TW504491B - Chip-type device for counting/classifying and analyzing the micro-fluid particle and manufacturing method thereof - Google Patents

Abstract

The present invention provides a chip-type device for counting/classifying and analyzing the micro-fluid particle and manufacturing method thereof which integrates an optical fiber structure and micro flow channel onto a chip by means of a fast and low cost processing manner for counting/classifying and analyzing the micro-fluid particle. The present invention uses a process of micro-electromechanical system to produce both micro-flow channel and optical fiber axial passage in a chip, and then, fills the optical fiber axial passage with material of high refractive coefficient, utilizing the difference between the refractive coefficient of the chip body and that of the filler, to make total reflection of the light occur at the interface between the aforementioned chip and the filler, so as to form an optical fiber structure. The inspecting light is transmitted via a set of optical fiber structure and passed through the sample flow, and the inspection of particle counting can be carried out through the absorption and scattering of the light by the micro-particles of the sample flow matching up with the variation of light intensity measured by an external photo detector. The chip of present invention further integrates a micro flow channel to make it possess the function of sheath flow, by which it can focus the fluid and to perform classification for various cell. The present invention is applicable to various particles, such as count, speed measurement, and classification of the cell and blood cell, and measurement of various bio-medical fluorescence analysis.

Description

504491 V. Description of the invention ^ ^ ----- [Technical Field of the Invention] ^ The invention provides a wafer-type microfluidic particle counting / sorting and classification method and its manufacturing method, which integrates light with a fast and low-cost process. And microfluidic channels on the wafer 'for counting / sorting and analyzing microparticles such as cells, blood cells, etc. [Background of the Invention] In recent years, due to the development of micro-electro-mechanical process technology, many of the original magic components have been miniaturized, and humans can gradually use the miniaturized equipment to perform experiments that were previously impossible, such as the manipulation of cells and Fast protein or DNA separation ... etc. In many fields of micro-electromechanical research, the application of components to biomedical testing is particularly valued. The microfluidic biomedical detection chip produced by the micro-electro-mechanical process technology has the advantages of high detection efficiency, low sample consumption, low energy consumption, small size, and low cost. Especially, it integrates the micro-fluid system and the detection mechanism in the same The design on the chip has the most development potential and market value. Therefore, in addition to the advantages of miniaturization, it also dispenses with complex and expensive testing equipment, so that a single chip has a complete testing function.

Counting and sorting of cells using a f cytometer has been widely used in biomedical testing, and the results are also one of the important basis for clinical diagnosis. The traditional cell counter is shown in Figure 1. The sample is injected through a central pipe, and the side sheath flow (sheathf 1 0w) injected from both sides is used to reduce the central sample flow to a single unit using the principle of hydrodynamic focusing. Cell width 'then let the sample flow through the electrode or optical detection mechanism

504491

Count. The detection mechanism is integrated in the chip, but for the development trend of solar tablets with a volume of only a few micrometers, the electrode through which the cells flow is not easy. The clothing can never resolve a single height of tens of micrometers. The direction must have an electroformed method to make the electrode. There are structural designs: trapped iUcheng; use of poor resolution and other issues. M and the detection space solution The typical design of the electrode method is, but whether it uses a capacitor or a meter-wide flow channel to detect cell flow, and in this design, the arrangement of the electrodes is very complicated. Not used to measure parallel electrode plates

The slight changes in electrical properties caused by dynamics and its insulation will make the process unskilled and should be fluorescently stained. Or, using a single material, the optical detection must be in the sample, and then the cells are used to light. < 5 = The optical fiber guides light into the sample for flow measurement. However, both of these two methods; ow = good measurement results of scattered light externally, C ': * Although the light staining method can damage the cells, it loses the original = so, ah, but the staining procedure Complex and possible calibration of optical substances is fine, but the optical fiber *: μ 2 fiber method is not necessary to use a fluorescent question, and i. To 2. The micron inner diameter connection is-大 问) # 炉 俨 姝 说 眚 ， 不 碥 ^) The early complaints (Slngie de) first dwarfed the shell, which is not suitable for commercial mass production. [Summary of the Invention] In view of the fact that the traditional technique of microfluidic cell counting from the 4+ to the 叹 # 叹 八 昍 日 日 / sigh has traditional process defects and disadvantages, the purpose of the present invention is to recover a trace of au, /

Particle counting

/ Knife and analysis of cracking and its manufacturing method. The main purpose of the Zhuang invention is to provide a wafer-type microfluidic particle. The knife counts one piece of clothing and at least includes: a pump for providing fluid drive; for the i-series to integrate microchannels and optical fiber structures on it A light source, which refers to 2, the optical fiber structure; a light detection unit; and a signal processing unit that processes the signals generated by the aforementioned light detection unit. ^ Said 'pu system is injection pump (Sy r i n g e p u m p). Road. The channel from G mainly includes the sample flow micropipes and side sheath flow micropipes. For example: laser (such as green light

The aforementioned light source may be various light emitting units (red light laser) or various LED light emitting diodes. The two light detection units include: a light detector and a signal amplifier. The light signal is used for amplification. The processor and signal processing unit mainly include: an analog / digital signal converter. The Keshu computer system can control the operation of the aforementioned pumps, and analyze the signals of the < prediction unit >. ... Another purpose of the invention is to provide an integrated optical fiber structure on the chip

The channel includes the following steps: buying a substrate; engraving the optical fiber on the substrate; combining two wafers with the same fiber channel into the aforementioned fiber channel; and forming an optical fiber structure. The method of Turismo may further include: coating the fiber channel surface after the etching first with a substance having a lower refractive index than the substrate, such as low-refractive material and organic materials to increase the interface between the substrate and the filler The refractive index difference 'and the surface inside the channel can be flattened. ^,

Page 7 504491 V. Photographs of sample sheath fiber made of materials. This diversion is used to edit the description of the invention. (4) The aforementioned wafers and the aforementioned fillings can be used. In the field of analysis, the method of the present invention is used, which can be the first product of the present invention. At least; and at least three pairs of non-computing micro-channels to detect substrates such as glass entering in a double beam line. ^ = Refers to the material whose refractive index is higher than the refractive index of the base material, like knife materials, organic solutions, etc. The method of optical fiber structure on the wafer, combined with the method of manufacturing, and using micropipes and optical fiber structures at the same time ~ for the counting / sorting of particles in microfluids or the above-mentioned manufacturing of various optical fiber structures and micropipes on the wafer The implementation of the above-mentioned wafer. A wafer according to an embodiment includes at least one structure serving as a sample as a lead wire transfer wheel. An embodiment has at least two: at least three of the two sides of the fiber are connected to the fiber at the same position as the optical fiber. The micropipe 4 is used for transportation and diversion; at least one group (two) of the side is used to draw fluid. For use; and at least one pair of lights. The aforementioned pair of optical fiber structures refers to the sheath-flow microport microtubule structure of two wafer types or more, and the structure is calculated by using a sample. It can be a multi-flow microfluidic flow microtubule pipeline. It is used to count the particles in the crystal as two rays of light are transmitted into a plurality of light by parallel tracking. The channel is used as a sample to introduce the edge sheath flow for the export of the sample. Beam lines and intersection lines for n AND "number of logical particles. Lido flow channel sample flow 504491

For example, the wafer of the third embodiment of the present invention is capable of detecting cells in parallel at the same time. Counting of micro-particles, dividing smoke, soil, soil, aa body, knife, and speed measurement, this chip contains at least: at least one sample flow micro-pipe, which is used as an inflammation pipe, and is used as a guide 2 :: ¥ to enter; at least The two sets of side sheath flow are slightly used for v-to-side sheath flow fluid; at least one of the outlet moaning tubes is used for sample derivation and u-line transmission. Use, and to V two pairs of Guangyi ,,,. Structure, as light and 隹: ΐ to 两组 two sets of edge sheath flow microchannels 'one of which is used as a sample flow', and the other is to transport different particles in the sample flow to different micro The two sets of edge-flow microchannels are arranged on the wafer in a back-and-forth arrangement. — The at least two pairs of optical fiber structures are arranged one behind the other for velocity measurement of particles and fluids. β The third embodiment of the present invention uses two parallel beam lines to perform particle flow measurement. The flow velocity of the particles in the fluid is calculated from the time difference between the particles passing through the two beam lines in the sample stream, and automatically calculated by the computer. After analyzing the parameters, the pump thrust of the second group of sheath flow is controlled to change the M velocity of the second group of sheath flow, and the particles in the sample stream are transported to different outlet microchannels according to their size or quantity, and various Required biomedical tests. The wafer according to the fourth embodiment of the present invention provides an analysis wafer for glory detection, including at least one sample flow microchannel for sample flow; and at least one optical fiber structure for light For transmission. The application of the fourth embodiment of the chip described above is characterized in that the optical fiber is bonded

$ 9Page V. Description of the invention (6) The structure is integrated in a & μ — " " " ",using; plus fluorescence ;: f :::, the sample is introduced into the aforementioned microchannel-through :: the optical fiber The structure excites the fluorescence of the sample :: =: J :. The present invention uses optical method to detect the fluorescence. Spheres or all kinds of tiny particles ... etc) Λ / edge (for example, · cells, blood fiber structure integrated in the chip, the solution of the structure of the structure, mainly defined, so you can easily mark 2 lithographic development program Therefore, the water flow focusing system 'completes a process and simply integrates the microfluidics of the optical alignment of the optical fiber detection mechanism; the refractive index between the inner and outer layers of the fiber structure is very different, so a polar body can be used as the detection light source, greatly Reduce the cost of detection equipment =. This kind of microfluidic particle counter can perform the function of detecting specific particles among many particles, and then distinguish their size, and cooperate with the sorting mechanism at the back end to quickly detect and collect specific Particles are very important sample pretreatment devices in biomedical wafers. [Comparison of main component symbols] Sample flow micropipes 1, 171, 172, 173 18 3 4 67 8 Edge sheath flow micropipe wafer substrate fiber structure particle light Sentiment unit pump < 1

Page 10 504491 V. Description of the invention (7) 9 --- Light source 10 --- Chip 11-Signal processing unit 12-Analog / digital signal converter 13 --- Computer 14 --- Intermediate layer 15- -Sample injection holes 1 6 and 1 6 'Edger flow holes 17 --- Outlet holes 19 --- Outlet microchannels 20, 20' --- One wafer 21 --- Fluorescence excitation light source 30 --- One wafer 31 --- Photoresist layer 32 --- Photomask 41 --- Fibre channel 7 0 --- Optical valence detector 71 --- Signal amplification 100 --- Chip-type microfluidic particle counting / sorting and analysis device 20 0 --- Fluid concentration mechanism 3 0 0 --- Optical fiber counting mechanism a, b, c, d --- Beamline [Detailed description of the invention] The present invention relates to the counting / classification and analysis of wafer-type microfluidic particles.

Page 11 丄 丄

V. Description of the invention (8)

The device 100 and its manufacturing method are explained and improved in accordance with the drawings. Many advantages and features will be understood from the following detailed description of the wafer type cold step of the present invention. It mainly consists of two major parts: particle counting / sorting and analysis device 100. Concentration mechanism 200, as shown in Figure 2. The first part is sample flow injection and fluid 3 0, and its operation principle is shown in Figure ::, The second part is the optical fiber counting mechanism. First, referring to FIG. 2 (A), the schematic diagram of the centralized mechanism "t ~", which is the sample stream injection and fluid sample stream micropipes of the present invention. The structure is two in the Λ structure 200 and contains at least one (two) edges. Sheath flow microchannel 2 ',] a: t 入 的 用' and at least -groups ^. ^, Ru, which are located in the aforementioned sample flow microtubes 1 \ are used as saddle fluid guides. This fluid concentration The mechanism uses = side sheath flow to reduce the central sample flow to the visibility of a single particle (such as a cell), so that the particles in the sample flow can be aligned as needed before the flow

Advance. Please refer to Fig. 2 (B) and Fig. 2 (C), where Fig. 2 (B) is an image diagram of the mechanism of Fig. 2 (A), and Fig. 2 (C) is a mechanism using the mechanism of Fig. 2 (B) for dye testing An image of the effect of water concentration. The test results in Figure 2 (C) show that the width of the dye can be reduced to 3 // m after focusing by the side sheath flow. After focusing the sample through the side sheath flow, it can enter the second part of the fiber counting mechanism 300, which will be described in detail through Figure 3. Next, referring to Fig. 3, Figs. 彡 (A) and (B) respectively show an upper view and a side view of the present invention using Snell's law to integrate an optical fiber structure into a wafer structure. The present invention uses a low-cost wafer substrate 3 (for example, glass), and the micro-pipe 1 and the channel of the optical fiber structure 4 of the sample stream are engraved on the aforementioned substrate 3 by wet etching, using refraction

504491 V. Description of the invention (9) A glass with a coefficient of about 1.5 is used as the material of the wafer substrate 3, and a material with a refractive index greater than 1.5 is inserted in the channel, such as: a refractive index of i 8 is different from _8 negative Photoresist, so that the fiber structure 4 can be formed. When the light travels inside the aforementioned filling material, because the outer surface is coated with a glass material with a smaller refractive index, if the incident angle of the light is greater than the critical angle, according to Sone's Law (S oe 11, s law), the light will be totally reflected In the medium with a high refractive index, = if the filling material is a SU-8 negative photoresistor and the sweep angle between the light and the interface is smaller than Tian 5. When,, the light will be totally reflected into the high-refractive index 讥 _8 negative light. Use light to travel through the filling material due to total reflection instead of denier principle 'design appropriate fiber path' and use the standard: the optical fiber structure 4 channel, and then fill the high refractive first / 釭 order clothes with the structure 4 'U incidence The light transmission to the sample stream micropipes: The difference in light intensity at the exit end caused by: or = different degrees, and an external light detection unit 7 was collected by an external light detection unit 7 to measure the passage of cells and calculate its number. ° k 'can be detected immediately as described above, the wafer-type micro-analysis device 100 of the present invention is mainly based on the combination of the aforementioned sample flow sampling number /: = 2000 and the optical fiber counting mechanism 3000, '; IL body centralized mechanism operation unit. Figure 4 shows: Sending :::: Measurement and signal counting / sorting and analysis device 1 2 00 Type 2 microfluidic particle pump 8, which is used to provide fluid-driven wide-return 1 at least: 1 4; a light detection unit 7; and one ton > old buckle first to month '' 'described fiber structure' 娆 processing unit 11, which processes the aforementioned light

504491 V. Explanation of the invention (ίο) Signal generated by the detection unit 7 The device 100 of the present invention introduces the sample flow and the edge sheath flow through the aforementioned pump 8 ^ Do not introduce the sample flow micropipe i and the edge sheath flow on the wafer 10 In the micro-channel 2, the sample stream is focused to a specific degree above 4 by controlling the flow velocity of the fluid and the sample stream on both sides of the sheath, for example, the width of a cell, and then the fiber structure light source 9 emits light, and the light is emitted from the wafer. " In the light, cut, and the entrance end of the mouth structure 4 enters and passes through the sample stream, at this time, i: 6 is made after receiving or scattering one light two or two ^ Detect the light center production structure 4 The output side outputs' and enters the "light-emitting" by the light detection unit 7 u and transmits the aforementioned optical signal to the signal processing unit 订 i for ordering analysis and counting. Heart stem d: 8, is an injection pump ( Syringe pump). Channel 3, ... "Forcing mainly includes: sample flow microchannel 2 and edge sheath flow microtubules or = various light-emitting units' for example: laser (such as: green light flying, first, the first series) or Various LED light emitting diodes. The aforementioned light detection unit 7 includes: a light detector Π. The aforementioned signal amplifier 71 is tested. . The 70 and-signals are released and can increase the sensitivity of debt measurement. The aforesaid signal processing unit n, which is said to be amplifying the measured optical signal, is mainly composed of a converter 12 and a computer 13. The aforementioned computer = may; several bits of signal conversion, the flow rate of the product flow and the edge sheath fluid, and the signal and analysis. The method for manufacturing the optical fiber structure 4 on the wafer 10 of the present invention will be described with reference to FIG.

Page 504491

And Figure 6 for a detailed description. FIG. 5 is a block diagram showing the manufacturing method of the optical fiber structure of the present invention including at least the following steps: providing a wafer substrate 3; etching the optical fiber through the channel θ on the aforementioned substrate 3; joining two pieces having the same meaning as the aforementioned optical fiber channel 41 Material 3; a filling material in the aforementioned optical fiber channel; and forming an optical fiber structure 4. The foregoing method may further include: coating a layer of a substance having a lower refractive index than the substrate 3 on the surface of the optical fiber channel after the etching, such as a low refractive index organic substance, to increase the distance between the substrate 3 and the filling material interface. Refraction is a refractive error, and can flatten the surface in the channel. The aforementioned wafer substrate 3 is made of glass or polymer materials, for example: acrylic (PMMA), polycarbonate (PC) or polydimethylsiloxane (pDMs). The aforementioned filling material means that the refractive index is higher than the wafer The material of the refractive index of the base material 3 may be, for example, a polymer material, an organic solution, etc. The aforementioned filling material is a SU-8 photoresist. Then, in each of the processes (A) to (G) in FIG. 6, The manufacturing process of the optical fiber structure of the present invention is described in detail as follows:

The manufacturing method of the present invention mainly includes the following steps: as shown in FIG. 6 (A), first, a wafer substrate 3 is provided, and a photoresist layer 3 1 (eg, AZ462 0 photoresist) is coated thereon, and Use a designed photomask 32 to perform the lithography (Lithography) step; then, as shown in Figure 6 (B), perform the PR developing step; then, as shown in Figure 6 (c), In the step of engraving the glass surname (G 1 ass et ch i ng), the wafer substrate 3 is etched into a fiber channel 41 of a predetermined size (for example, a width of 70 mm and a depth of 2 mm).

Page 15 504491

V. Description of the invention (12) # m); After that, as shown in FIG. 6 (β), a photoresist stripping (pR stripping) step is performed to remove the remaining photoresist; next, as shown in FIG. 6 (E) As shown in the figure, two wafer substrates 3 having the same optical fiber channel 41, which are also prepared through the steps of FIGS. 6 (a) to (D) above, are aligned and joined; it is increased to flatten the surface inside the optical fiber channel 41 Light transmission efficiency, reducing the light scattering effect caused by uneven surface, then the intermediate layer coating step can be performed to refract

A substance having a lower rate than the wafer substrate 3 is coated on the surface of the optical fiber channel 41 to form an intermediate layer 14, such as: an organic series of spin-on glass (SOG), as shown in FIG. 6 (F); finally The high-refractive-index material is filled in the aforementioned optical fiber channel 41 by a vacuum suction method to form the optical fiber structure 4 of the present invention, as shown in FIG. 6 (g). By using the above-mentioned manufacturing method of the optical fiber structure of the present invention in combination with the conventional manufacturing method of the micro-guan on the cymbal, various wafers can be made that integrate the optical fiber structure 4 and the micro-guan. Describe in detail. The first embodiment is shown in FIG. 7, which shows a wafer according to the first embodiment of the present invention, which includes a sample flow microchannel, which is used for sample diversion; a set of (two) side sheath flow microchannels 2. It is used to introduce the fluid in the side sheath; and a pair of optical fiber structures 4 is used to transmit light. Counting the particles in the microfluid can be performed by combining the wafer of the first embodiment with the device of FIG. 4 (such as: pump 8 light source 9, light detection unit 7, and signal processing unit 1 1). First A, separate the sample stream and the sheath fluid on both sides

504491 V. Description of the invention (13) Do not introduce from the sample injection hole 15 and side sheath flow hole 16 of the wafer 10, and focus the sample to the width of one particle by the flow rate of the sample stream and the side sheath flow on both sides. For example: the width of a cell, after which the particles flow through the optical fiber structure 4 sequentially and exit from the exit hole 17. An external light source 9 emits light, which enters from the entrance end of the optical fiber structure 4 in the wafer 10 and passes through the sample stream. At this time, the particles 6 in the sample stream absorb or scatter the light and cause the light intensity to change. The aforementioned light is output through the exit end of the fiber structure 4 at the other end, and the light detection unit 7 detects the change in light intensity, and transmits the aforementioned optical signal to the signal processing unit 丨 丨 for analysis and counting operations. The second embodiment is parallel: the wafer of the second embodiment of the present invention is a multi-flow channel = wafer: it can perform at least two or more microfluid medium particles at the same time. 1 71, 1 72, and ¥ are used for the flow into the side sheath fluid; nine outlets are micro-exposed :: used for the purpose; and four pairs of optical fiber junctions • (not shown in the figure) are formed by the process of multiple pairs of optical fiber structures Multiple beam lines (such as lines a, b, c, and d) intersect at different positions in each micro-pipe. $ 束 === 光线 为 “Logical operation” is used to measure micro :: The number of particles in the sample. Use Double beam, forcing; 7 simultaneous multi-channel sample flow (for example: fine movement measurement. For example, 'beam line_beam line b intersection shown in Figure 8; Road five, description of the invention (14) T, of / / 2 Top—Using the two sets of beam lines a and b for "AO" logic to detect that the particles pass through, it means that the sample is hungry for particles to flow through. Therefore, the counter of the sample flow is calculated. When the beam line a and the beam line 〇Number of QUs at the same time. When the # # 4 4 on the same micro-pipe 1 72 passes, it indicates the parallel detection of multiple channels. B. The buccal push can enter the third embodiment at the same time. As shown in FIG. 9 (A), the second to second aspects of the present invention are wafers 20 that can simultaneously execute the two consecutive embodiments of microparticles in microfluidics. The wafer 20 includes: a sample stream Micro-pipes} for classification and speed measurement. The two sets of side-sheath flow micro-pipes 2, 2 are used for sample introduction. The seven outlet micro-pipes 19 are used as samples. The use of side sheath flow fluid; the structure 4 'for light transmission. The use; and two pairs of optical fiber junctions. The first group (ie, the sheath microtubule 2 ') is a sample of different particles in the sample stream. The other group (ie, sheath sheath flow, 19; the two sets of sheath sheath microtubules described above reach different outlet microtubule configurations. On the wafer 20. 2 'is arranged in front and back, and it is described in the optical fiber structure of at least two pairs of 4 counting and fluid velocity measurement.', Phase parallel arrangement, as the particle diagram 9 (B) the third embodiment of the present invention As a result, the film 20 is compared with the image of the wafer 20 in Figure 9 (a), and the image of the chip 20 is different. First, the sample is flowed And the two sides = the sample injection hole 15 with three outlet micros and the first set of edges and holes are introduced from the wafer hole 16 respectively, and sample 504491 V. Description of the invention (15) ^ --- Focused on the width of one particle, the two parallel beam lines generated by flowing through the well fiber structure 4 pass through the middle = :::: difference, calculate the flow velocity of the particle in the fluid, and 4 2 1 I. After automatically analyzing the parameters through the computer, the sealing force of the second group of side sheath flow (side sheath flow micropipe 2,) is controlled to change the flow velocity of the second group of side sheath fluid. Sakata 1 / 丄 面 丄 σ, 士 丄 ^ t ^ Teeth nw μ, the method of switching water flow ~ to the different sample outlets in the official road 19, and send out the sample hole from the exit hole 17 to the particles in the sample stream for various purposes. Biomedical testing. The fourth embodiment is shown in FIG. 10. The wafer 30 of the fourth embodiment of the present invention provides: an analysis wafer 30 for fluorescent debt measurement, including: a sample flow microtube system for sample flow , And an optical fiber structure 4, for transmission purposes. The wafer 30 according to the fourth embodiment of the present invention is characterized in that an optical fiber structure is bonded to the wafer 30. Introduce the sample with the fluorescent cursor into the aforementioned microchannel 'excitation of the fluorescent dye in the sample with the additional fluorescent excitation light source 2 1 and excite the fluorescent radiation of the sample through the optical fiber structure 4 through the light detection Unit 7 performs detection. The application of this chip can be integrated with capillary electrophoresis analysis, or analysis of intracellular material after cell polarization destruction and detection of MA. As long as the embodiments of the present invention mentioned above do not depart from the gist of the present invention, ‘the various microfluidic pipelines and optical fiber structures may be modified in number and design’, the scope of protection is defined by the scope of patent applications below. [Effect of the invention]

504491 V. Description of the invention (16) The present invention provides a wafer-type particle counting / sorting and analysis device and a manufacturing method thereof, which have the following advantages: 1. The present invention can quickly, simply, and reliably produce an optical fiber integrated micro-particle ( For example: cells, blood cells) and bioassay analysis wafers. This process does not require any expensive vacuum coating or plasma etching equipment. It only requires standard fiber development procedures and chemical wet etching steps, and is supplemented with high-reliability fusion bonding technology to complete the aforementioned various aspects of the wafer of the present invention. 2. The present invention integrates the optical fiber structure in the wafer, so no optical alignment mechanism is required for operation, eliminating bulky and expensive optical equipment, and using cheap light sources (such as light emitting diodes or semiconductor lasers) ) As a detection light source, with the miniaturized light detection unit, the volume of the entire detection device can be greatly reduced, and made into a portable detection device. 3. The device of the present invention can be integrated with a computer detection system, and the measurement signals are processed by the computer in real time to perform real-time counting functions and data analysis. In summary, the present invention can be used in various analytical fields such as biochemical analysis, medical detection, etc. to improve the well-being of human society.

Page 504491 Brief description of the diagram Figure 1 is a schematic diagram of the principle of a typical cell counter. Figure 2 (A) is a schematic view of a sample stream sampling and fluid concentration mechanism of the present invention. Figure 2 (B) is an image of the mechanism of Figure 2 (A). Figure 2 (C) is an image of testing the effect of water flow concentration using the mechanism of Figure 2 (A). Figure 3 is a schematic diagram of the structure of an optical fiber on a wafer of the present invention. FIG. 4 is a schematic diagram of a wafer-type microfluidic particle counting device according to the present invention. FIG. 5 is a block diagram of the manufacturing method of the optical fiber structure of the present invention. FIG. 6 is a flowchart of the manufacturing method of the optical fiber structure of the present invention. Fig. 7 is an image diagram of a wafer for counting microfluidic particles according to the first embodiment of the present invention. Fig. 8 is a schematic diagram of a wafer for multi-channel parallel detection according to the second embodiment of the present invention. Figure 9 (A) is a schematic diagram of a wafer-type microfluidic particle counting / sorting and counting device according to a third embodiment of the present invention. Figure IX (B) is a wafer image of Figure IX (A). Fig. 10 is a schematic view of a fluorescence analysis wafer device according to a fourth embodiment of the present invention.

Page 21

Claims (1)

504491 6. Scope of patent application 1. A wafer-type microfluidic particle counting / sorting device, at least including: a pump for providing fluid driving; a chip for integrating microchannels and optical fiber structures on it; a light source for Providing light to the aforementioned optical fiber structure; a light detection unit; and a signal processing unit for processing signals generated by the light detection unit. 2. The wafer-type microfluidic particle counting / sorting device described in item 1 of the scope of patent application, wherein the aforementioned pump system is an injection pump. 3. The wafer-type microfluidic particle counting / sorting device described in item 1 of the scope of the patent application, wherein the aforementioned microchannels mainly include: sample flow microchannels and edge sheath flow microchannels. 4. The wafer-type microfluidic particle counting / classifying device described in item 1 of the patent application scope, wherein the aforementioned light source may be a laser or a light emitting diode. 5. The wafer-type microfluidic particle counting / sorting device as described in the first item of the scope of patent application, wherein the aforementioned light detection unit includes: a light detector and a signal amplifier, and the aforementioned signal amplifier will detect it. Optical signal amplification. 6. The wafer-type microfluidic particle counting / classifying device as described in item 1 of the scope of the patent application, wherein the aforementioned signal processing unit mainly includes: an analog / digital signal converter and a computer. 7. The wafer-type microfluidic particle counting / sorting device described in item 6 of the scope of patent application, wherein the aforementioned computer is capable of controlling the operation of the aforementioned pump. 8. — A method of integrating optical fiber structure on a wafer, including at least the following steps Page 22 Providing a wafer substrate;: = a fiber channel on the aforementioned wafer substrate; combining two wafer substrates with the same fiber channel; a filling material in the aforementioned fiber channel; and forming an optical fiber structure. 9. The integrated optical fiber structure as described in item 8 of the scope of the patent application: the method, wherein the method described above may further include: on the surface of the film after the last name is engraved, the coating coefficient of the A coating is lower than that of the substrate The thing ^ fibrous interlayer. F Ma Zhong <1 1 0 · Integrate the optical fiber structure on the wafer side as described in item 9 of the patent scope, where the aforementioned substance is 11 of the low refractive index · as described in item 8 of the patent scope The integrated optical fiber structure II. Method: Wherein the aforementioned wafer substrate is glass or high: 材料: The material system can be: acrylic (PMMA), polycarbonate (PC) or bin-siloxane (PDMS). -Said 12. The method of integrating the optical fiber structure and method as described in item 8 of the scope of the patent application, wherein the aforementioned filling material refers to a material having a refractive index above ^. Be sure to return to the base material. Integrate the optical fiber structure described in item 12 of the wm circle. Item 12: The aforementioned filler material in the liquid is polymer material or organic. 14. Fang Shen: Specialty The method of integrating the optical fiber structure on a wafer as described in Item 12, and the filling material described by Zhongke is a SU-8 photoresist. 504491 VI. Application patent scope 1 5. A kind of wafer, which can be used for microfluidic particles For counting, it includes at least: at least one sample microchannel used as a sample guide; at least one (two) side sheath flow microchannel used as a side sheath flow fluid; and at least one pair of optical fibers The structure is used for light transmission. 1 6. —A wafer, which can be used for multi-channel parallel detection of microfluidic particle counting, the wafer contains at least: at least two sample flow micro-pipes, which are used for sample introduction; At least three side flow micro-channels are used to introduce the flow into the side sheath flow; at least two outlet micro-channels are used to export the sample; and at least three pairs of optical fiber structures are used for light transmission. Apply for The wafers described in item 16 of the scope can be used to count particles in at least two or more microfluids at the same time. 18.-A wafer including at least one sample flow microchannel for sample introduction; At least two sets of side sheath flow microchannels are used to introduce side sheath flow fluid; at least one outlet micropipe is used for sample export; and at least two pairs of optical fiber structures are used for light transmission. The wafer described in item 18 of the patent scope is capable of performing counting, sorting, and speed measurement of fine particles in microfluidics at the same time. 20. The wafer described in item 18 of the patent scope, wherein at least two of the foregoing I SBli! Page 24 Scope of patent application ^ Set of side sheath flow microchannels, of which the 〆 group is used for focusing the sample stream, and one group is used to transport different coarse particles in the sample stream to different microchannels. 21 · The wafer according to item 8 of the scope of the patent application, wherein the at least two pairs of the optical fiber structures are arranged side by side 'are sufficient for particle counting and fluid.疋 之 2 2 · A wafer, including at least: at least one sample flow micro-pipe is used for sample flow; and at least one optical fiber structure is used for light transmission. 2 3 · If the wafer described in item 22 of the patent application scope, the aforementioned wafer is an analysis wafer that can be applied to optical detection. 2 4 · The chip according to item 23 of the scope of patent application, wherein the aforementioned optical detection is fluorescent detection. 2 5 · The wafer as described in item 5 of the patent application, item 16 or item 19, wherein the aforementioned particle system is a cell, a blood cell, or a variety of fine particles. Item 16, Item 16 or Item 18, wherein the aforementioned wafer is made of glass or a polymer material. 27. If the chip described in item 26 of the patent application scope, the aforementioned polymer material may be: acrylic (PMMA), polycarbonate (PC), or polydisiloxane (PDMS) . Page 25