KR102138344B1 - Cartridge and analyzer for fluid analysis - Google Patents

Abstract

A fluid cartridge and method of operation are described. The fluid cartridge includes a plurality of contact pads designed to electrically couple with the analyzer, a semiconductor chip with an array of sensors, and a substrate with a reference electrode. The fluid cartridge includes a first fluid channel having a first inlet and coupled to a second fluid channel, the second fluid channel being aligned such that the sensor array and the reference electrode are disposed within the second fluid channel. The first plug is disposed at the first inlet. The first plug comprises a compliant material that is configured to be perforated by capillaries without leaking fluid through the first plug.

Description

Cartridge and analyzer for fluid analysis {CARTRIDGE AND ANALYZER FOR FLUID ANALYSIS}

[Priority claim]

This application claims the benefit of U.S. Provisional Application No. 62/372,596, the entire contents of which are incorporated herein by reference, filed August 9, 2016 and entitled "CARTRIDGE AND ANALYZER FOR FLUID ANALYSIS".

A biosensor is a device for detecting and detecting biomolecules, and operates based on electronic, electrochemical, optical, and mechanical detection principles. A biosensor comprising a transistor is a sensor that electrically senses the mechanical properties of charges, protons and bio-entities or biomolecules. Detection may be performed by detecting the bio-entity or bio-molecule itself, or may be performed through interaction or reaction between a specific reactant and the bio-entity/bio-molecule. These biosensors can be manufactured using semiconductor processes, can quickly convert electrical signals, and can be easily applied to integrated circuits (ICs) and MEMS.

The interaction of the biological sample itself with the biosensor can be a challenge. Typically, the fluid containing the biological sample is metered with a pipette directly above the sensing portion of the biosensor. This method results in the majority of the fluid sample being unused, and it takes time to manually load each sensing area.

Aspects of the present disclosure are best understood when read in conjunction with the accompanying drawings from specific details for carrying out the invention that follows. It is noted that, in accordance with standard practice in the industry, various features are not drawn to scale. In fact, the dimensions of the various features can be arbitrarily increased or decreased for clarity of discussion.
1 is a diagram illustrating components of an exemplary bio-sensing cartridge.
2 is a cross-sectional view of an exemplary dual-gate back-side sensing FET sensor.
3 is a circuit diagram of a plurality of FET sensors constructed in an exemplary addressable array.
4 is a circuit diagram of an addressable dual gate FET sensor array and a heater.
5 is a cross-sectional view of an exemplary dual gate back sensing FET sensor configured as a pH sensor.
6A shows an example of the binding of ions to a receptor layer.
6B shows the change in threshold voltage in an exemplary FET sensor based on pH.
7 is a top view of an exemplary biosensor chip.
8 shows a series of cross-sectional views illustrating a manufacturing process for mounting an exemplary biosensor chip to a handle layer.
9 is a top view of a handle layer with an exemplary biosensor chip mounted on a substrate.
10 is a schematic diagram of an exemplary fluid cartridge with an integrated biosensor chip.
11 is a schematic diagram of a portion of a fluid channel in an exemplary fluid cartridge.
12 is a schematic diagram of an exemplary fluid cartridge coupled to an analyzer.
13 is a flow chart of an exemplary method using a fluid cartridge.
14 is a cross-sectional view of an exemplary dual gate back sensing bio FET (bioFET) for detecting DNA.
15A illustrates the mechanism of binding of DNA on the receptor surface.
15B shows the change in threshold voltage for an exemplary dual gate back sensing bio FET based on matched analyte binding.
16 is a cross-sectional view of an exemplary dual gate back sensing bio FET with antibodies immobilized on its sensing layer.
17 illustrates the binding mechanism of antigens and antibodies on the receptor surface.

The following disclosure provides many different embodiments or examples for implementing different features of the content provided. Specific examples of components and devices are described below to simplify the present disclosure. Of course, these are merely examples and are not intended to be limiting. For example, in the following description, the formation of the first feature above or on the second feature may include an embodiment in which the first and second features are in direct contact, and further features may include first and second features. Between features, the first and second features may include embodiments in which they may not be in direct contact. In addition, the present disclosure may repeat reference numerals and/or symbols in various examples. This does not refer to the relationship between the various embodiments and/or configurations discussed on their own.

Also, terms related to spaces such as “below”, “below”, “bottom”, “above”, “top”, and the like, may include other element(s) or feature(s) of one element or feature as shown in the figure. To explain the relationship to, can be used here for ease of explanation. Space-related terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. The device can be oriented differently (rotated at 90 degrees or other orientations), and the spatially relative descriptors used herein can be interpreted similarly accordingly.

Terms

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or identical to those described herein can be used in practice or in testing embodiments according to the present disclosure, the methods and devices and materials are now described. All patents and publications mentioned herein are incorporated herein by reference for the purpose of describing and disclosing materials and methods reported in publications that may be used in connection with the present invention.

As used herein, the two acronym "FET" refers to a field effect transistor. A very common type of FET is called a metal oxide semiconductor field effect transistor (MOSFET). Historically, MOSFETs have been flat structures constructed thereon on flat surfaces of substrates such as semiconductor wafers. However, recent advances in semiconductor manufacturing have provided tertiary fin-based MOSFET structures.

The term "bioFET" refers to an FET comprising a layer of immobilized capture reagent that acts as a surface receptor to detect the presence of a target analyte of biological origin. The bio FET is a field effect sensor having a semiconductor transducer, according to one embodiment. One advantage of biosensors is the possibility of label-free operation. Specifically, bio FETs enable avoidance of costly and time consuming labeling operations, such as labeling of analytes using, for example, fluorescent or radioactive probes. One particular type of bio-FET described herein is a dual-gate back-side sensing bioFET. The analyte for detection by a bio FET can usually be a biological source such as, but not limited to, proteins, carbohydrates, fats, tissue fragments, or parts thereof. However, in a more general sense, the bio FET is capable of detecting any compound (known as ChemFET in the art) or any other element including ions such as protons or metal ions (ISFET in the art) Known as) is part of a wider class of FET sensors. The present invention is intended to be applied to all types of FET based sensors (“FET sensors”). One specific type of FET sensor here is a Dual-Gate Back Side Sensing FET Sensor ("DG BSS FET Sensor").

"S/D" refers to the source/drain junction forming two of the four terminals of the FET.

The expression "high-k" refers to a high dielectric constant. In the field of semiconductor device structures and manufacturing processes, hi-k refers to a dielectric constant that is greater than the dielectric constant of SiO ₂ (ie, greater than 3.9).

The term "analysis" generally includes, but is not limited to, physical, chemical, biochemical or biological analysis including characterization, testing, measurement, optimization, separation, synthesis, addition, filtration, dissolution or mixing. Refers to a process or step that includes.

The term "analytic assay (assay)" generally refers to a process or step involving the analysis of a chemical or target analyte, cell-based assay assay, biochemical assay assessment, high-throughput assay assessment and screening, diagnostic assay assessment , pH determination, nucleic acid hybridization assay evaluation, polymerase activity assay evaluation, nucleic acid and protein sequencing, immunoassays (e.g. antibody-antigen binding assay evaluation, ELISA and iqPCR), gene Assessment of bisulfite methylation reaction assay to detect methylation patterns, protein assay assessment, protein binding assay assessment (e.g., protein-protein, protein-nucleic acid and protein-ligand binding assay assessment), enzyme assay assessment, Evaluation of binding enzyme assays, kinetic measurements (e.g., kinetics of protein folding and enzymatic reaction kinetics), screening of enzyme inhibitors and activators, chemiluminescence and electrochemiluminescence assay evaluation, fluorescence (fluorescent) analysis evaluation, fluorescence polarization and anisotropy analysis evaluation, absorbance and colorimetric analysis evaluation (e.g., Bradford analysis evaluation, Lowry analysis evaluation, Hartley -Harree-Lowry analysis evaluation, Burette analysis evaluation and BCA analysis evaluation], chemical analysis evaluation (eg, for detection of environmental pollutants and contaminants, nanoparticles or polymers) and drug discovery analysis evaluation It includes, but is not limited to. The devices, systems and methods described herein may employ or employ one or more of these analytical evaluations to be used with any FET sensor having the described design.

The term "liquid biopsy" generally refers to a biopsy sample obtained from a subject's bodily fluid compared to a subject's tissue sample. The ability to perform analytical evaluations using body fluid samples is often more desirable than using tissue samples. A less invasive approach using body fluid samples has a broad range in terms of the patient's welfare, the ability to perform longitudinal disease monitoring, and the ability to obtain expression profiles, for example, in the prostate, when tissue cells are not readily accessible. Have an effect. Analytical evaluations used to detect target analytes in liquid biopsy samples include, but are not limited to, those described above. As a non-limiting example, circulating tumor cell (CTC) assay evaluation can be performed on a liquid biopsy sample.

For example, a capture agent immobilized on a FET sensor (eg, an antibody) can be used for detection of a target analyte (eg, tumor cell marker) in a liquid biopsy sample using a CTC assay. CTCs are cells that flow from a tumor into a vasculature and circulate, for example, in the bloodstream. Generally, CTC is present in the circulation at extremely low concentrations. To analyze and evaluate CTCs, CTCs are concentrated from patient blood or plasma by various techniques known in the art. CTCs are stained for specific markers using methods known in the art, including, but not limited to, cytometry (e.g., flow cytometry) based methods and IHC based methods. Can be. For the devices, systems and methods described herein, CTCs can be captured or detected using a capture agent, or a nucleic acid, protein or other cellular environment from the CTC is a target for binding to or detection by a capture agent. It can be targeted as an analyte.

When a target analyte is detected on or from a CTC, an increase in the target analyte, e.g., expressing or including CTC, is likely to respond to a particular treatment (e.g., associated with the target analyte). It may help to identify a subject as having cancer, or it may enable optimization of an optimal treatment plan using, for example, antibodies against a target analyte. CTC measurement and quantification can, for example, provide information about the stage of the tumor, response to treatment, disease progression, or a combination thereof. Information obtained from detecting a target analyte on a CTC can be used, for example, as a prognostic, predictive or pharmacodynamic biomarker. In addition, CTC assay evaluations for liquid biopsy samples can be used alone or in combination with additional tumor marker analysis of solid biopsy samples.

The term "identification" generally refers to the process of determining the identity of a target analyte based on its binding to a known capture agent.

The term "measurement" generally refers to the process of determining the total amount, quantity, quality or properties of a target analyte based on binding to a capture agent.

The term "quantitation" generally refers to the process of determining the amount or concentration of a target analyte based on binding to a capture agent.

The term "detection" generally refers to the process of determining the presence or absence of a target analyte based on binding to a capture agent. Detection includes, but is not limited to, identification, measurement and quantitation.

The term "chemical" is a substance, compound, mixture, solution, emulsion, dispersion, molecule, ion, dimer, polymer or polymer such as a protein, biomolecule, Precipitates, crystals, chemical moieties or groups, particles, nanoparticles, reagents, reaction products, solvents, or any solid, liquid or gaseous state and typically subject to analysis Refers to the fluid.

The term "reaction" relates to at least one chemical and generally refers to the cleavage or formation of one or more bonds such as covalent bonds, non-covalent bonds, van der Waals bonds, hydrogen bonds or ionic bonds (chemical, In the case of biochemical and biological transformation) means physical, chemical, biochemical or biological transformation. These terms are common and non-covalent, with common chemical reactions such as synthetic reactions, neutralization reactions, decomposition reactions, substitution reactions, reduction-oxidation reactions, precipitation, crystallization, combustion reactions, and polymerization reactions. Bonding, phase change, color change, phase formation, crystallization, dissolution, luminescence, change in light absorption or luminescence properties, temperature change or heat absorption or exotherm, conformal change and folding or unloading of polymers such as proteins Includes folding.

A “capture reagent” as used herein is a molecule or compound capable of binding a target analyte or target reagent that can be attached directly or indirectly to a substantially solid material. The capture agent can be a chemical, specifically any naturally occurring target analyte (e.g., antibody, polypeptide, DNA, RNA, cell, virus, etc.) or any target analyte can be provided. , And the capture agent can bind to one or more target analytes in an analytical evaluation.

“Target analyte” as used herein is a substance to be detected in a test sample using the present invention. The target analyte may be a chemical, specifically, any naturally occurring target analyte (eg, antibody, polypeptide, DNA, RNA, cell, virus, etc.) or any capture agent may be provided. And the target analyte can bind to one or more capture agents in the assay assay. In addition, “target analyte” can include any antigenic material, antibody, and combinations thereof. Target analytes include proteins, peptides, amino acids, carbohydrates, hormones, steroids, vitamins, antibodies to drugs, bacteria, viruses and any of the aforementioned substances, including those registered for therapeutic purposes as well as those for illegal purposes. It may contain a metabolite of (metabolite).

As used herein, “test sample” means a composition, solution, substance, gas or liquid comprising a target analyte to be detected and assayed using the present invention. The test sample can include other components in addition to the target analyte, can have the physical properties of a liquid or gas, and can have any size or volume, including, for example, a moving stream of liquid or gas. The test sample can include any substance that is not the target analyte, unless another substance interferes with binding of the target analyte to a capture agent or binding of the first binding member to the second binding member. Examples of test samples include, but are not limited to, naturally occurring samples and non-naturally occurring samples, or combinations thereof. Naturally occurring test samples can be synthetic or synthetic. Naturally occurring test samples include blood, plasma, serum, urine, saliva or expectorant, spinal fluid, cerebrospinal fluid, pleural fluid, nipple aspirates, lymphatic fluid, respiratory, intestinal and Fluids of the genitourinary tract, tears, saliva, breast milk, fluid from the lymphatic system, semen, cerebrospinal fluid, intra-organ system fluid, ascitic fluid, tumor cyst fluid, Body fluids isolated from any location within or on the subject's body, including amniotic fluids and combinations thereof, and environmental samples such as groundwater or wastewater, soil extracts, air and pesticide residues, or food-related samples. do.

The substances detected can be, for example, nucleic acids (including DNA and RNA), hormones, other pathogens (viruses (eg H7N9 or HIV), protozoa (malaria caused by Plasmodium), or bacteria. Small organisms such as proteins, antibodies, hydrogen or other ions, nonionic molecules or compounds, polysaccharides, chemical binding library elements (including biological agents that cause disease or disorders against a host such as E. coli or Mycobacterium tuberculosis) Various drugs or therapeutic agents including compounds or other chemical or biological substances and the like. The detected or determined parameters may include, for example, changes in pH, lactose changes, varying concentrations, particles, such as particles per unit time the fluid flows to the device for a period of time and other parameters, for example, to detect rare particles. It can, but is not limited to.

As used herein, the term “immobilized” includes substantially attaching the capture agent at the molecular level to the surface, for example when used in connection with the capture agent. For example, the capture agent is an absorption technique that includes non-covalent interactions (e.g., electrostatic force, van der Waals and hydrophobic interface dehydration) and functional groups or linkers that facilitate the attachment of the capture agent to the surface. It can be immobilized on the surface of the substrate material using a covalent binding technique. Immobilization of the capture agent to the surface of the substrate material can be based on the properties of the substrate surface, the medium carrying the capture agent, and the properties of the capture agent. In some cases, the substrate surface can be first modified to have functional groups bound to the substrate. The functional groups can then bind to and immobilize biomolecules or biological or chemical substances thereon.

The term "nucleic acid" generally refers to a set of nucleic acids linked to each other through a phosphodiester bond, and adenine, guanine, cytosine and thymine linked to each other. Naturally present, such as DNA comprising deoxyribonucleotides having any of) and/or RNA comprising ribonucleotides having any of adenine, guanine, cytosine and uracil linked to each other And naturally occurring nucleic acids to which naturally occurring nucleotides are linked. In addition, non-naturally occurring nucleotides and non-naturally occurring nucleic acids are within the scope of the nucleic acids of the invention. Examples include peptide nucleic acid (PNA), peptide nucleic acids with phosphate group (PHONA), bridged nucleic acid/locked nucleic acid (BNA/LNA) ) And morpholino nucleic acids. As additional examples, chemically, such as methylphosphonate DNA/RNA, phosphorothioate DNA/RNA, phosphoramidate DNA/RNA and 2'-O-methyl DNA/RNA Modified nucleic acids and nucleic acid analogs. Nucleic acids include those that can be modified. For example, phosphate groups, saccharides and/or bases in nucleic acids can be labeled as needed. Any material for nucleic acid labeling known in the art can be used for labeling. Examples include radioactive isotopes (e.g., 32P, 3H and 14C), DIG, biotin, fluorescent dyes (e.g. FITC, Texas, cy3, cy5, cy7, FAM, HEX, VIC, JOE , Rox, TET, Bodipy493, NBD and TAMRA) and luminescent materials (eg, acridinium esters).

An aptamer as used herein refers to an oligo nucleic acid or peptide molecule that binds to a specific target molecule. The concept of using single-stranded nucleic acids (aptamers) as affinity molecules for semiconductor binding was first introduced in 1990 (Ellington and Szostak 1990, 1992; Tuerk and Gold 1990), and targets exist. It is based on the ability of a short sequence to fold into a unique three-dimensional structure that binds targets with high affinity and specificity. In 2006 Eugene W. M Ng et al. disclose that aptamers are oligonucleotide ligands selected for high affinity binding to molecular targets.

The term “antibody” as used herein refers to a polypeptide of the family of immunoglobulin antibodies capable of binding the corresponding antigen non-covalently, reversibly, and in a specific way. For example, naturally occurring IgG antibodies are tetramers comprising at least two heavy chains (H) and two light chains (L) interconnected by disulfide bonds. )to be. Each heavy chain consists of a heavy chain variable region (here abbreviated VH) and a heavy chain constant region. The heavy chain constant region consists of three domains (CH1, CH2, CH3). Each light chain consists of a light chain variable region (herein abbreviated VL) and a light chain constant region. The light chain constant region consists of one domain (CL). The VH and VL regions are further subdivided into hypervariability regions called complementarity determining regions (CDRs), and interspersed with more conserved regions called framework regions (FRs). Each VH and VL consists of 3 CDRs and 4 FRs arranged in the following order from amino-terminus to carboxy-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3 And FR4. The three CDRs make up approximately 15-20% of the variable domains. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant regions of an antibody bind the binding of an immunoglobulin antibody to a host tissue or factor comprising various cells of the immune system (e.g., effector cells) and the first component (Clq) of a traditional complement system. You can arbitrate. (Kuby, Immunology, 4th ed., Chapter 4. W.H. Freeman & Co., New York, 2000)

The term "antibody" refers to monoclonal antibodies, human antibodies, humanized antibodies, chimeric antibodies, and anti-idiotypic (anti-Id) antibodies. (Including, for example, anti-Id antibodies to the antibodies of the invention). Antibodies can be of any isotype/class (e.g., IgG, IgE, IgM, IgD, IgA and IgY) or lower class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) May be

The term "polymer" means any substance or compound consisting of one or more building blocks ('mers') that are repeatedly linked to each other. For example, "dimer" is a compound in which two constituents are bonded to each other. Polymers include both condensation polymers and addition polymers. Typical examples of condensation polymers include polyamide, polyester, protein, wool, silk, polyurethane, cellulose and polysiloxane. Examples of additive polymers include polyethylene, polyisobutylene, polyacrylonitrile, poly(vinyl chloride) and polystyrene. Other examples include polymers with enhanced electrical or optical properties (eg, nonlinear optical properties), such as electrically conductive or light refracting polymers. Polymers include both linear and branched polymers.

Biosensing( biosensing ) Cartridge Overview

1 illustrates an overview of various components integrated together to form an exemplary biosense cartridge 102. The biosense cartridge 102 can include a plurality of fluid channels configured to control fluid flow toward or away from a sensing location where the presence of the target analyte can be detected.

In this exemplary embodiment, the biosense cartridge 102 includes an array of FET sensors 104. The FET sensor 104 constitutes the transducer component of the biosense cartridge 102. The FET sensors 104 are arranged in an array and can be individually addressed to detect binding events at the surface of the FET sensor sensing layer. In one embodiment, FET sensor 104 includes a dual back FET sensor. In alternative embodiments, other types of FET sensor based sensors can be used.

The biosensing cartridge 102 includes a biological interface 106. Biological interface 106 can then be coupled to dual gate back sensing FET sensor 104 to facilitate binding reactions at the surface of dual gate back sensing FET sensor 104, which can then be detected. Various types of biomolecules can form part of biological interfaces 106, such as DNA or RNA aptamers and antibodies, to name a few. Additional details regarding biological interfaces and associated chemical and biological mechanisms will be discussed in detail herein.

The biosensing cartridge 102 includes various levels of chip packaging 108 to integrate the dual gate back sensing FET sensor chip into a fluid environment. In addition, the biosense cartridge 102 includes a fluid component 110 with microfluidic channels to manage the delivery of liquid to the FET sensor 104. In addition, the fluid component 110 includes a fluid inlet for interfacing with fluid delivered from outside of the biosense cartridge 102.

The integration of various components within the biosense cartridge 102 yields a compact, portable platform that can be used for a large number of different biosense applications. The use of FET sensors with integrated fluid components produces accurate results while using a low sample volume. Further, the biosense cartridge 102 can be configured to operate in a completely autonomous manner by the analyzer, and then discarded after use.

The description herein is divided into four main parts to describe the components of the biosense cartridge 102 in more detail. The first part will describe the arrangement and fabrication of the dual gate back bio FET sensor 104. The second part will describe the packaging process. The third part will describe the fluid component 110 and further describe the interaction between the biosense cartridge 102 and the analyzer. The final part will provide details on biological and various biosensing applications using the dual gate back FET sensor 104.

Dual Gate back FET sensor

The dual gate back FET sensor utilizes semiconductor manufacturing technology and biological capture agents to form a sensitive and easily arranged sensor. While a conventional MOSFET has a single gate electrode connected to a single electrical node, a dual gate back sensing FET sensor has two gate electrodes each connected to a different electrical node. The first gate electrode of the two gate electrodes is referred to herein as a front-side gate, and the second gate electrode of the two gate electrodes is referred to herein as a back-side gate. Both the front gate electrode and the rear gate electrode can be electrically charged and/or discharged in operation, whereby each is configured to affect the electric field between the source/drain terminals of the dual gate sensing FET sensor. The front gate is electrically conductive, separated from the channel region by the front gate dielectric, and is configured to be charged and discharged by the electrical circuit to which it is coupled. The back gate is typically separated from the channel region by a back gate dielectric and includes a biofunctionalized sensing layer disposed on the back gate dielectric. The amount of charge on the back gate is a function of whether a biorecognition reaction has occurred. In typical operation of a dual gate back sensing FET sensor, the front gate is charged to a voltage within a predetermined voltage range. The voltage on the front gate determines the corresponding conductivity of the channel region of the FET sensor. A relatively small change in charge to the charge at the back gate changes the conductivity of the channel region. This is such a change in conductivity that indicates a biometric response.

One advantage of FET sensors is the possibility of label-free operation. Specifically, FET sensors enable avoidance of costly and time consuming labeling operations, such as labeling of analytes using, for example, fluorescent or radioactive probes.

2, an exemplary dual gate back sensing FET sensor 200 is shown. The dual gate backside sensing FET sensor 200 includes a control gate 202 formed on the substrate 214 and separated from it by an intervening dielectric 215 disposed on the substrate 214. The substrate 214 further includes a source region 204, a drain region 206 and a channel region 208 between the source region 204 and the drain region 206. In one embodiment, the substrate 214 has a thickness between approximately 100 nm and approximately 130 nm. The gate 202, source region 204, drain region 206, and channel region 208 can be formed using suitable CMOS process techniques. Gate 202, source region 204, drain region 206, and channel region 208 form a FET. The insulating layer 210 is disposed on the opposite side of the substrate 214 from the gate 202. In one embodiment, the insulating layer 210 has a thickness of approximately 1 μm. In the present disclosure, the side of the substrate 214 on which the gate 202 is disposed is referred to as the “front-side” of the substrate 214. Similarly, the side of the substrate 214 on which the insulating layer 210 is disposed is referred to as the "back-side" of the substrate 214.

An opening 212 is provided in the insulating layer 210. The opening 212 can be substantially aligned with the gate 202. In other embodiments, opening 212 is larger than gate 202 and may extend over multiple dual gate back sensing FET sensors. An interface layer (not shown) can be disposed in the opening 212 on the surface of the channel region 208. The interface layer may be operable to provide an interface for positioning and immobilizing one or more receptors for detection of biomolecules or bio-entities. Additional details regarding the interface layer are provided here.

The dual gate back sensing FET sensor 200 includes a drain region 206 (Vd 206), a source region 204 (Vs 218), a gate structure 202 (front gate 220) and/or an active region 207. (Eg, rear gate 222). It should be noted that the rear gate 222 need not physically contact the substrate 214 or any interface layer over the substrate 214. Thus, while conventional FETs use gate contact to control the conductance of the semiconductor (e.g., channel) between the source and drain, the dual gate back sensing FET sensor 200 is a receptor formed on the opposite side of the FET device. Allows the conductance to be controlled, and the gate structure 202 provides another gate to control the conductance. Thus, the dual gate back sensing FET sensor 200 detects one or more special biomolecules or bio entities in the environment around and/or within the opening 212, as discussed in more detail using various examples herein. Can be used to

The dual gate back sensing FET sensor 200 includes additional passive components such as resistors, capacitors and/or fuses; Other active components such as P-channel field effect transistor (PFET), N-channel field effect transistor (NFET), metal-oxide-semiconductor field effect transistor (MOSFET), high voltage transistor and/or high frequency transistor; Other suitable components; And/or combinations thereof. Additional features can be added within the dual gate back sensing FET sensor 200 and some of the described features can be replaced or removed for additional embodiments of the dual gate back sensing FET sensor 200. For further details regarding exemplary manufacturing procedures for the dual gate back sensing FET sensor 200, see US Patent Application Publication No. 2013/0200438 and US Patent Application Publication No. It can be found at 2014/0252421.

Referring to FIG. 3, a schematic diagram of an addressable exemplary array 300 of FET sensors 304 coupled to bit lines 306 and word lines 308 is shown. Note that the terms bit line and word line are used herein to denote similarity to the array configuration in the memory device, but there is no indication that the memory device or storage array is necessarily included in the array. do. The addressable array 300 may have similarities to those employed in other semiconductor devices, such as dynamic random access memory (DRAM) arrays. For example, the dual gate back sensing FET sensor 200 described above with reference to FIG. 2 can be formed at a location that can be found in a capacitor DRAM array. It will be appreciated that the schematic diagram 300 is merely exemplary, and other configurations are possible,

The FET sensors 304 may be substantially similar to the dual gate back sensing FET sensors 200, respectively. FET 302 provides a connection between the drain terminal of FET sensor 304 and bit line 306. In this way, FET 302 is similar to an access transistor in a DRAM array. In this exemplary embodiment, the FET sensor 304 is a dual gate back sensing FET sensor, the sensing gate provided by the receptor material disposed on the dielectric layer overlying the FET active region disposed at the reaction site, and over the FET active region And a control gate provided by a gate electrode (eg, polysilicon) disposed on the dielectric layer.

Schematic 300 shows an array configuration that may be beneficial in detecting small biomolecules or small signal changes provided by bio-entities entering FET sensor 304. Formats arranged using bit lines 306 and word lines 308 enable a reduced number of input/output pads. An amplifier can be used to improve signal strength to improve the detection capability of devices with circuit arrangement of schematic diagram 300. In one embodiment, when a particular word line 308 and bit line 306 are asserted, the corresponding access transistor 302 will be turned on (eg, similar to a switch). If the gate of the associated FET sensor 304 (e.g., the rear gate 222 of the dual gate back sensing FET sensor 200) has a charge that is affected by the presence of biomolecules, the FET sensor 304 It will deliver electrons and induce charge of the field effect of the device, thereby modulating the current (eg, I _ds ). Changes in current (eg, I _ds ) or threshold voltage (V _t ) can serve to indicate the detection of related biomolecules or bio-entities. Thus, devices with schematic diagram 300 can achieve biosensor applications, including applications that use differential sensing for improved sensitivity.

4, an exemplary layout 400 is provided. The exemplary layout 400 includes an access transistor 302 and a FET sensor 304 arranged as an array 401 of individually addressable pixels 402. The array 401 may include any number of pixels 402. For example, the array 401 may include 128 x 128 pixels. Other arrangements may include non-square arrays such as 256 x 256 pixels or 128 x 256 pixels.

Each pixel 402 includes an access transistor 302 and a dual gate back sensing FET sensor 304 along with other components that may include one or more heaters 408 and a temperature sensor 410. In this example, the access transistor 302 is an n-channel FET. The n-channel FET 412 can also act as an access transistor for the temperature sensor 410. In this exemplary example, the gates of the FETs 302, 412, although not required, are commonly coupled. Each pixel 402 (and its associated components) can be individually addressed using a column decoder 406 and a row decoder 404. In one example, each pixel 402 has a size of approximately 10 micrometers x approximately 10 micrometers. In another example, each pixel 402 has a size of approximately 5 micrometers x approximately 5 micrometers, or approximately 2 micrometers x approximately 2 micrometers.

Column decoder 406 and row decoder 404 can be used to determine the ON/OFF state of n-channel FETs 302,412. Turning on the n-channel FET 302 provides current to the S/D region of the dual gate back sensing FET sensor 2304. When these devices are ON, the current I _ds flows through the FET sensor 304 and can be measured.

The heater 408 can be used to locally increase the temperature around the dual gate back sensing FET sensor 304. The heater 408 can be constructed using any known technique, such as forming a metal pattern through which a high current flows. Further, the heater 408 may be a thermoelectric heater/cooler such as a Peltier element. Heaters 408 can be used during certain biological tests, for example, to denature DNA or RNA or to provide a more ideal binding environment for certain biomolecules. The temperature sensor 410 can be used to measure the local temperature around the dual gate back sensing FET sensor 304. In one embodiment, a control loop may be formed to control temperature using feedback received from heater 408 and temperature sensor 410. In other embodiments, the heater 408 may be a thermoelectric heater/cooler that enables local active cooling of components within the pixel 402.

5, a cross-sectional view of an exemplary dual gate back sensing FET sensor 500 is provided. The dual gate back sensing FET sensor 500 is one implementation of the dual gate back sensing FET sensor 200, so the elements previously described in FIG. 2 are labeled with the element reference numerals in FIG. Is not repeated here. The dual gate back sensing FET sensor 500 includes a gate 202, a source region 204, a drain region 206, and a channel region 208, and the source region 204 and the drain region 206 include a substrate ( 214). Gate 202, source region 204, drain region 206, and channel region 208 form a FET. It should be noted that, as understood by those skilled in the art, the various components of FIG. 5 are not intended to be drawn to scale and are exaggerated for visual convenience.

In an exemplary embodiment, the dual gate back sensing FET sensor 500 is coupled to various sides of metal interconnects 502 that form electrical connections with various doped regions and other devices formed in the substrate 214. The metal interconnect 502 can be manufactured using a manufacturing process well known to those skilled in the art.

The dual gate rear FET sensor 500 may include a body region 504 separated from the source region 204 and the drain region 206. Body region 504 can be used to bias the carrier concentration in active region 208 between source region 204 and drain region 206. As such, a negative voltage bias may be applied to the body region 504 to improve the sensitivity of the dual gate backside FET sensor 500. In one embodiment, body region 504 is electrically connected to source region 205. In other embodiments, body region 504 is electrically grounded.

The dual gate back FET sensor 500 can be coupled to an additional circuit 506 fabricated within the substrate 214. Circuit 506 may include any number of MOSFET devices, resistors, capacitors, or inductors to form circuitry to assist in operation of dual gate backside sensing FET sensor 500. For example, column decoder 406 and row decoder 404 can be formed in circuit 506. Circuit 506 may include any amplifier, analog-to-digital converter (ADC), digital-to-analog converter (DAC), voltage generator, logic circuit, and memory elements, to name a few. All or part of the components of the additional circuit 506 can be integrated into the same substrate 214 as the dual gate back FET sensor 500. It should be understood that many FET sensors, each substantially similar to the dual gate back FET sensor 500, can be integrated on the substrate 214 and coupled to additional circuitry 506. In another example, all or part of the components of the additional circuit 506 are provided on another semiconductor substrate separate from the substrate 214. In another example, some of the components of the additional circuit 506 are integrated on the same substrate 214 as the dual gate backside FET sensor 500, and some of the components of the additional circuit 506 are separated from the substrate 214. It is provided on another semiconductor substrate.

With continued reference to the example example of FIG. 5, the dual gate back sensing FET sensor 500 includes an interface layer 508 formed over the insulating layer 210 and in an opening over the channel region 208. In one embodiment, the interface layer 508 has a thickness between approximately 20 mm 2 and approximately 40 mm 2. The interface layer 508 is hafnium silicate, hafnium oxide, zirconium oxide, aluminum oxide, tantalum pentoxide, hafnium dioxide-alumina )(HfO ₂ -Al ₂ O ₃ ) alloys or combinations thereof. The interface layer 508 can serve as a support for attachment of the capture agent, as discussed in more detail later in the section on biological sensing.

Exemplary operation of the dual gate back FET sensor 500 serving as a pH sensor will now be described. Briefly, a fluid gate 510 is used to provide electrical contact to the “second gate” of the dual gate back sensing FET sensor. A solution 512 with a given pH is provided over the reaction site of the dual gate back sensing FET sensor 500, and a fluid gate 510 is disposed within the solution 512. The pH of the solution is generally related to the concentration of hydrogen ions in the solution [H ⁺ ]. The accumulation of ions near the surface of the interface layer 508 over the channel region 208 causes the inversion layer in the channel region 208 to form a conductive path between the source region 204 and the drain region 206. Will influence formation. This can be measured by changing the conductivity of the FET sensor. In one embodiment, while gate 202 remains floating, fluid gate 510 is used as the gate of the transistor under sensing. In another embodiment, while gate 202 is biased to a given potential, fluid gate 510 is used as the gate of the transistor under sensing. For example, gate 202 can be biased to a potential between -2V and 2V depending on the application, and fluid gate 510 is swept between a wide range of voltages. In other embodiments, fluid gate 510 is biased (or grounded) to a given potential, and gate 202 is used as the gate of the transistor under sensing (eg, its voltage is swept across various potentials). . The fluid gate 510 can be formed of platinum or any other material(s) commonly used for reference electrodes in electrochemical analysis. The most common reference electrode is an Ag/AgCl electrode with a stable potential value of approximately 0.230V.

6A shows ions in solution bound to the surface of interface layer 508. The topmost atomic layer of the interface layer 508 is shown as various dangling [O ⁻ ], [OH] and [OH ₂ ⁺ ] bonds. As ions accumulate on the surface, the total surface charge affects the threshold voltage of the transistor. As used herein, the threshold voltage is the minimum potential between the gate and the source of the FET sensor required to form the conductive path of the minority carrier between the source and drain of the FET sensor. In addition, since higher accumulation of positive charges indicates a lower pH and higher accumulation of negative charges indicates a higher pH, the total charge is directly related to the pH of the solution. 6B shows the change in threshold voltage resulting from different pH values in the n-channel FET sensor. As can be observed in the figure, a 59 mV increase in threshold voltage represents an increase of approximately 1 in the pH of the solution. In other words, a pH change of 1 when measured as the voltage required to turn on the transistor gives 59mV and an equivalent total surface charge.

Chip packaging

Referring to FIG. 7, an exemplary top view of the semiconductor chip 702 is shown. Chip 702 includes sensor array 704, optional reference electrode 706, analog circuit 708 and I/O pad 716. The chip 702 may be silicon, gallium arsenide, or indium phosphide, to name a few. The chip 702 can have a dimension of approximately 3 mm x approximately 2.5 mm.

The sensor array 704 represents an array of dual gate back sensing FET sensors as illustrated above in FIGS. 2 and 5. The array can be arranged as a low-column matrix of pixels, for example, as illustrated in FIG. 4. Various FET sensors in the sensor array 704 can be functionalized with the same or different capture agents to perform biosensing for various analytes.

The reference electrode 706 can be patterned on the same chip 702 that includes the sensor array 704. The reference electrode 706 can be roughly aligned with the sensor array 704 along the X or Y direction so that the fluid channel can be disposed over both the sensor array 704 and the reference electrode 706.

The reference electrode 706 can include any material having a relatively stable potential. Exemplary reference electrode materials include platinum or Ag/AgCl. Manufacturing Ag/AgCl electrodes on a substrate surface is described, for example, by Moschou et al., "Surface and Electrical Characterization of Ag/AgCl Pseudo-Reference Electrodes Manufactured with Commercially Available PCB Technologies," Sensors, vol. 15(8), 2015, pp. It is well known in the art as described in 18102-18113.

The analog circuit 708 can include circuitry related to the operation of the sensor array 704. As such, the analog circuit 708 can be configured to provide signals to or measure signals from the array 704 while interfacing with various I/O pads 716. In one embodiment, the analog circuit 708 includes a serial peripheral interface (SPI) 712 and a sensor array circuit 714. In this embodiment, the distance between sensor array 704 and sensor array circuit 714 is not less than approximately 135 micrometers.

The SPI 712 can be a serial interface circuit to facilitate data communication between the sensor array circuit 714 and the analyzer unit described in more detail below. The general operation of the SPI will be well understood by those skilled in the art. The sensor array circuit 714 can include any number of reference voltage generators, operational amplifiers, low pass filters, ADCs and DACs to provide signals to and receive signals from the sensor array 704.

In one example, a biasing reference voltage can be generated using sensor array circuitry 714 to provide a negative voltage bias of approximately -0.24V to a given FET sensor of sensor array 704 or a body region of a set of FET sensors. Also, when performing sensing, a tunable voltage can be provided to a given FET sensor in the sensor array 704 or a fluid gate of a set of FET sensors.

When measuring a signal (such as Ids) received from a given FET sensor or set of FET sensors of the sensor array 704, the sensor array circuit 714 receives the measured signal so that the resulting signal is an I/O pad. Before being output to 716, one or more additional amplification stages, low pass filters, and ultimately the ADC can deliver them through a subsequent trans-impedance amplifier, i. In addition, before the measured signal is amplified, noise can be reduced from the measured signal by subtracting the background AC signal from the measured signal. In addition, a temperature signal (received from one or more temperature sensors in the sensor array 704) can be amplified, filtered and passed through the ADC before being output to the I/O pad 716.

In various embodiments, a plurality of I/O pads 716 may be patterned along the periphery of the chip 702. More I/O pads may be provided than the actual inputs and outputs used by the various components of the chip 702. In one embodiment, wire bonding technology may be used to couple the various I/O pads 716 to other substrates or packages bonded to the chip 702. In one particular embodiment, 32 I/O pads can be patterned around the periphery of chip 702. The size of a given I/O pad 716 may be approximately 80 micrometers x approximately 70 micrometers, and the pitch between I/O pads 716 may be approximately 150 micrometers. The spacing between the sensor array 704 and the closest I/O pad 716 may not be shorter than approximately 400 micrometers, and the spacing between the I/O pad 716 and the outermost edge of the chip 702 is It may not be shorter than approximately 177.5 micrometers.

Referring to FIG. 8, an exemplary packaging scheme is illustrated for chip 702. The chip 702 with the I/O pad 716 is bonded to the carrier layer 802. The carrier layer 802 may be another semiconductor substrate, such as a silicon substrate. In another example, the carrier layer 802 is an insulator such as a rigid plastic material. The chip 702 can be bonded to the carrier layer 802 using any known bonding technique, such as using solder or adhesive.

In one embodiment, carrier layer 802 includes a plurality of through holes filled with conductive material 804. The conductive material 804 can be any metal, such as, but not limited to, tin, copper, aluminum, gold, or any alloy thereof. The conductive material 804 may include solder bumps or solder balls on the lower surface 805 of the carrier layer 802. Solder may extend beyond surface 805.

In addition, the chip package, according to one embodiment, includes a first insulating layer 806 that contacts the side of the chip 702. Also, the first insulating layer 806 may be a plastic material or resin that fills the area around the chip 702 and helps to hold the chip 702 in place. In one exemplary embodiment, the first insulating layer 806 includes through holes that are also filled with a conductive plug 808. The conductive plug 808 may be the same material as the conductive material 804. The conductive plug 808 is substantially aligned over a corresponding area of conductive material such that an ohmic contact is formed between the conductive plug 808 and the conductive material 804.

When the chip 702 is fixed to the carrier layer 802 and has a first insulating layer 806 around it, an electrical connection 812 can be provided between the I/O pad 716 and the conductive plug 808. have. The electrical connections 812 can be formed using wire bonding techniques, as understood by one of ordinary skill in the art. In another example, electrical connections 812 are formed using lithographic patterning techniques to pattern conductive traces for electrically connecting I/O pads 716 with corresponding conductive plugs 808. When the electrical connection portion 812 is formed, the second insulating layer 810 may be attached to protect the electrical connection portion 812 from the surroundings. The second insulating layer 810 may be made of the same material as the first insulating layer 806. The second insulating layer 810 may be a resin material that flows around the electrical connection 812 and is then cured to form a protective shell. An opening 814 is formed in the second insulating layer 810 to form a path toward the sensor array present on the chip 702. In one embodiment where the reference electrode is also patterned on the chip 702, the opening 814 will form a path toward the sensor array and the reference electrode.

The final chip package 816 includes a chip 702 that is bound to the carrier layer 802 and electrically connected to various conductive solder points or metal pads on the lower surface 805 of the carrier layer 802. Further, the chip 702 is protected from the surroundings through the first insulating layer 806 and the second insulating layer 810. The chip package 816 can be easily handled and coupled to a larger substrate, such as a printed circuit board (PCB). In some embodiments, chip package 816 may be coupled to one or more heat sinks to provide a more efficient heat dissipation path from chip 702 to ambient air or whatever substrate the chip package 816 is attached to. Can. In other embodiments, chip package 816 may be coupled to a Peltier device to provide thermoelectric heating and/or cooling.

Referring to the exemplary embodiment of FIG. 9, the chip package 816 is bonded to the substrate 902. The substrate 902 may be a PCB that includes a conductive contact pad for forming electrical contact with a solder or conductive pad on the bottom surface of the carrier layer 802. Flip chip bonding technology can be performed to bond the chip package 816 onto the surface of the substrate 902. Briefly, solder or conductive pads along the bottom surface of the carrier layer 802 are aligned with corresponding conductive pads patterned on the substrate 902 to physically attach the chip package 816 to the substrate 902. Then, the I/O pads are bonded together to electrically couple from the chip 702 to the conductive traces present on the substrate 902. The conductive trace on the substrate 902 can be terminated at the edge connector 908.

One or more edge connectors 908 can provide electrical connections to the chip 702. One or more other edge connectors 908 can provide electrical connections to the reference electrode 906 patterned on the surface of the substrate 902. Using the reference electrode 906 can eliminate the need to provide a reference electrode on the chip 702. Each of the one or more edge connectors 908 may be patterned using a metal such as, but not limited to, copper, gold, or aluminum. The reference electrode 906 can be fabricated using techniques similar to those discussed above for the reference electrode 706 on the chip 702.

The dimensions of the exemplary chip package 816 may be approximately 1 to 2 centimeters x 1 to 2 centimeters or smaller, and the dimensions of the substrate 902 may be approximately 3 to 4 centimeters x 3 to 4 centimeters or smaller. .

An opening 814 is shown over the chip 702, exposing at least the sensor array of the chip 702. In one exemplary embodiment, the opening 814 is approximately aligned with the reference electrode 906 along the X or Y direction, such that a fluid channel can be disposed over both the opening 814 and the reference electrode 906.

Fluid design

10, a schematic diagram of an exemplary fluid cartridge 1000 is provided. It should be noted that the schematic diagram shows a top view of the fluid cartridge 1000, and not all elements shown are on the same horizontal plane. In addition, the specific dimensions and scales of the various fluid channels are purposefully drawn out of scale for improved visualization. The cartridge 1000 includes a housing 1002. The housing 1002 may be formed of any plastic material, such as polymethyl methacrylate (PMMA), using injection molding, casting or 3D printing techniques, to name a few. The housing 1002 can be formed of two or more segments that are connected together mechanically or through the use of an adhesive. In one embodiment, various fluid channels and chambers may be molded into one or more chambers of housing 1002. In other embodiments, various fluid channels and chambers are formed from other molded polymer materials, such as polydimethylsiloxane (PDSM). The overall dimensions of the housing 1002 may be between approximately 4 and approximately 7 centimeters x approximately 4 and approximately 7 centimeters. As technology advances, the housing 1002 can be much smaller. In one embodiment, a substrate 902 having a packaged chip 802 is disposed within the housing 1002. In one example, only a portion of the substrate 902 is enclosed within the housing 1002, and the edge connection 908 is exposed outside the housing 1002.

The fluid design of the exemplary housing 1002 includes at least a first channel 1004, a second channel 1006, and a third channel 1008. Each of the first channel 1004 and the second channel 1006 includes corresponding fluid inlets 1010a and 1010b, respectively. The fluid inlet provides an area for injecting fluid from the outside of the cartridge 1000 into the cartridge 1000. In addition, the fluid inlet provides an area for discharging fluid from the cartridge 1000 to the outside of the cartridge 1000. The third channel 1008 can be aligned over the packaged chip 802 bonded to the substrate 902. In one embodiment, the opening 814 over the sensor array is substantially within the third channel 1008. Further, the reference electrode 906 patterned on the substrate 902 is aligned to be within the third channel 1008, according to one embodiment.

Each of the first channel 1004, the second channel 1006, and the third channel 1008 may have a channel width between approximately 2 mm and 3 mm. The channel height can be approximately 1 mm. In other embodiments, one or more of the first channel 1004, the second channel 1006, and the third channel 1008 are microfluidic channels having a width and height dimension of less than 1 mm. Each of the first channel 1004, the second channel 1006, and the third channel 1008 may have a rectangular, square, or semi-circular cross section.

In some embodiments, at least one of the first channel 1004 and the second channel 1006 is connected to the third channel 1008. In this way, the fluid flowing through the first channel 1004 will eventually flow through the third channel 1008, and similarly the fluid flowing through the second channel 1006 will eventually flow through the third channel 1008. will be. In some embodiments, the third channel 1009 eventually flows into a waste chamber 1016 that collects all fluid flowing through the cartridge 1000. The waste chamber 1016 may include vents (not shown) to the atmosphere to prevent back pressure from forming in the fluid system.

In some embodiments, each inlet 1010a, 1010b includes plugs 1012a, 1012b, respectively. The plug 1012a/1012b may be a soft compliant material that fits comfortably within the inlet 1010a/1010b to seal the inlet from any fluid leakage. The plug 1012a/1012b may be a polytetrafluoroethylene (PTFE) or polymeric material such as cork. The plug 1012a/1012b can seal the inlet 1010a/1010b, allowing the capillary to puncture the plug 1012a/1012b without damaging the fluid seal. The capillary may be a needle-like tube, such as an injection needle. The capillary may include a hard rigid material such as metal or hard plastic. The coupling of the capillary tube to the cartridge 1000 will be described in more detail later when discussing the coupling of the cartridge 1000 with the analyzer.

The cartridge 100 includes a sample inlet 1014 arranged to introduce a sample into the first channel 1004 (as shown in FIG. 10) or the second channel 1006. In one example, a blood sample is placed through the sample inlet 1014 into the fluid system. Once the sample is introduced, the sample inlet 1014 can be sealed using a cap or other similar structure to provide a leak-tight seal around the sample inlet 1014. In the illustrated channel arrangement of FIG. 10, fluid flowing from inlet 1010a through first channel 1004 will be mixed with the sample introduced through sample inlet 1014, and the mixture is within third channel 1008. It will flow over opening 814 and reference electrode 906. When the sample is delivered through the aperture 814 to the exposed sensor array, interactions between biomolecules occur, and the FET sensor can be used to detect the presence of a particular analyte in the sample or to measure its concentration. The fluid can be moved along and between the various channels using pressure driven flow. Pressure may be generated, for example, by a syringe pushing liquid or air through the cartridge 1000, or pressurized air pushing the liquid. Other examples of techniques for transporting liquid through the cartridge 1000 include using an electro-wetting or on-chip peristaltic pump. In some embodiments, fluid mixing can occur within cartridge 1000 using any one of a variety of on-chip mixing methods known in the art. The dimensions of the fluid channel of the cartridge 1000 may be large enough to occur due to turbulence of the liquid as some fluid mixing moves through the channel. It should be understood that the location of the sample inlet 1014 can be varied. For example, the sample inlet 1014 can be positioned directly above the opening 814 such that samples introduced into the sample inlet 1014 are also introduced over the sensor array exposed through the opening 814.

Once the substrate 902 is integrated into the housing 1002, the sensor array accessed through the opening 814 can be functionalized with various capture agents, according to one embodiment. This process involves flowing a liquid buffer containing the capture agent through the third channel 1008 so that the capture agent has the opportunity to bind to various FET sensors in the sensor array. In another example, when the sample inlet 1014 is positioned over the opening 814, the capture agent is placed just above the opening 814. After the capture agent is immobilized, the sample inlet 1014 can be sealed so that the cartridge 1000 can be stored until ready to perform a biological detection test. The capture agent may remain in the initial buffer solution, or a new buffer solution may be introduced to preserve the capture agent while the cartridge 100 is waiting for the test. Examples of different capture agents and tests performed with the capture agents are provided herein.

11, another design for various fluid channels of the cartridge 1000 is illustrated. In this design, a first channel 1104 having a first inlet 1102a and a second channel 1106 having an inlet 1102b converge in an area having a sample inlet 1110. A third channel 1108 having an opening 814 aligned therein is connected to the first channel 1104 and the second channel 1106 at the sample inlet 1110. Opening 814 provides a path down to the chip to expose at least the sensor array on the chip to fluid in the third channel 1108. Fluid flowing from the first channel 1104 or the second channel 1106 through the third channel 1108 is ultimately collected in the waste chamber 1112. Fluid may be directed to the waste chamber 1112 based on the geometry of various channels or by using a valve to close a given channel. Also, the sample inlet 1110 may be located over the opening 814.

One or more of the first channel 1104, the second channel 1106, and the third channel 1108 may include a bubble trap 1114. Bubble trap 1114 is a region of a fluid channel that suddenly has a larger cross-section (or higher "ceiling") so that any air present in solution can rise into the additional space formed in bubble trap 1114. Can represent Other bubble trap designs can also be used, as would be understood by those skilled in the art. Removing air bubbles from the solution before reaching the sensor array under the aperture 814 can be important to ensure accurate detection results.

Referring to FIG. 12, a cartridge 1000 coupled to the analyzer 1200 is illustrated to perform biological detection. The cartridge 1000 may be in physical contact with the analyzer 1200, for example, by pressing the cartridge 1000 against the receiving portion of the analyzer 1200. The receptacle of the analyzer 1200 may include electrical pads for forming ohmic contacts to some or all of the edge connectors 908. The edge of the substrate 902 can be comfortably fitted into the receptacle of the analyzer 1200 such that the edge connector 908 presses against the corresponding conductive pad of the analyzer 1200. Other methods of assembling the cartridge 1000 and the analyzer 1200 include, in particular, snapping them together and plugging one to the other. The analyzer 1200 can be small enough to be easily portable and fit into the palm of an adult's hand.

In some embodiments, the analyzer 1200 includes at least a first syringe 1202a and a second syringe 1202b. Each of the first syringe 1202a and the second syringe 1202b may include a buffer or other solution used during operation of the cartridge 1000. The syringes 1202a/1202b each include needles 1204a/1204b that can be aligned to extend into a space remote from the rest of the analyzer 1200. In some embodiments, the needles 1204a/1204b are inserted through the corresponding plugs 1012a/1012b by the needles 1204a/1204b by pressing the cartridge 1000 against the receptacle of the analyzer 1200 and the inlet. (1010a/1010b). In this embodiment, the needle 1204a/1204b is an example of a capillary tube punching the corresponding plug 1012a/1012b. Thus, a leak-tight seal is formed to deliver the solution from each syringe 1202a/1202b into the corresponding inlet 1010a/1010b of the cartridge 1000. Although this description only describes two syringes aligned with two input ports, it is understood that any number of syringes and fluid input ports can be used, including examples where only one syringe is used to engage a single inlet. Should be. Each syringe 1202a/1202b can be preloaded with a solution for use in various tests. In another example, each syringe 1202a/1202b can be easily removed and replaced by a different syringe by the user.

Each syringe 1202a/1202b can have an associated plunger controlled via a corresponding actuator 1206a/1206b. Examples of the actuators 1206a/1206b include a stepper motor and an induction motor. The speed at which the actuators 1206a/1206b push the plunger of the syringe 1202a/1202b will directly affect the flow rate of the solution in the fluid channel of the cartridge 1000. The actuators 1206a/1206b can be controlled through the motor control module 1208a/1208b. The motor control module 1208a/1208b includes circuitry necessary to generate a voltage for controlling the speed and operation of the actuator 1206a/1206b, as understood by those skilled in the art.

All electrical connections formed with respect to the edge connector 908 of the cartridge 1000 may be routed to the sensing electronic device 1210. The sensing electronics 1210 can include any number of individual circuits, integrated circuits, and individual analog circuit components designed to provide and receive many different electrical signals between the sensing electronics 1210 and the edge connector 908. . For example, the sensing electronics 1210 can provide the edge connector 908 with power, ground, and clock signals that can then be used to power and operate sensor arrays and other electronic devices on the chip 702. Can be configured. The sensing electronics 1210 can also provide various voltage bias levels to activate the gates of specific FET sensors in the sensor array. The sensing electronic device 1210 may receive a signal indicating the drain current measured from the specific FET sensor and a signal indicating the output from the temperature sensor on the chip 702. The sensing electronics 1210 can store this received data in memory, use the received data to change the voltage bias level, or change the amount of heat generated by the heater on the chip 702. have. In general, the sensing electronics 1210 controls all signal processing related to biosensing performed by the sensor array of the cartridge 1000.

In some embodiments, the analyzer 1200 also includes a processor 1212 that controls the timing and function of each of the other modules of the analyzer 1200, such as the motor control module 1208a/1208b and the sensing electronics 1210. do. The processor 1212 may be any type of central processing unit (CPU) or microcontroller, and may be programmable by a user to perform certain functions related to the operation of the analyzer 1200. The processor 1212 may be configured to analyze a signal received from the sensing electronic device 1210 to determine a concentration level of an analyte given from a sample in the cartridge 1000. Data related to the determined concentration level may be stored in the memory of the analyzer 1200. In another example, the sensing electronic device 1210 may be further configured to determine a concentration level of an analyte given from a sample in the cartridge 1000 and to store data related to the determined concentration level in the memory of the analyzer 1200. .

In some embodiments, the analyzer 1200 includes a communication module 1214 designed to communicate data to an external processing device. The processor 1212 may be electrically coupled with the communication module 1214 to control data transmission. The communication can be wired or wireless. Examples of wired communication include data transmission via a network cable or universal serial bus (USB) cable. Wireless communication may include wireless RF transmission, Bluetooth, WiFi, 3G or 4G. The communication module 1214 can also be designed to receive data from an external processing device. For example, a program on how to operate various components of the analyzer 1200 may be transmitted to the communication module 1214 and executed by the processor 1212. The communication module 1214 can include any number of well-known hardware elements to facilitate the transmission and reception of analog and/or digital data.

After the biosensing test is performed, the cartridge 1000 can be removed from the analyzer 1200 and discarded. Also, the syringe 1202a/1202b can be removed from the analyzer 1200 and discarded. Thus, all reagents remain contained within the cartridge 1000 or syringe 1202a/1202b, and no contamination of any other portion of the analyzer 1200 occurs. In this way, a single analyzer 1200 can be reused to test any number of additional cartridges, and each cartridge can be individually functionalized with a different capture agent to perform different biosensing tests.

In another embodiment, the syringe 1202a/1202b is integrated on the cartridge 1000, and the coupling between the cartridge 1000 and the analyzer 1200 causes the associated plunger of the syringe 1202a/1202b to actuate on the analyzer 1200. (1206a/1206b). In this embodiment, the analyzer 1200 has no reagent transport container at all.

In another embodiment, cartridge 1000 includes one or more capillaries that are perforated through corresponding plugs 1012a/1012b. In this embodiment, when coupling occurs between the cartridge 1000 and the analyzer 1200, the capillary fluidly couples with the rest of the syringes 1202a/1202b in the analyzer 1200. After the biosensing test has been performed, the cartridge 1000 can be removed from the analyzer 1200 and discarded along with its capillaries.

13, an exemplary method 1300 is provided. The method 1300 can be performed by the analyzer 1200 after the cartridge 1000 is coupled to the analyzer 1200. Other operations related to fluid handling and electrical measurements that are not illustrated in method 1300 can be performed before, during, or after the illustrated operations of method 1300. Various operations of method 1300 may be performed in a different order than illustrated. In one embodiment, the method 1300 is performed after the capture agent has already been immobilized in the cartridge 1000.

In block 1302, a first solution flows through the first channel of the cartridge. The first solution can enter the cartridge through an inlet coupled to the first channel. The first channel can be provided by a syringe having a needle punching a plug disposed at the inlet of the first channel. The first solution may include a buffer solution to provide a stable pH environment.

In block 1304, the dual gate back sensing FET sensor of the sensor array is calibrated in the first solution. Calibration can be performed to measure noise or background signals from various FET sensors. These measurements can be stored and then subtracted from the measured signal when detecting biomolecules to test for noise reduction and achieve a cleaner detection signal. In order to perform calibration, the first solution must be present on the reference electrode and sensor array patterned in the main detection channel. In some embodiments, the first solution does not flow during calibration measurements. In some embodiments, calibration measurements represent a baseline threshold voltage for the FET sensor.

At block 1306, a sample is entered through the sample inlet into the fluid network of the cartridge. The sample can be any liquid sample, including a blood sample. In some examples, the sample is a semi-solid sample that dissociates in solution. After the sample is entered through the sample inlet, the sample inlet can be sealed by a cap or other similar structure.

In block 1308, a second solution flows through the second channel of the cartridge. The second solution may be the same solution as the first solution. The second solution can be mixed with the sample across the sample entered into the fluid system at block 1306. The mixture of sample and second solution can then flow through the second channel and into the main detection channel where the sensor array is located. The second solution can be a buffer solution. In one example, the second solution is a lysing buffer solution. The second solution can be moved along or between the various channels using pressure driven flow. Pressure may be generated, for example, by a syringe pushing liquid or air through the cartridge, or pressurized air pushing the liquid. Other examples of techniques for transporting the second solution through the cartridge include electrowetting or using an on-chip peristaltic pump.

In block 1310, biomolecules present in the sample are incubated on the sensor array. Incubation can last for any given amount of time, for example between 30 seconds and 10 minutes. During incubation, the sample mixed with the second solution may not flow, or it may be flowing at a very low flow rate. The flow rate can be designed such that a new solution is present over the sensor array over time, but the flow is not too strong to cause damage to the capture agent or no binding reaction.

In block 1312, after the incubation time has expired, a third solution flows through the first channel of the cartridge and through the main detection channel, pushing substantially all of the sample mixed with the second solution into the waste chamber. The third solution can be injected through the main detection channel for a given period to ensure that the sample is removed from the main detection channel. The third solution used in block 1312 should ideally be the same solution as the first solution. In other embodiments, the third solution is different from the first solution. The third solution can be a buffer solution.

At block 1314, the output from the sensor array is measured to determine if any binding reaction has occurred. The sensor output can be a drain current measured from one or more dual gate back sensing FET sensors in the sensor array. The measured drain current can be compared to the drain current measured during calibration of the same sensor in block 1304. If the threshold voltage (e.g., roughly corresponds to the voltage required to turn on the FET to allow the drain current to flow) changes from when the sensor was calibrated, it can be determined that a binding reaction has occurred and the target analyte is present in the sample. . The amount and sign of change in threshold voltage depends on many factors such as whether the dual gate back sensing FET sensor is an n-channel device or a p-channel device, the type of analyte being detected and the amount of positive or negative charge associated with the analyte. can do. In another example, the measured output from the sensor array is its own threshold voltage, which can be compared to the threshold voltage measured during calibration of the same sensor at block 1304.

Chemical, biological and interface

The devices, systems and methods of the present invention as described in this application can be used to detect and/or monitor interactions between various entities. Interactions include biological and chemical reactions to detect target analytes in a test sample. As an example, reactions involving physical conversion, chemical conversion, biochemical conversion or biological conversion can be monitored to detect the production of intermediate products, by-products, products, contaminants or combinations thereof. In addition, the devices, systems and methods of the present invention include circulating tumor cell assays used in liquid biopsies and chelation assays for detecting the presence of heavy metals and other surrounding contaminants. It can be used to detect these reactions in various analytical evaluations as described herein, but not limited to. These assay evaluations and responses can be monitored in a single format, or in an array format for detecting multiple target analytes, for example.

DGBSS Biological detection example using FET sensor

Referring to FIG. 14, an exemplary biosense test is performed using the dual gate back sensing FET sensor described above. Probe DNA 1404 (an example of a capture agent) is bound to the interface layer 508 through a linking molecule 1402. The link molecule 1402 may have a reactive chemical group that binds to a portion of the interface layer 508. Examples of link molecules include thiols. In addition, link molecules may be formed by exposing the surface of the interface layer 508 to ammonia (NH ₃ ) through silanization of the surface of the interface layer 508 or to form reactive NH ₂ groups on the surface. Can. The silanization process sequentially exposes the surface of the interface layer 508 to different chemicals to form covalently bonded molecules on the surface of the interface layer 508, as understood by those skilled in the art. Includes prescribing. Probe DNA 1404 represents single stranded DNA. According to one embodiment, the link molecule 1402 is bound to the interface layer 508 before any steps of the method 1300 are performed. In addition, probe DNA 1404 is bound to link molecule 1402 before any steps in method 1300 are performed. In another example, probe DNA 1404 is bound to link molecule 1402 at block 1302 of method 1300.

The dual gate back sensing FET sensor shown in FIG. 14 is, according to one embodiment, one FET in the sensor array that may be on the same chip as the chip 702 described above. The link molecule 1402 can be bound to the interface layer 508 before the wafer containing the chip 702 is diced to separate the chip 702 from the wafer.

The probe DNA 1404 can be immobilized on the interface layer 508 before applying the FET sensor to the sample 1401. The sample 1401 may include a matching single-stranded DNA sequence 1406 that strongly binds to the matching probe DNA 1404. The binding of additional DNA increases the negative charge present on the interface layer 508 and directly above the channel region 208 of the FET sensor.

DNA binding is conceptually illustrated in Figure 15A. Here, the probe DNA having the nucleic acid sequence TCGA is bound to a complementary matching strand having the nucleic acid sequence AGCT. Any unmatched sequence will not hybridize to the probe DNA sequence. The binding of the matching DNA increases the negative charge formed at the interface of the interface layer 508. In the example illustrated in FIG. 15A, the interface layer 508 is hafnium oxide.

15B illustrates the shift in threshold voltage of the dual gate back sensing FET sensor when matching DNA is bound to the surface of the interface layer 508. Briefly, voltage is applied to the fluid gate 510 until the FET sensor “turns on” and current flows between the drain region 206 and the source region 204. When more negative charge is present in the interface layer 508 due to complementary DNA binding, a higher voltage is needed to form a conductive reversal layer in the channel region 208. Thus, according to one embodiment, a higher voltage may be applied to the fluid gate 510 before the FET sensor conducts and the I _ds current flows. This difference in threshold voltage can be measured and used to determine the concentration as well as the presence of a target matching the DNA sequence. It should be understood that the net positive accumulated charge in the interface layer 508 will decrease, not increase, the threshold voltage. Also, the change in threshold voltage will have the opposite sign for the n-channel FET compared to the p-channel FET.

16, another exemplary biosense test is performed using a dual gate back sensing FET sensor. Probe antibody 1604 (another example of a capture agent) is bound to interface layer 508 through link molecule 1602. The link molecule 1602 can have reactive chemical groups that bind to a portion of the interface layer 508. A sample solution 1601 may be provided over the probe antibody 1604 to determine if a matching antigen is present in the sample solution 1601. According to one embodiment, link molecule 1602 is bound to interface layer 508 before any steps of method 1300 are performed. In addition, probe antibody 1604 can be bound to link molecule 1402 before any steps of method 1300 are performed. In another example, probe antibody 1604 is bound to link molecule 1402 at block 1302 of method 1300.

Referring to FIG. 17, a binding process of the matching antigen to the probe antibody 1604 is illustrated. Here, the matching antigen will bind to the immobilized probe antibody, and the unmatched antigen will not bind. Similar to the DNA hybridization process described above, the matching antigen will change the accumulated charge present in the interface layer 508. The shift in threshold voltage due to the accumulated charge from the matching antibody binding to the probe antibody is measured in substantially the same manner as already described above with reference to FIG. 15B.

According to one aspect of the present invention, a fluid cartridge is provided, the fluid cartridge comprising a plurality of contact pads configured to be electrically coupled to an analyzer, a semiconductor chip having a sensor array, and a reference electrode. A substrate comprising; A first fluid channel having a first inlet and coupled to a second fluid channel, the second fluid channel being arranged such that the sensor array and reference electrode are disposed within the second fluid channel; A sample inlet for placing a sample in the path of the first fluid channel or the second fluid channel; And a first plug disposed at the first inlet, the first plug comprising a compliant material configured to be perforated by capillaries without leaking fluid through the first plug. According to another aspect, the fluid cartridge further comprises a third fluid channel having a second inlet. According to another aspect, the third fluid channel is coupled to the second fluid channel. According to another aspect, the fluid cartridge further comprises a second plug disposed at the second inlet, the second plug comprising a flexible material configured to be perforated by the capillaries without leaking fluid through the second plug. According to another aspect, the first plug and the second plug are configured to align with the first capillary and the second capillary coupled to the analyzer, and the plurality of contact pads engage the analyzer when the fluid cartridge and the analyzer are in physical contact . According to another aspect, the first capillary and the second capillary perforate the first plug and the second plug, respectively, when the fluid cartridge and the analyzer are in physical contact. According to another aspect, the substrate is a printed circuit board. According to another aspect, the fluid cartridge further comprises a waste chamber coupled to the second fluid channel. According to another aspect, one or more sensors of the sensor array include a plurality of probe molecules configured to bind target molecules present in the sample. According to another aspect, the sensor array includes a dual gate back sensing FET sensor array.

According to one aspect of the invention, a fluid cartridge is provided, the fluid cartridge having a first inlet and a first fluid channel coupled to a second fluid channel, wherein the second fluid channel has a sensor array and a reference electrode as a second fluid channel Arranged to be placed within -; A sample inlet for placing a sample in the path of the first fluid channel or the second fluid channel; And a first plug disposed at the first inlet, the first plug comprising a compliant material configured to be perforated by the capillaries without leaking fluid through the first plug, the capillaries comprising Combined, the capillary punctures the first plug when the fluid cartridge and the analyzer are in physical contact. According to another aspect, the sensor array includes an array of dual gate back sensing FET sensors. According to another aspect, one or more sensors of the sensor array include a plurality of probe molecules configured to bind target molecules present in the sample. According to another aspect, the plurality of probe molecules comprises one or more of DNA, RNA and antibodies. According to another aspect, the fluid cartridge further includes a substrate comprising a plurality of contact pads configured to electrically couple with the analyzer, a semiconductor chip with a sensor array, and a reference electrode.

According to one aspect of the present invention, an analyzer configured to engage a fluid cartridge is provided, which is a syringe-syringe, when the fluid cartridge is physically coupled to the analyzer, the needle of the syringe is connected to the corresponding input port of the fluid cartridge. Arranged to sort -; An actuator configured to control the operation of the syringe; A sensing module configured to send and receive signals to and from the fluid cartridge through the plurality of conductive pads contacting the corresponding plurality of conductive pads on the fluid cartridge when the fluid cartridge is physically coupled to the analyzer; And a processor electrically coupled to the sensing module and configured to determine a concentration level of the analyte given from a sample in the fluid cartridge based on a signal received from the fluid cartridge. According to another aspect, the analyzer further includes an actuator controller configured to control the operation of the actuator. According to another aspect, the processor is also electrically coupled to the actuator controller. According to another aspect, the analyzer further comprises at least one other syringe, the at least one other syringe, when the fluid cartridge is physically coupled to the analyzer, the needle of the at least one other syringe is the corresponding input port of the fluid cartridge It is arranged to align with. According to another aspect, the analyzer further comprises a memory configured to store data related to the concentration level of a given analyte determined by the processor.

Last comment

It should be understood that portions of [the specifics for carrying out the invention] rather than the [summary] portion are intended to be used to interpret the claims. The [Summary] section may describe exemplary embodiments of one or more of the invention as contemplated by the invention(s), but not all of them, thus limiting the invention and the appended claims in any way. It is not intended to.

It should be understood that the phraseology or terminology herein is for the purpose of description and not limitation, so that the terminology or phraseology herein is understood by a person skilled in the art in light of the teaching and guidance.

The breadth and scope of the invention should not be limited by any of the foregoing exemplary embodiments, but should be defined only by the appended claims and equivalents thereof.

Claims (10)

As a fluid cartridge,
A substrate including a plurality of contact pads configured to be electrically coupled to the analyzer, a semiconductor chip having a sensor array, and a reference electrode;
A first fluid channel having a first reagent inlet and coupled to a second fluid channel, wherein the second fluid channel is arranged such that the sensor array and the reference electrode are disposed within the second fluid channel;
A sample inlet for placing a sample in the path of the first fluid channel or the second fluid channel; And
A first plug fitted within the first reagent inlet of the fluid cartridge, the first plug comprising a compliant material configured to be perforated by capillaries without leaking fluid through the first plug-
It characterized in that it comprises a fluid cartridge. The fluid cartridge of claim 1, further comprising a third fluid channel having a second reagent inlet. According to claim 1,
The substrate is a printed circuit board, characterized in that the fluid cartridge. According to claim 1,
And a waste chamber coupled to the second fluid channel. According to claim 1,
Wherein at least one sensor in the sensor array comprises a plurality of probe molecules configured to bind to target molecules present in the sample. According to claim 1,
Wherein said sensor array comprises a dual gate back sensing FET sensor array. As a fluid cartridge,
A first fluid channel having a first reagent inlet and coupled to a second fluid channel, wherein the second fluid channel is aligned such that a sensor array and a reference electrode are disposed within the second fluid channel;
A sample inlet for placing a sample in the path of the first fluid channel or the second fluid channel; And
A first plug fitted within the first reagent inlet of the fluid cartridge, the first plug comprising a compliant material configured to be perforated by capillaries without leaking fluid through the first plug-
Including,
The capillary tube is coupled to the analyzer, the capillary fluid cartridge, characterized in that to puncture the first plug when the fluid cartridge and the analyzer are in physical contact. An analyzer configured to engage a fluid cartridge,
Syringe, wherein the syringe is arranged so that when the fluid cartridge is physically coupled to the analyzer, the needle of the syringe punches a plug that fits into the corresponding reagent input port of the fluid cartridge;
An actuator configured to control the operation of the syringe;
A sensing module configured to, when the fluid cartridge is physically coupled to the analyzer, send and receive signals to and from the fluid cartridge through a plurality of conductive pads contacting a corresponding plurality of conductive pads on the fluid cartridge; And
A processor electrically coupled to the sensing module and configured to determine a concentration level of an analyte given from a sample in the fluid cartridge based on a signal received from the fluid cartridge
Analyzer comprising a. The method of claim 8,
And an actuator controller configured to control the operation of the actuator. The method of claim 8,
And further comprising at least one other syringe, wherein the at least one other syringe comprises, when the fluid cartridge is physically coupled to the analyzer, the needle of the at least one other syringe and the corresponding reagent input port of the fluid cartridge. Analyzer arranged to be arranged.

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