JP2006078475A - Reactor and manufacturing method therefor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a chip type reactor having a flow passage manufactured precisely on a substrate, capable of exhibiting thereby an aiming device function sufficiently, and suitable for mass production. <P>SOLUTION: This chip type reactor 1 is provided with a glass substrate 11 and a resin film layer 12. The resin film layer 12 is arranged on the substrate 11, and forms on the substrate 11 a plurality of vessels 21, 22 for introducing a fluid for a sample, and the flow passage 23 for communicating the plurality of vessels. The resin film layer 12 is formed of a photosensitive resin. The plurality of vessels 21, 22 comprise a reaction vessel, a waste liquid vessel and a silica vessel for adsorbing a DNA. The reaction vessel 21 is communicated each other with the waste liquid vessel 22 via the flow passage 23, and the silica vessel 25 is arranged in a midway of the flow passage 23 to be subjected to DNA extraction. Electrodes 14 for impressing a voltage are arranged respectively in the reaction vessel 21 and the waste liquid vessel 22. The silica vessel 25 is covered with a cover glass 13. <P>COPYRIGHT: (C)2006,JPO&amp;NCIPI

Description

  The present invention relates to a reactor used for control of a biochemical reaction such as general chemical reaction and DNA (deoxyribose nucleic acid) extraction from cells, and a method for producing the same. The present invention relates to a microchip-type reactor having a flow path to be connected and a manufacturing method thereof.

  In recent years, as seen in gene therapy and the like, research on DNA substances has become active. Experiments with substance-to-material reactions are essential for this study. By performing this intersubstance reaction in a reactor having a fine structure, the amount of reagents and waste liquid can be reduced, and process control such as temperature control and reaction rate becomes easy. Further, by dropping the reaction process into a fine structure, not only the apparatus can be downsized but also the processing can be automated and speeded up easily. Therefore, when a reactor having a fine structure is used, the reaction product can be recovered more safely and efficiently than when a large capacity cell is used.

  As such a reactor, a reactor having a structure in which a minute flow path of several hundred μm or less is formed on a glass or resin substrate is known. A technique for introducing a test solution into the flow path of this reactor to cause a reaction such as mixing and separation and efficiently controlling a reaction between substances to obtain a reaction substance or the like is a microreactor, μ-TAS, laboratory It is called on-chip and is used for applied research and development in various fields.

  Specifically, for example, in Patent Document 1 (Japanese Patent Laid-Open No. 2004-033901), a microreactor having a channel width of 500 μm or less is used as a method for efficiently producing a metal colloid solution having a dispersed particle diameter of 5 nm or less. A method has been proposed. Further, Patent Document 2 (Japanese Patent Laid-Open No. 2003-307521) proposes a water quality analyzer that performs a water quality survey quickly and easily by combining a microreactor that prepares a sample for water quality analysis and a spectroscopic analyzer. ing.

  Further, Patent Document 3 (Japanese Patent Laid-Open No. 2003-280126) discloses a method for obtaining a silver halide photographic emulsion having uniform emulsion performance by using a microreactor having a fine processing region to correct the reaction. A method for improving efficiency has been proposed. In Patent Document 4 (Japanese Patent Application Laid-Open No. 2003-210956), ozone gas is dissolved in a fine groove-shaped flow path (microchannel) in order to efficiently dissolve ozone in water and obtain high-concentration ozone water. A high-concentration ozone water production apparatus equipped with a microreactor that absorbs water is shown.

  On the other hand, a reactor having such a fine structure is applied to a biochemical experimental process, and an electric field is applied to a mixed solution of biological substances such as cells, DNA, RNA, proteins, etc. Biological material separation technology applying electrophoretic chromatography to separate substances has been actively researched and developed. In this field, chip technology is particularly advanced. Also known is a microreactor that performs operations such as cell decomposition and separation on a chip to extract and detect biological materials.

  As a specific example, Patent Document 5 (Japanese Patent Laid-Open No. 08-233778) is cited. This document proposes a capillary electrophoresis chip in which a flow path and electrodes are formed on the surface of a transparent flat plate, the periphery is surrounded by an insulating material frame, and the solution is electrophoretically separated in the flow path. In Patent Document 6 (Japanese Patent Laid-Open No. 10-132783), for the purpose of improving detection sensitivity, an electrophoresis function is configured on a chip, and at the same time, an optical detection function is provided on a separation channel to separate a solution. An electrophoresis chip that simultaneously performs detection has been devised.

  Further, Patent Document 7 (Japanese Patent Laid-Open No. 10-337173) discloses a chip type for biochemical reaction that can simultaneously process a large number of biochemical reactions and perform substance synthesis reactions such as protein synthesis on the chip. A reactor is disclosed.

  As a material constituting these reactors, glass, a silicon wafer, a resin material, or the like is used, and the method for obtaining a fine structure is also different.

  When glass is used as the substrate material, direct machining with a tiremond tool or the like is common to obtain a fine structure. However, a technique for obtaining a fine pattern more efficiently is proposed in, for example, Japanese Patent Application Laid-Open No. 2003-299944. According to this, a glass-containing layer that is easy to be removed is provided on the surface of the glass substrate, and a flow path pattern is produced by sandblasting using a hard mask having an arbitrary shape.

  In addition, when silicon is used for a substrate, a flow path pattern forming method using anisotropic etching of silicon or a mask process is generally used, as in the conventional method of forming a fine semiconductor pattern. .

Further, when the substrate is made of a resin material, a more mass production method can be used. For example, as proposed in Patent Document 9 (Japanese Patent Laid-Open No. 2003-159696), the flow path pattern is transferred to a resin substrate by using injection molding, and a low-cost chemical micro plate for a resin substrate in consideration of mass productivity. A device is obtained. In addition, a method of producing a fine channel on a resin substrate by using an imprinting technique used for transferring a signal recording pattern of a recording medium such as a CD (compact disc) or a DVD (Digital Versatile Disc) is also used. It has been established as a method for replicating a large number of microchannels of the same shape. Also known is a method of obtaining a substrate on which mold irregularities are transferred by curing a resin material such as PMMA (polymethylmethacrylate) or PDMS (Polydimethylsiloxane) on a mold.
JP 2004-033901 A JP 2003-307521 A JP 2003-280126 A JP 2003-210956 A Japanese Patent Laid-Open No. 08-233778 Japanese Patent Laid-Open No. 10-132783 JP 10-337173 A JP 2003-299944 A JP 2003-159696 A

  However, in the case of various conventional chip reactors described above, there is a problem in that there are restrictions on the channel design depending on the respective manufacturing processes.

  For example, the direct processing of a glass substrate has a problem of processing surface that it is difficult to create a portion having a small curvature due to brittleness peculiar to glass. In addition to the difficulty in creating this small curvature part, it is necessary to design the flow path so as not to cause chipping in the corner and edge shapes, which greatly limits the degree of flow path integration in the flow path structure. There is also the problem of being. In particular, in the case of a chip reactor that controls the reaction by rapidly expanding the channel width or merging multiple channels, a shape with a small curvature (acute angle) is required. It is difficult to realize.

  In addition, when using a silicon process such as anisotropic silicon etching, the direction to be removed by etching or the angle of the wall surface (etching angle) is determined on the material side, so the flow path structure must be designed accordingly. There is a restriction that it cannot be obtained. In addition, when performing wet etching as is generally done in the laboratory, it is difficult to strictly control the depth and width of the flow path, and it is difficult to produce a fine pattern with high accuracy. is doing.

  Furthermore, when adopting injection molding, imprinting, or a method of curing a resin, it is necessary to produce a fine pattern as a mold first, which complicates the production process and further increases the flow path. There is also the inconvenience that it is difficult to change the pattern after designing and producing the mold.

  In this way, regardless of which manufacturing method is used, the design process of the flow path pattern is facilitated, the pattern is highly accurate, the flow path integration is improved, the manufacturing process is simplified, the microstructure is miniaturized, etc. There are drawbacks, and a chip reactor that can sufficiently satisfy these problems cannot be provided.

  For this reason, in the past, a commercially available DNA collection kit using a test tube-shaped container (cylindrical container having a length of about 3 cm) generally called a microtube is usually used for taking out DNA. Has been. In general, steps such as cell membrane lysis, protein removal, residual separation, pick-up / fixation, etc. are required as pre-treatment until DNA analysis. It becomes complicated and takes manpower. For this reason, even if it is desired to identify and analyze an extract from a single cell, since a column is used, it is impossible to determine from which cell a large amount of samples are processed and extracted. Furthermore, although the extracted sample is desired to be analyzed as it is, there is a disadvantage that the sample is crushed or decomposed before the sample in the column is fixed to the sample stage.

  In such a situation, especially in the case of a chip-type reactor, the device function is determined by the variety of flow channel designs, so it has a highly accurate flow channel on the substrate, and analysis such as sample separation, extraction, and fixation. There is a demand for efficient and labor saving the previous series of basic operations. At the same time, there is also a demand to mass-produce chip reactors that can improve the efficiency of such basic operations, support DNA analysis, and promote biotechnology more.

  The present invention is intended to meet the above-mentioned demand from the field of experiments for inter-substance reactions such as DNA analysis, and has a highly accurate flow path on a substrate, thereby achieving a desired device function. It is an object of the present invention to provide a chip reactor suitable for mass production.

  In order to achieve the above-described object, according to the present invention, there is provided a reactor used for intersubstance reaction. The reactor includes a substrate, a plurality of tanks that are arranged on the substrate and that introduce a sample fluid, and a resin layer that forms a flow path that connects the plurality of tanks on the substrate. The resin layer is formed of a photosensitive resin.

  Preferably, the substrate is formed of a glass material, and the resin layer is formed of a film-like photosensitive resin. The plurality of tanks include, for example, a cell introduction tank (for example, a reaction tank), a waste liquid tank, and an adsorption tank (for example, a silica powder tank) (more preferably, in addition to these, an extraction liquid tank and a sample collection tank (also referred to as an extraction tank). The reaction tank and the waste liquid tank (in the case of including the extraction liquid tank and the sample collection tank, both tanks) are communicated with each other via the flow path, and in the middle of the flow path. The silica powder tank is disposed in the structure, and is used for, for example, DNA extraction. In addition, the plurality of tanks include, for example, a cell introduction tank and a waste liquid tank, and the cell introduction tank and the waste liquid tank communicate with each other through the flow path, and a cell capture trap is provided in the middle of the flow path. Are arranged to be used for extraction of DNA or other intracellular constituents from a single cell. As an example, the flow path is a straight flow path that allows the reaction tank and the waste liquid tank to communicate with each other in a straight line through the silica powder tank. The flow path is a straight flow path that linearly connects the cell introduction tank and the waste liquid tank via the cell capture trap. More preferably, a voltage application electrode is provided in each of the cell introduction tank and the waste liquid tank, and a voltage application electrode is provided in each of the cell introduction tank and the waste liquid tank. Further, at least the silica powder tank may be covered with a cover member.

  On the other hand, according to the present invention, there is also provided a method for producing a reactor for use in an intersubstance reaction. In this manufacturing method, an electrode is formed at a predetermined position on a substrate, a photosensitive resin layer is formed on the substrate on which the electrode is formed, and a plurality of tanks for introducing a fluid for a sample and a space between the plurality of tanks are formed. Forming the plurality of tanks and the flow paths on the substrate by exposing the photosensitive resin layer with a pattern mask on which a flow path to be communicated is drawn and developing the exposed photosensitive resin layer. Features.

  According to the present invention, the resin layer is formed with a plurality of tanks for introducing a sample fluid and a flow path communicating between the plurality of tanks on the substrate, and the resin layer is formed of a photosensitive resin. As in the prior art, it is not necessary to directly process glass, etch process, or use a special imprint mold, and a chip reactor can be easily manufactured. In addition, by providing an adsorption tank (for example, a silica powder tank) for adsorbing intracellular constituents such as DNA in the middle of a flow path connecting a plurality of tanks, a chip type for extracting intracellular constituents that requires a small and simple structure A reactor can be provided. In addition, by providing a trap for capturing cells in the middle of the flow path connecting a plurality of tanks, a chip-type reactor for extracting intracellular constituents such as DNA for a single cell with a small and simple structure. Can be provided.

  That is, an arbitrary fine pattern can be transferred onto the substrate by direct drawing using a laser beam or the like, or selective exposure using a mask process on a substrate having at least a surface formed of a photosensitive resin. It can be formed easily. Therefore, it is not necessary to produce a mold like imprinting or injection molding. Further, by using a photosensitive resin capable of optically producing a fine structure, a flow channel structure necessary for a chip reactor can be created accurately and easily. At the same time, by selecting a material having alkali resistance as the photosensitive resin, a chip reactor that can be suitably used in the biochemical field can be produced.

  Hereinafter, an embodiment of the chip reactor according to the present invention will be described with reference to FIGS. In this embodiment, a simple chip-type reactor used for DNA extraction is illustrated, but the chip-type reactor according to the present invention is not necessarily limited to DNA extraction and is widely used between substances. It can be implemented as a small chip reactor used for reaction experiments.

  1 and 2 schematically show a plan view and a cross-sectional view of a simple chip reactor 1 used for DNA extraction.

  As shown in these drawings, the chip reactor 1 according to this embodiment includes a glass substrate 11, a photosensitive resin film layer 12 laminated on the surface of the substrate 11, and a surface of the resin film layer 12. It has a three-layer structure with a partially placed cover glass 13 and a laminated structure in which film electrodes 14 and 14 are partially inserted between the glassy substrate 11 and the resin film layer 12. It has become.

  The size of the chip reactor 1 is 10 mm wide and 20 mm long in FIG. The substrate 11 is formed to have a width of 10 mm, a length of 20 mm, and a thickness of 1 mm. After the film electrodes 14 and 14 are partially formed on the surface of the substrate 11, a photosensitive resin film is formed on the entire surface of the substrate 11. The layer 12 is laminated, and the central portion of the surface of the resin film layer 12 is partially covered with a cover glass 13 as a cover member.

  By exposing the resin film layer 12 to the surface of the glass substrate 11, a reaction tank 21, a waste liquid tank 22, a fine channel 23, and a silica tank 25 are formed along the film layer as shown in FIG. Is formed.

  That is, a rectangular reaction tank 21 is formed at a predetermined position near one end of the substrate 11 in the longitudinal direction, and a rectangular waste liquid tank 22 reacts at a predetermined position near the other end opposite to the reaction tank 21. It is formed so as to face the tank 21 at a predetermined distance. The reaction vessel 21 is used for cell degradation and purification. The waste liquid tank 22 is formed to collect the waste liquid. The fine channel 23 is a straight channel having a width of 100 μm and a predetermined length, and is formed so as to linearly connect the reaction tank 21 and the waste liquid tank 22 to each other. That is, the reaction tank 21 and the waste liquid tank 22 communicate with each other through the fine flow path 23.

  In addition, a silica tank (silica powder tank) 25 for adsorbing DNA is formed in an intermediate portion in the longitudinal direction of the fine channel 23. The silica tank 25 has a structure in which DNA can be trapped and taken out from the cell decomposition solution flowing in the fine flow path 23 from the reaction tank 21 toward the waste liquid tank 22.

  Furthermore, the membrane electrode 14 is arrange | positioned in the position of the bottom face of the reaction tank 21 and the waste liquid tank 22, respectively. Each of the membrane electrodes 14 occupies the bottom surface area of each tank 21 (22), and has a surface shape in which a part of the membrane electrode 14 protrudes toward the fine flow path 23. For example, platinum (Pt) is used as the material of the film electrode 14. A voltage is applied between the two film electrodes 14. As a result, an electric field is applied to the solution, and a driving force for transporting the solution in the reaction tank 21 and the reactant in the silica tank 25 can be applied to the solution.

  On the other hand, the cover glass 13 is a rectangular protective glass and is disposed so as to cover a partial region centering on the surface of the silica tank 25. For this reason, the remaining part of the fine channel 23, the reaction tank 21 and the waste liquid tank 22 at both ends are exposed.

  Next, materials and the like that can be used for each component constituting the chip reactor 1 described above will be described.

  As the glass material used for the substrate 1, commercially available borosilicate glass (Pyrex (registered trademark), code 7740) can be used. However, it is not necessarily limited to this. When performing a biochemical reaction as in the present embodiment, it is desirable to use a material with as few impurities as possible in consideration of application to chemical cleanliness of the surface, optical analysis, and the like. In addition, the surface treatment of the target sample may be applied to the surface of the substrate 1 in consideration of the adsorption characteristics of the reaction product to the substrate surface. The substrate 1 is not necessarily made of glass, and may be formed of other materials such as a resin material or a silicon wafer suitable for the purpose of use.

  Further, as the cover glass 13, a commercially available cover glass (Matsunami Co., Ltd., micro cover glass thickness: 0.12 to 0.17 mm) is used. In this case as well, any other glass material or resin material may be used. You can select and use one.

  Further, the photosensitive resin film layer 12 may be a commercially available negative-type dry film resist (photosensitive layer film thickness 30 μm, alkali development type) that is generally used for semiconductor processes. As described above, for biochemical applications, it is desirable to select a resist material having excellent alkali resistance after the photosensitive process.

  Further, the film electrode 14 can be formed of a sputtered film of platinum (Pt). The film electrode 14 causes an electrochemical reaction on the electrode surface due to a direct current potential applied when the chip is driven, and the electrode is consumed due to electric field corrosion. Therefore, as an electrode material, Pt, Au, Ag or the like is preferably used. It is desirable to select a material with a high standard electrode potential (having a positive value). In addition, if a transparent electrode such as ITO (Indium Tin Oxide) is used, the transparency of the chip reactor is maintained, and this is desirable depending on the application.

  A biochemical operation performed on the chip when DNA extraction is performed using the chip reactor 1 described above will be described.

  First, target cells are placed on the reaction tank 21, and a denaturant and a cell lysate are added to produce a cell disruption solution. While rinsing this with a TAE buffer as appropriate, low molecular weight RNA, lipids, cell walls, etc. are transported to the waste liquid tank 12. The driving force for transporting the solution at this time is based on electrophoresis or electroosmotic flow generated by an electric field applied between the reaction tank 21 and the waste liquid tank 22. At that time, mass transport is performed under conditions (pH, specific salt concentration, etc.) satisfying the silica and DNA binding conditions, and only the DNA is adsorbed in the silica tank 25. Thereafter, the DNA is eluted from the silica tank 25 under conditions satisfying the dissociation conditions of silica and DNA, and the DNA is extracted.

(Manufacturing method of chip reactor)
With reference to FIG. 4, the manufacturing method of a chip-type reactor is demonstrated.

  First, a cleaned glass substrate 1 is prepared, and a film electrode 14 having a thickness on the order of submicron is formed on the substrate 1 (see FIGS. 5A and 5B).

  Although the film electrode 14 is produced by physical vapor deposition or chemical vapor deposition, as described above, an electrochemical reaction (electric field corrosion or generation of microbubbles) occurs on the electrode surface when a voltage is applied. It is desirable to produce a film with a small number of micropores). In addition, these electrodes do not need to be in the form of a film, and bulk electrodes having a shape and size that do not impair the structure of the chip can be used. In this case, a long electrode life can be secured. For this electrode production, pattern transfer and mask process (lithography) can be used. Further, a part of these film electrodes 14 are exposed so that they can be electrically connected to external electrodes. A preferred embodiment of this film-like electrode is a Ti / Pt two-layer sputtered film having a thickness of about 1 μm.

  A photosensitive film (resin) 12 is attached to the surface of the glass substrate 1 on which the membrane electrode 14 is produced using a laminator (see FIG. 5C). For example, a negative type is used for the photosensitive resin material of the photosensitive film 12, but a positive type may also be used.

  As an example of a standard for laminating conditions when laminating this photosensitive resin (resin film layer 12), a laminating temperature is 100 ° C., a pressure is 0.5 MPa, and a speed is about 1.0 m / min. At this time, if dust or the like is present between the glass substrate 11 and the photosensitive film 12, the flow path may be damaged or the film may be peeled off. Therefore, it should be performed in a clean environment such as a clean room or a clean bench. Is desirable. In addition, since the photosensitive film 12 may be sensitive to ultraviolet rays contained in natural light, the work should be performed quickly in an environment where measures against ultraviolet light are taken or under natural light. It is desirable to take care not to expose it. In order to avoid air bubbles from being mixed between the film and the substrate, it is desirable to laminate the film with tension.

  Next, the photosensitive film 12 on the substrate 11 is exposed using a mask exposure process (see FIGS. 5D and 5E).

  More specifically, first, a pattern mask 31 is affixed to the film 12 ((FIG. 5 (d)). This pattern mask 31 is a mask made of, for example, PET (polyethylene terephthalate). As shown in Fig. 6, the mask area is printed in black (shaded area in Fig. 6), and the other area (the remaining area other than the hatched area in Fig. 6) is transparent. This is because the resin material 12 is a negative type, and when the material is a positive type, the black / transparent relationship of the pattern mask 31 shown in FIG.

  Next, the pattern mask 31 is exposed with an exposure apparatus and then developed (FIG. 5E). In the example of this explanation, since a negative photosensitive resin is used for the photosensitive film 12, the resin portion corresponding to the light-irradiated region, that is, the transparent region of the pattern mask 31, is crosslinked by photosensitivity. It remains undissolved even after development. On the other hand, since the resin portion corresponding to the black region of the pattern mask 31 that is not irradiated with light by exposure is not cross-linked by photosensitivity, it is removed by development processing. As a result, as shown in FIG. 3, portions corresponding to the reaction tank 21, the waste liquid tank 22, the fine channel 23, and the silica tank 25 are removed, and the surrounding portions remain as wall portions. That is, each tank and flow path are reliably formed on the glass substrate 2 along the resin film layer 12 by exposure. The preferred size of the flow path connecting the reaction tank 21 and the waste liquid tank 22 is a flow path width of 100 μm and a depth of 60 μm.

  As an example, an MJB3 manufactured by Karl Suss is used as the exposure apparatus, and an ultra-high pressure mercury lamp USH-350DP (generated wavelength region: 300 to 600 nm) manufactured by USHIO is used as the exposure light source. The mask and the film surface are brought into close contact, and after exposure for 1 minute, alkali development is performed with a 1 wt% aqueous sodium carbonate solution. At this time, if the exposure time is long, the light may be exposed to the inside of the flow path due to the surrounding light. In this case, since the flow path is blocked by the exposed resin and inhibits the chip resin, it is necessary to appropriately set the exposure conditions in order to obtain a fine flow path shape accurately.

  After this, although not shown in FIG. 5, the silica tank 25 is filled with silica to collect DNA in the silica tank 25 on the flow path. Specifically, an appropriate amount of spherical silica powder is inserted into the silica tank 25. At this time, in order to prevent the outflow of silica, a filter in which silica is fixed by an insoluble property such as a resin may be used. A porous material mainly composed of silica can also be suitably used. By adjusting the amount and particle diameter of the silica powder at this time, the DNA collection characteristics can be changed.

  Finally, the cover glass 13 is attached to the upper surfaces of the fine channels 23 and the silica tank 25 so as to cover them (see FIG. 5F). Thereby, the chip reactor 1 is manufactured.

  In the above embodiment, the silica tank is used as the nucleic acid adsorption tank. However, the present invention is not limited to this, and any other one can be suitably used as long as the adsorption / dissociation of nucleic acid can be controlled. For example, magnetic particles can be used instead of silica powder.

  A second embodiment according to the present invention will be described with reference to FIG.

  The components and the creation process of this embodiment are the same as those of the first embodiment.

  Differences in structure and function are described below.

A reaction tank 51 for decomposing and purifying cells is provided on one side of the substrate by a photosensitive resin film layer 42 on a glass substrate 41, and a waste liquid tank 52 for collecting waste liquid is installed on the other side. . The reaction tank 51 and the waste liquid tank 52 are connected by a fine flow channel 53 having a width of 100 μm. In addition, an extraction liquid tank 54 and a sample collection tank 55 are provided in a direction orthogonal to the fine flow path 53 via a similar fine flow path 56, and a silica is formed at the intersection of the fine flow path 53 and the fine flow path 56. A tank 57 is provided. The four tanks, that is, the reaction tank 51, the waste liquid tank 52, the extract liquid tank 54, and the sample collection tank 55 are provided with thin film electrodes 81, 82, 83, and 84 so that voltages can be individually applied. Each thin film electrode is integrated at the right end of the chip reactor 100 by each lead pattern, and has a structure in which a voltage controller (not shown) is connected to apply a voltage between the electrodes.
Although not shown, a cover member made of glass, for example, is attached on the resin film layer 32 so as to cover at least the silica tank 57 and expose the other tanks.

  An operation performed on the chip reactor 100 will be described. Place target cells on reaction tank 51, add denaturant and cell lysate, prepare cell disruption solution, and rinse low-molecular-weight RNA, lipid, cell wall, etc. with appropriate TAE buffer. Transport to 52. The transport driving force at this time is due to electrophoresis or electroosmotic flow generated by an electric field applied between the reaction tank 51 and the waste liquid tank 52, that is, between the thin film electrodes 81 and 82. At this time, mass transport is performed under conditions (pH, specific salt concentration, etc.) that satisfy the binding conditions of silica and nucleic acid, and only the target substance (for example, DNA) is adsorbed to the silica tank 57. That is, a low molecular weight substance and a high molecular compound such as a target substance such as DNA can be separated by an electrotransport means such as electrophoresis or electroosmotic flow.

  After that, a solution that satisfies the dissociation conditions of silica and nucleic acid is put into the extract solution tank 54, and this time, an electric field is applied between the extract solution tank 54 and the sample collection tank 55, that is, between the thin film electrode 83 and the thin film electrode 84. The target substance adsorbed on the silica tank 57 is eluted into the sample collection tank 55 and collected.

  At this time, in order to stabilize the flow of the solution from the extraction liquid tank 54 to the sample collection tank 55, the thin film electrodes 81 and 82 attached to the reaction tank 51 and the waste liquid tank 52 have a certain potential with the thin film electrode 83. It is desirable to grant. By doing so, the flow of the extraction liquid from the extraction liquid tank 54 can reach the sample collection tank 55 without spreading in the direction of the reaction tank 51 and the waste liquid tank 52. This improves sample extraction efficiency.

  A third embodiment according to the present invention will be described with reference to FIG. In this example, one set of DNA or other intracellular constituents is extracted from one isolated cell, and the preparation process is the same as in Example 1. Differences in structure and function are described below.

  As shown in FIG. 8, the chip reactor 200 according to the third embodiment of the present invention includes a glass substrate 61 and a photosensitive resin material layer 62 for forming a flow path on the glass substrate 61. It is configured. A film electrode 64 is partially interposed between the glass substrate 61 and the photosensitive resin material layer 62. In this case, a part or the whole of the upper part of the photosensitive resin material layer 62 can be covered with a cover glass (cover plate) 63 as necessary.

  The chip-type reactor 200 has a width of 10 mm and a length of 20 mm, and the glass substrate 61 has a thickness of 1 mm and is made of commercially available Pyrex (registered trademark) glass. The photosensitive resin material layer 62 is configured using a negative photoresist (trade name SU-8-3050) manufactured by Kayaku Microchem. The film electrode 64 has a two-layer structure in which Ti is a base and the surface is covered with a Pt film. Adhesion with a glass substrate can be improved by using Ti as a base. Moreover, an electrode reaction can be suppressed by using Pt as a film | membrane provided on it.

  In the photosensitive resin material layer 62, as shown in FIG. 9, a cell introduction tank 71, a waste liquid tank 72, a main flow path 73, an extraction liquid tank 74, an extraction tank 75, and a sub flow path 76 are formed. The main flow path 73 is formed as a flow path connecting the cell introduction tank 71 and the waste liquid tank 72, and this main flow path 73 includes one sub flow path from an intersection 78 in the middle of the flow path of the main flow path 73. The extract tank 74 is connected via the 76, and the extract tank 75 is connected from the intersection 78 via the other sub-flow channel 76. A cell capture trap 77 is installed near the cell introduction tank 71 in the main channel 73. In addition, a substance for adsorbing a specific intracellular constituent substance can be installed at the intersection 78 between the main channel 73 and the sub channel 76. For example, when extracting DNA, a filter material mainly composed of silica may be provided.

  As shown in FIG. 10, the film electrode 64 interposed between the glass substrate 61 and the photosensitive resin material layer 62 is composed of six types of electrode patterns 141 to 146. Each of the electrode patterns 141 to 146 is gathered in a state where a part thereof is exposed on the left end side of the photosensitive resin material layer 62 so that the left end portion of the chip reactor 200 can be in contact with the external terminal. Among these electrode patterns 141 to 146, the electrode pattern 141 supplies the waste liquid tank 72, the electrode pattern 142 supplies the extraction liquid tank 74, the electrode pattern 144 supplies the cell injection tank 71, and the electrode pattern 146 supplies the extraction tank 75. The electrode patterns 143 and 145 are configured as electrode patterns for applying electrical stimulation to the cells of the cell trapping trap 77. 14 is a plan view in which the electrode pattern of FIG. 10 is superimposed on the pattern of each tank of FIG. As shown in FIG. 14, the electrode pattern is aligned with the pattern of each tank, and the two overlap each other in plan view, so that the electrode pattern can selectively apply a voltage to each tank.

  Next, the procedure for using the chip reactor 200 will be sequentially described.

  First, each solution tank and flow path in the chip reactor 200 are filled with a working fluid (for example, pH buffer solution), and the isolated cells are put into the cell introduction tank 71 by an appropriate method such as pipetting. Thereafter, an electric field is generated between the electrode pattern 144 connected to the cell introduction tank 71 and the electrode pattern 141 connected to the waste liquid tank 72, and the fluid moves from the cell introduction tank 71 to the waste liquid tank 72 as a working fluid. (Electroosmotic flow) is generated. This flow guides the cells to the cell capture trap 77. After the cells are introduced into the cell capture trap 77, an electric field is generated in the electrode patterns 143 and 145 to disrupt the cell shell. After crushing the cell shell, the intracellular material passes through the cell capture trap 77 and is introduced into the intersection 78 of the main flow path 73. In the process of introducing the intracellular substance into the intersection point 78, the intracellular substance is separated by its flow characteristics, and the extraction pattern 75 and the electrode pattern 142 connected to the extraction liquid tank 74 at the timing when the nucleic acid part passes through the intersection point 78. An electric field is generated between the electrode pattern 146 connected to the electrode pattern 146, a flow from the extract solution tank 74 toward the extract tank 75 is generated, and the nucleic acid portion is extracted in the extract tank 75. At this time, when a silica filter is installed at the intersection point 78 to adsorb and dissociate the nucleic acid, the pH adjusted extract is injected into the extract tank 74 before the nucleic acid extraction. The nucleic acid extracted to the extraction tank 75 is collected together with the solution, and subjected to analysis after being subjected to treatments such as purification and PCR as necessary. Alternatively, a mica sheet that can be attached to and detached from the extraction tank 75 is installed, and can be directly visualized and analyzed with an intermolecular force microscope or the like.

  Next, the cell capture trap 77 that temporarily captures cells will be described in more detail.

  In the cell capture trap 77, in order to prevent the cells from passing without being crushed, the width of the flow path is partially smaller than at least the minimum diameter of the cells to be captured. However, here, in order to efficiently capture the cells, the flow path width of the capture unit is 90% or less of the minimum diameter of the cells to be captured so as to cope with the deformation of the cells at the time of capture. Preferably, it is desirable that the content be 70% or less. When the flow path width of the capture unit is larger than this, the cells may pass through the capture unit while being deformed by being suppressed by the movement of the fluid. On the other hand, if the flow path width is too small, it will not only capture minute foreign substances, but also make it difficult to pass intracellular substances (nuclei (DNA, RNA), proteins, etc.) that are target substances. It is desirable that the minimum cell diameter is 10% or more, preferably 20% or more. In addition, when the flow path width is stepped, the continuous flow of fluid is inhibited and a local vortex is generated, which may inhibit the capture of cells.

  Therefore, it is desirable that the capturing part has a continuous shape so as not to disturb the fluid flow as much as possible. Possible typical channel shapes are shown in FIGS. Using the cell trap having the triangular cell capture section 77a shown in FIG. 11 (a) as a model, the flow path width of the cell capture section 77a is changed, and Examples (1) to (5) shown in Table 1 below and Comparative examples (1) to (2) were prepared. Zucchini isolated cells (minimum diameter of about 100 μm) were used as cells to be captured. The isolated cells can be obtained by collecting cells of an appropriate size with a micropipette from a cell solution obtained by enzymatic treatment of zucchini pulp. This protoplast was introduced into a reaction vessel (a cell introduction vessel in the embodiments described later).

  In these Examples (1) to (5) and Comparative Examples (1) to (2), the cell capture probability (●) and the cell internal constituent passage rate (▲) when the target cells were introduced were summarized. This is shown in FIG. The horizontal axis of FIG. 12 represents the ratio between the width of the cell flow path and the cell size (diameter) as a percentage (%), and the vertical cell capture probability indicates that a plurality of cells flow to the cell capture part (trap part) 77a. The ratio of the number of cells flowed to the number of captured cells was expressed as a percentage (%) as the cell capture probability. As shown in FIG. 12, by setting the ratio between the width of the flow path and the cell size (diameter) to 90% or less, more preferably 70% or less, the passage of cells is drastically reduced, and the cell capture probability is reduced. It is high. On the other hand, if the ratio between the width of the flow path and the cell size (diameter) is 10% or less, it becomes difficult to pass the cell internal constituent material to be extracted, which may hinder the function of the chip.

  If the cell size distribution is wide and it is difficult to set the channel width, a different size can be obtained by using a cell capture section 77b having a funnel-type channel as shown in FIG. Can also be used for other cells. At this time, in order to use electrical stimulation for cell disruption, electrodes are installed on both sides of the funnel-type channel. By doing so, electrical stimulation can be applied to any position of the cell capture portion 77b where the cell is captured. At this time, it is necessary to set the dimensions of the funnel inlet and outlet according to the nucleus dimensions of the largest cell and the smallest cell of the target cell group. The inlet dimension is set to the maximum cell diameter and the outlet dimension is 90% or less, desirably 70% or less of the smallest cell. In addition, for the purpose of purifying (separating) the internal material of the crushed cells after capture, as shown in FIG. 11 (c), the channel minimum portion of the cell capture portion (trap portion) 77c is extended in the longitudinal direction. You can also. In this case, the intracellular material is separated by ease of flow in the extension and is convenient for subsequent processing. Although not shown in the drawing, the upstream side of the trap portion may have a funnel shape as shown in FIG. 11B, and the minimum portion of the flow path may be extended as shown in FIG.

  As shown in FIG. 15, the electrode portion corresponding to the cell trap 77 in FIG. 10 has a configuration in which the electrode 143 and the electrode 145 are spaced apart from each other. The tip of the electrode 143 is divided into a plurality (two in FIG. 15) of small rectangular electrode bodies 143A. Similarly, the tip of the electrode 145 may be divided into small rectangular electrode bodies 145A. These electrode bodies enter the electrode body side of the other electrode. That is, the facing portion 200A between the electrode 143 and the electrode 145 is configured such that the comb-shaped electrodes approach each other without being in contact with other electrodes. Thereby, it is comprised so that the electric field strength of this opposing part can be enlarged.

  FIG. 16 shows the positional relationship between the cell trap portion 77 and the facing portion of the electrode 143 and the electrode 145 (shown as 200A in FIGS. 16A and 16C). The facing portion is located at least in the upstream portion of the trap portion 77 where the width is the narrowest, and can apply an electric field to the cell 200B captured in the upstream portion. In addition, the opposing part of electrodes can be changed suitably according to the shape of a trap part. In FIG. 16 (b), in accordance with the fact that the trap portion is formed in a funnel shape (tapered shape), the opposing portion of the end portion 143A of the electrode 143 and the end portion 145A of the electrode 145 has the same shape, a similar shape, or A similar shape may be used.

  A direct current, an alternating current, or a pulsed voltage is applied to an opposing portion between the electrodes from an external power source. An alternating voltage is desirable to suppress bubble generation and electrode reaction. The cells are electrically crushed by the electric field generated between the electrodes. Intracellular substances are separated by flow characteristics when flowing through the flow path. A substance having a lower molecular weight is easier to flow, and a substance having a higher molecular weight is less likely to flow. Therefore, the nucleic acid can be selectively separated by forming a working fluid flow from the extraction liquid layer to the extraction layer at the timing when the target substance (for example, nucleic acid) passes through the intersection of the flow paths (see FIG. 9). it can. In order to make this separation operation easier to confirm, DNA is fluorescently stained. Nucleic acids can be separated and extracted while checking with a microscope. Since the nucleic acid can be separated from other cell contents based on the flow characteristics of the nucleic acid in the flow path, the aforementioned silica powder or the like as a nucleic acid capturing agent is not necessarily essential. In addition, the value of the electric field applied to the facing part between the electrodes and the intensity of the electric field applied to each tank are appropriately selected as suitable values suitable for cell disruption and transport of cell contents.

  In addition, as shown in FIG.11 (c), the reason for making the cell trap part funnel-shaped is to ensure that cells having various diameters can be trapped. This is also because the cells are smoothly introduced into the trap portion.

  Next, the electric field strength necessary for driving the fluid in the chip will be described in more detail. The fluid in this chip mainly uses an electroosmotic flow (a flow generated when a localized layer (electric double layer) of electric charges generated at the interface between a glass wall surface and a liquid is driven by an external electric field). Accordingly, fluid drive characteristics such as flow rate and flow velocity change depending on the strength of the external electric field generated between the electrodes. That is, the intensity of the electric field generated between the electrodes is an important parameter in controlling the function of the chip.

  In the chip reactor 200 in this embodiment, the DC voltage applied between the electrodes is 10 V or more and 70 V or less, more preferably 30 V or more and 50 V or less. When the applied voltage exceeds 70 V, bubbles are generated from the electrode surface, impeding fluid flow. On the other hand, when the applied voltage is less than 10 V, a sufficient driving force cannot be applied to the hydraulic fluid in the flow path, and the fluid flow becomes unstable.

  The preferable voltage range (10 V or more and 70 V or less) in the chip reactor 200 in this example is calculated as the electric field strength of about 7 to 90 V / cm when calculated from the electrode spacing of the chip (longitudinal direction 15 mm, short side direction 8 mm). Equivalent to.

  In order to confirm the effect of the invention, a cell driving test on a chip was performed under the conditions of Examples (1) to (5) and Comparative Examples (1) to (2) shown in Table 2.

  A summary of the results of the cell drive test is shown in FIG. The horizontal axis in FIG. 13 is the applied interelectrode voltage, and the vertical axis is the cell moving speed. The cell moving speed is standardized with 1 in Example (4). Under the condition of Comparative Example (1) where the interelectrode voltage was 5 V, the cells did not move. Further, under the condition of Comparative Example (2) of 80 V, cells did not flow because bubbles were generated in the liquid and the flow of the hydraulic fluid itself was inhibited. In the range of Examples (1) to (5), the cell moving speed was changed according to the change of the voltage between the electrodes, and a good practical correlation was obtained.

  Here, the photosensitive resin that can be used in the process of laminating the resin film layer 12 or 42 on the substrate 11 will be described from various angles.

  As the photosensitive resin that can be used, various generally known photosensitive polymers can be applied, and a multi-component system including a reactive monomer, a reactive oligomer, a photoinitiator, a sensitizer, and the like. It is desirable that it can maintain chemically and physically stable characteristics through structural changes caused by light irradiation.

  For example, when classified from reaction forms, those using photo-crosslinking with photo-duplex photosensitive polymers such as polyvinyl cinnamate, bifunctional monomers (reactive radicals) such as diazo groups and azide groups, and azides・ Novolak, o-nitrobenzyl cholate ester ・ P (MMA-MAA) photo-modification (especially change in alkali solubility), monomers having acryloyl group such as acrylamide, acrylonitrile, acrylic acid, acrylate ester, etc. Suitable are those by radical polymerization with a photopolymerization initiator or by photopolymerization using an aryldiazonium salt or sulfonic acid ester to initiate photocationic polymerization. On the other hand, as described above, like photo-degradable polymers such as PMMA (polymethyl methacrylate), a positive type utilizing photodegradation of a polymer by light and low molecular weight by photodepolymerization (light irradiation part is possible). (Solubilized) photosensitive polymer can also be used.

  In particular, as a photoresist material used in semiconductor manufacturing processes, etc., a photocrosslinking agent is added to polyvinyl cinnamate, polyisoprene, and polybutadiene that are insolubilized by forming a crosslinked structure by photodimerization of cinnamoyl groups. Thus, a rubber-based resist, a novolak resin, an azide-based resist, or the like that generates an insolubilization reaction by adding an azide compound to a novolak resin can be used.

  Moreover, when using a resist as a permanent structural material (resin film layer 12) like a present Example, it is necessary to select the raw material excellent in the weather resistance. Moreover, it is desired to have resistance to the solution used, and chemical resistance is also important. In view of this, it is possible to suitably use a photosensitive polyimide that is particularly excellent in these characteristics.

  Most photoresists are commercially available in the form of a solution, which is applied onto a substrate by a method such as spin coating, and then formed as a resist film through a process such as drying or baking (annealing). There is also a method in which the resin is formed into a sheet and laminated (heated and compressed) on the substrate. These are commercially available as photosensitive resist films, and by laminating this film on the substrate 11, a fine pattern can be created.

  In the biochemical field, in order to handle environmentally sensitive biological substances (cells, proteins, DNA, RNA, etc.), it is often operated in a pH buffer such as TAE buffer (mixed solution of Tris acetate and EDTA). Is done. These buffers are usually set to be weakly alkaline (pH of about 8.0) in many cases, and in the case of a chip reactor that executes these biochemical processes, the flow path is formed of an alkali-resistant material. Is desirable. However, many of the above-mentioned photosensitive resins have highly active molecular chains that cause photocrosslinking and photopolymerization reactions, and chemical resistance is not superior to other resin products. . In addition, many photosensitive resins intended for use in semiconductor processes have the purpose of passivation in the strong acid etching of silicon, and conversely may be soluble on the alkali side.

  In particular, as a resist film having resistance in an alkali application, a rubber resist obtained by adding a bisazite compound as a photocrosslinking agent to the polyisoprene or polybutadiene, a photosensitive polyimide based on polyimide, or the like can be used.

  Japanese Patent Application Laid-Open No. 08-311130 discloses a photosensitive resin composition excellent in alkali resistance, comprising a vinyl ester resin having a tricyclodecane skeleton, tricyclodecane dimethylol diacrylate and a photopolymerization initiator as essential components. It is disclosed and can be used.

  An epoxy-based permanent resist represented by SU-8 as used in the third embodiment can also be suitably used.

  In the above-described embodiment, since a biological material such as a cell is strongly influenced by pH, a pH buffer solution whose pH does not easily change is used as the working fluid. TAE is a typical example. Furthermore, it is desirable to adjust the osmotic pressure of a plant cell (protoplast) that is easily crushed by osmotic pressure. In the embodiment using the single cell of the plant (Zucchini) described above, an aqueous solution of sugar called d-mannitol was used as an osmotic pressure adjusting component.

  As described above, in the chip reactor 1 according to the present embodiment, the photosensitive resin is used as the structural material of the fine flow path 23. For this reason, there is no need to use conventional glass direct processing, etching processes, and special imprint molds, making it easier to design flow path patterns, improving pattern accuracy, and improving flow path integration. In addition, it is possible to provide a chip reactor that can sufficiently satisfy demands such as simplification of the manufacturing process and miniaturization of the fine structure. That is, it is very difficult to form a fine pattern that forms the above-mentioned constricted part by machining, but the pattern part of each tank or flow path is formed by photolithography using a photosensitive resin as in the present invention. By forming, a fine pattern such as a constricted portion can be formed accurately and in a short time, which is suitable for mass production.

  In particular, when the chip-type reactor 1 is used for DNA extraction, it exhibits a significantly superior effect compared to the case where a conventional container called a microtube is used. In other words, it is not limited to simply simplifying the operation and promoting labor saving, and as in this embodiment, by making a chip as a reactor, a small amount of sample can be processed, and from which cell It can also be extracted, that is, an extract from a single cell can be identified and subjected to DNA analysis. Moreover, the extracted sample can be fixed to the sample stage as it is, that is, the whole reactor, and the sample can be prevented from being crushed and decomposed. That is, a series of basic operations such as separation, extraction, and fixation necessary for DNA analysis can be easily performed with high accuracy (corresponding to a small amount). Further, since the chip reactor 1 is small, it is easy to handle and can be mass-produced, so that it can be provided at low cost.

  Furthermore, in the third embodiment described above, in the introduction tank for introducing the cells, the cells are not crushed, and instead, a constricted part for trapping the cells is provided in the flow path, and the trap is trapped in the flow path. Since the electrode for crushing the cells is provided, it is not necessary to add a chemical solution for decomposing the cells to the introduction layer. Thereby, there is no possibility that the chemical solution as an impurity is mixed into the extracted nucleic acid, and it is not necessary to be careful in handling the chemical solution.

  Moreover, the timing of cell disruption can be controlled from the outside, and the extraction operation of the target intracellular constituent substance can be speeded up.

  Note that 62a and 62b in FIG. 9 and 64a and 64b in FIG. 10 align the mask at the time of pattern exposure of each tank of resin formed on the electrode pattern and the flow path. This is an alignment mark.

The top view which shows typically the chip-type reactor which concerns on one Embodiment of this invention. Sectional drawing which shows typically the cross section along the II-II line | wire in FIG. The top view of the board | substrate and resin film layer which show the positional relationship between each tank, flow path, and electrode which were formed on the board | substrate. FIG. 4 is a cross-sectional view schematically showing a cross section taken along line IV-IV in FIG. 3. The figure explaining the manufacturing process of the chip-type reactor which concerns on embodiment. The top view which shows the example of a pattern mask. The top view which shows typically the chip-type reactor which concerns on 2nd embodiment of this invention. Sectional drawing of the chip-type reactor which concerns on 3rd embodiment of this invention. The top view of the photosensitive resin material layer which shows the positional relationship between each tank, flow path, and electrode which were formed on the board | substrate. The top view of the electrode pattern which comprises a membranous electrode. The top view which shows the other Example of the cell capture trap provided in the chip | tip. The characteristic view which shows the relationship between the cell capture probability when an object cell is introduce | transduced into an Example and a comparative example, and the flow path width of a capture part. The figure which shows the cell drive test result with respect to an Example and a comparative example, Comprising: The characteristic view which shows the relationship between the voltage between electrodes, and the moving speed of a cell. 10 is a plan view in which the electrode pattern of FIG. 10 is superimposed on the pattern of each tank of FIG. It is an enlarged view of the opposing part of electrodes located in a cell trap part. It is a top view explaining the positional relationship of the cell trap part of a flow path, and the opposing part of electrodes.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Chip type reactor 11 Glass substrate 12 Resin film layer 13 Cover glass 14 Membrane electrode 21 Reaction tank 22 Waste liquid tank 23 Fine flow path (flow path)
25 Silica tank 31 Pattern mask

Claims (19)

A substrate,
A plurality of tanks arranged on the substrate and introducing a fluid for a sample and a resin layer forming a flow path communicating between the plurality of tanks on the substrate;
A reactor characterized in that the resin layer is formed of a photosensitive resin.   The reactor according to claim 1, wherein the substrate is formed of a glass material, and the resin layer is formed of a film-like photosensitive resin.   The plurality of tanks include a cell introduction tank, a waste liquid tank, and an adsorption tank. The cell introduction tank and the waste liquid tank communicate with each other via the flow path, and the middle of the flow path The reactor according to claim 2, wherein an adsorption tank is arranged to obtain an intracellular constituent material to be extracted.   The reactor according to claim 3, wherein the cell introduction tank comprises a reaction tank that performs a reaction between a cell and a chemical solution having a crushing action for the cell.   The reactor according to claim 3, wherein the channel is a linear channel that linearly connects the cell introduction tank and the waste liquid tank via the adsorption tank.   The reactor according to any one of claims 3 to 5, wherein a cover member is attached to at least the adsorption tank.   The plurality of tanks include a cell introduction tank and a waste liquid tank. The cell introduction tank and the waste liquid tank communicate with each other through the flow path, and a cell capture trap is disposed in the middle of the flow path. The reactor according to any one of claims 1 to 3, which is configured to be used for extraction of DNA, other intracellular constituent substances.   The reactor according to claim 7, wherein an electrode portion for electrically crushing the cells captured in the trap is provided corresponding to the trap.   The reactor according to claim 7 or 8, wherein the trap is formed so as to have a constricted portion, and the size of the constricted portion is 10% or more and 90% or less of the size of the cells introduced into the cell introduction tank.   The reactor according to any one of claims 7 to 9, wherein the trap has a constricted portion extended.   The reactor of Claim 9 or 10 formed in the funnel shape or taper shape from which the diameter becomes small gradually toward the said waste liquid tank from the said cell introduction tank side of the said constriction part.   The reactor according to claim 7, wherein the flow path is a straight flow path that linearly connects the cell introduction tank and the waste liquid tank via the cell capture trap.   The reactor according to any one of claims 1 to 12, wherein electrodes for applying a voltage are respectively disposed in the cell introduction tank and the waste liquid tank.   A second flow path is crossed in the middle of the first flow path that allows the cell introduction tank and the waste liquid tank to communicate with each other via the flow path, and the intracellular target substance is placed in the second flow path. The reactor of any one of Claims 3 thru | or 13 provided with the extraction liquid tank and extraction tank for isolate | separating from one flow path. In a method for producing a reactor for intersubstance reaction,
Forming electrodes at predetermined locations on the substrate,
Forming a photosensitive resin layer on the substrate on which the electrode is formed;
A pattern mask on which a plurality of tanks for introducing a sample fluid and flow paths communicating between the plurality of tanks are drawn is placed on the photosensitive resin layer and exposed,
The method for producing a reactor is characterized in that the plurality of tanks and the flow path are formed on the substrate by developing the exposed photosensitive resin layer.   The method for producing a reactor according to claim 15, wherein the substrate is made of a glass material, and the resin layer is made of a film-like photosensitive resin.   The plurality of tanks include a cell introduction tank, a waste liquid tank, and an adsorption tank. The cell introduction tank and the waste liquid tank communicate with each other via the flow path, and the adsorption is performed in the middle of the flow path. The method for producing a reactor according to claim 15, wherein a tank is provided for extraction of intracellular constituent substances.   The method for producing a reactor according to any one of claims 15 to 17, wherein a cover glass is attached to at least the adsorption tank.   The plurality of tanks are composed of a cell introduction tank and a waste liquid tank. The cell introduction tank and the waste liquid tank communicate with each other through the flow path, and a cell capture trap is disposed in the middle of the flow path. The method for producing a reactor according to claim 15, which is used for extraction of DNA, DNA or other intracellular constituents.

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