JP2008026078A - Method of manufacturing recessed chip - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a recessed chip having a cover member closely bonded and fixed thereto and capable of effectively preventing the leak of a sample liquid. <P>SOLUTION: A photosensitive material layer 12 is formed to a substrate 11 by coating the substrate 11 with a photosensitive material and cover glass 13 is laminated to the photosensitive material layer 12 to be closely bonded thereto. Thereafter, the photosensitive material layer 12 is exposed to light through the cover glass 13 and developed to form a sample charging tank 21, a waste liquid tank 22, a flow channel 23 and a silica tank 25. If the flow channel 23 or the like is formed to the photosensitive material layer 12 as is conventional and covered with the cover glass 13 after cured, a gap is formed between the cover glass 13 and the photosensitive material layer 12 by the irregularity of the thickness of the photosensitive material layer 12 and a sample solution is leaked from the flow channel 23 to be penetrated in the gap. The photosensitive material layer 12 is cured by this manufacturing method after the cover glass 13 is placed, the cover glass 13 and the photosensitive material layer 12 can be closely bonded. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a method for manufacturing a recess-formed chip in which recesses such as fine flow paths and reaction liquid tanks are formed on a chip.

  In recent years, in biochemical analysis of cells, DNA (deoxyribonucleic acid), RNA (ribonucleic acid), and the like, a chip formed with a fine channel having a width of several hundred μm or less may be used (see Patent Document 1). ). Electrodes are formed at both ends of the flow channel in the extending direction, and a voltage is applied to both electrodes to move DNA or cells in the sample solution introduced into the flow channel by electroosmotic flow or electrophoresis. Thus, operations such as mixing and separation of the DNA and cells can be performed. Further, by laminating the cover member so as to cover the flow path, it is possible to prevent the sample solution from drying and the sample solution from leaking out from the flow path. As a method for forming such a flow path, there is a method using a lithography technique as shown in Patent Document 1 in addition to machining (injection molding, cutting, etc.). If lithography technology is used, extremely fine channels can be made with high accuracy and mass production is possible.

  When manufacturing using a lithography technique, first, a photosensitive material is applied onto a substrate such as glass with a spin coater, etc., and heated at a predetermined temperature to volatilize organic components (soft bake), and the photosensitive material layer is applied to the substrate. Let it settle. Then, the flow pattern is transferred to the photosensitive material layer by exposing the photosensitive material layer with a pattern mask on which the flow pattern is printed. Thereafter, the photosensitive material layer in the flow path forming region is removed by development to complete the flow path.

  Here, when the cover member is provided, the cover member is fixed to the photosensitive resin material layer after the flow path is completed. In this case, in order to prevent leakage of the sample solution and the like, it is necessary to bring the cover member and the photosensitive material layer into close contact. For this purpose, the surface of the photosensitive material layer needs to be flattened. However, when the photosensitive material is applied by a spin coater, unevenness is generated on the surface, and the end of the photosensitive material layer is raised, so that it is difficult to ensure the required flatness.

On the other hand, in patent document 1, the flat surface is obtained by forming the photosensitive material layer by affixing the photosensitive resin film on a board | substrate. Patent Document 2 discloses a technique for making the surface uniform by controlling the number of rotations stepwise when a photosensitive material is applied by a spin coating method.
JP 2004-286449 A JP 2000-082647 A

However, when a photosensitive resin film is used as in Patent Document 1, the surface can be flattened, but the composition of the photosensitive resin material is limited and the adhesion between the photosensitive resin film and the substrate is poor. There is a problem to say.
In addition, the method of Patent Document 2 can achieve a certain degree of effect in applications where the photosensitive material layer is thin. However, in applications where flow paths are formed, it is necessary to make the photosensitive material layer relatively thick. In this case, a photosensitive material having a high viscosity is often used. It is difficult to form a conductive material layer. On the other hand, it is possible to cut and remove the raised edge after curing of the photosensitive material layer or to dissolve and remove it with an organic solvent. However, if the substrate size is small, only the edge is removed after curing. It ’s difficult. For example, when the cover member is pressure-bonded to the photosensitive material layer for close contact, the flow path is crushed and the shape cannot be maintained.

  The present invention has been made in view of the above-described problems, and an object of the present invention is to provide a recess-forming chip in which a cover member is closely attached and fixed to effectively prevent leakage of a sample solution.

  In order to solve the above-described problem, a method for manufacturing a recess-forming chip according to claim 1 of the present invention includes stacking a photosensitive material layer on a substrate and partially forming the photosensitive material layer on the surface. A method for producing a recess-forming chip formed by forming a recess and laminating a cover member so as to block at least a part of the recess in the photosensitive material layer, and applying a photosensitive material to the substrate, A step of forming the photosensitive material layer, a step of laminating the cover member made of a translucent material on the photosensitive material layer, and attaching the cover member; and exposing the photosensitive resin layer through the cover member; And developing the photosensitive resin layer after the exposure to form the concave portion.

The manufacturing method of the recessed part formation chip | tip by Claim 2 of this invention is characterized by laminating | stacking the said cover member before baking processing of the said photosensitive material layer in Claim 1.
Here, the baking process includes a pre-bake process performed before the exposure process.
According to a third aspect of the present invention, there is provided a method for manufacturing a recess forming chip according to the first or second aspect, wherein the developing process is performed while the recess forming chip is swung in a direction along the plate surface.

  According to the manufacturing method of the present invention, it is possible to manufacture a recessed portion-forming chip that allows the cover member and the photosensitive material layer to be in close contact to effectively prevent leakage of the sample solution.

Next, embodiments of the present invention will be described with reference to the drawings.
(Configuration of recessed portion forming chip)
1 is a plan view of a recess forming chip 1 according to the present embodiment, FIG. 2 is a cross-sectional view taken along the line II of FIG. 1, and FIG. 3 is a cross-sectional view taken along the line II-II.
The recess forming chip 1 includes a glass substrate 11, film electrodes 14 and 14 partially laminated on the substrate 11, a photosensitive material layer 12 laminated on the substrate 11 or film electrodes 14 and 14, and a photosensitive property. And a cover glass 13 partially laminated on the material layer 12.

  Then, by partially removing the photosensitive material layer 12, a sample charging tank 21, a waste liquid tank 22, a flow path 23, and a silica tank 25 are formed. The sample charging tank 21, the waste liquid tank 22, the flow path 23, and the silica tank 25 correspond to the recesses of the present invention. In FIG. 1, the sample introduction tank 21 and the waste liquid tank 22 have substantially the same shape, and are formed in a rectangular shape on both the left and right sides in plan view, and a flow path 23 that communicates these is formed to extend linearly. . A silica tank 25 having a width wider than that of the flow path 23 is disposed in the center of the flow path 23 in the extending direction.

  The sample loading tank 21 and the waste liquid tank 22 are used for loading or collecting the sample solution, and are exposed to the outside so that the sample solution can be loaded or discarded from the outside. In addition, film electrodes 14 and 14 are laminated in the formation region of the sample charging tank 21 and the waste liquid tank 22, respectively, and constitute the bottom surfaces of the sample charging tank 21 and the waste liquid tank 22. The film electrodes 14 and 14 form an electric field for driving the sample solution between the sample introduction tank 21 and the waste liquid tank 22. In the present embodiment, a DC voltage is applied with the membrane electrode 14 on the sample loading tank 21 side as the positive electrode and the membrane electrode 14 on the waste liquid tank 22 side as the negative electrode, thereby generating an electroosmotic flow in the sample solution. It moves through the flow path 23 from the charging tank 21 toward the waste liquid tank 22. The channel 23 has a channel width larger than the diameter of the detection target substance so that the detection target substance contained in the sample solution can pass through. For example, when detecting cells having a diameter of about 20 μm contained in a sample solution, the flow path width needs to be set to be at least larger than this, but when detecting DNA or biomolecules from a cell disruption solution. Can be set smaller than this.

  In the silica tank 25, silica particles (not shown) for adsorbing the detection target substance are arranged, and the detection target substances (cells, DNA, RNA, protein) contained in the sample solution that moves through the flow path 23. Trap molecules). For example, if an oligonucleotide having a base sequence complementary to a part of DNA to be detected is synthesized on the silica particle surface by a solid phase method, the target DNA can be trapped from the sample solution. In this case, for example, if all the DNA in the sample solution is fluorescently labeled, the target DNA concentration in the sample solution can be measured by measuring the fluorescence intensity of the silica particles after the sample solution has flowed through the flow path. can do. A cover glass 13 is laminated on the photosensitive material layer 12 in the area where the silica tank 25 is formed and the area around it. Thus, by covering the silica tank 25 and the flow path 23 formed in the periphery thereof with the cover glass 13, evaporation of the sample solution in the silica tank 25, mixing of impurities, leakage of the sample solution to the outside of the tank, etc. Can be prevented.

  Note that the present invention is not limited to the application to the recess forming chip 1 in which the flow path 23 and the like are arranged as shown in FIG. Is possible. For example, a configuration in which two or more flow paths intersect, a configuration in which a silica tank is not formed, a configuration in which a sample solution is flowed with a pump, or a configuration in which only substances in the sample solution are flowed by electrophoresis The present invention can be applied even if it is a thing.

Next, materials for the above-described constituent members will be described.
The substrate 11 is a support that supports the photosensitive material layer 12, and a material having a necessary strength as the support can be used. For example, plastic (polystyrene) is used in addition to glass (borosilicate glass, quartz glass, etc.). , Polymethyl methacrylate, polysulfone, polyester, etc.) and glass fiber and plastic composites can also be used. Among these, when glass or translucent plastic is used, reflection at the time of exposure can be suppressed, and an experimental operation can be performed while observing the recessed portion-forming chip 1 with a microscope, or optical measurement such as measurement of the fluorescence intensity can be performed. It is also suitable for analysis.

  In addition, an electrode material generally used for forming a film electrode can be used for the film electrode 14, but in the application of the present embodiment, since it is exposed to a sample solution, Pt is used from the viewpoint of preventing electrolytic corrosion and the like. It is preferable to select a material having a relatively high standard electrode potential (such as a positive value) such as Au, Ag, or the like. Further, when a transparent electrode such as ITO (Indium Tin Oxide) is used, the transparency of the recessed portion forming chip 1 can be maintained, which is preferable when performing optical analysis of the recessed portion forming chip 1 as described above. Further, it is not always necessary to integrally form the recess-forming chip 1 as a film-like electrode, and a driving electrode may be formed by immersing a separately formed electrode bar in the sample solution.

The photosensitive material includes a photosensitive resin as a main component, and subcomponents (photosensitive agent, acid generator, etc.) for causing the photosensitive resin to undergo a photosensitive reaction (crosslinking reaction, main chain cleavage, deprotection reaction, etc.) Containing. As the photosensitive material, a commonly used material can be used, and either a positive type or a negative type can be used.
The cover glass 13 corresponds to the cover member of the present invention. In the present invention, as will be described later, since the photosensitive material layer 12 laminated on the lower portion is exposed through the cover member, the cover member has translucency that transmits the irradiation light during the exposure process. There must be. If it has such a property, the plastics etc. which were enumerated as a material used for the said board | substrate 11 other than glass can also be used. Moreover, by using a translucent material, the optical analysis of the recessed portion forming chip 1 becomes possible as described above.

  From the above viewpoint, in this embodiment, a commercially available borosilicate glass (trade name: Pyrex (registered trademark)) is used for the substrate 11. By using a hard borosilicate glass excellent in heat resistance, damage or the like in the manufacturing process of the recess forming chip 1 can be prevented. Further, a negative photoresist (trade name: SU-8) manufactured by Kayaku Microchem Corporation is used as the photosensitive material. The film electrode 14 has a two-layer structure in which Ti is a base and the surface is covered with a Pt reaction protective film. Adhesion with the substrate 11 can be improved by using Ti as a base, and electrode reaction can be suppressed by covering Pt. A commercially available cover glass 13 is used. In the example of FIG. 1, the size of the recess forming chip 1 is 20 mm in length and 10 mm in width, and the thickness of the substrate 11 is 1 mm. The cover glass 13 has a length of 10 mm, a width of 5 mm, and a thickness of 0.15 mm. The cover glass 13 is arranged on the silica tank 25 with its longitudinal direction aligned with the short direction of the substrate 11.

(Manufacturing method of recessed part forming chip)
Next, a manufacturing method of the recess forming chip 1 having the configuration according to the present embodiment will be described with reference to FIG. 4 is a cross-sectional view taken along line III-III in FIG.
First, the surface of the substrate 11 is cleaned with acetone (FIG. 4A).
Next, the film electrodes 14 are formed on the substrate 11 at predetermined positions (FIG. 4B). The film electrode 14 can be formed by physical vapor deposition (electron beam vapor deposition, ion plating, sputtering, etc.) or chemical vapor deposition.

  Next, a photosensitive material is applied to the entire surface of the substrate 11 to form a photosensitive material layer 12 (FIG. 4C). Specifically, in the present embodiment, the substrate 11 is attached to a spin coater and rotated at a predetermined rotational speed, thereby forming the photosensitive material layer 12 having a predetermined thickness. The coating may be performed by spray coating instead of spin coating. However, in the application of the present invention, it is preferable to use spin coating because the layer thickness is relatively thick.

  Next, the cover glass 13 is placed on the uncured photosensitive material layer 12 and the surfaces thereof are brought into close contact with each other (FIG. 4D). Thereafter, soft baking is performed at a predetermined temperature to volatilize the solvent in the photosensitive material layer 12. Thereby, the photosensitive material layer 12 is cured while the cover glass 13 is in close contact, and the cover glass 13 is fixed to the photosensitive material layer 12.

Next, the substrate 11 is exposed, and a flow path pattern is baked on the photosensitive material layer 12 (FIG. 4E). Here, the flow path pattern in this embodiment includes not only the flow path 23 formation area but also the entire removal part (formation area of the sample introduction tank 21 and the waste liquid tank 22, the flow path 23 and the silica tank 25), and other parts. It is a pattern that represents. In FIG. 4E, the proximity exposure is performed using the pattern mask 4 and the negative photosensitive material is used as described above. Therefore, the pattern mask 4 corresponds to the removed portion of the photosensitive material layer 12. The area to be used is a light shielding area, and the other part is a light transmitting area. The photosensitive material layer 12 is partially insolubilized by the irradiation light transmitted through the light transmitting region. At this time, the cover glass 13 is laminated in the area where the silica tank 25 is formed. Since the translucent cover glass 13 is used as described above, the irradiated light passes through the cover glass 13 and the laminated portion. The flow path pattern is also baked on the photosensitive material.
In addition, in consideration of the level difference between the region where the cover glass 13 is laminated and other regions, if more accuracy is required, for example, the portion where the cover glass 13 is laminated and the other portion are divided into two steps. You may expose. Moreover, it is not limited to the case where it exposes using the pattern mask 4 as mentioned above, You may draw directly with an electron beam.

  Next, the substrate 11 is developed (FIG. 4F). As a result, the region of the photosensitive material layer 12 that has received the irradiation light in the exposure process remains without being dissolved, and the light-shielded region is dissolved and removed, so that the concave sample feeding tank 21 and the waste liquid tank 22 are flown. A passage 23 and a silica tank 25 are formed. At this time, if development is performed while the substrate 11 is swung in the extending direction of the flow path 23, the developer or dissolved material flows through the flow path 23, and development of the area where the cover glass 13 is laminated can be promoted. preferable.

Although not shown, the silica particles are arranged inside the silica tank 25 by flowing the silica particles into the flow path 23 after development.
As described above, the translucent cover glass 13 is brought into close contact with the photosensitive material layer 12 before curing, and the photosensitive material layer 12 is exposed and developed in this state, so that the cover glass 13 is also laminated. A flow path 23 is formed. Accordingly, the cover glass 13 can be adhered and fixed without crushing the flow path 23 by pressing the cover glass 13 in order to flatten the photosensitive material layer 12, and the flow path of the sample solution Leakage from 23 can be prevented.

It is a top view of the recessed part formation chip | tip which concerns on this embodiment. It is the II sectional view taken on the line of FIG. It is the II-II sectional view taken on the line of FIG. It is a figure for demonstrating the manufacturing method of this embodiment (III-III sectional view taken on the line of FIG. 1).

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Recessed chip | tip, 11 Substrate, 12 Photosensitive material layer, 13 Cover glass, 14 Film electrode, 21 Sample input tank, 22 Waste liquid tank, 23 Channel, 25 Silica tank, 4 Pattern mask

Claims (3)

A photosensitive material layer is laminated on the substrate, and a concave portion is formed on the surface by partially forming the photosensitive material layer, and at least a part of the concave portion is blocked by the photosensitive material layer. A method for manufacturing a recess-forming chip formed by laminating cover members,
Applying a photosensitive material to the substrate to form the photosensitive material layer;
Laminating the cover member made of a light-transmitting material on the photosensitive material layer and bringing it into close contact;
Exposing the photosensitive resin layer through the cover member, and developing the exposed photosensitive resin layer to form the concave portion; and
The manufacturing method of the recessed part formation chip | tip characterized by the above-mentioned.   The method for manufacturing a recess-forming chip according to claim 1, wherein the cover member is laminated before baking the photosensitive material layer.   The method for manufacturing a recess forming chip according to claim 1, wherein the developing process is performed while the recess forming chip is swung in a direction along a plate surface.

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