KR100788313B1 - Bio-sensor Chip of having multi-channel - Google Patents

Abstract

There is provided a multi-channel biochip having a plurality of inlets and a plurality of inlets for simultaneous analysis of different biological samples. The coupling between the upper substrate and the lower substrate is easily achieved without using a separate fastening means. To this end, the upper substrate and the lower substrate is made of a polydimethyl-siloxane (PDMS) material. In addition, the thin film layer of the PDMS material is provided between the upper substrate and the lower substrate, the thin film layer is elastic, the upper portion is provided with a valve. The elasticity of the membrane layer allows the valve to control the flow of biological samples or buffered solutions.

Description

Bio-sensor Chip of having multi-channel

1 is an exploded view showing a multi-channel biochip according to a preferred embodiment of the present invention.

2A and 2B are plan views illustrating upper substrates according to a preferred embodiment of the present invention.

3 is a plan view showing a lower substrate according to a preferred embodiment of the present invention.

4 is a plan view showing a reaction substrate according to a preferred embodiment of the present invention.

5A and 5B are perspective views illustrating an upper case and a lower case of a biosensor chip according to a preferred embodiment of the present invention.

6 is an exploded view showing that the case is provided in the biosensor chip, in accordance with a preferred embodiment of the present invention.

7A and 7B are cross-sectional views illustrating the operation of a valve according to a preferred embodiment of the present invention.

8A to 8C are cross-sectional views illustrating an upper substrate, a lower substrate, a thin film layer, and a method of combining them according to a preferred embodiment of the present invention.

9 is a cross-sectional view of a biosensor chip for explaining that a biological sample or a buffer solution is injected and discharged through a recovery port according to a preferred embodiment of the present invention.

Explanation of symbols on the main parts of the drawings

100: lower substrate 200: thin film layer

250: valve 300: upper substrate

400: reaction substrate 500: metal thin film layer

600: Prism 700: upper case

800: lower case

The present invention relates to a microfluidic delivery and analysis device for detecting and analyzing protein samples, and more particularly, to a device capable of comparatively analyzing a plurality of protein samples using surface plasmon resonance (SPR). It is about.

The biosensor chip is a tool for confirming the existence of various biological samples such as DNA, RNA, protein, and cells, or performing qualitative or quantitative analysis thereof. In particular, the classical method of performing analysis on biological samples is a blot hybridigation method.

This is a method of attaching DNA, RNA or protein to a membrane filter and forming hybrid molecules using specific intermolecular binding on the filter. According to the blot hybridization method, Southern blotting for transferring DNA to a membrane filter and fixing it, Northern blotting for fixing RNA, and protein are fixed according to the type of biological sample probe to be fixed. Western blotting and the like. All of these methods basically utilize a binding reaction with a ligand that selectively binds to the ligand.

However, the above-described methods have to be performed for each sample, and despite the very repeated experiments, it is difficult to perform the experiments under the same conditions, and requires a lot of manpower, time and huge resources.

An indirect method for analyzing ligand and ligand binding of the biosensor chip described above is a fluorescence assay using a secondary antibody reaction. However, fluorescence assays require the development of a secondary antibody that binds to a specific protein (primary antibody), and a protein containing fluorescent material capable of binding to the secondary antibody. This can only be met if a great deal of labor, time and capital is required. Therefore, much research has been made on analytical methods that do not use fluorescent materials.

Surface plasmon resonance refers to a phenomenon of optically inducing vibration of electrons occurring on a thin metal thin film. The reason why the surface plasmon resonance is used as a bioreaction detector is that the surface plasmon resonance is very sensitive to physicochemical changes occurring on the metal thin film surface (or 300 nm or less from the surface) (particularly, the change of the resonance angle or wavelength). Because. Therefore, the interaction of various types of biomolecules can be analyzed at the nano level by constructing various sensing interfaces based on the chemical surface structure of the metal thin film. In addition, surface plasmon resonance can suppress damage or modification of the sample at the time of measurement compared to the other methods described above, and does not require the development of a secondary antibody that is a marker like fluorescence analysis, and can measure biochemical reactions with high sensitivity. It has an excellent advantage.

A device for detecting protein bioreaction using surface plasmon resonance described above has already been commercialized, and the most advanced type is equipped with a microfluidic system to minimize the initial amount of the biological sample and the amount of loss due to the reaction experiment. can do.

U.S. Patent 5313264 discloses a biosensor chip incorporating a micro pneumatic valve. The patent has a structure in which compressed air is stored in a small internal pressure reservoir, and compressed air is introduced using a solenoid valve. In addition, it has a structure in which one injection port for injecting a sample, etc. and one discharge port for discharging the sample which has been measured are provided, respectively, and the upper plate and the lower plate for closing the flow path from the outside have a firm and constant hardness such as plastic, metal or ceramic. Use the materials you have. In order to combine them, appropriate fastening means for fastening the upper plate and the lower plate in addition to the housing surrounding the outside should be used. However, the patent does not disclose such fastening means. In particular, since the inlet and outlet are respectively provided inlet and outlet, the measurement of one sample is performed, there is a hassle that must clean the inlet and outlet.

The first object of the present invention for solving the above problems is to provide a multi-channel biochip that can fasten the upper substrate and the lower substrate without a separate fastening means, and at the same time can be injected or discharged different types of biological samples To provide.

In addition, a second object of the present invention is to provide a method for manufacturing a multi-channel biochip according to the first object.

The present invention for achieving the first object, the lower substrate having a pneumatic / hydraulic flow path which is supplied with pneumatic or hydraulic pressure from the outside, and made of a polydimethyl-siloxan (PDMS) material; The biological sample or the buffer solution is formed on the lower substrate and is made of polydimethyl siloxane material, and the biological sample or the buffer solution is injected through the injection channels located therein, and the biological sample or the buffer is discharged through the discharge channels spaced apart from the injection channel. An upper substrate on which the solution is discharged, each of the injection channels having a pneumatic / hydraulic control corresponding to each of the pneumatic / hydraulic flow paths; A thin film layer interposed between the lower substrate and the upper substrate and made of a polydimethyl siloxane material and having a valve positioned on a surface interposed between the pneumatic / hydraulic flow path and the pneumatic / hydraulic control part, the valve Is controlled by the flow of the biological sample or the buffer solution is elevated by the pneumatic or hydraulic pressure delivered to the thin film layer through the pneumatic / hydraulic flow path to the pneumatic / hydraulic control unit; A reaction substrate provided on the upper substrate and made of the polydimethyl siloxane material and having a reaction flow path communicating with the injection channel and the discharge channel to flow the biological sample or the buffer solution; A metal thin film layer having an antibody formed as a single layer, for analyzing a biological sample of the reaction flow path; And it provides a multi-channel biochip comprising a prism for utilizing the optical properties of the biological sample bound to the antibody by irradiating light to the metal thin film layer.

In addition, the present invention for achieving the second object is provided with a lower substrate, a thin film layer having a valve, an upper substrate, a reaction substrate, a metal thin film layer to inject or discharge a biological sample or a buffer solution using multiple channels. A method of forming a multi-channel biochip, the method of forming the upper substrate comprising: applying a photoresist on a ceramic or metal substrate; Selectively irradiating ultraviolet rays to the photoresist to form a photoresist pattern; Coating a polydimethyl siloxane film on the photoresist pattern; And separating the polydimethyl siloxane film from the substrate and the photoresist pattern.

The second object of the present invention is to provide a multi-channel biochip including a lower substrate, a thin film layer having a valve, an upper substrate, a reaction substrate, and a metal thin film layer to inject or discharge a biological sample or a buffer solution using a multi-channel. A method of forming the lower substrate and the thin film layer, the method comprising: applying a photoresist on a ceramic or metal substrate; Selectively irradiating ultraviolet rays to the photoresist to form a photoresist pattern; Coating a polydimethyl siloxane film on the photoresist pattern to form the thin film layer and the valve; Curing the lower substrate formed of polydimethyl siloxane in contact with the thin film layer; And separating the lower substrate and the thin film layer from which the silanization bond is formed by the curing, from the substrate.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings.

Example

1 is an exploded view showing a multi-channel biochip according to a preferred embodiment of the present invention.

Referring to FIG. 1, a multi-channel biochip has a lower substrate 100, a thin film layer 200, an upper substrate 300, a reaction substrate 400, a metal thin film layer 500, and a prism 600.

The lower substrate 100 includes a plurality of flow paths, and controls the micro pneumatic valve by pneumatic or hydraulic pressure. In addition, the biological sample and the buffer solution are introduced through the flow path formed in the thin film layer 200 and the upper substrate 300. The reaction substrate 400 is positioned on the upper substrate 300, and a plurality of reaction passages are formed on the reaction substrate 400. The biological sample supplied to the reaction channel reacts with a monolayer or antibody of the biological sample formed on the metal thin film layer 500. That is, the biological sample coupled to the antibody changes the reflection angle of the light source incident through the prism 600, and the components of the biological sample are analyzed by the changed reflection angle.

That is, when the specific protein and the biological sample included in the metal thin film layer 500 do not adsorb, the incident angle and the reflection angle of the light source incident and reflected through the prism 600 will be the same, but the biological sample is adsorbed to the specific protein. When the reflection angle is different from the incident angle, the reflection angle is used to analyze the components of the biological sample.

In addition, the lower substrate 100 is provided with a pneumatic / hydraulic inlet 110, and the upper substrate 300 is provided with a biological sample / buffer solution inlet 310. The biological sample and the buffer solution introduced through the upper substrate 300 are discharged to the recovery port 330 through the reaction substrate 400.

2A and 2B are plan views illustrating an upper substrate 300 according to a preferred embodiment of the present invention.

Referring to FIG. 2A, the upper substrate 300 may include a biological sample / buffer solution inlet 310, a first pneumatic / hydraulic controller 312, an injection channel 314, a second pneumatic / hydraulic controller 316, and a second substrate. The first reaction substrate connector 320, the second reactor plate connector 322, the recovery passage 324, and the recovery port 330 are provided.

A plurality of biological samples / buffer solution injection holes 310 are provided in the upper substrate 300, and two or more biological samples may be introduced at the same time. In addition, the first pneumatic / hydraulic control unit 312 controls the movement of the biological sample or buffer solution introduced into the biological sample / buffer solution inlet 310 to the injection flow path. This is due to the operation of the first pneumatic / hydraulic controller 312 to block the flow of the biological sample or the buffer solution or to continue the flow by the operation of the valve formed on the thin film layer 200. In addition, the second pneumatic / hydraulic controller 316 performs the same operation as the first pneumatic / hydraulic controller 312.

An injection passage 314 is provided between the first pneumatic / hydraulic controller 312 and the second pneumatic / hydraulic controller 316. That is, the biological sample or the buffer solution selected by the first pneumatic / hydraulic controller 312 is introduced into the first reaction substrate connector 320 by the second pneumatic / hydraulic controller 316.

The first reaction substrate connector 320 and the second reaction substrate connector 322 are connected to the reaction substrate 400. Accordingly, the biological sample or the buffer solution introduced into the first reaction substrate connector 320 is introduced into the reaction substrate 400, and the biological sample or the buffer solution passing through the flow path provided in the reaction substrate 400 is the second reaction substrate. It flows into the connecting portion 322.

A recovery flow path 324 is connected to the second reaction substrate connector 322, and the introduced biological sample or buffer solution is discharged to the recovery port 330 through the recovery flow path 324.

In FIG. 2A, four biological sample / buffer solution injection holes 310 are provided, and three recovery ports 330 are provided. In addition, the plurality of biological sample / buffer solution inlets and recovery ports are shown to be listed at regular intervals from one another. However, if two or more biological sample / buffer solution inlets are provided according to the use example, they do not depart from the spirit of the present invention, and the same applies to the recovery port. In addition, the interval between the biological sample / buffer solution inlets and the recovery ports may be irregularly set as necessary.

Referring to FIG. 2B, another configuration example of the upper substrate 300 is shown. In the upper substrate 300 of FIG. 2B, an injection passage 314 is provided for each of the biological sample injection holes 310a. In addition, the buffer solution inlet 310b and the biological sample inlet 310a are provided separately. Therefore, the biological sample may be simultaneously supplied to the reaction substrate 400 through the injection passage 314, and at the same time, the biological sample or the buffer solution may be washed through the injection passage 314.

2A and 2B, two types of pneumatic / hydraulic controllers, such as a first pneumatic / hydraulic controller 312 and a second pneumatic / hydraulic controller 316, are provided. Accordingly, one type of pneumatic / hydraulic control unit may be provided.

Preferably, the width of the injection flow path and the recovery flow path is about 250 um and the depth is set to about 50 um. In addition, the material of the upper substrate is a polydimethyl siloxane (polydimethyl-siloxane; hereinafter referred to as PDMS) that is a transparent and elastic elastomer (elastomer). Since the PDMS has excellent elasticity and mutually high adsorption property, in the case of using the same PDMS material for the thin film layer and the lower substrate in FIG. 1, the upper substrate 300 and the lower substrate 100 without introducing a separate fastening means. Can be assembled.

In addition, in FIGS. 2A and 2B, since a plurality of biological samples / buffer solution inlets and a plurality of recovery ports are provided, a plurality of biological samples may be injected at the same time, and a plurality of biological samples may be simultaneously recovered.

3 is a plan view illustrating a lower substrate 100 according to a preferred embodiment of the present invention.

Referring to FIG. 3, the lower substrate 100 includes a plurality of pneumatic / hydraulic inlets 110 and a pneumatic / hydraulic flow path 130 corresponding to each pneumatic / hydraulic inlet 110.

In addition, a thin film layer 200 is formed on an upper surface of the lower substrate 100, and a valve is provided on the thin film layer 200.

Pneumatic or hydraulic pressure is delivered to the pneumatic / hydraulic flow path 130 through the pneumatic / hydraulic inlet 110, the pneumatic or hydraulic pressure controls the operation of the corresponding valve. In addition, the number of valves corresponds to the pneumatic / hydraulic control unit provided in the upper substrate of FIGS. 2A and 2B. That is, the number of the pneumatic / hydraulic control unit and the number of the valves are preferably provided the same.

In addition, the lower substrate 100 may further include a biological sample / buffer solution injection hole 150. In FIG. 3, the width of the pneumatic / hydraulic flow path 130 is about 250 μm and the depth is set to 50 μm. In addition, the lower substrate 100 is in contact with the upper substrate 300 through the thin film layer 200, the flow path is provided so as not to overlap with the upper substrate 300. Therefore, even if pneumatic or hydraulic pressure is applied to the lower substrate 100, the flow paths provided in the upper substrate 300 are not affected, and the hydraulic or pneumatic pressure is provided only to the thin film layer 200 of the valve.

In addition, the lower substrate 100 is manufactured using PDMS which is a transparent and elastic elastomer, so that the lower substrate 100 can be easily combined with the upper substrate 300 without a separate fastening means.

4 is a plan view illustrating a reaction substrate 400 according to a preferred embodiment of the present invention.

Referring to FIG. 4, the reaction substrate 400 has a first upper substrate connection part 410, a reaction flow path 430, and a second upper substrate connection part 450.

The first upper substrate connecting portion 410 is connected to the first reaction substrate connecting portion 320 shown in FIGS. 2A and 2B, and the second upper substrate connecting portion 450 is the second of FIGS. 2A and 2B. It is connected to the reaction substrate connector 322. In addition, a reaction flow path 430 is positioned between the first upper substrate connecting portion 410 and the second upper substrate connecting portion 450.

In FIG. 4, it is preferable that the material of the reaction substrate 400 uses a transparent and elastic elastomeric chain PDMS. In addition, the reaction flow path 430 is in close contact with the metal thin film layer 500 of FIG. 1. When the biological sample or the buffer solution is introduced into the biological sample / buffer solution inlet 310 of the upper substrate 300, the reaction flow path 430 is provided in an intaglio form so that the biological sample or the buffer solution does not leak, and the metal thin film layer It takes the form that the flow path is formed in contact with 500. In addition, an opposite surface of the surface on which the metal thin film layer 500 is formed is disposed to contact the prism 600. In addition, a refractive index matching oil is applied to a surface in contact with the prism 600 to correct a change in the optical refractive index.

In FIG. 4, the width of the reaction flow path 430 is about 300 μm, and the depth is about 35 μm. In addition, the edge of the reaction substrate 400 is in a concave shape so that the pressure applied to the prism 600 is concentrated and transmitted to the reaction flow path 430. The reaction flow path 430 abuts the metal thin film layer 500, and a monolayer for detecting a biological sample is fixed to the metal thin film layer 500.

The single layer for detecting a biological sample is made of a biological material such as an antibody, and the like, and the metal thin film layer 500 may be formed of gold, silver, copper, aluminum, or the like. Preferably, the metal thin film layer 500 is formed of gold. In order to coat a single layer on the above-described metal thin film layer, a method of fixing using a self assembly monolayer on a gold thin film may be used. In addition, the method of forming a single layer may be a method using a polymer such as nylon, cellulose.

5A and 5B are perspective views illustrating an upper case 700 and a lower case 800 of a biosensor chip according to a preferred embodiment of the present invention.

Referring to FIG. 5A, an upper case 700 of a biosensor chip is shown.

The upper case 700 is provided with a plurality of fasteners 710, the prism mounting portion 730 formed intaglio from the surface of the upper case 700 is provided.

In addition, the lower case 800 is shown in Figure 5b, the lower case 800 is a plurality of fasteners 810, the pneumatic / hydraulic connection for coupling with the fasteners 710 of the upper case 700 830 and biological sample / buffer solutions connections 850 are provided.

The pneumatic / hydraulic connections 830 are connected to the pneumatic / hydraulic inlet 110 of the lower substrate 100. In addition, the biological sample / buffer solution connection part 850 is connected to the biological sample / buffer solution inlet 150 of the lower substrate 100.

However, according to the exemplary embodiment, the biological sample / buffer solution connecting part 850 illustrated in FIG. 5B may be provided in the upper case 700 illustrated in FIG. 5A. That is, when the biological sample / buffer solution inlet 310 is formed on the upper substrate 300, the biological sample / buffer solution connection part 850 is provided in the upper case 700 which is in direct contact with the upper substrate 300. Can be.

The upper and lower cases illustrated in FIGS. 5A and 5B protect the biosensor chip from the external environment. In addition, the upper and lower cases are made of a hydrophilic polycarbonate material to form a coupling with PDMS without intervening an adhesive with the upper or lower substrate. That is, the upper or lower case is attached without using a separate coupling means with the upper or lower substrate, preferably a biological sample, only by locking to the fastener provided in the upper or lower case with a suitable pressure using a screw, The leakage of a fluid such as a buffer solution or oil can be effectively prevented.

Figure 6 is an exploded view showing that the case is provided in the biosensor, in accordance with a preferred embodiment of the present invention.

Referring to FIG. 6, the upper case 700 is coupled to an upper portion of the biosensor 900 illustrated in FIG. 1 and formed of a lower substrate 100, a thin film layer 200, an upper substrate 300, and a reaction substrate 400. The lower case 800 is coupled to the bottom of the biosensor 900. In addition, a prism 600 is provided at the prism seating portion 730 of the upper case 700 to provide optical characteristics of the biological sample in which the biological sample on the reaction substrate 400 is combined with a single layer provided in the metal thin film layer 500. The biological sample can be analyzed.

7A and 7B are cross-sectional views illustrating the operation of a valve according to a preferred embodiment of the present invention.

In FIG. 7A, the lower substrate 100 is coupled to the upper substrate 300, and the thin film layer 200 is provided between the lower substrate 100 and the upper substrate 300. In addition, the biological sample or the buffer solution flows through the injection passage 314 through the injection passage 314 provided on the upper substrate 300.

The valve 250 is provided on the thin film layer 200, and a pneumatic / hydraulic flow path 130 of the lower substrate 100 is formed below the valve 250. In addition, the first pneumatic / hydraulic providing unit 312 of the upper substrate 300 may be positioned above the valve 250 to correspond to the valve 250.

In FIG. 7B, when pneumatic or hydraulic pressure is applied through the pneumatic / hydraulic flow path 130 provided in the lower substrate 100, the thin film layer 200 raises the valve 250, and the raised valve 250 moves upward. The injection passage 314 provided on the substrate 300 is closed.

Through the above-described operation, the valve 250 may control the inflow of the biological sample or the buffer solution flowing from the biological sample / buffer solution inlet 310.

7A and 7B, the lower substrate 100 and the upper substrate 300 are made of PDMS of the same material, and the thin film layer 200 and the valve 250 are also made of PDMS.

In particular, the thin film layer 200 and the lower substrate 100 are made of silanization bonding. That is, when energy such as plasma is applied to the surface of the PDMS, silanization is performed in which a silane group on the surface becomes a silanol group. In addition, when these are brought into contact with each other for a predetermined time or more, the bonds are firmly formed by the chain linkage between molecules.

In addition, the aforementioned silanization bond may be generated between the lower substrate 100 and the upper substrate 300.

As shown in FIGS. 7A and 7B, the thin film layer 200 made of PDMS has excellent elasticity and has a valve 250 protruding from the top. Therefore, when pneumatic or hydraulic pressure is transmitted from the lower substrate 100, the valve 250 may be raised to the upper substrate 300 to block the inflow of the biological sample. In addition, when the pneumatic or hydraulic pressure is removed due to the elasticity, the thin film layer 200 and the valve 250 are returned to their original position to facilitate the flow of the biological sample and the buffer solution through the flow path.

8A to 8C are cross-sectional views illustrating an upper substrate, a lower substrate, a thin film layer, and a method of combining them according to a preferred embodiment of the present invention.

First, referring to FIG. 8A, a photoresist 352 is coated on a substrate 350. The substrate 350 may have a ceramic material such as silicon or a substrate having a material such as metal.

Subsequently, ultraviolet rays are irradiated to the photoresist 352 formed on the substrate 350. Irradiation of the ultraviolet rays is selectively performed on the photoresist 352 by the mask 354 or the like. In addition, although the photoresist 352 illustrated in FIG. 8A has been disclosed as a negative type in which a portion to which light is irradiated is cured and remains due to development, the photoresist 352 may be light according to an embodiment. This irradiated part may be of a positive type removed by the developing process.

Subsequently, a developing process is performed on the photoresist 352 irradiated with ultraviolet rays selectively. When the photoresist pattern 356 is formed by the development of the photoresist 352, the PD resist is molded on the photoresist pattern 356, and the substrate 350 and the photoresist pattern 356 are removed. An upper substrate 300 is formed.

In FIG. 8A, the PDMS portion corresponding to the photoresist pattern 356 becomes the injection passage 314 or the pneumatic / hydraulic control unit of the upper substrate 300.

Referring to FIG. 8B, a photoresist 162 is coated on the substrate 160. The substrate 160 may have a ceramic material such as silicon or a substrate having a material such as metal.

Subsequently, ultraviolet rays are irradiated to the photoresist 162 formed on the substrate 160. Irradiation of the ultraviolet rays is selectively performed on the photoresist 162 by the mask 164 or the like. In addition, although the photoresist 162 illustrated in FIG. 8B has been disclosed as a negative type in which a portion to which light is irradiated is cured and remains due to development, the photoresist 162 may be light. This irradiated part may be of a positive type removed by the developing process.

Subsequently, a developing process is performed on the photoresist 162 irradiated with ultraviolet rays selectively. The photoresist pattern 166 is formed by the development of the photoresist 162, and the PDMS 168 is applied to the photoresist pattern 166.

Application of the PDMS is preferably using spin coating.

In addition, the lower substrate 100 formed by using the same method as FIG. 8A on the substrate to which the PDMS 168 is applied is covered. When curing is performed on the substrate on which the lower substrate 100 and the PDMS 168 are applied, the PDMS 168 on the lower substrate 100 and the substrate 160 are bonded by a silanization bond.

Subsequently, when the thin film layer 200, which is combined with the lower substrate 100 and made of PDMS, is removed from the substrate 160, a valve 250 formed in a shape protruding on the thin film layer 200 and the thin film layer 200 is obtained. Can be.

Referring to FIG. 8C, the upper substrate 300 formed by FIG. 8A is bonded onto the lower substrate 100 and the thin film layer 200 formed by FIG. 8B. In the coupling, the valve 250 of the lower substrate 100 and the pneumatic / hydraulic control unit of the upper substrate 300 are coupled to correspond.

9 is a cross-sectional view of a biosensor chip for explaining that a biological sample or a buffer solution is injected and discharged through a recovery port according to a preferred embodiment of the present invention.

9, the biological sample or the buffer solution is introduced through the biological sample / buffer solution injection hole 310 of the upper substrate 300 and flows through the injection flow path 314. The biological sample / buffer solution injection hole 310 may be provided on the lower substrate 100 according to the embodiment.

The valve 250 is provided on the injection passage 314. The valve 250 is formed on the thin film layer 200 and is controlled by pneumatic or hydraulic pressure supplied by the pneumatic / hydraulic inlet 110 of the lower substrate 100. In addition, the upper substrate 300 is provided with pneumatic / hydraulic controllers 312 and 316 corresponding to the valve 250. By the operation of the valve 250 provided in the pneumatic / hydraulic controllers 312 and 316, the biological sample or the buffer solution flows into or is blocked from the reaction substrate 400. The biological sample introduced into the reaction substrate 400 is combined with an antibody provided in the metal thin film layer 500 and changes the reflection angle characteristic of the light irradiated through the prism 600.

The biological sample and the buffer solution passing through the reaction substrate 400 are discharged to the outside through the recovery port 330 provided on the upper substrate 300.

According to the present invention as described above, it comprises a plurality of biological sample / buffer solution inlet and recovery ports. Therefore, a plurality of biological samples can be introduced or discharged simultaneously. In addition, an upper substrate, a lower substrate, a thin film layer or a valve is formed using a PDMS material. Therefore, adhesion of the upper substrate and the lower substrate can be easily performed without a separate fastening means, and the valve can be controlled using the elasticity of the thin film layer.

Therefore, two or more unknown biological samples can be compared and analyzed, and after the reaction with the detection antibody formed on the metal thin film, the residual amount of the biological sample can be recovered without loss by selecting a specific recovery port from a plurality of recovery ports. have.

Although described above with reference to a preferred embodiment of the present invention, those skilled in the art will be variously modified and changed within the scope of the invention without departing from the spirit and scope of the invention described in the claims below I can understand that you can.

Claims (11)

A lower substrate 100 having pneumatic / hydraulic flow paths 130 provided with pneumatic or hydraulic pressure from the outside and made of polydimethyl-siloxan (PDMS) material; The biological sample or the buffer solution is formed on the lower substrate 100 and is made of polydimethyl siloxane, and the biological sample or the buffer solution is injected through the injection channels located therein, and the biological samples are disposed through the discharge channels spaced apart from the injection channel. An upper substrate 300 through which a sample or buffer solution is discharged, each of the injection channels having a pneumatic / hydraulic control section 312 and 316 corresponding to each of the pneumatic / hydraulic flow paths 130; A surface interposed between the lower substrate 100 and the upper substrate 300, and is made of a polydimethyl siloxane material, and is interposed between the pneumatic / hydraulic flow path 130 and the pneumatic / hydraulic control parts 312 and 316. And a thin film layer 200 having a valve 250 positioned thereon, wherein the valve 250 is elevated by pneumatic or hydraulic pressure delivered to the thin film layer 200 through the pneumatic / hydraulic flow path 130. To control the flow of biological samples or buffer solutions to the pneumatic / hydraulic controls 312 and 316, Reaction substrate 400 provided on the upper substrate 300, made of the polydimethyl siloxane material, and having a reaction flow path 430 in communication with the injection channel and the discharge channel flows the biological sample or buffer solution ; A metal thin film layer 500 for analyzing a biological sample of the reaction flow path 430 with an antibody formed as a single layer; And And a prism (600) for irradiating light onto the metal thin film layer (500) to use optical characteristics of the biological sample bound to the antibody. delete The multi-channel biochip according to claim 1, wherein the lower substrate (100) further comprises biological sample / buffer solution inlets (150) for supplying the biological sample or the buffer solution. The multi-channel biochip of claim 1, wherein the thin film layer is coupled with the lower substrate by silanization coupling with the lower substrate. The method of claim 1, wherein the upper substrate 300, Biological sample / buffer solution inlets 310 into which a biological sample or a buffer solution is introduced; An injection passage 314 through which the biological sample or the buffer solution introduced from the biological sample / buffer solution injection holes 310 is transferred; First reaction substrate connections 320 for supplying the biological sample or the buffer solution delivered through the injection passage 314 to the reaction substrate 400; Second reaction substrate connections 322 for recovering the biological sample or the buffer solution from the reaction substrate 400; Recovery flow paths 324 to which the biological sample or the buffer solution delivered to the second reaction substrate connections 322 is transferred; And Further comprising recovery ports 330 for discharging the biological sample or buffer solution from the recovery flow paths 324, wherein the pneumatic / hydraulic control unit 312, 316 from the biological sample / buffer solution inlet 310 The first pneumatic / hydraulic control unit 312 for controlling the flow of the biological sample / buffer solution flows into the injection flow path 314 and the flow introduced into the first reaction substrate connecting portion 320 from the injection flow path 314 And a second pneumatic / hydraulic controller (316) for controlling the flow of the biological sample / buffer solution. The method of claim 1, wherein the multi-channel biochip, Upper case 700 and lower case 800 for protecting the upper substrate 300, the metal thin film layer 500, the lower substrate 100, the reaction substrate 400, and the thin film layer 200 from the outside. Further comprising, wherein the upper case 700 or the lower case 800 is a multi-channel biochip, characterized in that made of a hydrophilic polycarbonate material. 7. The multi-channel biochip of claim 6, wherein the upper case (700) has a prism seat (730) engraved from a surface to receive the prism (600). The multi-channel biochip of claim 1, wherein the metal thin film layer comprises gold, silver, copper, or aluminum. The lower substrate 100, the thin film layer 200 having the valve 250, the upper substrate 300, the reaction substrate 400, and the metal thin film layer 500 may be provided to inject a biological sample or a buffer solution using multiple channels. In the method of forming a multi-channel biochip provided to discharge, The method of forming the upper substrate 300, Applying the photoresist 352 to the substrate 350 of ceramic or metal material; Selectively irradiating the photoresist 352 to form a photoresist pattern 356; Applying a polydimethyl siloxane film on the photoresist pattern (356); And And separating the polydimethyl siloxane film from the substrate (350) and the photoresist pattern (356). The polydimethyl siloxane film portion corresponding to the photoresist pattern 356 is an injection channel 314 or a pneumatic / hydraulic controller 312 and 316 of the upper substrate 300. Method of forming a multi-channel biochip. The lower substrate 100, the thin film layer 200 having the valve 250, the upper substrate 300, the reaction substrate 400, and the metal thin film layer 500 are injected or discharged into the biological sample or the buffer solution using multiple channels. In the method of forming a multi-channel biochip provided, The method of forming the lower substrate 100 and the thin film layer 200, Applying the photoresist 162 on the substrate 160 of ceramic or metal material; Selectively irradiating ultraviolet rays to the photoresist 162 to form a photoresist pattern 166; Applying a polydimethyl siloxane film (168) on the photoresist pattern (166) to form the thin film layer (200) and the valve (250); Curing the lower substrate 100 formed of polydimethyl siloxane in contact with the thin film layer 200; And And separating the lower substrate (100) and the thin film layer (200) from which the silanic bond is formed by the curing from the substrate (160).

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