micro vials has a unique identifier for individually identifying each of the micro vials, the unique identifier being transmitted to the control processor, GCA002.7, preferably each The micro vial has an electrical contact to receive a firing pulse for the droplet dispenser and allows the control processor to interrogate the integrated circuit. GCA002.8 Preferably, the apparatus also has a shelf, wherein the microcapsules are removably mounted to the shelf for electromechanical control of the micro vials. GCA002.9 Preferably, the mounting surface is a platform that is configured to move along two orthogonal axes that extend parallel to one of the orthogonal axes. GCA002.1 0 Preferably, the droplet dispenser has a piezoelectric actuator - 180 - 201209402 GCA002.1 1 Preferably, the droplet dispenser is configured to have an ejection volume of between 50 picoliters and 150 The droplets of the skin rise. GCA002.1 2 Preferably, the microfluidic device is a LOC device for gene analysis of biological samples. The LOC device has a polymerase chain reaction (PCR) portion, and the reagent list has one or more of the following: water, polymerase, Primer' buffer, anticoagulant, deoxyribonucleoside triphosphate (dNTP), lysing reagent, ligase, linker and restriction enzyme.

GCA002.1 3 Preferably, the apparatus also has means for applying a film to the LOC device to cover a reagent reservoir formed on the outer surface. GCA002.14 preferably 'the LOC device is one of an array of LOC devices built on a germanium wafer, the platform being configured to removably mount the germanium wafer to load reagents to all LOC devices of the array . GCA002.1 5 Preferably, the LOC device is one of an array of LOC devices mounted on a printed circuit board (PCB) that is configured to detachably mount the PCB to load reagents to all of the array LOC device. GCA002.1 6 Preferably, the oligonucleotide spotting device has a reservoir array and an emitter array for comprising an oligonucleotide probe, and the LOC device has an acceptor for receiving an oligonucleotide probe An array of hybridization chambers having a nucleic acid sequence complementary to a target nucleic acid sequence to be detected in a biological sample. The hybridization chamber array is organized to accommodate a complete array of oligonucleotide probes required for predetermined analysis of biological samples. The reservoir array is configured to contain a complete series of oligonucleotide probes required for predetermined analysis by the LOC device, the control processor being configured to operate the correct location of the injector - 181 - 201209402 The hybrid chamber array and the oligonucleotide probes from each reservoir are downloaded and positioned in an array position of the hybridization chambers spotted by each reservoir. GCA002.1 7 Preferably, each of the injectors has more than a plurality of actuators comprising a plurality of oligonucleotide probes from the corresponding reservoir, one of the actuators corresponding to each of the nozzles The chamber ejects droplets via the corresponding nozzle, the control being configured to individually operate each of the actuators. GCA002.1 8 Preferably, the control processor is spotted on the LOC device via the array of emitters at 8 probe spotting oligonucleotides per square millimeter. GCA002.1 9 Preferably, the control processor is spotted on the LOC device via the array of emitters at 60 probe points per square millimeter." GCA002.20 Preferably, The control processor is passed through an array of emitters that are sampled on the LOC device at a density ranging from 1 square inch per square millimeter to 1 500 probe spots per square millimeter. The combined apparatus for reagent dispensing, loading of oligonucleotides and oligonucleotide spotting is used as part of a cost effective manufacturing environment for dispensing reagents contained therein to the micro a reagent reservoir of the fluid device, dispensing the oligonucleotide in the acid-packed micro-bottle to the oligonucleotide spotting device reservoir, and ejecting the oligonucleotide to the specification data of the surface needle to be spotted The nozzles, the chambers with liquids, and the densities of the actuators are organized to operate at a density that manipulates the fabric to manipulate 500 probe oligonucleotides. The spotting device and the automated large reagent micro vial are contained on the oligonucleotide of the oligonucleoside. By the design -182- 201209402

The data provided by the preparation includes automated computer controlled dispensing of the reagent to the reagent reservoir of the microfluidic device, dispensing of the oligonucleotide to the oligonucleotide reservoir of the oligonucleotide spotting device, and dispensing of the oligo Nucleotide to the surface to be spotted, and the reagent data stored in the micro-bottle of the micro-bottle is checked for comparison with the reagent specification list that must be loaded on the microfluidic device, and the sample is stored in the micro-bottle. The oligonucleotide data in the memory is aligned with the oligonucleotide specification list that must be loaded on the oligonucleotide spotting device, and the reagent data is stored in the memory of the microfluidic device, and the storage is performed. The oligonucleotides are specified in the cells of the device to be spotted, and the reagents and oligonucleotides are delivered to other departments of the automated manufacturing environment. The combined reagent dispensing, loading oligonucleotide spotting device and oligonucleotide spotting device provide automated, volumetric and positional accurate, fast and high density reagent dispensing and oligonucleotide spotting techniques To simplify the complexity of the automated manufacturing environment and increase the reliability of the automated manufacturing environment 'increasing the security of the automated manufacturing environment, increasing the safety of the automated manufacturing environment and reducing the cost of the automated manufacturing environment. Automated, secure, secure, and inexpensive data monitoring in the automated manufacturing environment is automated by the combined data provided for reagent dispensing, loading oligonucleotide spotting devices, and oligonucleotide spotting equipment And management technology. GCA003 · 1 This aspect of the invention provides an apparatus for loading an oligonucleotide spotting device and a spotting oligonucleotide probe, the device comprising: a plurality of oligonucleotide bottles, each of which has a droplet dispenser; a mounting surface for detachably mounting an oligonucleotide point-183-201209402-like device: a fixture for detachably mounting the oligonucleotide spotting device adjacent to the mounting surface And a control processor for operating the oligonucleotide vial, the oligonucleotide spotting device mounted on the fixture, and the movement of the mounting surface relative to the oligonucleotide vial and the oligonucleotide a spotting device; wherein the control processor is configured to activate the drop dispenser and move the oligonucleotide spotting device to register the oligonucleotide vial. GCA003.2 Preferably, the control processor is configured to operate an oligonucleotide spotting device on the fixture to spot the oligonucleotide probe to the microfluidic device on the mounting surface. GCA003.3 Preferably, each of the oligonucleotide vials has an integrated circuit for storing oligonucleotide specification data, and the control processor is configured to download the oligonucleotide specification data to the oligonucleoside Digital Memory in Acid Spotting Devices GCA003.4 Preferably, the device also has a camera to optically feedback the registration between the bottle selected by the control processor and the oligonucleotide spotting device. GCA003.5 Preferably, the oligonucleotide bottle is in a micro vial having a volume ranging from 282 microliters to 400 microliters. GCA003.6 Preferably, the integrated circuit of each of the micro vials has a unique identifier for individually identifying each of the micro vials, the unique identifier being transmitted to the control processor. GCA003.7 Preferably, each of the micro vials has an electrical contact for receiving a start pulse of the drop dispenser from -184 to 201209402 and allows the control processor to interrogate the integrated circuit. GCA003.8 Preferably, the apparatus also has a rack, wherein the micro vials are removably mounted to the rack for electromechanical control of the micro vials. GCA003.9 Preferably, the mounting surface is a platform that is configured to move along two orthogonal axes that extend parallel to one of the orthogonal axes. GCA003.1 0 Preferably, the droplet dispenser has a piezoelectric actuator

GCA003.il Preferably, the droplet dispenser is configured to eject droplets having a volume between 50 picoliters and 150 picoliters. GCA003.1 2 Preferably, the apparatus also has a reagent bottle containing a reagent for processing a biological sample, wherein the microfluidic device is a LOC device for genetic analysis of a biological sample, the LOC device having a polymerase chain reaction (PCR) And the reagent list has one or more of the following: water, polymerase, primer, buffer, anticoagulant 'deoxyribonucleoside triphosphate (dNTP), φ lysing reagent, ligase, linker and restriction enzymes》 GCA003.1 3 Preferably, the apparatus also has means for applying a film to the LOC device to cover a reagent reservoir formed on the outer surface. GCA003.14 Preferably, the LOC device is one of an array of LOC devices built on a germanium wafer, the platform being configured to removably mount the germanium wafer to load reagents to all LOC devices of the array . GCA003.1 5 Preferably, the LOC device is one of an array of LOC devices mounted on a printed circuit board (PCB) that is configured to detachably mount the PCB to load reagents to all of the array LOC installed -185- 201209402 set. GCA003.1 6 Preferably, the oligonucleotide spotting device has a reservoir array and an emitter array for comprising an oligonucleotide probe, and the LOC device has an acceptor for receiving an oligonucleotide probe A hybridization chamber array having a nucleic acid sequence complementary to a target nucleic acid sequence to be detected in a biological sample, the hybridization chamber array being organized to accommodate a complete array of oligonucleotide probes required for predetermined analysis of the biological sample The reservoir array is configured to contain a complete series of oligonucleotide probes required for predetermined analysis by the LOC device, the control processor being configured to operate the injector in the correct location for the hybridization The chamber array and the specification data of the oligonucleotide probes from each reservoir are downloaded and the correlation between the array position data of the hybridization chambers spotted by each reservoir is located. GCA003.1 7 Preferably, each of the injectors has a plurality of nozzles, a chamber for suspending liquid comprising oligonucleotide probes from the corresponding reservoir, and a plurality of actuators, one of the actuators Each of the nozzles is adapted to cause the actuator to eject droplets from the chamber via the corresponding nozzle, and the control processor is configured to individually operate each of the actuators. GCA003.1 8 Preferably, the control processor is configured to operate the array of emitters to spot the oligonucleotide on the LOC device at a density greater than 8 probe spots per square millimeter. GCA003.1 9 Preferably, the control processor is configured to operate the array of emitters to spot the oligonucleotide on the LOC device at a density greater than 60 probe spots per square millimeter. GCA003.20 Preferably, the control processor is configured to operate an array of -186-201209402 injectors with a range of 10,000 probes per square millimeter to 10 points per square millimeter. The density of 0 probe spottings occupies oligonucleotide 0 on the LOC device.

The combined apparatus for loading oligonucleotide spotting devices and spotting oligonucleotides is used as part of a cost-effective automated mass production environment for distribution to oligonucleosides The oligonucleotide in the acid microbottle is transferred to the oligonucleotide reservoir of the oligonucleotide spotting device and the oligonucleotide is emitted onto the surface to be spotted. Automating the data provided by the device includes automated computer controlled dispensing of the oligonucleotide to the oligonucleotide reservoir of the oligonucleotide spotting device, dispensing the oligonucleotide to the surface to be spotted Aligning and checking the oligonucleotide data stored in the memory of the micro-bottle to compare with the specification list of the oligonucleotides that must be loaded into the oligonucleotide spotting device, and storing the oligonucleotide specification In the memory of the device to be spotted, and transmitting the oligonucleotide specifications to other departments of the automated manufacturing environment. The combined oligonucleotide spotting device and oligonucleotide spotting device provide automated, volumetric and positionally accurate, fast and high density oligonucleotide spotting techniques to simplify automated manufacturing The complexity of the environment, increasing the reliability of the automated manufacturing environment, increasing the security of the automated manufacturing environment, increasing the safety of the automated manufacturing environment, and reducing the cost of the automated manufacturing environment. Automated, secure, secure, and inexpensive data monitoring and management techniques in the automated manufacturing environment, automated by the combined data provided for loading oligonucleotide spotting devices and oligonucleotide spotted devices . -187-201209402 [Embodiment] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS OF THE INVENTION The general description of the main components of the molecular diagnostic system of the embodiment of the present invention is described in detail below. Discussed in the manual. Referring to Figures 1, 2, 3, 104 and 010, the system has the following most important components: The test modules 1 and 1 are the same size as the conventional u S B flash drive, which can be produced very cheaply. Test modules 1 and 1 each comprise a microfluidic device, typically in the form of a lab-on-lab (LOC) device 30, which preloads reagents and typically more than one probe for molecular diagnostic analysis ( See Figures 1 and 1 04). The test module 10 illustrated in Figure 1 uses a fluorescent substrate detection technique to identify the target molecule, whereas the test module 11 of Figure 104 uses the detection technique of the electrochemiluminescence substrate. The LOC device 30 has an integrated light sensor 44 (described in detail below) for fluorescence or electrochemiluminescence detection. Test modules 1 and 1 1 use standard micro-USB connector 14 for power supply, data and control. Both test modules have printed circuit board (PC B) 5 7 and external power supply capacitor 3 2 and inductor. 1 5. Test modules 1 0 and 1 1 are for single use only, and are mass-produced and distributed for aseptic packaging for immediate use. The outer casing 13 has a large container 24 that accepts a biological sample and a removable sterile sealing tape 22, which preferably has a low viscosity adhesive to cover the large container before use -188-201209402. The membrane seal 408 having the membrane shield 410 forms part of the outer casing 13 to reduce the reduction in humidity within the test module while providing a pressure relief when the pressure changes. Membrane guard 410 protects the membrane seal 408 from damage. Test module reader 1 2 supplies power to the test module 1 〇 or 1 1 via the micro-USB 埠1 6 . The test module reader 12 can be in many different forms, the choice of which is described later. The reader 12 φ version shown in Figures 1, 3 and 104 is an implementation of the smart phone. The block diagram of this reader 12 is shown in Figure 3. The processor 42 executes the application software from the program storage 43. The processor 42 is also interfaced with a display screen 18 and a user interface (UI) touch screen 17 and buttons 19, a cellular radio 21, a wireless network connection 23, and a satellite navigation system 25. Honeycomb radio 21 and wireless internet connection 2 3 are used for communication. The satellite navigation system 25 is used to update the epidemiological database with location information. The location data can optionally be manually entered via touch screen i 7 or button 19. The data store 27 stores genetic and diagnostic information φ, test results, patient information, analysis and probe data for identifying the position of each probe and its array. The data store 27 and the program store 43 can be shared by a common memory device. The application software installed in the Test Module Reader 1 2 provides results analysis and other test and diagnostic information. To perform a diagnostic test, the test module 1 〇 (or test module 1 i ) is inserted into the micro-USB port 16 on the test module reader 12. The sterile sealing tape 22 is torn back and the biological sample (in liquid form) is loaded into the sample large container 24. Press the start button 20 to start the test via the application software. The sample flows into the LOC device 30 and the device is subjected to on-board analysis to extract -189-201209402, culture, amplification and pre-synthesized hybrid-reactive oligonucleotide probes with the sample nucleic acid (target) Hybrid. In the case of test module 10 (which uses detection of a fluorescent substrate), the probes are fluorescently labeled and provide the necessary excitation light by LEDs 26 mounted in the housing 13 to induce hybridized probes. Fluorescent emission (see Figures 1 and 2). In the case of a test module (which uses electrochemical luminescence (ECL) detection), the LOC device 30 is loaded with an ECL probe (as described above) and the LED 26 is not required to produce luminescent emissions. In fact, the excitation current is supplied by electrodes 8 60 and 870 (see Fig. 1〇5). The emission (fluorescence or illumination) is detected by a light sensor 44 integrated into the CMOS circuitry of each LOC device. The detection signal is amplified and converted to a digital output for analysis by the test module reader 12. The reader then displays the results. This information can be stored locally and/or uploaded to a network server containing patient records. The test module 10 or 1 1 is removed from the test module reader 1 2 and processed appropriately. 1, 3 and 104 show a test module reader 12 designed as a mobile phone/smartphone 28. Other forms of test module readers may be laptops/notebooks 101, dedicated readers 103, e-book readers 107, tablets 109 or desktop computers 105 for use in hospitals, private clinics or laboratories ( See Figure 106). The reader can interface with additional applications such as patient records, accounting, online databases and multi-user environments. It can also interface with some local or remote peripherals, such as printers and patient smart cards. Referring to FIG. 1 07, the data generated by the test module 1 can be used to update the epidemiological data host system 1 1 1 through the reader 1 2 and the network 1 25 -190- 201209402

The epidemiological database, the genetic data host system 1 1 3 stored in the gene database, the electronic health record (EHR) host system 1 15 stored electronic health record, electronic medical record (EMR) host system 121 The stored electronic medical record, as well as the personal health record stored by the personal health record (PHR) host system 123. Conversely, the epidemiological data stored in the epidemiological data host system 1 1 1 , the genetic data stored in the genetic data host system 113, and the electronic data stored in the electronic health record (EHR) host system 1 1 5 The health record, the electronic medical record stored in the electronic medical record (EMR) host system 121, and the personal health record stored in the personal health record (PHR) host system 123 can be used to read and read via the network 125 The device 12 updates the digits in the LOC 30 of the test module 10 to count the body. Referring to Figures 1, 2, 104 and 010, the reader in the mobile phone configuration uses battery power. The mobile phone reader contains all pre-loaded test and diagnostic information. Data can also be downloaded or φ transmitted via some wireless or contact interface to communicate with peripheral devices, computers or online servers. The micro USB port 16 is used to connect a computer or a primary power supply for charging the battery. Figure 77 shows an embodiment of a test module 10 for a positive or negative test result requiring only a particular target, such as for testing whether a human is infected with, for example, influenza A virus H1N1. Only the USB power/indicator module 47 built for a particular purpose is required. No other readers or application software is required. The indicator 45 on the USB power/indicator module 47 alone displays a positive or negative result. This configuration is ideal for large screenings. Other items that may be provided by the system may include test tubes containing reagents for pretreatment of specific -191 - 201209402 samples and spatula and lancets for sample collection. Figure 7 7 shows an embodiment of a test module for the convenience of a spring-loaded telescopic lancet 309 and a lancet release button 3 92. Satellite phones can be used in remote areas. Test module electronics

Figures 2 and 105 are block diagrams of the electronic components in test modules 10 and 1 respectively. The CMOS circuit integrated in the LOC device 30 has a USB device driver 36, a controller 34, a USB compatible LED driver 29, a clock 3 3, a power conditioner 3 1 , a RAM 3 8 and a program and data flash memory 40. These components provide for the entire test module 10 or 1 1 including a light sensor 44, a temperature sensor 170, a liquid sensor 174 and various heaters 152, 154, 182' 234 and associated drivers 37 and 39 and a temporary storage Control and memory of devices 35 and 41. Only LED 26 (with test module 10 as an example), external power capacitor 32 and micro USB connector 14 are located outside of LOC device 30. LOC device 30 includes pads for connection to these external components. The RAM 38 and the program and data flash memory 40 have application software and diagnostic and test information (flash/security storage, such as encryption) of more than 1 000 probes. In the test module 1 1 designed for ECL detection, the test module 11 does not contain LEDs 26 (see Figures 104 and 105). The data is encrypted by the LOC device 30 for secure storage and secure communication with external devices. The LOC device 30 is loaded with electrochemiluminescent probes each having a pair of ECL excitation electrodes 860 and 870. Many types of test modules 1 are manufactured in several test formats for use in stock from -192 to 201209402. The difference between the different test formats is that the reagents and probes of the boardn board assay can be quickly identified by this system. Some examples of infectious diseases include: • Influenza-influenza virus A, B, C, infectious salmon anemia virus (Isavirus), tohogovirus (Thogotovirus) • pneumonia-respiratory fusion virus (RSV), adenovirus, interstitial pneumonia virus, pneumococci, Staphylococcus aureus

• Tuberculosis · Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium africana, Mycobacterium vaccae and Mycobacterium vaccae • Plasmodium falciparum, Toxoplasma gondii and other parasitic protozoa • Typhoid-cold typhoid • Ebola virus • Human immunodeficiency virus (HIV) • Dengue fever – Yellow fever virus • Hepatitis (A to E) • Iatrogenic infections – such as Clostridium difficile, vancomycin-resistant enterococci and drug-resistant golden yellow Staphylococcus • Herpes simplex virus (HSV) • Cytomegalovirus (CMV) • Epstein-Barr virus (EBV) • Encephalitis – Sputum encephalitis virus, Zhangdipula virus • Pertussis- pertussis • Hemp - Paramyxovirus • Meningitis - Streptococcus pneumoniae and Neisseria meningitidis (Neisseria -193- 201209402 meningitidis) • Examples of some genetic diseases that anthrax - anthrax can use this system to identify include: • Cystic fibrosis • Blood Friendship 4 • Sickle cell anemia • Black Mongolian idiot

• Hemochromatosis • Cerebral artery disease • Crohn's disease • Polycystic kidney disease • Congenital heart disease • Rett's disease A few examples of cancer identified by this diagnostic system include:

• Ovarian cancer • Colon cancer • Multiple endocrine neoplasms • Retinal blastoma • Turcot syndrome The above list is not complete and the diagnostic system can be configured to detect more samples using nucleic acid and proteomic analysis Diseases and conditions. Detailed structure of system components LOC device -194- 201209402

The LOC device 30 is the heart of the diagnostic system. The device rapidly performs the four major steps of molecular diagnostic analysis of nucleic acid substrates on a microfluidic platform, namely sample preparation, nucleic acid extraction, nucleic acid amplification and detection. The L Ο C device also has a selective use' and will be described in detail below. As discussed above, test modules 1 and 11 can take many different configurations to detect different targets. Similarly, the LO C device 30 can also be used to create a variety of different embodiments for the target of interest. One form of LOC device 30 is used for fluorescent detection of the L Ο C device 310 of the target nucleic acid sequence of the pathogen in a whole blood sample. For the purpose of explanation, the structure and operation of the LOC device 301 will now be described in detail with reference to Figs. 4 to 26 and 27 to 57. Figure 4 is a representation of the structure of the LOC device 301. For the sake of convenience, the processing stages shown in Fig. 4 are denoted by the component symbols corresponding to the functional portions of the LOC device 301 that implements the processing stage. The processing stages associated with each of the major steps in the molecular diagnostic analysis of the nucleic acid substrate are also shown: sample input and preparation 288, extraction 2 90, culture 291, amplification 2 92 and detection φ 294. The various reservoirs, chambers, valves and other components of the LOC device 301 will be described more closely below. Figure 5 is a perspective view of the LOC device 301. The device is fabricated using high volume CMOS and MST (microsystem technology) fabrication techniques. The layered structure of the LOC device 301 is illustrated in a schematic (not to scale) partial cross-sectional view of Fig. 12. The LOC device 301 has a germanium substrate 84 that supports a COMS + MST wafer 48 that includes a CMOS circuit 86 and an MST layer 87 and has a cover 46 overlying the MST layer 87. For the purposes of this patent specification, the term "MST layer j" refers to a collection of structures and layers of a sample treated with various reagents. -195- \\ 201209402 Accordingly, these structures and components are organized to define flow paths having characteristic dimensions. The feature size will support the capillary action to drive the flow of liquid similar to the physical properties of the sample at the processing stage. In view of this, the MS T layer and assembly are typically fabricated using surface micromachining techniques and/or stereo micromachining techniques. However, other manufacturing methods can also produce structures and components that are sized for capillary drive flow and for processing very small samples. The specific embodiments described in this specification show that the MST layer is a structure and active component supported by CMOS circuitry 86. However, it does not have the features of the upper cover 46. However, those skilled in the art will understand that the MST layer does not require the CMOS or even the upper cover to process the sample. The overall size of the LOC device shown in the following figure is 1. 760 microns X 5824 microns. Of course, LOC devices manufactured for different applications can have different sizes. Figure 6 shows the MST layer 87 The features are overlapped with the features of the upper cover. The AA to AD, AG, and AH regions shown in Figure 6 are enlarged in Figures 13, 14, 35, 56, 55, and 67, respectively, and are described in detail below to fully understand the LOC device. The various structures within 301. Figures 7 through 10 show the features of the upper cover 46 independently, while Figure 11 shows the structural layered structure of the CMOS + MST device 48 independently. Figures 12 and 22 illustrate the CMOS + MST device 48, the upper cover 46, and both. The hierarchical structure of the interaction between the fluids is not drawn to scale for the purpose of illustration. Figure 12 is a schematic view through section 196-201209402 of sample inlet 68, and Figure 22 is a schematic cross-sectional view through reservoir 54. Figure 12 clearly shows that the CMOS + MST device 48 has a germanium substrate 84 supporting a CMOS circuit 86 that operates the active components within the MST layer 87 thereon. The passivation layer 88 seals and protects the CMOS layer 86 from flowing through the MST layer. The liquid of 8 7 flows in. The liquid flows through the upper cover layer 46 and the upper cover of the MST channel layer 100, respectively.

Both channel 94 and MST channel 90 (see, for example, Figures 7 and 16). Cell transport occurs in the larger channel 94 made in the upper cap 46, while biochemical treatment takes place in the smaller MST channel 90. The size of the cell delivery channel is designed to deliver cells in the sample to a predetermined location in the M S T channel 90. Cells that transport greater than 20 microns (e.g., certain white blood cells) require channel sizes greater than 20 microns, so the cross-sectional area across the flow must be greater than 400 square microns. The MS Τ channel can be significantly smaller, especially in the case where it is not necessary to transport cells. It will be understood that the 'top cover channel 94 and the MST channel 90 are generic, and the particular MST channel 90 may also be referred to as, for example, a heated microchannel or a dialysis MST channel due to its particular function. The MST channel 90 is formed by etching a MST channel layer 1 deposited on the passivation layer 88 and patterned by a photoresist. The MST channel 90 is closed by a top layer 66 which forms the top of the CMOS + MST device (shown in the figure direction). Although sometimes shown in separate layers, the upper cover channel layer 80 and the reservoir layer 78 are formed from a single sheet of material. Of course, the sheet material can also be non-unitary. Both sides of the sheet material are etched to form an upper cover channel layer and a reservoir layer 78'. The upper cover channel 94 is etched in the upper cover channel layer 8 and the reservoirs 5 4, 5 are etched in the reservoir layer 197-201209402. 6, 5 8, 6 0 and 6 2. Alternatively, the reservoir and the upper cover channel are formed by micromolding. Both etching and micromolding techniques are used to fabricate channels having a cross-sectional area across the fluid of up to 20,000 square microns and a minimum of 8 square microns. A variety of suitable channel cross-sectional areas across the fluid can be selected for different locations in the LOC device. When a large number of samples or samples having a large composition are accommodated in the channel, a cross-sectional area of up to 20,000 square micrometers (e.g., a channel having a width of 200 micrometers in a layer having a thickness of 100 micrometers) is suitable. When the channel contains a small amount of liquid or a mixture free of large cells, the cross-sectional area of the traversing fluid is preferably very small. The lower sealing layer 64 encloses the upper cover passage 94, and the upper sealing layer 82 closes the reservoirs 54, 56, 58, 60 and 62 » The five reservoirs 54, 56, 58, 60 and 62 preload the reagents for the particular analysis. In the embodiments described herein, the reservoirs are preloaded with the following reagents, but can be easily replaced with other reagents: • Reservoir 54: Anticoagulant, optionally including red blood cell lysate • Reservoir 5 6 : Dissolving Cytokines • Reservoirs 5 8 : Restriction enzymes, ligases and linkers (for linker-primed PCR (see Figure 76, excerpt from T. Stachan et al, Human Molecular Genetics 2, Garland Science, NY and London, 1 999)) Reservoir 60: amplification mixture (d NTP, primer, buffer), and • reservoir 62: DNA polymerase. Upper cover 46 and CMOS+ MST layer 48 are in fluid communication via corresponding openings in lower seal 64 and top layer 60. These openings are referred to as riser 96 and descending port 92 depending on the flow system flowing from the 198-201209402 MST channel 90 to the upper cover channel 94 or vice versa. LOC device operation

The operation of the LOC device 301 is described step by step as an example of the analysis of pathogenic DNA in a blood sample. Of course, other types of biological or non-biological fluids can also be analyzed using appropriate combinations of reagents, test protocols, LOC variants, and detection systems. Referring to Figure 4, the analytical biological sample is largely divided into five steps, including sample input and preparation 288, nucleic acid extraction 290, nucleic acid culture 291, nucleic acid amplification 292, and detection and analysis 294. The sample input and preparation step 288 involves mixing the blood with the anticoagulant 116, followed by separation of the pathogen from the pathogen dialysis unit 70 with white blood cells and red blood cells. As best seen in Figures 7 and 12, the blood sample enters the device via sample inlet 68. Capillary action draws the blood sample along the upper cover channel 94 to the reservoir 54. When the sample blood flow opens the surface tension valve 1 18 of the reservoir 54, the anticoagulant is released from the reservoir 54 (see Figures 15 and 22). Anticoagulants prevent clot formation from obstructing flow. As best shown in Fig. 22, the anticoagulant 116 is aspirated from the reservoir 54 by capillary action and enters the MST channel 90 via the descending port 92. The drop port 92 has a capillary initiation feature (CIF) 1〇2 to control the geometry of the meniscus such that the meniscus is not fixed at the edge of the drop port 92. The venting opening 12 in the upper sealing layer 82 allows air to replace the anticoagulant that is aspirated from the reservoir 54. The MST channel 90 shown in Figure 22 is part of the surface tension valve 118 -199 - 201209402. The anticoagulant 116 fills the surface tension valve 118 and secures the meniscus 120 to the meniscus anchor 98 of the riser 96. The meniscus 120 remains fixed to the riser 96 prior to use so that the anticoagulant does not flow into the upper cover passage 94. As blood flows through the upper cover passage 94 to the ascending port 96, the meniscus 120 is removed and the anticoagulant is drawn into the flow.

Figures 15 through 21 show the AE zone, which is part of the AA zone shown in Figure 13. As shown in Figures 15, 16 and 17, surface tension valve 118 has three separate MST passages 90 extending between respective lower and upper risers 92, 96. The number of MST channels 90 in the surface tension valve can vary to change the flow rate of reagent into the sample mixture. When the sample mixture and reagents are mixed by diffusion, the flow rate out of the reservoir determines the concentration of the reagent in the sample stream. Thus, the surface tension valves of each reservoir are configured to meet the desired reagent concentration.

The blood enters the pathogen dialysis section 70 (see Figures 4 and 15) where the target cells are concentrated from the sample using a well-sized array of holes 1 64 according to a predetermined valve. Cells smaller than the valve plaque pass through the well, while large cells cannot pass through the well. The undesired cells are either larger cells that are blocked by well array 1 64 or smaller cells that pass through the well, which are transduced to waste unit 76, whereas the target cells are still part of the assay. In the pathogen dialysis section 70 described herein, the pathogen system from the whole blood sample is concentrated for microbial DNA analysis. The array of holes is formed by a plurality of 3 micron diameter holes 1 64 that fluidly communicate with the input in the upper cover channel 94 to the target channel 74. The 3 micron diameter aperture 164 and the dialysis riser aperture 168 in the target channel 74 are connected by a series of dialysis MST channels 204 -200 - 201209402 (best shown in Figures 15 and 21). The pathogen is small in volume and can therefore be filled through the 3 micron diameter well 164 and filled through the target channel 74 via the dialysis MST channel 204. Cells larger than 3 microns, such as red blood cells and white blood cells, remain in the waste channel 72 of the upper cover 46, which leads to the waste receptacle 76 (see Figure 7).

Other pore shapes, sizes, and aspect ratios can be utilized to isolate specific pathogens or other target cells, such as white blood cells for human DNA analysis. Detailed instructions for the dialysis section and dialysis variants are provided below. Referring again to Figures 6 and 7, fluid is drawn through target channel 74 to surface tension valve 128 of lysis reagent reservoir 56. The surface tension valve 128 has seven MST channels 90 that extend between the lysis reagent reservoir 56 and the target channel 74. When the meniscus is removed by the sample stream, the flow rate from all seven MS T channels 90 will be greater than the flow rate from the anticoagulant reservoir 54, where the surface tension valve 118 of the reservoir 54 has three MST channels 9 (Assume that the physical properties of the liquids are approximately equal). Thus the ratio of lytic reagent to the sample mixture is greater than the ratio of anticoagulant to the sample mixture. The lysis reagent and the target cells are mixed by diffusion in the target channel 74 in the chemical lysis unit 130. The boiling start valve 126 stops the flow until sufficient time for diffusion and lysis has occurred to release the genetic material from the target cells (see Figures 6 and 7). The structure and operation of the boiling start valve are described in detail below with reference to Figures 31 and 32. Other active valve types (as opposed to passive valves such as surface tension valve 118) have also been developed by the applicant, and other types of active valves can be used herein to replace the boiling start valve. These alternative valve designs are also described below -201 - 201209402. When the boiling start valve 1 26 is opened, the lysed cells flow into the mixing portion 1 3 1 to perform restriction enzyme cleavage and linker ligation before amplification. Referring to Figure 13, the restriction enzyme, linker and ligase are released from the reservoir 58 when the fluid is removed from the meniscus of the surface tension valve 132 at the beginning of the mixing portion 131. The mixture flows through the length of the mixing section 133 to diffusely mix. The end of the mixing portion 131 is a descending port 134 (see Fig. 13) leading to the incubator inlet passage 133 of the culture portion 114. The incubator inlet channel 133 feeds the mixture into a heated microchannel 210 in a crimped configuration that provides a culture chamber for holding the sample during restriction enzyme cleavage and linker ligation (see Figures 13 and 14). » Figures 23, 24, 25, 26, 27, 28 and 29 show the layers of the LOC device 301 in the AB zone of Figure 6. The figures show successive layers of addition to form the structure of CMOS + MST layer 48 and upper cover 46. The AB area shows the end point of the culture unit 114 and the starting point of the amplification unit 112. As best shown in Figures 14 and 23, the fluid fills the microchannel 2 1 0 of the culture portion 1 1 4 until it reaches the boiling start valve 1〇6' where the fluid stops to allow diffusion to occur. As discussed above, the microchannels 210 in the upstream of the boiling start valve 106 become the culture chamber containing the sample, restriction enzyme, ligase, and linker. The heater crucible 54 is then activated and maintains a stable temperature for a specific period of time to allow for restriction enzyme cleavage and linker engagement to occur. Those skilled in the art will appreciate that this culturing step 29 (see Figure 4) is optional. Some types of nucleic acid amplification assays are required. In addition, in some cases, it may be desirable to provide a heating step at the end of the incubation period to allow the temperature to rise sharply to a temperature of -202 - 201209402 » the temperature rises to cause the restriction enzyme and ligase to enter the amplification section. Go to live before Π2. Deactivation of restriction enzymes and ligases is particularly important when using isothermal nucleic acid amplification. After the incubation, the boiling start valve 106 is activated (opened) and the fluid continues to enter the amplifying portion 112. Referring to Figures 31 and 32, the mixture is filled with heated microchannels 158 in a curved configuration until reaching a boiling start valve 108 which forms one or more amplification chambers. As shown in the schematic cross-sectional view of Fig. 30, the amplification mixture (dNTP, primer, buffer) is released from the reservoir 60 and the polymerase is then released from the reservoir 62 to enter the junction culture section and the amplification section (114 and 112) The intermediate MST channel 212. Figures 35 through 51 show the layers of the LOC device 301 in the AC region of Figure 6. The figures show the layers of successive additions to form the structure of the CMOS + MST device 48 and the upper cover 46. The end of the AC zone amplification unit 1 12 and the start of the hybridization and detection unit 52. The cultured sample, amplification mixture, and polymerase flow through microchannel 158 to boiling start valve 108. After sufficient diffusion mixing between φ, the heater 154 in the microchannel 158 is activated for thermal cycling or isothermal amplification. The amplification mixture undergoes a predetermined number of thermal cycles or a predetermined amplification time to amplify sufficient target DNA. After the nucleic acid amplification procedure, the boiling start valve 108 is opened and the fluid continues to enter the hybridization and detection portion 52. The operation of the boiling start valve is described in detail below. As shown in Fig. 52, the hybridization and detection unit 52 has a hybridization chamber array 110. Figures 52, 53, 54, and 56 show the hybridization chamber array 110 and the single-hybridization chamber 180 in detail. A diffusion barrier 175 is provided at the entrance of the hybridization chamber 180 to prevent the target nucleic acid, the probe strand and the hybridized probe from diffusing between the -203-201209402 hybridization chambers during the hybridization to prevent erroneous hybridization. Test results. The diffusion barrier 1 75 represents a flow path that is long enough to prevent the target sequence and probe from diffusing out of one chamber and contaminating another chamber during the time required for the probe and nucleic acid hybridization and signal detection to be detected. Wrong result. Another mechanism to prevent erroneous results is to include the same probe in multiple hybridization chambers. CMOS circuit 86 derives a single result from photodiode 184 corresponding to hybridization chamber 180 containing the same probe. When extrapolating a single result, the anomalous results can be ignored or given different weights. The thermal energy required for hybridization is provided by a CMOS controlled heater 182 (described in more detail below). Hybridization occurs between the complementary target-probe sequences after the heater is activated. The LED driver 29 in the CMOS circuit 86 transmits a message to the LED 26 located in the test module 1 to illuminate it. These probes only fluoresce when hybridization occurs, so the cleaning and drying steps typically required to remove unbound strands are omitted. The hybrid forced FRET probe 1 86 has a stem-loop structure that allows the fluorophore to emit fluorescent energy in response to the LED excitation light, as described in more detail below. Fluorescence is detected by photodiode 184 located in CMOS circuit 86 below each hybridization chamber 180 (see description of hybridization chamber below). The photodiode 184 and associated electronics for all of the hybridization chambers together form a light sensor 44 (see Figure 68). In other embodiments, the light sensor can be an array of charge coupled devices (CCD arrays). The detection signal from the photodiode 1 84 is amplified and converted to a digital output for analysis by the test module reader 12. Further details of the detection method are described below. -204- 201209402 Other Detailed Description of LOC Device The modular design L〇C device 301 has many functional parts (including reagent reservoirs 54, 56, 58, 60 and 62, dialysis unit 70, lysis unit 13〇, culture department). 114 and amplification unit 112), valve type, humidifier and humidity sensor. In other embodiments of the LOC device, these functional portions may be omitted, but additional functional portions may be added or the functional portions may be used for different purposes than those described above.

For example, the culture portion 114 can be used as the first amplification portion 112 of the tandem amplification analysis system, and the chemical lysis reagent reservoir 56 is used to add the first amplification primer, dNTP, and buffer. The mixture, and reagent reservoir 58 are used to add reverse transcriptase and/or polymerase. If the sample is to be chemically lysed, a chemical lysing reagent (along with the amplification mixture) may also be added to the reservoir 56' or alternatively, the sample may be heated for a predetermined period of time to cause thermal lysis in the culture section. . In some embodiments, if chemical lysis is desired and it is desirable to separate the chemical lysis reagent from the primer, dNTP, and buffer mixture, an additional reservoir can be introduced upstream of the reservoir 58 immediately adjacent to the mixture. In some cases it may be desirable to omit steps such as incubation step 2 1 1 . In this case, the L0C device may be specially manufactured to avoid the reagent reservoir 58 and the culture portion 114, or may simply not be loaded with the reagent, or if an active valve is present, the active valve is not activated to release the reagent to In the sample stream, the culture portion becomes a channel for transferring only the sample from the lysis portion 1 30 to the amplification portion 112. The heater can be operated independently, so when the reaction depends on heating, such as hot lysis, setting the heater not to start during this step -205- 201209402 ensures that hot lysis does not occur in L0C devices that do not require hot lysis in. The dialysis unit 70 can be located at the beginning of the fluid system within the microfluidic device as shown in Figure 4 or can be located at any other location within the microfluidic device. By way of example, in some cases, it may be advantageous to dialyze after amplification stage 292 to remove cell debris prior to hybridization and detection step 294. Alternatively, two or more dialysis sections can be inserted at any location in the LOC device. Similarly, it is possible to break in additional amplifications 1 1 2 so that the multiplex targets can be amplified simultaneously or sequentially before detection of the specific nucleic acid probes in the hybridization chamber array 110. Since dialysis is not required for analysis of a sample such as whole blood, the dialysis portion 70 is simply omitted from the sample input and preparation portion 288 of the LOC design. In some cases, it is not necessary to omit the dialysis section 70' of the L〇c device even if the analysis does not require dialysis. If the presence of the dialysis section does not cause a geometrical impediment to the analysis, the sample input and preparation section can still be used to have the LOC of the dialysis section 70 without losing the desired function. Further, the detection portion 294 can include a protein body chamber array that is identical to the hybrid chamber array but carries a probe that is designed to conjugate or hybridize to a sample target protein present in the non-amplified sample, rather than a design. A nucleic acid probe for hybridization to a target nucleic acid sequence. It will be appreciated that the LOC devices manufactured for use in this diagnostic system are different combinations of functional components selected for a particular LOC application. The vast majority of functional units are the same for many LOC devices, and the design of additional LOC devices for new applications relates to the combination of appropriate functional components from the various functional options used in existing LOC devices. Only a few LOC devices are shown in this description, and more LOC devices are illustrated in the -206-201209402 schema to illustrate the design flexibility of the LOC devices manufactured for this system. Those skilled in the art will readily appreciate that the LOC devices described in this specification are not exhaustive, and that many additional LOC designs are related to integrating appropriate functional combinations. Sample type

LOC variants can accept and analyze a variety of nucleic acid or protein contents in a liquid form, including but not limited to blood and blood products, saliva, cerebrospinal fluid, urine, semen, amniotic fluid, cord blood, breast milk , sweat, pleural effusion, tears, pericardial fluid, peritoneal fluid, environmental water samples and dip samples. Amplicon obtained from macroscopic nucleic acid amplification can also be analyzed using the LOC device; in this case, all reagent reservoirs will be empty or configured to not release their contents, and the dialysis, dissolution The cell, culture and amplification sections will only be used to deliver the sample from sample inlet 68 to hybridization chamber 180 for nucleic acid detection as described above. Some sample types require a pre-treatment step prior to input into the LOC device. For example, semen may require liquefaction and mucus may require pre-treatment with enzyme to reduce viscosity. Sample Input Referring to Figures 1 and 12, the sample is added to the large container 24 of the test module 1 . The large container 24 is a truncated cone that is fed by capillary action into the inlet 68 of the LOC unit 301. The sample is here flowed into a 64 μιη wide X 6 Ομηι deep cap channel 94 and is also attracted to the anti-207-201209402 coagulant reservoir 54 by capillary action. Reagent Reservoirs A small amount of reagents required for the analysis system of a microfluidic device, such as LOC device 301, are used to allow the reagent reservoirs to contain all of the reagents required for biochemical treatment in each of the reagent reservoirs having a small volume. This volume must be less than 1,000,000,000 cubic microns, mostly less than 300,000,000 cubic microns, typically less than 70,000,000 cubic microns, and less than 20,000,000 cubic microns in the LOC device 301 shown. Dialysis section Referring to Figures 15 to 21, 3 3 and 3 4, the pathogen dialysis section 70 is designed to concentrate the pathogen target cells from the sample. As previously described, a plurality of wells 164 having a 3 micron diameter opening in the top layer 66 filter the target cells from the sample body. As the sample flows through a 3 micron diameter well 1 64, the microbial pathogen enters a series of dialysis MST channels 204 through the opening and flows back into the target channel 74 via the 16 μιη dialysis riser 168 (see Figures 33 and 34). The remaining sample (red blood cells, etc.) remains in the upper cover channel 94. Downstream of the pathogen dialysis section 70, the upper cover passage 94 becomes a waste passage 72 to the waste reservoir 76. For a biological sample type that produces a large amount of waste, a foam insert or other porous member 49 within the outer casing 13 of the test module 10 is configured to be in fluid communication with the waste reservoir 76 (see Figure 丨). The function of the pathogen dialysis unit is completely dependent on the capillary action of the fluid sample. A 3 micron diameter bore 64 at the upstream end of the pathogen dialysis section 70 has a capillary initiation feature (CIF) 166 (see Figure 33) -208-201209402 to allow fluid to be drawn downwardly to the dialysis MST channel 204 below it. Among them. The first rising aperture 198 of the target channel 74 also has a CIF 202 (see Figure 15) to prevent fluid from easily forming a meniscus on the dialysis upwell 168.

The small component dialysis section 682 shown in Fig. 81 may have a structure similar to that of the pathogen dialysis section 70. The small component dialysis section separates any small target cells or molecules from the sample by the size of the well (and shape if desired) which is adapted to allow small target cells or molecules to pass through the target channel and continue to be further analyzed. Large size cells or molecules are removed to waste receptacle 766. Thus, the LOC device 30 (see Figures 1 and 104) is not limited to pathogens having a size of less than 3 μηη, and can be used to isolate cells or molecules of any desired size. Lysis section Referring again to Figures 7, 1 1 and 13 , the gene material in the sample is released from the cell by chemical φ lysis treatment. As described above, the lysing reagent from lysis vessel 56 is mixed with the sample stream in target channel 74 downstream of surface damper valve 128 of lysis vessel 56. However, some diagnostic assays are more suitable for use with hot lysis, or even a combination of chemical and thermal lysis of target cells. The LOC device 301 cooperates with the heating microchannel 210 of the culture portion 114. The sample stream is filled with the culture portion 1 14 and stopped at the boiling start valve 106. The culture microchannel 210 heats the sample to a temperature at which the cell membrane ruptures. The enzyme reaction in the chemical lysis unit 130 is not required in some hot lysis applications, and the enzyme reaction in the chemical lysis unit 130 is completely replaced by the hot lysis. • 209- 201209402 Boiling Start Valve As mentioned above, the LOC unit 301 has three boiling shocks, 106 and 108. The position of these valves is shown in Figure 6. Figure 31 shows an enlarged plan view of the boiling start valve 108 at the end of the augmented heating microchannel 158. The sample stream 1 1 9 is attracted by the capillary through the heating 1 58 until the boiling start valve 108 is reached. The sample flow guide 120 is fixed to the meniscus anchor 98 of the valve inlet 146. The geometry of the bender 98 stops the advancing meniscus to prevent capillary movement. As shown in Figures 31 and 32, the meniscus anchor 98 is a hole provided by the passage 90 to the raised opening of the upper cover passage 94. The surface tension of the 120 keeps the valve closed. The annular heater 152 is surrounded by the inlet 146. The ring heater 152 is CMOS controlled via a boiling heater contact 53. To open the valve, the CMOS circuit 86 delivers an electrical pulse to the heater junction 153. The ring heater 152 is heated by resistance until the sample 1 19 is boiled. This boiling removes the meniscus 110 from the valve inlet and begins to wet the upper cover passage 94. Once the upper cover is started, the capillary action can be restored. The fluid sample 1 1 9 is filled with a cap and flows through the valve lower port 150 to the valve outlet 1 4 8 , where the capillary flow continues to advance along the amplifying outlet channel 1 60 to the hybrid and 52. The liquid sensor 1 74 is placed before and after the valve for diagnosis] valve 1 26 is a separate part 112 microchannel meniscus liquid surface solid flow from the MST meniscus at the valve valve plus the valve plus the liquid 146 shift I channel 94 I channel 94 with the drive detection section off -210- 201209402 It will be understood that once the boiling start valve is opened, it can no longer be closed. However, since the LOC device 301 and the test module 1 are single-use devices, there is no need to close the valve. % 4 ZJ 11 is increased by 3' expansion 1 acid 7' core, and 6 parts are cultured 2 3 to 1 Λ 5 and the culture portion 114 and the amplification portion 112 are shown. The culture portion 114 has a single strip of hot culture microchannels 210 which are etched into a meandering pattern in the MST channel layer 1 'starting at the lowering opening 134 and finally boiling the starting valve ι 6 (see Figure I: 1 4). Controlling the temperature of the culture section 1 14 allows the enzyme reaction to occur with higher efficiency. Similarly, the amplifying portion 112 has a heating amplifying microchannel 158 (see Figs. 14 and 14) which is a bouncing structure that starts from the boiling start valve 1〇6 to the boiling start valve 108. When mixed 'culture and nucleic acid amplification occurs, these valves stop the flow to retain the target cells in the heated culture or amplification microchannel 210 or 158. The curved pattern of the microchannels also promotes (to some extent) the mixing of the target cells with the reagents. In the culture unit 1 14 and the amplification unit 1 1 2, the sample cells and reagents are heated by a heater 154 controlled by a pulse width modulation (PWM) CMOS circuit 86. Each curved corner of the heated culture microchannel 210 and the expanded microchannel 158 has three independently operable heaters 154 extending between the individual heater contacts 156 (see Figure 14). It provides a two-dimensional control of the input heat flux density. As best shown in Figure 51, the heater 154 is supported by the top layer 66 and embedded in the lower sealing layer 64. The heater The material is TiAl, but many other conductive -211 - 201209402 metals are also suitable. The long heater 154 is parallel to the longitudinal length of each channel portion forming a wide curve of the curved structure. In the amplifying portion 112, Each wide curve can be controlled via a separate heater as a separate PCR chamber.

The use of a small volume of amplicons required for the analysis system of a microfluidic device, such as LOC device 301, allows for the use of a small volume of amplification mixture in amplification portion 1 1 2 for amplification. This volume must be less than 4,000 liters, mostly less than 170 liters, typically less than 70 nanoliters, and the LOC device 301 is exemplified by 2 nanoliters to 30 nanoliters. Increased heating rate and better diffusion mixing

The small cross-sectional area of each channel portion increases the heating rate of the amplification fluid mixture. The distance between all fluids and heaters 154 is quite short. Reducing the cross-sectional area of the channel (i.e., the cross section of the augmented microchannel 158) to less than 1 〇〇, the squared micrometer can achieve a significantly higher heating rate than the "large scale" device. The lithography manufacturing technique allows the amplifying microchannel 158 to have a cross-sectional area of less than 1 6,000 square micrometers across the flow path, which provides a substantially higher heating rate. Dimensional features of 1 micron are easily obtained using lithography manufacturing techniques. If only a very small amount of amplicons are required (in the case of LOC device 301), the cross-sectional area can be reduced to less than 2,500 square microns. For a diagnostic analysis performed on a LOC device with 1, 〇〇〇 to 2,000 probes and requiring "input of the sample, yielding results" within 1 minute, a cross between 400 square microns and 1 square micron The cross-sectional area of the fluid is appropriate. The heater element in the amplification microchannel 158 heats the nucleic acid sequence at a rate of greater than 80 absolute temperatures (K) per second, in most cases at a rate of -212 to 201209402 per second greater than 100 K. Typically, the heater element heats the nucleic acid sequence at a rate greater than 1,000 psi per second, and the heater element often heats the nucleic acid sequence at a rate greater than 10,000 Torr per second. Typically, depending on the requirements of the analytical system, the heater element is greater than 100,000 sec per second, greater than 1,000,000 sec per second, greater than 10,000,000 sec per second, greater than 20,000,000 sec per second, greater than 40,000,000 sec per second, greater than 80,000,000 sec per second, and The nucleic acid sequence is heated at a rate of greater than 160,000,000 ticks per second.

The small cross-sectional area channel is also beneficial for the diffusive mixing of any reagent with the sample fluid. The phenomenon of one liquid diffusing to another liquid is most pronounced near the interface between the two liquids before diffusion mixing is completed. The concentration decreases as the distance from the interface increases. The use of microchannels having a relatively small cross-sectional area across the fluid direction causes the two fluids to flow closer to the interface for faster diffusion mixing. Reducing the cross-sectional area of the channel to less than 100,000 square microns provides a significantly higher mixing rate than "large scale" equipment. The lithography manufacturing technique allows the microchannel to cross the flow path with a cross-sectional area of less than 16,000 square Φ microns, which provides a significantly higher mixing rate. If only a very small volume is required (in the case of LOC unit 30 1), the cross-sectional area can be reduced to less than 2500 square microns. Cross-fluid section between 400 square microns and 1 square micron for diagnostic analysis on a LOC device with 1000 to 2000 probes and requiring "input sample, result" in 1 minute The area is appropriate. Rapid Thermal Cycle Time Keep the sample mixture close to the heater and use a very small amount of fluid -213-201209402 to perform a rapid thermal cycle during nucleic acid amplification. For a target sequence of up to 1 50 base pairs (bp) in length, each thermal cycle (i.e., denaturation, adhesion, and extension of the primer) is completed in less than 30 seconds. In most diagnostic analyses, the individual thermal cycle time is less than 11 seconds and most are less than 4 seconds. For LOC device 30, which performs some of the most common diagnostic assays, the thermal cycle time of a target sequence of up to 150 base pairs (bp) is between 0.45 seconds and 1.5 seconds. This rate of thermal cycling allows the test module to complete the nucleic acid amplification process in as little as 10 minutes: usually less than 220 seconds. For most analyses, the amplification produces enough amplicons within 80 seconds of the sample fluid entering the sample inlet. Many analyses produce enough amplicons in 30 seconds. When a predetermined number of amplification cycles are completed, the amplicon is fed to the hybridization and detection portion 52 via the boiling start valve 108. Hybridization Chamber v Figures 52, 53, 54, 56 and 57 show the hybridization chamber 180 in the hybridization chamber array 110. The hybridization and detection section 52 has a 24x45 array 1 10 of hybrid chambers 180 each having a hybrid-reactive FRET probe 186, a heater element 182, and an integrated photodiode 184. The photodiode 1 8 4 is introgressed to detect fluorescence produced by hybridization of the target nucleic acid sequence or protein to the FRET probe 186. Each photodiode 184 is independently controlled by a CMOS circuit 86. Any substance between the FRET probe 186 and the photodiode 1 84 must be transparent to the emitted light. Therefore, the wall portion 97 between the probe 186 and the photodiode 184 can also be optically penetrated by the emission -214 - 201209402. In the LOC device 301, the wall portion 97 is a thin layer of cerium oxide (about 0.5 μm).

Direct incorporation of photodiode 184 beneath each hybridization chamber 180 allows for a detectable fluorescent signal to be produced by the extremely small probe-target hybrid volume (see Figure 5 4). This small amount allows the use of a small volume of hybridization chamber. The amount of probe before hybridization required for the amount of detectable probe-target hybrid must be less than 2 70 pg 〇 1 <: (^^111) (corresponding to 900,000 cubic microns), in most cases less than 60 picograms (corresponding to 200,000 cubic micrometers), typically less than 12 picograms (corresponding to 40,000 cubic micrometers), and less than 2.7 picograms (corresponding to a chamber of 9,000 cubic micrometers) using the LOC device 301 as shown in the accompanying drawings. volume). Of course, reducing the size of the hybridization chamber allows for a higher density chamber, thus allowing the LOC device to have more probes. In the LOC device 301, the hybrid has more than 1,000 chambers in an area of 1,500 micrometers by 1,500 micrometers (i.e., less than 2,250 square micrometers per chamber). The smaller volume also reduces reaction time, thus hybridization. And testing can be faster. Another advantage of requiring a small number of probes in each chamber is that only a very small amount of probe solution needs to be spotted into each chamber during manufacture of the LOC device. Embodiments of the LOC device of the present invention can be spotted using a probe solution having a volume of 1 liter or less. After nucleic acid amplification, the boiling start valve 108 is activated and the amplicon flows along the flow path 176 and flows into each of the hybridization chambers 180 (see Figures 52 and 56). Endpoint liquid sensor 178 displays the point at which hybridization chamber 180 is filled with amplicon and priming heater 182. After a sufficient hybridization time, LED 26 (see Figure 2) is activated. -215- 201209402

The opening in each hybrid chamber 180 is provided with a light window 136 to expose the FRET probe 186 to the excitation radiation (see Figures 52, 54 and 56). The LED 26 emits light for a time long enough to induce a high intensity fluorescent signal from the probe. During the excitation, the photodiode 184 is shorted. After pre-programmed delay 300 (see Figure 2), the photodiode 184 is enabled and detects fluorescent emissions in the absence of excitation light. The incident light on the active region 185 (see FIG. 54) of the photodiode 184 is converted into a photocurrent that can then be measured by the CMOS circuit 86.

Hybridization chambers 180 each carry a probe for detecting a single target nucleic acid sequence. Each hybridization chamber 180 can carry probes that detect more than 1,000 different targets, if desired. Alternatively, many or all of the hybridization chambers may carry the same probe to repeatedly detect the same target nucleic acid. Copying the probes in the hybridization chamber array 110 in this manner results in increased confidence in the results obtained, if desired to provide a single result by combining the results of the photodiodes adjacent to the hybridization chambers. Those skilled in the art will appreciate that there may be from 1 to more than 1,000 different probes on the hybrid chamber array 110, depending on the analytical specifications. Humidifier and Humidity Sensor The AG zone of Figure 6 indicates the location of the humidifier 1 96. The humidifier prevents the reagents and probes from evaporating during operation of the LOC device 301. Preferably, as shown in the enlarged view of Figure 55, the reservoir 188 is in fluid communication with the three evaporators 1 90. The water reservoir 1 8 8 holds the molecular bio-grade water and is sealed during manufacture. As best seen in Figures 5 and 74, water is drawn by capillary action to three lowering ports 194 and along the individual water supply passages 192 to three of the evaporators 190 - 216 - 201209402 ascending port 193 sets. The meniscus is fixed to each of the risers 193 to retain water. The evaporator has a ring heater 191 that surrounds the riser 193. The ring heater 191 is connected to the top metal layer 195 of the CMOS circuit 86 by a conductive post 376 (see Figure 37). At startup, the ring heater 191 heats the water to evaporate and wet the surrounding devices.

Figure 6 also shows the location of the humidity sensor 232. However, as shown in the enlarged view of the AH zone of Fig. 67, the humidity sensor has a capacitive comb structure. The lithographically etched first electrode 296 and the lithographically etched second electrode 298 are opposite each other' such that their teeth are interleaved. The opposing electrode forms a capacitor having a capacitance that can be monitored by CMOS circuit 86. As the humidity increases, the permittivity of the air gap between the electrodes increases, causing the capacitance to increase. The humidity sensor 232 is adjacent to the array of hybridization chambers 1 1 where the humidity measurement is important to slow the evaporation of the solution containing the exposed probe. The feedback sensor temperature and liquid sensors are shunted into the LO C unit 310 to provide feedback and diagnostics during operation of the unit. Referring to Fig. 35, nine temperature sensors 1 7 〇 are assigned to the amplification unit 1 1 2 everywhere. Similarly, the culture unit 1 14 also has nine temperature sensors 170. These sensors each use a 2 X 2 array of bipolar junction transistors (BJT) to monitor fluid temperature and provide feedback to CMOS circuitry 86. The CMOS circuit 86 utilizes this to accurately control thermal cycling during nucleic acid amplification as well as thermal lysis and any heating during incubation. In hybrid chamber 180, CMOS circuit 86 uses hybridization heater 182 as a temperature sensor (see Figure 56). The resistance of the hybrid heater 182 is -217 - 201209402 degree dependent, and the CMOS circuit 86 utilizes this to derive temperature readings for each 180. The LOC device 301 also has a number of MST channel liquids 174 and an upper cover channel liquid sensor 208. Figure 35 shows a row of MST channel liquids 174 at one of the other corners of the heating 158. As best seen in Figure 37, the MST channel liquid sensor is opposed to one of the exposed regions of the top metal layer 195 in the CMOS structure 86. The liquid closes the current between the electrodes to indicate the location of their presence. Figure 25 shows an enlarged cross-sectional view of the upper lid channel liquid sensor 208. The TiAl electrode pairs 218 and 22 are deposited between the top layer 66 electrodes 21 8 and 220 as a gap 222 for holding the circuit in the absence of a circuit. Open. The electrical CMOS circuit 86 is turned off when the liquid is present to monitor the flow. Non-gravity dependent test module 1 is non-directional dependent. These modules do not require a smooth surface to operate. The fluid flow driven by the capillary action is connected to the external piping of the auxiliary device, so that the module is simply portable and can be easily inserted into a similar portable handheld reader. Operations such as non-gravity-dependent operations on behalf of these test modules also contribute to all practical areas. They can withstand shock and vibration, they can operate on the carrier, or when the mobile phone is moving. The hybrid chamber sensor microchannel sensor 174 is formed in the sensing view. on. In the case of less liquid, it is fixed and not required and can be telephone. Independence on the move -218- 201209402

Nucleic acid amplification variant

Conventionally, PCR requires extensive purification of the target DNA prior to preparation of the reaction mixture. However, with appropriate changes in chemical properties and sample concentrations, it is possible to perform nucleic acid amplification or direct amplification with minimal DNA purification. When the nucleic acid amplification program is PCR, this method is called direct PCR. When the nucleic acid amplification is carried out in a controlled constant temperature LOC unit, the method is directly thermostated. Direct nucleic acid amplification techniques have significant advantages when used in LOC devices, especially with regard to simplifying the fluid design required. Adjustments to the amplification chemistry of direct PCR or direct isothermal amplification include increasing buffer strength, using high activity and processivity polymerases and additions to potential polymerase inhibitors. It is also important to dilute the inhibitor in the sample. To take advantage of direct nucleic acid amplification techniques, the LOC device design incorporates two additional features. The first feature is a reagent reservoir (e.g., reservoir 58 in Figure 8) that is suitably sized to supply a sufficient amount of amplification reaction mixture or diluent to cause the sample component that may interfere with the amplification chemistry. The final concentration is low enough to allow for successful nucleic acid amplification. The desired dilution factor of the non-cellular sample component is between 5 and 20 times. Different L0C structures are used where appropriate to confirm that a sufficiently high target nucleic acid sequence concentration is maintained for amplification and detection, such as the pathogen dialysis section 70 of Figure 4. In this embodiment (further illustrated in Figure 6), a concentration that effectively concentrates the pathogens that are small enough to enter the amplification section 292 and discharges the larger cells to the dialysis section of the waste container 76 is used upstream of the sample extraction section 290. . In another embodiment -219 - 201209402, a dialysis section is used to selectively remove proteins and salts in plasma while retaining cells of interest.

A second LOC structural feature that supports direct nucleic acid amplification is the channel aspect ratio design to adjust the mixing ratio between the sample and the amplified mixed components. For example, to ensure that the inhibitor associated with the sample is diluted to a preferred range of 5 to 20 times via a single mixing step, the length and cross-section of the sample and reagent channels are designed to be at the beginning of the mixing The upstream sample channel has a flow impedance that is 4 to 19 times higher than the flow impedance of the channel through which the reagent mixture flows. The flow impedance of the microchannel can be easily controlled by controlling the design geometry. In terms of a fixed cross-sectional area, the flow impedance of the microchannel increases linearly with the length of the channel. Important for hybrid designs, the flow impedance in the microchannel is most dependent on the smallest cross-sectional area. For example, when the aspect ratio is extremely non-uniform, the flow resistance of a microchannel having a square cross section is inversely proportional to the cube of the smallest vertical dimension.

Reverse Transcriptase PCR (RT-PCR) When the sample nucleic acid species analyzed or extracted are RNA, such as from RNA viruses or messenger RNAs, the RNA must first be reverse transcribed into complementary DNA (cDNA) before PCR amplification. The reverse transcription reaction can be carried out in the same chamber as the PCR (one-step RT-PCR), or it can be an initial reaction (two-step RT-PCR). In the LOC variants described herein, a one-step RT-PCR can be performed by simply adding a reverse transcriptase to a reagent reservoir 62 containing the polymerase and staging the heater 1 54 to perform the reverse transcription step. The nucleic acid amplification step is carried out. The second step RT-220-201209402 PCR can also use the reagent reservoir 58 to store and distribute the buffer, primer, dNTP and reverse transcriptase, and use the culture unit 114 to perform the reverse transcription step, followed by amplification. Amplification in the section 112 in a conventional manner is simply accomplished. Thermostatic Nucleic Acid Amplification For some applications, thermophilic nucleic acid amplification is preferred for nucleic acid amplification.

By the method, therefore, the reaction components do not need to be subjected to repeated cycles of different temperatures, but the amplifying portion is maintained at a constant temperature of usually about 37 ° C to 41 ° C. Some thermostated nucleic acid amplification methods have been described, including strand-substituted amplification (SDA), transcription-mediated amplification (TMA), nucleic acid sequence basal amplification (NASBA), recombinase polymerase amplification (RPA), unwinding Any of these methods, or other isostatic amplification methods, for enzyme-dependent isothermal DNA amplification (HDA), rolling cycle amplification (RCA), branched-type amplification (RAM), and circular media constant temperature amplification (LAMP) It can be used in specific implementations of the LOC devices described herein. In order to perform a thermostatic nucleic acid amplification, the reagent reservoirs 6A and 62 of the adjacent amplification section will carry an appropriate reagent for a specific constant temperature method instead of carrying the PCR amplification mixture and the polymerase. For example, in the case of s D A , the reagent reservoir 60 contains an amplification buffer, an primer, and dNTP, and the reagent reservoir 62 contains an appropriate endonuclease and an exo-DN A polymerase. In the case of RPA, the reagent F'T0 0 contains amplification buffer, primer, d Ν τ P and recombinase protein, and the reagent reservoir 62 contains a strand-substituted DNA polymerase, such as Bsu 〇, similarly to HDA. For example, the reagent reservoir 6 contains an amplification buffer, -221 201209402 primer and dNTP, and the reagent reservoir 62 contains a suitable DNA polymerase and helicase to unwind the double strand DNA instead of using heat. Those skilled in the art will appreciate that 'the necessary reagents can be dispensed into the two reagent reservoirs in any manner suitable for the nucleic acid amplification method to amplify viral nucleic acids from RNA viruses such as HIV or hepatitis C virus. NASBA or TMA is appropriate because it does not require the transcription of RNA into cdNA. In this example, the reagent reservoir 60 is filled with an expansion buffer' primer and dNTP, and the reagent reservoir 62 is filled with an RNA polymerase, a reverse transcriptase, and an arbitrary RNase. Some forms of thermostatic nucleic acid amplification must have an initial denaturation cycle to separate the double stranded DNA template before maintaining the temperature suitable for constant temperature nucleic acid amplification for the reaction to proceed. This can be easily achieved in embodiments of all of the LOC devices described herein because the temperature of the mixing in the amplification section 112 can be carefully controlled by the heater 154 in the amplification microchannel 158 (see Figure 14). . Thermostatic nucleic acid amplification is more tolerant to potential inhibitors in the sample and is therefore generally suitable for situations where direct nucleic acid amplification from the sample is desired. Therefore, thermostatic nucleic acid amplification can sometimes be used for LOC variant XLIII 673, LOC variant XLIV 674 and LOC variant XLVII 677, etc., respectively, shown in Figures 82, 83 and 84. Direct thermostatic amplification can also be combined with one or more of the pre-amplification dialysis steps 70, 686 or 682 as shown in Figures 82 and 84, and/or the pre-hybridization dialysis step 682 as shown in Figure 83, respectively. Part of the target cells in the sample are concentrated prior to nucleic acid amplification, or unwanted cell debris is removed before the sample enters the hybridization chamber array 110. Those skilled in the art will appreciate that any combination of pre-amplification dialysis and pre-hybridization dialysis can be used. 0 Constant temperature nucleic acid amplification can also be performed in parallel amplifications such as those shown in Figures 78, 79 and 80. Multiplex and some methods of constant temperature nucleic acid amplification such as the LAMP line are compatible with the initial reverse transcription step to amplify rnA. Other details of the fluorescent detection system

Figures 58 and 59 show hybridization-reactive FRET probes 236. These probes are often referred to as molecular beacons, which are stem-loop probes produced by single-stranded nucleic acids that emit fluorescence when hybridized to complementary nucleic acids. Figure 58 shows a single FRET probe 23 6 prior to hybridization to the target nucleic acid sequence 23 8 . The probe has a ring 240, a stem 242, a fluorophore 246 at the 5' end, and a quencher 248 at the 3' end. This loop 240 is composed of a sequence complementary to the target nucleic acid sequence 238. The complementary sequences flanking the probe sequence are bonded together to form stem 242. When the complementary target sequence is absent, the probe is closed as shown in Figure 58. The stem 242 maintains the fluorophore-quenching agent pairs in close proximity to each other such that significant resonance energy transfer can occur between them, substantially eliminating the ability of the luminaire to emit fluorescence upon exposure to the excitation light 244. Figure 59 shows FRET probe 23 6 in an open or hybridized configuration. Upon hybridization to the complementary target nucleic acid sequence 238, the stem-loop structure is disrupted and the fluorophore and quencher are spatially separated, thereby restoring the ability of the fluorophore 246 to emit fluorescence. The fluorescent emission 250 is optically detected as an indicator that the probe has hybridized. The probe hybridizes to the complementary target with very high specificity because the stem helix of probe 223-201209402 is designed to be more stable than the probe-target helix with a non-complementary single nucleotide. Since the double-stranded DNA is quite robust, the probe-target helix and the stem helix cannot coexist in the stereoscopic space. The probe-attached probe and primer-attached stem-loop probe and the primer-connected linear probe (also known as the scorpion-type probe) are alternative molecular beacons that can be used in LOC devices for immediate and quantitative Nucleic acid amplification. Immediate amplification can be performed directly in the hybridization chamber of the LOC device. An advantage of using a probe coupled to a primer is that the probe element is actually linked to the primer, so that only a single hybridization event is required during nucleic acid amplification without the need for separate primer hybridization and probe hybridization. This ensures an immediate and efficient response, resulting in a stronger signal than the use of separate primers and probes, 'shorter reaction time and better recognition. The probes (and polymerase and amplification mixture) will be deposited in the hybrid chamber 180 during manufacture without the need to provide separate amplifications on the LOC device. Alternatively, the amplification portion is not used or used for other reactions. Linear probes linked to the primers. Figures 85 and 86 show linear probes 692 linked to the primer during the first round of nucleic acid amplification, respectively, and subsequent nucleic acid amplification. A linear probe that is hybridized to the primer during the hybrid configuration. Referring to Figure 85, the linear probe 692 coupled to the lead has a double stem section 242. One of the probes 696, which is homologous to the region 696 on the target nucleic acid, is linked to the primer, and is labeled with a fluorophore 246 at the 5' end thereof, and amplified at the 3' end of the other. Blocker-224-201209402 6 94 was ligated to oligonucleotide primer 700. The 3' end of the other strand of the stem 242 is labeled with a quencher group 248. After completing the first round of nucleic acid amplification, the probe can be rolled up and extended with a stretch of strands having sequence 698 (now complementary)

cross. During the first round of nucleic acid amplification, the oligonucleotide primer 700 is affixed to the target DNA 23 8 (Fig. 85) and then extended to form a DNA strand containing both the probe sequence and the amplification product. The amplification blocker 694 prevents the polymerase from reading and replicating the probe region 696. Upon subsequent denaturation, the extended oligonucleotide primer 700/template hybridization system is separated, and the double stem 242 of the linear probe linked to the primer is also separated, thereby releasing the quencher 248. When the temperature is lowered for the bonding and extension step, the primer-linked probe sequence 696 of the linear probe linked to the primer is rolled up and hybridized with the amplified complementary sequence 698 on the extended strand, and the display can be detected. Fluorescence of the target DNA. The unexpanded linear probe attached to the primer retains the double stem and the fluorescence remains quenched. This test method is especially suitable for fast detection systems because it requires only a single molecule treatment.

Stem ring probe attached to the primer Figure 87A to 87F show the operation of the stem probe 704 attached to the primer. Referring to Fig. 87A, the primer-connected stem-loop probe 76 has a stem 242 of complementary double-stranded DNA and a loop 240 having a probe sequence. The 5' end of one of the stems 708 is labeled with a fluorophore 246. Another 710 line is labeled with a 3'-end quencher 248 and carries both amplification blocker 694 and oligonucleotide primer 700. During the initial denaturing phase (see Figure 87B), the strands of the target nucleic acid 238 are separated, and the stem 242 of the stem-loop probe 704 attached to the primer is also separated -225-201209402. When the temperature is cooled to effect the binding phase (see Figure 87C), the oligonucleotide primer 700 on the stem loop probe 704 attached to the primer hybridizes to the target nucleic acid sequence 2 3 8 . During extension (see Figure 8 7D), the complementary sequence 706 of the target nucleic acid sequence 23 is synthesized to form a DNA strand containing both the probe sequence 704 and the amplification product. The amplification blocker 694 prevents the polymerase from reading and replicating the probe region 704. When the probe is bonded after denaturation, the probe sequence of the loop segment 240 of the stem-loop probe attached to the primer (see Figure 8 7F) is bonded to the complementary sequence 706 on the stretched strand. This configuration causes the fluorophore 246 to be at a great distance from the quencher 248, resulting in significantly enhanced fluorescence emission. Control Probes Hybridization chamber arrays 1 1 〇 include some hybridization chambers 180 with positive and negative control probes for analytical quality control. Figures 100 and 101 schematically illustrate negative control probes without fluorophore 796, and Figures 〇2 and 103 show positive control probes without quencher 79 8 . The positive and negative control probes have a stem-loop structure as described above for the FRET probe. However, regardless of whether the probes hybridize to an open configuration or remain closed, the positive control probe 79 will emit a fluorescent signal 250, and the negative control probe 796 will never emit a fluorescent signal 2 50 » Referring to Figures 1A and 101, the negative control probe 796 does not have a fluorophore (which may or may not have a quencher 248). Thus, regardless of whether the target nucleic acid sequence 23 8 hybridizes to the probe (see Figure 1-1) or the probe maintains its stem-loop configuration (see Figure 1A), the reaction to excitation light 244 can be ignored. . Alternatively, the negative control probe 796 can be designed to keep quenching forever. For example, by synthesizing a loop 240 having a probe sequence that does not hybridize to any of the nucleic acid sequences in the study sample, the stem 242 of the probe molecule will self-hybridize itself, allowing the fluorophore and quencher Stay close together without emitting visible fluorescent signals. This negative control signal corresponds to a small amount of emission from the hybridization chamber 180 where the probes are not hybridized but the quencher does not quench all of the emission from the reporter.

Conversely, the positive control probe without the quencher was constructed as shown in Figures 102 and 103. Regardless of whether the positive control probe 798 is hybridized to the target nucleic acid sequence 238, no material will quench the fluorescent emission 250 from the fluorophore 246 back to the luminescent 244. Figure 52 shows the possible distribution of positive and negative control probes (3 78 and 380, respectively) in hybridization chamber array 110. The control probes 3 78 and 380 were placed in a hybridization chamber 1 80 located on the line across the hybridization chamber array 110. However, the configuration of the control probes within the array is arbitrary (like the configuration of the hybrid chamber array 1 1 )). The fluorophore design requires a fluorophore with a long fluorescent lifetime, which is to allow the excitation light to have sufficient time to decay to a low intensity compared to the intensity of the fluorescent emission when the photosensor 44 is enabled. This provides a sufficient signal to noise ratio. Moreover, a longer fluorescent lifetime represents a larger integrated fluorescence photon count. The fluorescence lifetime of the fluorophore 246 (see Figure 59) is greater than 100 nanoseconds, often greater than 200 nanoseconds, more typically greater than 300 nanoseconds, and greater than 4000 in most cases of -227-201209402. second. Metal-coordination complexes based on transition metals or lanthanide metals have long lifetimes (from hundreds of nanoseconds to milliseconds), appropriate quantum yields, and high thermal, chemical, and photochemical stability. Both are advantageous for the needs of fluorescent detection systems. In particular, the metal-coordination complex based on the transition metal ion ruthenium (Ru(ii)) is ginseng (2,2'-bipyridyl) ruthenium (11) ([1111〇?7). 3] 2 + ), the life of the latter is about Ιμδ. This complex is commercially available from Biosearch Technologies under the trade name Pulsar 650. Table 1: Photophysical properties of Pulsar 6 5 0 (钌 chelate)

Parameter symbol 値 unit absorption wavelength ^abs 460 nm emission wavelength ^em 650 nm absorption coefficient E 14800 M-1 cm·1 fluorescence lifetime Tf 1.0 μδ quantum yield Η 1 (deoxidized) N/A 铽 chelate is A lanthanide metal-coordination complex that has been shown to be successful as a fluorescent reporter for the FRET probe system and has a long lifetime of 1 600 μβ. -228- 201209402 Table 2: Photophysical properties of ruthenium chelate parameters Symbol 値 Unit absorption wavelength ^abs 330-350 nm Emission wavelength λέπι 548 nm Absorption coefficient Ε 13800 (with Us and ligand dependence, up to 30,000 @ Xe=340nm) Fluorescence lifetime Xf 1600 (hybridized probe) μδ quantum yield Η 1 (coordination dependent) Ν/Α

The fluorescent detection system used by the LOC device 310 does not utilize filters to remove unwanted background fluorescence. Therefore, if the quencher 248 has no natural emission in order to increase the signal to noise ratio, it is advantageous. The quencher 248 without natural emission does not contribute to background fluorescence. High quenching efficiency is also important, so there is no fluorescence before the hybridization event occurs. Black Hole Cleaner (BHQ), available from Biosearch Technologies, Inc. of Novato, Calif., does not have a natural emission and has high quenching efficiency and is suitable for use in the quenching agent of this system. The maximum absorption enthalpy of BHQ-1 occurs at 534 nm and the quenching range is 480-580 nm, making it suitable for quenching Tb-chelating fluorophores. The maximum absorption enthalpy of BHQ-2 occurs at 579 nm and the quenching range is 560-670 nm, making it suitable for quenching Pulsar 650. Iowa Black FQ and RQ from Integrated DNA Technologies, Coralville, Iowa, are suitable alternative quenchers with little or no background emission. The Iowa Black FQ has a quenching range from 420 to 620 nm with a maximum absorption enthalpy at 531 nm, which is suitable for use as a quencher for Tb-chelating fluorophores at -229-201209402. The maximum absorption enthalpy of Iowa Black RQ occurs at 656 nm and the quenching range is from 500 to 700 nm, making it an ideal quencher for Pulsar 6 50. In the embodiment described herein, the quencher 248 is a functional group attached to the probe initially, but in other embodiments, the quencher may be separated from the solution. molecule.

Excitation source In the implementation of the fluorescence detection as the substrate described in this paper, the LED is selected as the excitation source instead of the laser diode, high power lamp or laser because of the low power consumption, low cost and small size of the LED. . Referring to Fig. 88, LEDs 26 are disposed directly above hybrid array array 110 of the outer surface of LOC device 301. Opposite the array of hybridization chambers 110 is a light sensor 44 consisting of an array of photodiodes 184 for detecting fluorescent signals from the chambers (see Figures 53, 54 and 68).

Figures 89, 90 and 91 schematically illustrate other embodiments for exposing the probe to excitation light. In the LOC device 30 shown in Fig. 89, the excitation light 2 44 generated by the excitation LED 26 is directed by the lens 254 onto the hybridization cell array 110. The excitation LED 26 is pulsed and the fluorescent sensor 44 detects the fluorescent emission. In the LOC device 30 as shown in Fig. 90, the excitation light 244 generated by the excitation LED 26 is directed by the lens 254, the first optical stop 712 and the second optical stop 714 over the hybridization chamber array 110. The excitation LED 26 is pulsed and the fluorescent sensor 44 detects the fluorescent emission. Similarly, in the LOC device 30 shown in FIG. 91, the excitation light 244 generated by the excitation LED 26 is directed to the hybridization chamber array 11 by the lens 254, the first mirror 716, and the second mirror 718. on. Furthermore, the excitation [ED 26 is pulsed and the fluorescence emission is detected by the light sensor 44.

The excitation wavelength of the LED 26 is dependent on the choice of egg light dye. Philips LXK2-PR14-R00 is the appropriate source of excitation for Pulsar 650 dyes. SET UVT0P3 3 5T039BL LED is the appropriate excitation source for the ruthenium chelate. Table 3: Philips LXK2-PR14-R00 LED Specifications

Parameter Symbol 値 ρα — Single 兀 Wavelength λβχ 460 nm Transmit frequency Vem 6.52(10)14 Hz Output power Pi 0.515(min)@ ΙΑ W Transmit mode Lambertian data graph N/A Table 4 : SET UVTOP33 4T039BL LED Specifications

Parameter Symbol 单元 Element Wavelength λβ 340 nm Transmit frequency Ve 8.82(10)14 Hz Power Pi 0.000240(min)@ 20mA W Pulse forward current I 200 mA Emission mode Lambertian N/A UV excitation 矽 Absorbs a small amount of UV light. Therefore, it is advantageous to use ultraviolet excitation light. The LED excitation source can be used with ultraviolet light, but the wide spectral drop of LED 26 -231 - 201209402 is lower than this method. To solve this problem, a filtered UV LED can be used. Optionally, an ultraviolet laser can be used as an excitation source unless the relatively high cost of the laser is not practical for a particular test module market. LED Driver The time required for the LED driver 29 to drive the LED 26 at a fixed current. The low-power USB 2.0-certified unit can supply up to 1 unit load (! 〇〇 mA) with a minimum operating voltage of 4.4 volts. Standard power conditioning circuits are used for this purpose. Photodiode Figure 54 shows a photodiode 184 that breaks into the CMOS circuit 86 of the LOC device 301. The photodiode 184 is part of the CMOS circuit 86 without additional masking or steps. This is a significant advantage of CMOS photodiodes over CCDs, an alternative sensing technique that can be integrated on the same wafer or fabricated on adjacent wafers using non-standard processing steps. On-wafer inspection is inexpensive and reduces the size of the array system. This shorter optical path length reduces noise from the surrounding environment to effectively collect fluorescent signals and reduce the need for conventional optical assemblies for lenses and filters. The quantum efficiency of the photodiode 184 is the fraction of photons that are effectively converted into photoelectrons in the photons that collide with their active regions 185. In terms of standard enthalpy treatment, the quantum efficiency of visible light ranges from 0.3 to 0.5, depending on processing parameters such as the amount of cover layer and absorption characteristics. -232- 201209402 The detection valve of photodiode 184 determines the minimum intensity of the fluorescent signal that can be detected. The detection valve 値 also determines the size of the photodiode 184 and therefore the number of hybridization chambers 180 in the hybridization and detection section 52 (see Figure 52). The size and number of such chambers are limited by the size of the LOC device (in the case of the LOC device 301, which is 1760 microns X 5 8 24 microns) and into other functional modules such as the pathogen dialysis unit 70 and The technical parameters of the size of the available space after the addition 112.

The photodiode 184 detects a minimum of 5 photons in terms of standard 矽 processing. However, to ensure reliable detection, the minimum chirp can be set to 10 photons. Thus, in the quantum efficiency range of 0.3 to 0.5 (as discussed above), the fluorescence emission from the probes should be a minimum of 17 photons, but 30 photons will contain an appropriate margin of error for reliable detection. Calibration Chamber The inhomogeneity of the electrical characteristics of the photodiode 184, autofluorescence, and residual excitation photon flux that is not fully attenuated by φ 导入 introduces background noise and offset into the output signal. This background is removed from each output signal using one or more calibration signals. The calibration signal is generated by exposing one or more of the calibrated photodiodes 184 in the array to individual calibration sources. The low calibration source is used to determine the negative result of the target 尙 not reacting with the probe. The high calibration source is indicative of a positive result from the probe-target complex. In the embodiment described herein, the low calibration source is provided by a calibration chamber 382 in the hybrid chamber array 110, the calibration chamber 382: does not contain any probes; -233-201209402 contains no fluorescence a probe for the reporter; or a probe having a reporter and a quencher configured to cause quenching to occur forever. The output signal from the calibration chamber 382 is very close to the noise and bias in the output signals from all of the hybrid chambers in the LOC device. The output signal generated by the other hybridization chamber minus the calibration signal substantially removes the background, leaving the signal generated by the fluorescent emission (if any). Signals generated by ambient light in the area of the chamber array are also removed. It will be appreciated that a negative control probe as described above with reference to Figures 1A through 103 can be used in the calibration chamber. However, as shown in Figures 94 and 95 (which is an enlarged view of the DG and DH regions of the LOC variant X 728 as described in Figure 93), another option is to fluidly isolate the calibration chamber 382 from the amplicons. . The background noise and bias can be determined by maintaining the fluid isolation chamber clearance or by using a probe containing no reporter or indeed any "standard" probe having both a reporter and a quencher, as hybridization Blocked by fluid isolation. The calibration chamber 382 can provide a high calibration source to produce a high signal at the corresponding photodiode. This high signal corresponds to all probes that have been hybridized in the chamber. Spotting a probe with a reporter but no quencher, or a probe with only a reporter will consistently provide a signal that approximates that most of the probes in the hybridization chamber have been hybridized. It will also be understood that the 'calibration chamber 382 can be used in place of or in addition to the control probe. The number and arrangement of calibration chambers 382 throughout the array of hybrid chambers is arbitrary. However, if the photodiode 184 is calibrated by a relatively close calibration chamber 382, the calibration will be more accurate. Referring to Figure 56, the hybridization chamber array 11 --234-201209402 has one calibration chamber 382 for every eight hybridization chambers 180. That is, the calibration chamber 382 is placed in the middle of each of the three-by-three hybrid chambers 180 squares. In this configuration, the hybridization chambers 180 are calibrated by the immediately adjacent calibration chamber 382.

Figure 99 shows a differential imager circuit 78 8 for subtracting the signal from the fluorescent signal of the surrounding hybridization chamber 180 corresponding to the photodiode 184 of the calibration chamber 382 due to the excitation light. The differential imager circuit 788 samples the signal from the pixel 790 and the "virtual" pixel 792. In the embodiment - the "virtual" pixel 792 is shielded from light illumination, so its output signal provides a dark reference. Alternatively, the "virtual" pixel 792 can be exposed to the excitation light along with other portions of the array. In the embodiment in which the "virtual" pixel 792 is exposed to light, the signal generated by ambient light in the area of the array of cells is also subtracted. The signal from pixel 790 is very weak (i.e., close to the dark signal), and it will be difficult to distinguish between background and very weak signals without reference to the dark signal level. During use, the "read_column" 794 and "read_column_(1" 795 are activated, and the M4 797 and MD4 80 1 transistors are turned on. The switches 807 and 809 are turned off to enable the pixel from The outputs of the 79" and "virtual" pixels 792 are stored in pixel capacitor 803 and virtual pixel capacitor 805, respectively. After the pixel signal is stored, switches 807 and 809 are disabled. Next, the "read-line" switch 8 1 1 And the virtual "read_row" switch 8 1 3 is turned off 'the switched capacitor amplifier 815 at the output amplifies the differential signal 8 17 « suppression and enabling of the photodiode - 235 - 201209402 The photodiode 184 LED 26 must be suppressed during excitation and must be enabled during fluorescence. Figure 69 is a circuit diagram of a single photodiode 丨 84, and Figure 70 is a timing diagram of a photodiode control signal. The circuit has a photodiode 184 and six MOS transistors Mshunt 3 94, Mtx 396, Mreset 3 98, Msf 400, Mread 402 and Mbias 404. At the beginning of the excitation cycle t1, by pulling up the Mshunt gate 3 84 and resetting the gate 3 8 8 and turn on the crystal Mshunt 3 94 and Mreset 3 98. During this period, The excitation photons generate a carrier in the photodiode 184. These carriers must be removed because the amount of carrier generated is sufficient to saturate the photodiode 184. During this cycle, Mshunt 3 94 directly removes the photodiode The carrier generated in body 184, and Mreset 398 resets any carrier that accumulates at node 'NS' 406 due to transistor leakage or carrier diffusion due to excitation in the substrate. After excitation, the capture cycle begins at t4» During the cycle, the emission reaction from the fluorophore is captured and integrated into the circuit on the node 'NS' 406. This is achieved by pulling up the tx gate 3 86, which turns on the transistor M, x 396 and accumulates in the photodiode Any carrier on pole 184 is transferred to node 'NS' 406. The length of the capture cycle can be as long as the fluorophore emission. The output from all of the photodiodes 184 in hybrid array 110 is simultaneously captured. There is a delay between the end of cycle t5 and the beginning of read cycle t6. This delay is due to the need to separately read each photodiode 184 in hybrid cell array 110 after the capture cycle (see Figure 52). The photodiode 184 will have the shortest read cycle delay, and the last photodiode 184 will have the longest read cycle delay. During the read cycle, the read gate is pulled high. 393 and turn on the transistor -236- 201209402

Mread 402. The voltage of the 'NS' node 406 is buffered and read using the source follower transistor Msf 400. Other optional methods for activating or suppressing the photodiode are discussed below: 1. Suppression method

Figures 96, 97 and 98 show three possible configurations of Mshunt transistor 3 94 77 8, 780, 7 82. The Mshunt transistor 394 has a very high turn-off ratio at maximum |FCS| = 5 V, which is enabled during excitation. As shown in FIG. 96, the Mshunt gate 3 84 is configured at the edge of the photodiode 184. Optionally, as shown in FIG. 97, the Mshunt gate 384 is configured to surround the photodiode 184. The third option is to configure the Mshunt gate 3 84 within the photodiode 184, as shown in FIG. In the third option, the active region 185 of the photodiode will be smaller. These three configurations 778, 780, and 782 reduce the average path length from all locations in the photodiode 184 to the Mshunt gate 3 84. In Figure 96, the Mshunt Gate 3 84 is located on one side of the photodiode 1 84. This configuration is the easiest to fabricate and does not occupy the active region of the photodiode 1 85. However, any carrier remaining on the far-end side of the photodiode 1 84 takes a long time to pass through the Mshunt gate 3 84. In FIG. 97, the Mshunt gate W4 surrounds the photodiode 184. This further reduces the average path length of the carrier in the photodiode 184 to the Mshunt gate 3 84. However, the Mshunt gate 384 extending around the periphery of the photodiode 184 causes the larger photodiode active region 185 to be reduced from -237 to 201209402. Configuration 782 in Figure 98 positions Mshunt Gate 3 84 in main area 185. This provides the shortest average path to the Mshunt gate 384, so the transition time is the shortest. However, the encroachment of the active zone 185 is large. It also creates a wider leak path. 2. Enable method a. Trigger the photodiode to drive the shunt transistor with a fixed delay. b. Triggering the photodiode to drive the shunt transistor with a programmable delay c. The shunt transistor is driven by the LED drive pulse with a fixed delay. d. Drive the shunt transistor as a 2c but with a programmable delay. Figure 75 is a schematic cross-sectional view showing the cross-cutting cell 180 showing the photodiode 184 and the trigger photodiode 1 embedded in the CMOS circuit 86. The small area of the corner of the photodiode 184 is replaced by the trigger photodiode 187. Triggering photodiode 187 with a small area is sufficient, and the intensity of the excitation light will be higher than that of the fluorescent emission. The trigger photodiode 187 is sensitive to excitation light 244. The trigger photodiode 187 temporarily extinguishes the excitation light 2 44 and activates the electrical diode 184 after a short delay of Δί 300 (see Fig. 2). This delay allows the fluorescent photodiode 1 to detect the fire emission from the FRET probe 186 when there is no excitation light 244. This enables detection and enhancement of signal to noise ratio. Both photodiode 184 and trigger photodiode 187 are located in CMOS circuitry 86 under each hybrid cell 180. The combination of photodiodes is combined with appropriate electronic components to form a photosensor 44 (see Figure 68). The electric diode 184 is a pn junction made during the manufacture of the CMOS structure. The maximum length of the light is 84. The light is stored in the light - 238-201209402. No additional mask or step is required. During MST fabrication, the dielectric layer (not shown) over the photodiode 184 is arbitrarily thinned using standard MST photolithography techniques to allow more phosphor to illuminate the active region 185 of the photodiode 184. . The photodiode 184 has a field of view such that a fluorescent signal from the probe-target hybrid within the hybridization chamber 180 is incident on the surface of the sensor. The fluorescence is converted to a photocurrent which can then be measured using CMOS circuitry 86.

Alternatively, one or more of the hybridization chambers 180 can be configured to only trigger the photodiode 187. These choices can be used in combination with 2a and 2b above. Fluorescence Delay Detection The following derivation illustrates the use of long-lived fluorophores for delayed detection of the above-described LED/fluorescent combination of fluorescence. The fluorescence intensity is derived from the time function by the ideal pulse of the fixed intensity Ie after excitation between time Μ and h, as shown in Fig. 60. Let [Sl](t) be equal to the intensity of the excited state at time t, then the number of excited states per unit volume per unit time during and after excitation is described by the following differential equation _· d[Sl] dt ( 0 + _) τρ

Hve (1) where c is the molar concentration of the fluorophore, ε is the molar quenching factor, ve is the excitation frequency, and h = 6.62606896(1 0)_34 JS is the Planck constant. This differential equation has the general formula: -239- 201209402 ^- + p(x)y = g(x) αχ There is a solution: y(x) = jpWdx q(x)dx + k jpU)dx (2) Now used This solution (1)

[called W = ^_ + fe-"v...(3) hvM at time h, [we](7))=〇, and from (3) (4)

,1e^T/ /ι; k =--e / ...

hvB Substitutes (4) into (3): [s\m-

Iescrf Iescrf hve hve

At time t2: when hvo κ ... (5) the excited state is exponentially decayed and described by equation (6) -240 - 201209402 [magic] (〇=[幻]Ο—(ί—...(6) will (5 Substitute (6): [51](〇= Ie£CTf [1 -]e—(,_,2)/r' ...(7 ) hye

The fluorescence intensity is obtained by the following equation: (8) //(0 = - αχ where ν/ is the fluorescence frequency, η is the quantum yield, and 1 is the optical path length. Then from (7):

Dt kVe ... (9) Substituting (9) into (8):

If{t) = Ie£Clv^-[le~{,2'l')lTf ]e-(,-i2)/r/ ve ...(1 0) When ^14 〇〇,々(7)-> / #/;/立,-,2)/r,

Tf Ve therefore 'we can write the following approximation, which describes the intensity of the fluorescence -241 - 201209402 after a sufficiently long excitation pulse ((2-h>>rf) decays: when ikt2,1 gossip = 1 fine\"

Ve ...(1 1) In the previous section, we summarized the situation, when tkt2, If(t) = Iescl7j丄e””.

Ve From the above equation, we can derive the following formula: heart (,) => y £ c / 77e_ (, w:) / r / ... (1 2) where = ^ is the fire per unit area per unit time The photon number and hvf 弋=·^- are the number of excitation photons per unit area per unit time. Hye Therefore, 00 nf{t) = \nf{t)dt- (13) h where & is the number of fluorescent photons per unit area and ί3 is the time point at which the photodiode is turned on. Substituting (12) into (13): 00

Hf=jyie€chje-(HWr/dt ...(14) h -242- 201209402 At present, the number of fluorescent photons per unit area per unit area of the photodiode is obtained ^(0 is obtained by the following formula: genus...(15 )

Among them is the light collection efficiency of the optical system. Substituting (12) into (15) we find 氓_((_<2)/, (16) Similarly, the number of fluorescent photons per unit of fluorescent area reaching the photodiode will be as follows: And substitute (16) and integrate:

=φ0ηΐεοΙητ/ε'((ί~,ι),Τί Therefore, η^φ^εοίητ^'^ · (17) The ideal 値 of t3 is the electron generated in the photodiode 184 due to the photon photons The rate becomes equal to the electron rate generated by the excitation photon -243-201209402 in the photodiode 184' because the excitation photon flux decays more rapidly than the fluorescence photon flux. The sensor per unit of fluorescent area is The electron ratio of the fluorescent output is: έ > (〇 = ^η; (〇 where " is the quantum efficiency of the sensor at the fluorescent wavelength.

Substituting (17) we get: = (b '2) / r / ... (1 8 ) Similarly, the electron rate per unit of fluorescent area of the sensor due to excitation of photons is: έ; (0 = ^>'("2)/Γ,- (19)

Nine of them are the quantum efficiencies of the sensor at the excitation wavelength, and % is the time constant corresponding to the "off" characteristic of the LED. After time t2, the attenuated photon flux of the LED will increase the intensity of the fluorescent signal and extend the decay time of the fluorescent signal, but we assume that the effect on If(t) can be ignored, so we take a conservative method. Currently, as mentioned earlier, the ideal system of ^ is:

-244- 201209402 Therefore, we get from (18) and (19): and after the reorganization we get:

\η{εοΙη^^~) h~h = —i~^——(20) -—一丨.

Tf ^ From the above two paragraphs, we get the following two expressions: ' =iWr/e_Ai/r/...(2 1 ) △,=”“V-... (22)

Tf Te where f = £c/;7 and Δί = ί3-ί2. We also know that in fact /2_ii>>iV. The ideal time for fluorescence detection and the number of fluorescent photons detected using Philips LXK2-PR 14-ROO LED and Pulsar 650 dye are as follows. The ideal detection time is determined using equation (2 2): recalling the concentration of the amplicon' and assuming that all amplicons are hybridized, the fluorescing fluorophore concentration is: c = 2.89 (l〇)-6 mol/ L. The height of the chamber is the optical path length 1 = 8 (10) · 6 m. -245- 201209402 The fluorescent region has been regarded as equivalent to the photodiode region, but the actual fluorescent region is substantially larger than the photodiode region; therefore, it can be assumed that the optical system is optically collected. effectiveness. From the characteristics of the photodiode, the ^1 = 10 series is extremely conservative. The ratio of the <Pe quantum efficiency of the photodiode at the fluorescence wavelength to the quantum efficiency at the excitation wavelength. With a typical LED decay life & = 〇.5 nanoseconds and using the Pulsar 65〇 specification, the heart can be determined: F = [\ .48(10)6 ][2.89(10)-6 ][8(10)- 6 ](1) = 3.42(10)-5 1η([3.42(1〇Γ5](10)(0.5)) Δ^= 1 Γ ια 0)-6 ~ 0.5(10)-9 = 4.34(10) The number of photons detected by -9 s is determined using equation (2 1). First, the number of excitation photons emitted per unit time is determined by examining the illumination geometry.

Philips LXK2 -PR 1 The 4-R00 LED has a Lambertian emission mode, so: W/〇C_)...(2 3) where Η•, the angle to the direction of the axis of the LED is 每 per unit solid angle per unit time The number of photons emitted, and &. For Κ, in the direction of the forward axis. The total number of photons emitted by the LED per unit time is: -246- 201209402

=J ril0 cos(0)dQ ω ..· (24) Now,

ΔΩ = 2<1 - cos(0+A0)] - 2;r[l - cos(0)] ΔΩ = 2^[cos(0) - cos(0 + Δ0)] Δ0, = 4^sin( 0)cos dCl = 2π siii(^)d Θ Substituting this into (24): ΑΘ + 4^cos(0)sin2 ΑΘΎ. ή; = J 27fnl0 cos(e)sm(6)de 〇= ^10 Rearrange ,we got:

·ή·丨. ...(26) π LED output power is 0.515 watts and ve = 6.52 (1 Ο) 14 Hz, therefore:

Pi hve ...(27) __0515_ ~ [6.63(10)-34][6.52(10)14] =1.19(10)18 photons/sec. Bring this 値 into (26) we get: -247- 201209402 ...1.19( 10) 18 n,o=~ir~ = 3.79(10)17 photons/sec/sphericity Referring to Fig. 61, the optical center 252 and lens 254 of the LED 26 are schematically shown. The photodiode system is 16 micrometers x 16 micrometers and for the photodiode in the middle of the array, the solid angle (Ω) of the light cone emitted from the LED 26 to the photodiode 184 is approximately: Ω = sensor area / R2 [16 (10)-6][16(10)-6] 2.825(10)-3]2 = 3.21(10)_5 Sphericality It will be understood that the central photodiode 184 of the photodiode array 44 is used. For the purposes of these calculations. Sensors located at the edge of the array receive only less than 2% of photons when using L a m b e r t i a η to excite the hybridization event of the source intensity distribution. Number of excitation photons emitted per unit time: he = η, Ω ... (28) = [3.79(10),7][3.21(10)"5] = 1.22(10)13 Photons/sec Reference now Equation (29): -248 - 201209402 η!=Φ〇ΚΡτ^'Τ' ns = (0.5)[1.22(10)13][3.42(10)-5][l(10)^]e^34( , 〇r9/1(10)'< = 208 Photon/Sensor So, with Philips LXK2-PR14-R00 LED and Pulsar 65 0 fluorophore, we can easily detect any photons that are emitted Hybrid event.

The SET LED illumination geometry is shown in Figure 62. When ID = 20 mA, the LED has a minimum optical power output of Pl = 240 microwatts and a wavelength center at λβ = 340 nm (absorption wavelength of the ruthenium chelate). Driving the LED with ID = 200 mA will linearly increase the output power to Pl = 2.4 mW. By placing the optical center 252 of the LED about 17.5 mm from the array of hybridization chambers 11, we concentrated this output flux on a dot size having a maximum diameter of 2 mm.

The photon flux in the 2 mm diameter dots of the hybrid array plane is obtained from Equation 27. h =1l. 1 hve 2.4(10)-3 _ [6.63(1〇Γ34][8·82(10)14] = 4.10(10)15 photons/sec Using equation 2 8 we get:

= 4.10(10)15 [16(10)6]2 41(1〇)·3]2 -249- 201209402 = 3.34(10)11 Photons/sec Now, return to Equation 22 and use the previously listed Tb chelate Compound property ln[(6.94(l 0)-5)(10)(0·5)] Δ/= 1 1- 1(10)-3 ~ 0.5(10)-9 =3.98(10)_9 seconds now Self-condition 21: n3 = (0.5)[3.34(10)n][6.94(10)-5][l(10)-3]e-398(10)', /1(,〇r5 = 11,600 photons The sensor is simple to detect by the theoretical number of photons emitted by the hybridization event using the SET LED and the ruthenium chelate system, and far exceeds the 30 photons required for reliable detection of the photosensor as described above. The lower limit between the probe and the photodiode The detection of hybridization on the wafer does not require detection with a conjugated focal microscope (see prior art). This difference from conventional detection techniques represents a savings in this system. An important factor in time and cost. Traditional inspection requires the use of lens and curved mirror imaging optics. By using non-imaging optics, the diagnostic system avoids the need for complex and cumbersome strings of optical components. Place the photodiode very Close to the probe for extremely high collection efficiency The advantage: when the material thickness between the probe and the photodiode is 1 micron 'the hair -250-

201209402 The collection angle of the light is as high as 173. . This angle is calculated by considering the light emitted from the probe closest to the center of the surface of the hybridization chamber of the electric diode, the electric diode having a planar active surface area parallel to the surface of the chamber. Light within the pyramid can be absorbed by the photodiode, which has an emission at its apex and at the sensor corners around its plane. Taking a 16 micron X 16 micron sensor as an example, the apex angle of the pyramid is; in the case where the photodiode is expanded such that its area conforms to the 29th. The angle is 173. The spacing between the surface of the chamber and the active surface of the photodiode is as small as 1 micron, which is easily achieved. The application of non-imaging optical methods does require that the photodiode 184 be placed close to the hybridization chamber to collect a sufficient amount of fluorescent emission photons. The maximum spacing between the photodiode probes is determined as described below with reference to Fig. 54. Using the ruthenium chelate fluorophore and SET UVT0P 3 3 5T039BL, we calculated that 1,600 photons were from individual hybridization chambers 180 to 16 turns of 16 micron photodiodes 184. In carrying out this calculation, the light collection region of our pseudo hybrid chamber 180 has the same bottom area as the active region of the photodiode, and half of the total number of hybrid photons reaches the photovoltaic body 184»that is, the optical system The light collection efficiency is nine = 0.5. More precisely, we can write nine = [(the bottom area of the light collection in the hybrid chamber) / (photodiode area)] [Ω / 4 7:], where Ω = The solid angle of the photodiode of the representative point on the hybrid substrate. In terms of the geometry of the square cone: Q = 4arcsin(a2/(4d〇2 + a2)), where d〇 = the chamber and the photodiode. The light is defined as probe 170. Ϊ́ί米 X. In the more or more body and LED intersection meter X set the distance between the 185 two-pole area intersection room -251 - 201209402, and a is the size of the photodiode. Each hybrid chamber releases 232 00 photons. The selected optoelectronics has a detection low limit of 17 photons, so the minimum required: ? = 17/23200 = 7.33 x 10 The bottom area of the light collection region of the hybridization chamber 180 is 29 1 9.75 microns. Solving dQ will result in a maximum limiting distance between the hybridization chamber and the photodiode 184 of d 〇 = 249 μm. Under this limitation, the collecting cone angle as above is only 0.8°. It should be noted that this analysis ignores the effects of refraction. Test module for dialysis device, LOC, and microfluidic device with attached caps The test module for analyzing sample fluids containing the target molecules is shown in Figure 109. The test module 1 1 includes a housing 13 having a container 24 for receiving the body, a sterile sealing tape 22 covering the container 24 before use, and a membrane seal having a membrane guard 410 as a partial housing 13 To reduce the humidity in the test module and provide pressure relief by the membrane guard 410 to prevent damage to the seal 408, the printed circuit board (PCB) 57, the microfluidic device porous member 49, and the standard micro USB connector. 14 for power supply, control, external power supply capacitor 3 2 and inductor 15. The defined negligible test module between the diode optical effect micrometer X" is a sample stream that can be removed at 408 when the film is dense 7 83 > data and -252-201209402 the microfluidic device 783 has The container 24 is in fluid communication and is configured to separate the target molecule from the other components of the sample, the dialysis device 784, the LOC device 785 for analyzing the target molecule, and the LOC device 785 and the dialysis device 784. The upper cover 51 is used to establish fluid communication between the LOC device 785 and the dialysis device 784. Reagent loading and probe spotting system

Reagent reservoirs 54, 56, 58, 60, and 62 (see Figure 6) are filled with reagents and water via a robotic droplet ejection system as shown in Figures 63-66. The robotic system also spots the oligonucleotide FRET probe 186 or ECL probe 237 into the hybridization chamber 180. Droplet dispensing technology is an inexpensive spotting technique that delivers small volumes of reproducible volume and droplets of many different solutions can be dispensed simultaneously. This allows the LOC device to be mass produced with extremely high throughput and low cost. The reagent and probe spotting system includes three robotic subsystems: 1. Reagent dispensing robot 256 (see Figure 63) - each vial 258 (see Figure 64) having a droplet dispenser 262 dispenses reagents to the reservoir 54, 56, 58 '60 and 62 and distribute water to the reservoir 188 (see Figure 6). The robot then applies the patterned upper sealing layer 82 (if desired) to the upper cover 46. The 2.0 NEC refill robot 274 (see Figure 65) - a micro vial 25 8 with a drop dispenser 262 dispenses a probe to the reservoir 278 of the oligonucleotide emitter wafer (ONEC) 272 (see Figures 71 and 72). ). The ONEC receptacle 278 is fed to an array of thermal droplet generators 271. The ONEC is then used for the third robotic subsystem, the LOC spotting robot. - 253 - 201209402 3. LOC Staking Robot 289 (illustrated schematically in Figure 66) - ONEC 272 uses thermal droplet generator 271 to spot probes to each of the hybrid chambers 180 of the LOC device 30 (see Figure 72). Micro vial The reagent dispensing robot 2 56 and the ON EC refilling robot 274 both use a micro vial 2 5 8 as shown in FIG. Probes and reagents are ordered directly from the supplier in the large bottle (not shown). The liquid is drawn from the large vial to a container 25 9 on each of the micro vials 25 8 to form a small aliquot (typically between 28 2 microliters and 400 microliters), which can be refrigerated with the micro vials until When needed. Each micro vial 2W has a piezoelectric droplet dispenser 262 and an associated quality assurance wafer (i.e., integrated circuit) 266 having flash memory and electrical contacts 2 64 for power and data transfer. The droplet dispenser 26 2 has a piezoelectric actuator 26 1 configured to emit droplets having a volume between 50 picoliters and 150 microliters for a reasonably fast reagent loading while maintaining correct Droplet position. Probe and Reagent Identification System The quality assurance wafer 266 (see Figure 64) has a digital memory for storing, identifying, and tracking specification data that characterizes reagents or oligonucleosides in the microbottle 258. Acid probe solution. At the end of the spotting and loading process, the data from each of the micro vials 2·5 8 and other loading and spotting data are downloaded and stored to the control microprocessor 263 which controls the reagent dispensing robot or probe dispensing robot. The program of the LOC device 30 and the resource - 254 - 201209402 flash memory 40. This information is used for diagnostic information and processing tasks, quality control and auditing. Referring to Fig. 73, ONEC 272 also has a digital memory such as flash memory 281 in ONEC CMOS structure 285 to store oligonucleotide specification data such as probe identification data, lot number, and the like. As with the LOC device, the ONEC refill robot 274 downloads the specification material from the quality assurance wafer 266 on the micro vial 258 to the ONEC flash memory.

Automated information transfer minimizes the possibility of errors that can be identified by the test module reader 12 or other system components when processing the diagnostic information if the wrong micro vial is used. Reagent Dispensing Robot The simplified top view and side view of the reagent dispensing robot 256 are shown in Figures 63 and 108. It includes:

• Micro vials 25 8 containing reagents and molecular bio-grade water (only partial micro-bottles are displayed) • Mechanical/electrical racks 286 (only outlines shown), which hold the micro vials 258 and provide electrical connection to the micro vials 258 • XY platform 268, which provides a surface for detachably mounting a portion of the deep cut wafer 260 or other fixed array such as a detachable PCB wafer 720. • A registration camera 270 that provides feedback to the control microprocessor 263 for positioning The exact position of the piezoelectric droplet dispenser 262 - 255 - 201209402 The piezoelectric droplet dispenser 262 on the micro vial 258 is used to directly dispense the reagent and water to the LOC device reservoir 54, 56, respectively. , 58, 60 and 62 and the humidifier reservoir 188. ONEC Refill Robot The ONEC Refill Robot 274 is shown in Figure 65. It is similar to the reagent dispensing robot 256 and includes:

• 1080 micro-bottles 258 containing solutions of oligonucleotide probes (for illustration only, not all micro-bottles) • Mechanical/electrical rack 286 (profile only) - it holds the micro-bottle 258 and provides electrical connection To micro vial 258 • Oligonucleotide emitter wafer (〇NEC) 272 - a total of 1 080 ONEC reservoirs 27 8 supply respective emitters 2 8 7 , each of which has four ONEC thermal droplets Generator 271 (see Figures 71 and 72)

• XY stage 268: immobilizing the oligonucleotide emitter wafer (ONEC) 2 72 • registration camera 270 that provides feedback to the control microprocessor 263 to locate the exact location of the thermal droplet generator 2 7 1 The ONEC 2 72 is moved under the mechanical/electrical frame 286. A unique probe solution is dispensed from each vial 258 to each of the ONEC reservoirs 278. The ONEC 2 72 is then used in the probe spotting robot 273 to spot the LOC device hybridization chamber 180 with a single drop of probe solution.

ONEC

Figure 71 '72 and 73 show the details of the ONEC 2 72. The ONEC -256 - 201209402

2 72 is an oligonucleotide spotting device for contactless spot-like probes onto a surface, such as an array of hybridization chambers in any of the L〇c devices. Its overall size is 23,296 micrometers xi, 760 micrometers, and is manufactured using sophisticated, high-quality photo-etching manufacturing techniques. Each ONEC has 1,080 reservoirs 278 etched on the reservoir side 2 77 of the monolithic substrate 2 75 (see Figure 73). With more than one reservoir 278, each ONEC has a complete series of probes required to spot the LOC device described herein. This allows the LOC spotting process to be a single step because there is no need to use more than one ONEC to spot the LOC for each particular analysis. The ONEC receptacle 278 has a rectangular base (96 microns χ 208 microns) and a depth of 200 microns. Each ONEC receptacle 27 8 feeds the probe suspension to a respective injector 28 7 . The liquid suspension of the probe fills the common chamber 282 via a pair of chamber inlets 284 (see Figure 72). The chamber inlet 2 84 is a two-micron diameter hole from the reservoir 27 8 to the common chamber 282. One of the four thermal droplet generators 271 generates a vapor bubble by heating the actuator 28 to eject probe droplets to the hybrid chamber 180 via a nozzle 283 at the emitter side 279. Four hot drop generators 271 allow for backup to prevent the drop generator from malfunctioning. LOC Probe Staking Robot The LOC Probe Staking Robot 289 is shown in Figures 66 and 92. For the sake of clarity, components that are not on the PCB wafer 720 that are not the LOC device 30 are not shown. It includes the following: • ONEC 2 72 - an oligonucleotide emitter wafer with 1080 reservoirs 278, each reservoir containing a probe solution (see Figures 71 and 72) -257- 201209402 • XY Stage 268: Fixed A partially deep cut LOC wafer 260 (see FIG. 66) or the detachable PCB wafer 720 (see FIG. 92) • a registration camera 270 that provides feedback to the control processor 263 to locate the ONEC thermal droplet The exact location of the generator 271 The LOC wafer 260 or the detachable PCB wafer 720 is detachably mounted on a platform that is movable along two orthogonal axes. The ONEC 272 is removably secured by a clamp 265 that is in close proximity to the platform 'the injector 287 faces the platform (see Figure 66). The LOC wafer 260 or the detachable PCB wafer 720 is moved by the control processor 26 3 to the ONEC 272. Each LOC device hybridization chamber 180 is spotted by the injector under the operational control of the control processor 263. Using a volume of less than 1 皮 picoliter reduces the reaction time and increases the density of the array of hybrid chambers. Spotted low volume probe droplets have not been previously used, as it is difficult to accurately and reliably eject very small droplets. Drops that are misdirected cannot be spotted into the correct room and may contaminate adjacent chambers. TheONEC 2 72 can be driven to produce a series of droplet volumes. For proper dispensing, the droplets produced by the ONEC 272 will be less than 100 picolitres. To improve the correctness of the dispensed probe and reagent (in terms of volume and position on the LOC device), droplets produced by the ONEC can be reduced to less than 25 picoliters, and preferably less than 6 pics. The ONEC 272 dispenses the probe solution in droplet form to the 1080 hybridization chambers 180, the volume of which varies from 〇_1 skin to 1.6 picoliters and is highly positionally correct. The hybrid chamber array 11 〇 system is organized into 24 rows, each row has 45 -258- 201209402

Adjacent rooms (see Figure 52). The sample flow path 176 passes between every two rows such that the overall array has a substantially square shape for the LED 26 to illuminate approximately. Since the hybrid array 1 1 is limited to an area of less than 1 500 μm by 1500 μm, the correctness of the ONEC 272 must be very high. The registration camera 270 is used by the control processor 263 to determine the exact position of the ONEC thermal droplet generator 271, and the drive pulse of the droplet generator is via the ONEC pad 276 to the XY Platform 268 is synchronized. The LOC probe spotting robot 273 using the ONEC 272 and camera 270 can easily spot the probe onto the surface at a rate of more than 100 probes per second (such as the hybrid chamber array 11); in most cases The rate is higher than 1,400 probes per second. Typically, the array of droplet generators spot the probe onto the surface at a rate of more than 20,000 probes per second, and in many cases, the array of droplet generators is at 30,000,000 per second. Spotting the probe to a rate of 1, 〇〇〇, 探针 probes per second. The probe onto the surface. The droplet generator array constructed on the ruthenium substrate allows the ONEC 272 to be much higher. The spotting density of the existing probe spotter spots the oligonucleotide to the surface. ONEC 272 easily spotted at a density of more than one probe per square millimeter. In the vast majority of cases, the spot density is more than 8 probes per square millimeter. In most cases, the spot density is more than 60 probes per square millimeter, and typically the density is between 50,000 probes per square millimeter and 1,500 probes per square millimeter. The LOC probe spotting using the ONEC 272 as a biochemical deposition device - 259 - 201209402 The robot 273 can easily deposit biochemical substances onto the surface at a rate higher than loo droplets per second 'in most cases, the rate is higher than 1, 4 drops per second. Typically, the array of droplet generators spot the droplet onto the surface at a rate of more than 20 per second per droplet, and in many cases, the array of droplet generators is interposed The droplets are spotted onto the surface at a rate of 300,000 droplets per second to 1,000,000 droplets per second. The LOC probe spotting robot 273 using the ONEC 272 as a biochemical deposition device can easily deposit biochemical substances onto the surface at a density of more than one droplet per square millimeter. In the vast majority of cases, the spot density is more than 8 droplets per square millimeter. In most cases, the spot density is more than 60 droplets per square millimeter, and typically the density is between 500 droplets per square millimeter to 1,500 droplets per square millimeter. Conclusion The devices, systems, and methods described herein facilitate rapid, inexpensive, and suitable molecular diagnostic testing for local care. The above-described system and its components are merely illustrative, and many variations and modifications will be readily apparent to those skilled in the art without departing from the scope of the invention. BRIEF DESCRIPTION OF THE DRAWINGS The preferred embodiments of the present invention will now be described by way of example only with reference to the accompanying drawings in which: FIG. 1 shows a test module and test module configured for fluorescence detection. Reader; -260- 201209402 Figure 2 is a schematic diagram of the electronic components assembled in the test module for fluorescence detection; Figure 3 is a diagram of the electronic components in the test module reader槪Figure 4 is a schematic view of the structure of the LOC device; Figure 5 is a perspective view of the LOC device; Figure 6 is a plan view of the LOC device, with features and structures of all layers overlapping each other;

Figure 7 is a plan view of the LOC device, showing the structure of the upper cover separately. Figure 8 is a top view of the upper cover, showing the internal passage and the reservoir in broken lines; Figure 9 is an exploded top view of the upper cover, showing the internal passage and the reservoir in broken lines; 10 is the lower cover view, which shows the configuration of the upper channel:

Figure 1 is a plan view of the LOC device, which independently shows the structure of the CMOS + MST device;

Figure 12 is a schematic cross-sectional view of the sample inlet of the LOC device; Figure 13 is an enlarged view of the AA region shown in Figure 6; Figure 14 is an enlarged view of the AB region shown in Figure 6; Figure I5 is the AE region shown in Figure U Figure 16 is a partial perspective view showing the layered junction of the LOC device in the AE area. Figure 17 is a perspective view showing the layered junction of the LOC device in the AE area. Figure 18 is a perspective view showing the LOC in the AE area. The layered structure of the device is -261 - 201209402; the partial perspective view of Figure 19 illustrates the layered structure of the LOC device in the AE area; the partial perspective view of Figure 20 illustrates the layered setback of the LOC device in the AE area. Figure 21 is a partial perspective view showing the layered structure of the LOC device in the AE area; Figure 22 is a schematic cross-sectional view of the lysis reagent reservoir shown in Figure 21; and a partial perspective view of Figure 23 illustrating the LOC device in the AB area Layer structure; Fig. 24 is a partial perspective view showing the layered structure of the LOC device in the AB zone; Fig. 25 is a partial perspective view showing the layered structure of the LOC device in the AI zone; The layered structure of the LOC device; a partial perspective view of Figure 27 illustrates the layered frustration of the LOC device in the AB zone, the portion of Figure 28 The view illustrates the layered structure of the LOC device in the AB zone; the partial perspective view of FIG. 29 illustrates the layered nomenclature of the LOC device in the AB zone, and FIG. 30 is a schematic cross-sectional view of the amplification mixing reservoir and the polymerase reservoir; 31 shows the characteristics of the boiling start valve independently; -262- 201209402 Figure 32 is a schematic cross-sectional view of the boiling start valve taken along line 33-33 shown in Figure 31; Figure 33 is the AF area shown in Figure 15. Figure 34 is a schematic cross-sectional view of the upstream end of the dialysis section taken along line 3 5 - 3 5 shown in Figure 33; Figure 35 is an enlarged view of the AC zone shown in Figure 6; A re-enlarged view of the AC area, which shows an amplification section;

Figure 37 is a re-enlarged view of the AC region, which shows an amplification portion; Figure 38 is a re-enlarged view of the AC region, which shows an amplification portion; Figure 39 is a re-enlarged view of the AK region shown in Figure 38; FIG. 41 is a re-enlarged view of the AC region, and FIG. 42 is a re-enlarged view of the AC region, which shows an amplification chamber; Figure 43 is a re-enlarged view of the AL region shown in Figure 42; Figure 44 is a re-enlarged view of the AC region, showing the amplification portion; Figure 45 is a re-enlargement view of the AM region shown in Figure 44; A re-enlarged view of the AC region, which shows an amplification chamber: Figure 47 is a re-enlarged view of the AN region shown in Figure 46; Figure 48 is a re-enlarged view of the AC region, which shows an amplification chamber; Figure 49 is an AC A re-enlarged view of the area, the figure shows an amplification chamber; FIG. 50 is a re-enlarged view of the AC area, which shows an amplification section; FIG. 51 is a schematic cross-sectional view of the amplification section; FIG. 52 is an enlarged plan view of the hybridization section; A re-enlarged plan view of two independent hybridization chambers of 53; -263- 201209402 Figure 54 is a schematic cross-sectional view of a single hybridization chamber; Figure 55 is an enlarged view of the humidifier in the AG region shown in Figure 6; Figure 52 is an enlarged perspective view of the LOC device of the AD region; Figure 58 is a schematic view of the FRET probe in a closed configuration; Figure 59 is an open and hybridized configuration. Figure of the FRET probe; Figure 60 is a time-intensity diagram of the excitation light;

Figure 61 is a diagram of the excitation illumination geometry of the hybrid chamber array; Figure 62 is a diagram of the sensor electronics technology LED illumination geometry; Figure 63 is a schematic diagram of the reagent dispensing robot: Figure 64 is a reagent with a built-in droplet generator Perspective view of the micro vial Figure 6 5 is a schematic plan view of an oligonucleotide injector robot for loading a selected probe to a probe injector wafer;

Figure 66 is a plan view of a probe spotting robot used in a LOC device for loading a probe onto a partially deep cut germanium wafer; Figure 67 is an enlarged plan view of the humidity sensor shown in the AH region of Figure 6; Figure 68 is a schematic view showing a portion of a photodiode array of a photosensor; Figure 69 is a circuit diagram of a single photodiode; Figure 70 is a timing diagram of a photodiode control signal; and Figure 71 is an illustration of an oligonucleotide emitter wafer. (ONEC); Figure 72 shows an array of droplet generators for the ONEC shown in the AO region of Figure 71: -264 - 201209402 Figure 73 is a schematic cross-sectional view of the array of droplet generators, the section along Figure 72 Figure 74 is an enlarged view of the evaporator shown in the AP area of Figure 55; Figure 75 is a schematic cross-sectional view of the hybridization chamber having the detecting photodiode and the triggering photodiode; Figure 76 shows the PCR initiated by the linker; Figure 77 is a representative view of the test module with the lancet;

Figure 78 is a representation of the structure of LOC variant VII; Figure 79 is a representative representation of the structure of LOC variant VIII; Figure 80 is a representative representation of the structure of LOC variant XIV; Figure 81 is representative of the structure of LOC variant XLI Figure 82 is a representative view of the structure of the LOC variant XLIII; Figure 83 is a representative view of the structure of the LOC variant XLIV; Figure 84 is a representative representation of the structure of the LOC variant XLVII; Figure 8 is a first round of amplification Figure of linear fluorescent probe connected to the primer during the period; Figure 86 is a diagram of the linear fluorescent probe connected to the primer during the subsequent amplification cycle; Figure 87A to 87F illustrate the fluorescent stem probe connected to the primer Figure 88 is a schematic diagram of an excitation LED relative to a hybridization chamber array and a photodiode; Figure 89 is a schematic diagram of an excitation LED and an optical lens for directing illumination to an array of hybridization chambers of a LOC device; -265- 201209402 Figure 90 is a schematic illustration of an excitation LED, an optical lens, and an optical raft for directing illumination to an array of hybrid chambers of a LOC device; Figure 91 is an excitation LED, optical lens, and mirror arrangement for an array of hybridization chambers for directing illumination to the LOC device Schematic diagram; Figure 92 is for loading A schematic plan view of the probe spotting robot in the LOC device on the detachable PCB; Fig. 93 is a plan view showing that all the features overlap each other, and the position of the DA to the DK area is not indicated; Fig. 94 is Fig. 93 An enlarged view of the DG region shown; Fig. 95 is an enlarged view of the DH region shown in Fig. 93; Fig. 96 shows an embodiment of the shunt transistor of the photodiode t. Fig. 97 shows the shunt of the photodiode One embodiment of the crystal» Figure 98 shows an embodiment of a shunt transistor for a photodiode. Figure 99 is a circuit diagram of a differential imager. Figure 100 is a schematic illustration of the stem-loop configuration of a negative control fluorescent probe: 1 〇1 schematically illustrates the open configuration of the negative control fluorescent probe of Fig. 1; Fig. 1 02 schematically illustrates the stem loop configuration of the positive control fluorescent probe; Fig. 1 03 schematically illustrates the positive control fluorescing of Fig. 102 The open configuration of the optical probe; Figure 1 04 shows the test module configured for ECL detection and the test -266-201209402 module reader; Figure 1 05 is configured for ECL detection The schematic diagram of the electronic components in the test module; Figure 1 06 shows the test Test groups and optionally reader module; FIG. 107 shows the host system test module and the test module reader and stores various materials library resources; schematic side view 108 of the robot-based spotting agents;

Figure 109 is a representation of an electrochemiluminescent substrate test module with a multi-device microfluidic device. [Main component symbol description] 1 〇: Test module 1 1 : Test module 1 2 '· Test module reader 13 : Shell 14 : Micro USB connector 15 : Inductor 16: Micro USB 埠 1 7 : Touch screen 18 : Display screen 19 : Button 20 : Start button - 21 : Honeycomb radio 22 : Aseptic sealing tape -267- 201209402 2 3 : Wireless network connection 24 : Large container 2 5 : Satellite navigation system 26 : Light-emitting diode 2 7: Data Storage 29: USB Compatibility LED Driver 30: Laboratory Chip (LOC) Device

3 1 : Power conditioner 32 : Capacitor 33 : Clock 34 : Controller 35 : Register 36 : USB device driver 37 : Driver

38 : RAM

39 : ECL excitation driver 40 : program and data flash memory 41 : register 42 : processor 43 : program memory 44 : light sensor 45 : indicator 46 : upper cover 47 : USB power / indicator module only -268- 201209402 48: COMS + MST Wafer 49: Porous Element 51: Upper Cover 52: Hybridization and Detection Section 54: Reservoir 56: Reservoir

57: Printed Circuit Board (PCB) 5 8 : Reservoir 60: Reservoir 62: Reservoir 64: Lower Sealing Layer 66: Top Layer 68: Sample Inlet 70: Pathogen Dialysis Section 72: Waste Channel 74: Target Channel 76: Waste unit (reservoir) 78: reservoir layer 80: upper cover channel layer 8 2: upper sealing layer 84: 矽 substrate 86: CMOS circuit 87: MST layer 8 8 : passivation layer - 269 - 201209402 90 : MST channel 92: descent Port 94: Upper cover channel 96: Ascending port 97: Wall 98: Meniscus anchor 1 00 : MST channel layer

101: laptop/notebook 102: capillary start feature (CIF) 103: dedicated reader 105: desktop computer 106: boiling start valve 107: e-book reader 108: boiling start valve 109: tablet

1 1 〇: hybridization chamber array 1 1 1 : epidemiological data host system 1 1 2 : amplification unit 1 1 4 : culture unit 1 1 5 : electronic health record (EHR) host system 1 1 6 : anticoagulation Agent 1 1 8 : Surface tension valve 1 20 : Meniscus 121 : Electronic Medical Record (EMR) Host System - 270 - 201209402 122 : Vent 123 : Personal Health Record (PHR) Host System 125 : Network

126: boiling start valve 1 28 : surface tension valve 1 3 0 : chemical lysis unit 1 3 1 : mixing portion 132: surface tension valve 1 3 3 : incubator inlet passage 1 34 : lowering port 136: light window 146 : valve Inlet 1 4 8 : Valve outlet 1 50 : Drop port 1 5 2 : Heater 153 : Valve heater contact 1 5 4 : Heater 1 5 6 : Heater contact 1 5 8 : Amplification microchannel 160 : Expansion Addition exit channel 164: aperture array 166: capillary initiation feature 168: dialysis riser aperture 170: temperature sensor - 271 - 201209402 174: liquid sensor 1 7 5 : diffusion barrier 1 7 6 : flow path 1 7 8 : Endpoint liquid sensor 1 80 : Hybridization chamber 1 8 2 : Heater 1 84 : Photodiode 18 5: Active region 186: Oligonucleotide FRET probe 187: Trigger photodiode 1 8 8 : Water storage 190: evaporator 1 9 1 : heater 192 : water supply passage 1 93 : rising port 1 94 : lowering port 1 9 5 : top metal layer 196 : humidifier 198 : first rising hole 202 : capillary starting feature 204 : Dialysis MST channel 208: Liquid sensor 2 1 0 : Culture microchannel 2 1 2 Intermediate MST channel 201209402 218 : TiAl electrode 220 : TiAl electrode 222 : Gap 232 : Humidity sensor 2 3 4 : Heat 236: FRET Probe 237: ECL Probe

23 8 : Target nucleic acid sequence 240 : Ring 242 : Stem 244 : Excitation light 246 : Fluorescent group 2 4 8 : Quencher 250 : Fluorescence emission 252 : Optical center

256: Reagent dispensing robot 25 8 : Micro vial 2 5 9 : Container 260 : Partially deep cut 矽 wafer 2 6 1 : Piezoelectric actuator 262 : Droplet dispenser 263 : Control microprocessor 264 : Electrical contact -273- 201209402 265 : Fixture 266 : Quality Assurance Wafer 26 8 : XY Stage 270 : Registration Camera 2 7 1 : Thermal Drop Generator 272 : Oligonucleotide Injector Wafer (ONEC) 273 : Probe Spotting Robot

274 : ONEC refill robot 275 : Monolithic 矽 substrate 276 : ONEC pad 2 7 7 : Reservoir side 2 7 8 : Reservoir 279 : Emitter side 2 8 0 : Actuator 2 8 1 : Flash memory body

28 2 : Common room 28 3 : Nozzle 2 8 4 : Chamber entrance 2 8 5 : ONEC CMOS structure 286: Mechanical / electrical frame 2 8 7 : Injector 2 8 8 : Sample input and preparation 2 8 9 : LOC spotting robot 290: Extraction - 274 - 201209402 2 9 1 : Culture 292 : Amplification 294 : Detection 296 : First electrode 298 : Second electrode 300 : Preprogrammed delay 301 : Laboratory wafer (LOC) device

3 76 : Conductive column 3 78 : Positive control probe 3 80 : Negative control probe 3 82 : Calibration chamber 3 8 4 : Gate 3 8 6 : Idle 3 8 8 : Gate 3 9 0 : Lancet 3 92: lancet release button 3 9 3 : gate 3 94 : Mshunt transistor 3 96 : MOS transistor 398: MOS transistor 400: MOS transistor 402: MOS transistor 404 : MOS transistor 4 0 6 : node -275- 201209402 408: Membrane seal 4 1 0 : Membrane guard 6 8 2 : Small component dialysis section 6 8 6 : Dialysis section 692 : Linear probe 694 connected to the primer: Amplification blocker 696 : Probe sequence

6 9 8 : Sequence 700 : Oligonucleotide primer 704 : Stem loop probe linked to primer 706 : Complementary sequence 708 : Stem strand 710 : Another strand

7 1 2 : first optical 稜鏡 7 1 4 : second optical 稜鏡 7 1 6 : first mirror 7 1 8 : second mirror 720 : PCB wafer 728 : LOC variant X 766 : waste receptacle 778 : Configuration 780: Configuration 7 82: Configuration 783: Microfluidic Device - 276 - 201209402 784: Dialysis Device 78 5 : LOC Device 78 8 : Differential Imager Circuit 7 9 0 : Pixel 7 9 2 : Virtual Pixel 794: Read Take _column 795: read_column_d

796: Negative control probe 797 : M4 798 : Positive control probe 801 : MD4 8 0 3 : Pixel capacitor 8 05 : Virtual pixel capacitor 8 0 7 : Switch 809 : Switch 811 : "Read - line" switch 8 13 : Virtual "read_row" open 8 1 5: switched capacitor amplifier 817: differential signal 8 60 : ECL excitation electrode 8 70 : ECL excitation electrode

Claims (1)

201209402 VII. Patent application scope: 1. A device for loading an oligonucleotide spotting device and a spotting oligonucleotide probe, the device comprising: a plurality of oligonucleotide bottles, each of which has a liquid a drip dispenser; a mounting surface for detachably mounting an oligonucleotide spotting device; a jig for detachably mounting the oligonucleotide spotting device adjacent to the mounting surface; and a control processor The control processor is configured to operate an oligonucleotide spotting device for controlling the oligonucleotide bottles, mounted on the jig, and movement of the mounting surface relative to the oligonucleotide bottles, and the oligonucleoside An acid spotting device; wherein the control processor is configured to activate the droplet dispenser and move the oligonucleotide spotting device to register the oligonucleotide vials. 2. The device of claim 1, wherein the control processor is configured to operate an oligonucleotide spotting device on the fixture to spot the oligonucleotide probe onto the mounting surface Microfluidic device. 3. The device of claim 1, wherein the oligonucleotide vials each have an integrated circuit for storing oligonucleotide specification data, and the control processor is configured to download the oligonucleotide Specification data to the digital memory in the oligonucleotide spotting device. 4. The device of claim 1, further comprising a camera optically feeding back the registration between the bottle selected by the control processor and the oligonucleotide spotting device. -278- 201209402 5. The device of claim 3, wherein the oligonucleotide bottle is in a volume of from 282 microliters to 400 microliters. 6. The device of claim 5 The integrated circuit of each of the micro vials has a unique identifier for individually identifying each of the micro vials, the unique identifier being transmitted to the control processor. 7. The apparatus of claim 6 wherein each of the micro vials has an electrical contact for receiving a start pulse of the drop dispenser and allowing the control processor to interrogate the integrated circuit. 8. The apparatus of claim 6 further comprising a shelf, wherein the micro vials are detachably mounted to the shelf for mechanically controlling and electronically controlling the micro vials. 9. The apparatus of claim 8 wherein the mounting surface is a platform that is configured to move along two orthogonal axes extending parallel to one of the orthogonal axes. 10. The device of claim 1, wherein the droplet dispenser has a piezoelectric actuator. The apparatus of claim 1, wherein the droplet dispenser is configured to emit droplets having a volume of between 50 picoliters and 150 microliters. 1 2 . The device of claim 2, further comprising a reagent bottle containing a reagent for processing a biological sample, wherein the microfluidic device is a LOC device for genetic analysis of a biological sample, the LOC device having The polymerase chain reaction (PCR) section, the reagent list has one or more of the following water; -279- 201209402 polymerase, primer; buffer; anticoagulant; deoxyribonucleoside triphosphate (dNTP); Ligase and linker; and Restriction enzymes. 13. The apparatus of claim 12, further comprising means for applying a film to the LOC device to cover a reagent reservoir formed on the outer surface. 14. The device of claim 12, wherein the LOC device is one of an array of LOC devices built on a germanium wafer, the platform being configured to removably mount the germanium wafer to load reagents to Within all LOC devices in the array. 15. The device of claim 12, wherein the LOC device is one of an array of LOC devices mounted on a printed circuit board (PCB) that is configured to detachably mount the PCB to load reagents To all LOC devices in the array. The device of claim 2, wherein the oligonucleotide spotting device has a reservoir array and an emitter array for comprising an oligonucleotide probe, and the LOC device has an acceptor A hybridization chamber array of nucleotide probes having a nucleic acid sequence complementary to a target nucleic acid sequence to be recognized in a biological sample, the array of hybridization chambers being organized to accommodate a raw-280-201209402 sample preparation A complete oligonucleotide probe sample is required for analysis, the reservoir array being configured to contain the complete oligonucleotide probe sample required for the predetermined analysis to be performed by the LOC device, the control treatment The apparatus is configured to operate the array of the hybrid chamber and to download the specification data of the oligonucleotide probes from the respective reservoirs and to locate the array position data of the hybridization chambers sampled by the respective reservoirs. Association. 17. The apparatus of claim 16, wherein each of the injectors has a plurality of nozzles, a chamber, and a plurality of actuators for containing the suspension liquid from the oligonucleotide probes of the corresponding reservoirs, One of the actuators corresponds to each nozzle, such that the actuator ejects droplets from the chamber via corresponding nozzles, and the control processor is configured to individually operate the actuators. 18. The apparatus of claim 17, wherein the control processor is configured to operate the array of emitters and spot the oligonucleotides at a density of greater than 8 probes per square millimeter. On the LOC device. 19. The apparatus of claim 17, wherein the control processor is configured to operate the array of emitters and spot the oligonucleotides at a density of greater than 60 probes per square millimeter at the LOC. On the device. The apparatus of claim 17, wherein the control processor is configured to operate the array of emitters and to spot the oligonucleotides at 500 probes per square millimeter to 1500 per square millimeter. The density of the probe spots was spotted on the LOC device. -281 -