JP2006006287A - Method for producing microchip and microchip produced by the method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a chip requiring no tube addition in after processing, which tube is for injecting or collecting samples such as cells and chemotactic factors. <P>SOLUTION: The microchip 1 is made of a thermosetting silicone and has wells 31 and 32 for filling chemotactic factors or cells having chemotaxis, a path 4 having micro-paths communicating the well 31 with the well 32, and tubes 21 and 22 for injecting or collecting samples such as cells or chemotactic factors. The method for producing a microchip 1 comprises at least a step for inserting tubes 21 and 22 to molds 11 and 12 for molding chips, a step for injecting a silicone material for forming chips into molds 11 and 12 for forming chips, a step for baking the injected material to cure and a step for releasing the cured molded products from the mold. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to a microchip manufacturing method and a microchip manufactured by the manufacturing method.

  Various techniques for mixing and analyzing liquid specimens such as medicines and biological substances have been proposed and put into practical use. For example, blood viscosity measurement and cell state observation are evaluated, and cell chemotaxis is observed. Chemotaxis is the property that cells move in response to a concentration gradient of chemical substances called chemotaxis factors. There is a movement to apply this cell chemotaxis to the development of therapeutic drugs. In particular, it is thought that by using this chemotaxis, a new approach can be found in the development of therapeutic agents for various inflammations, allergies, and cancers, and the importance of chemotaxis research is increasing. As a technique for mixing and analyzing such a liquid specimen, a flow-type microchip has attracted attention (see, for example, Patent Document 1 and Non-Patent Document 1).

  Here, a chip for observing chemotaxis will be described as an example of a microchip. A microchip for observing chemotaxis has at least a well that is filled with a chemotactic factor and a well that is filled with a chemotactic cell or the like. These wells communicate with each other through a flow path, and a number of fine passages separated by barriers are provided in the flow path. A cell moves through the separated minute passage.

  FIG. 5 is a cross-sectional view for explaining the overall configuration of a unit 100 including a conventional microchip 1. In the figure, the microchip 1 is provided with a well 31 and a well 32 which are regions filled with cells or chemotactic factors. A tube 21 and a tube 22 are connected to the wells 31 and 32, respectively. The tube 21 and the tube 22 are not particularly limited, but are made of, for example, a silicon material.

  The tube 21 and the tube 22 are cylindrical bodies having minute through holes provided for injecting and collecting a sample. Further, between the well 31 and the well 32, a flow path portion 4 that communicates between them is provided. The channel portion 4 is formed with fine passages separated by a barrier. The size of such a passage is made slightly smaller than the normal size of the cell, and if a concentration gradient of the chemotactic factor is created, the cell moves to the higher concentration through the passage, so that the observation of chemotaxis is possible. it can.

  The microchip 1 is used in close contact with a substrate 10 having transparency. The cells passing through the fine passages of the flow path portion 4 from the substrate 10 side are observed. The conventional microchip 1 mainly uses a method of processing a silicon wafer (single crystal silicon) using a micromachining technique such as anisotropic dry etching or PDMS (Polydimethylsiloxane) which is a kind of silicon rubber. It is manufactured by the molding method. The tube 21 and the tube 22 were attached to the microchip 1 by post-processing after the microchip 1 was molded.

  FIG. 6 shows a state in which the microchip 1 is manufactured by PDMS. In this case, PDMS that has been stirred and degassed and vacuum degassed is cast into a region P surrounded by at least the first mold 11 and the second mold 12 and baked and cured. After that, the mold is released, and in some cases, the chip is separated from the runner and inspected for the target shape.

Next, FIG. 7 shows a state in which the tubes 21 and 22 are attached to the microchip 1 formed by PDMS. Here, only the left half of the microchip 1 shown in FIG. 5 is shown, but the right half is assumed to be line-symmetrical. First, as shown in FIGS. 7A, 7 </ b> B, and 7 </ b> C, a hole 21 ′ is formed in the microchip 1 using a puncher 200. Next, as shown in FIG. 7D, the tube 21 is inserted into the hole 21 ′. Finally, as shown in FIG. 7 (e), the tube 21 is bonded by an adhesive 201 to fix the tube 21.
JP 2002-159287 A Nikkei Biobusiness November 2001, p. 48-50

  Since the microchip 1 is molded by PDMS, the microchip 1 is elastic, and the surface is distorted when drilling as shown by a Q portion in FIG. Then, the hole 21 'becomes concave as shown in the R part of FIG. When the tube 21 is inserted and fixed in the hole 21 ′ as shown in FIG. 7D in this state, the connection surface with the tube is not flat, so that the sealing performance remains questionable. In addition, regarding the adhesion by the adhesive 201 shown in FIG. 7 (e), if the adhesive 201 flows out into the well 31 or the tube 21, the path of the cell may be blocked and cannot be used for observation of chemotaxis. It is also possible to become.

  The operation of attaching the tube 21 to the microchip 1 is a manual operation. It is very precise work, such as controlling the position where the hole 21 'is opened, inserting the tube 21, and applying the adhesive 201. In addition to requiring the skill of the operator, there is a limit to the production speed. is there.

  The present invention has been made to solve such a problem, and provides a microchip that does not require post-processing attachment of a tube for injecting and collecting a sample such as a cell or a chemotactic factor. The purpose is to do.

  The method for manufacturing a microchip according to the present invention includes a step of inserting a tube for injecting or collecting a sample into the microchip into a mold for microchip molding, and using the microchip molding material as the microchip molding material. A step of casting into the mold, a step of baking and curing the cast material, and a step of releasing the cured molded product. With such a configuration, it is possible to provide a microchip that does not require the tube to be attached by post-processing.

  The microchip manufacturing method according to the present invention includes at least one of the first and second regions, a flow path having a fine passage communicating between the regions, and at least one of the regions. A method for producing a microchip made of thermosetting silicon comprising a tube for injecting or collecting a sample, the step of inserting the tube into at least the microchip molding die, and the silicon for microchip molding The method includes a step of casting a material into the mold for microchip molding, a step of baking and curing the cast material, and a step of releasing the cured molded product. With such a configuration, it is possible to provide a microchip that does not require the tube to be attached by post-processing.

  Here, the microchip molding die is provided with a projection having a diameter larger than the inner diameter of the tube. In the step of inserting the tube into the microchip molding die, the tube is inserted into the projection. You may make it fit. As a result, the silicon material can be prevented from flowing into the tube, and the generation of burrs can be suppressed.

  Specifically, the diameter of the protrusion is 1.1 to 1.2 times the inner diameter of the tube, and when the tube is fitted into the protrusion, the diameter is expanded to be 1.1 to 1.2 times. It is preferable.

  Further, the protrusion provided on the microchip mold for the previous period may be a pin driven into the mold.

  On the other hand, the microchip according to the present invention is configured to inject a sample into at least one of the first and second regions, a flow path having a fine passage communicating between the regions, A microchip made of thermosetting resin having a tube for sampling, and the tube is inserted into the microchip mold when the microchip is molded, and is molded integrally with the microchip. It is a thing. With such a configuration, it is possible to provide a microchip that does not require the tube to be attached by post-processing.

  The mold for forming a microchip according to the present invention includes at least one of the first and second regions, a flow path portion having a fine passage communicating between the regions, and at least one of the regions. A thermosetting resin microchip molding die provided with a tube for injecting or collecting a sample into the hole, and a hole for inserting the tube into the molding die, and fitting the tube inside the molding die Protrusions,

It is characterized by having. With such a configuration, it is possible to provide a microchip that does not require the tube to be attached by post-processing.

  According to the present invention, it is possible to provide a microchip that does not require attaching a tube for injecting and collecting a sample by post-processing.

  Hereinafter, embodiments to which the present invention can be applied will be described. The following description is to describe the embodiment of the present invention, and the present invention is not limited to the following embodiment. For the sake of clarity, the following description is omitted and simplified as appropriate. Moreover, those skilled in the art can easily change, add, and convert each element of the following embodiments within the scope of the present invention. Further, in each of the following embodiments, a case where a chip is used for observing the chemotaxis of cells will be described for concreteness, but the liquid specimen is not limited to this, for example, protein, nucleic acid, DNA, It is a liquid containing a measurement object such as RNA, peptide, hormone, antigen, antibody, ligand, receptor, enzyme, substrate, low molecular organic compound, cell, ion, etc., and also includes a dispersion system such as a suspension or a colloid. That is, the microchip according to the present invention can also be used for mixing and analysis of liquid specimens as described above.

Embodiment 1
FIG. 1 is a cross-sectional view of a unit 100 including at least a tube 21, a microchip 1 integrated with the tube 22, and a substrate 10 having transparency. The microchip 1 is provided with a well 31 and a well 32. For the sake of simplicity, two wells, a well 31 and a well 32, will be described, but a plurality of wells may be provided. In addition, the tube may not be provided in all wells, and it is sufficient that at least one well has.

  The wells 31 and 32 form spaces when the microchip 1 and the transparent substrate 10 are in close contact with each other, and the spaces are filled with cells or chemotactic factors. The wells 31 and 32 according to this example are formed to be recessed about 100 μm from one surface of the microchip 1. There is no particular limitation on the volume of the wells 31 and 32, and it is sufficient to have a minimum volume necessary for filling samples such as cells and chemotactic factors. For example, the depth may be about 100 μm, and the width and depth may be about 1 to 2.5 mm.

  Tubes 21 and 22 are connected to wells 31 and 32. Tubes 21 and 22 are cylindrical bodies provided with minute through holes for injecting and collecting samples of cells or chemotactic factors. . The diameters of the tubes 21 and 22 are not particularly limited as long as the sample can be injected using, for example, a microsyringe having a diameter of 0.5 mm. Therefore, the inner diameters of the tubes 21 and 22 in this embodiment are about 1 mm and the outer diameter is about 2 mm. Moreover, the tubes 21 and 22 do not need to be cylindrical, and may be rectangular.

  Between the well 31 and the well 32, the flow path part 4 which connects both is provided. A minute passage separated by a barrier is formed in the flow path section 4, and when a concentration gradient of the chemotactic factor is formed, the cells move to the higher concentration through the passage. Can do.

  Chemotactic observation is possible by detecting an image of a cell moving in the passage of the flow path section 4 from the outer observation surface through the substrate 10 having transparency. Specifically, a moving cell is detected by a detector (not shown) such as a microscope (not shown) or a video camera (not shown) installed on the observation surface. In this case, the cell image can be sufficiently confirmed. Brightness is required. For this reason, a light source (not shown) is installed on the observation surface side to illuminate the vicinity of the flow path portion 4 with illumination light.

  In the present embodiment, when the microchip 1 is formed, the tubes 21 and 22 are inserted into a mold in advance and formed. Then, when the microchip 1 molding is completed, the microchip 1 and the tubes 21 and 22 are integrated, so that it is possible to improve the mass productivity by eliminating post-processing such as drilling and bonding after molding.

  FIG. 2 is a cross-sectional view showing a state in which the microchip 1 and the silicon tubes 21 and 22 according to the present embodiment are integrally formed. In FIG. 2, 11 and 12 are molds for molding the microchip 1. Here, the molds are divided into an upper mold 11 and a lower mold 12. However, the present invention is not limited to this and is divided into three or more. It may be. A P portion which is a cavity inside the upper die 11 and the lower die 12 is filled with a silicon material which is a material of the microchip 1. The silicon material used here is, for example, PDMS (Polydimethylsiloxane: silicone-based two-mixed thermosetting resin).

  The material of the tubes 21 and 22 is not particularly limited, but is preferably silicon. This is because the tubes 21 and 22 must be inserted into the mold during baking of the PDMS cast in the upper mold 11 and the lower mold 12 and are therefore preferably made of a material that does not thermally deform. Further, if the tubes 21 and 22 are made of silicon, the tubes 21 and 22 are closely adhered to the microchip 1 formed by PDMS.

  The silicon tubes 21 and 22 may be twisted due to their elasticity. Straightening pins 211 and 221 are inserted into the tubes 21 and 22 in order to eliminate problems caused by twisting of the tubes 21 and 22 during molding. The tubes 21 and 22 are extended by the correction pins 211 and 221 and pressed against the S portion. This prevents the silicon material from flowing into the tube.

  FIG. 3 is a flowchart showing a process for manufacturing the microchip 1. First, PDMS required for molding is weighed (S101). Next, PDMS is defoamed by stirring (S102). Further, PDMS is completely degassed by evacuating (S103). Further, the upper mold 11 and the lower mold 12 for molding PDMS are assembled, and the silicon tubes 21 and 22 are inserted into the assembled mold (S104). Straightening pins 211 and 221 are inserted into the inserted tubes 21 and 22. When materials and molds are ready in this way, PDMS is cast into a combined mold of upper mold 11 and lower mold 12 (S105). When the inside of the mold is filled with PDMS, it is baked and cured (S106). When it is cured and can be taken out as a molded product, it is released (S107).

  Here, S104 is arranged after S101 to 103, but S104 may be performed before S101 to S103 or may be performed simultaneously. By forming the microchip 1 by inserting the tubes 21 and 22 into the mold in the above-described procedure, a microchip excellent in mass productivity that does not require post-processing after chip formation can be provided. .

Embodiment 2

  Next, Embodiment 2 of the present invention will be described with reference to FIG. The basic contents of the present embodiment are the same as those of the first embodiment, and a description thereof will be omitted. The same reference numerals are the same as those in the first embodiment. FIG. 4 is a cross-sectional view showing a state in which the microchip 1 and the silicon tubes 21 and 22 according to the present embodiment are integrally formed. 4A, when the tubes 21 and 22 are inserted into the upper mold 11 and the lower mold 12, the tubes 21 and 22 are placed on the driving pins 210 and 220 driven into the mold 12 when the tubes are pressed against the S portion. Try to plug in. The diameter of the driving pins 210 and 220 is not particularly limited, but is 1.1 to 1.2 times the inner diameter of the tubes 21 and 22, for example. Accordingly, when the tubes 21 and 22 are inserted into the driving pins 210 and 220, the tubes 21 and 22 are expanded 1.1 to 1.2 times.

  FIG. 4B is an enlarged cross-sectional view of the S portion. In FIG. 4B, only the S portion on the tube 21 side is shown enlarged, but the same applies to the tube 22 side. In FIG. 4 (b), the tube 21 which is an elastic body is fitted into a cylindrical driving pin 210 having an outer diameter 1.1 to 1.2 times its inner diameter, due to its properties as an elastic body. It shows how it is. Regarding the T portion, the side surface of the driving pin 210 and the inner surface of the tube 21 are pressed to such an extent that the tube 21 is elastically expanded. For this reason, PDMS can be more effectively prevented from flowing into the tube 21 from below. In addition, since the tube 21 is securely fixed by being fitted into the driving pin 210 rather than simply being pressed, it is possible to prevent molding defects due to displacement or inclination of the tube 21.

It is sectional drawing of the microchip concerning this invention. It is sectional drawing showing the shaping | molding method of the microchip concerning Embodiment 1 of this invention. It is a flowchart showing the manufacturing process of the microchip concerning this invention. It is sectional drawing showing the shaping | molding method of the microchip concerning Embodiment 2 of this invention. It is sectional drawing of the microchip concerning a prior art. It is sectional drawing showing the shaping | molding method of the microchip concerning a prior art. It is sectional drawing showing the connection method of the microchip and tube concerning a prior art.

Explanation of symbols

1 microchip, 100 units,
21, 22, tube, 21 'hole,
200 hole punch, 201 adhesive, 211, 221 straightening pin,
210, 220 Driving pins 31, 32 wells, 4 channels,
10 substrate, 11 upper mold, 12 lower mold,
P cavity, Q strained part, R drilling strained part, S pressing surface,
T expansion section

Claims (10)

Inserting a tube for injecting or collecting a sample into a microchip into a mold for forming a microchip in the previous period;
Casting the microchip molding material into the microchip molding die;
Baking and curing the cast material;
Releasing the cured molded product; and
A method for producing a microchip, comprising:   The microchip mold is provided with a protrusion having a diameter larger than the inner diameter of the tube, and the tube is inserted into the protrusion in the step of inserting the tube into the microchip mold. The method of manufacturing a microchip according to claim 1.   The diameter of the projection is 1.1 to 1.2 times the inner diameter of the tube, and when the tube is fitted into the projection, the tube is expanded and fitted to 1.1 to 1.2 times. The method for producing a microchip according to claim 2.   4. The method of manufacturing a microchip according to claim 2, wherein the protrusion provided on the mold for forming the microchip in the previous period is a pin driven into the mold. At least a first and a second two or more regions, a flow path portion having a fine passage communicating between the regions, and a tube for injecting or collecting a sample into at least one of the regions. A method of manufacturing a thermochip made of thermosetting silicon,
Inserting the tube into at least the microchip mold;
Casting the microchip molding silicon material into the microchip molding die; and
Baking and curing the cast material;
Releasing the cured molded product; and
A method of manufacturing a microchip having   The microchip mold is provided with a protrusion having a diameter larger than the inner diameter of the tube, and the tube is inserted into the protrusion in the step of inserting the tube into the microchip mold. The method of manufacturing a microchip according to claim 5.   The diameter of the projection is 1.1 to 1.2 times the inner diameter of the tube, and when the tube is fitted into the projection, the tube is expanded and fitted to 1.1 to 1.2 times. A method for manufacturing a microchip according to claim 6.   8. The method of manufacturing a microchip according to claim 6, wherein the protrusion provided on the mold for forming the microchip in the previous period is a pin driven into the mold. At least a first and a second two or more regions, a flow path portion having a fine passage communicating between the regions, and a tube for injecting or collecting a sample into at least one of the regions. A microchip made of thermosetting resin,
The tube is inserted into the mold for forming the microchip when the microchip is formed, and is formed integrally with the microchip. At least a first and a second two or more regions, a flow path portion having a fine passage communicating between the regions, and a tube for injecting or collecting a sample into at least one of the regions. A mold for thermochip molding made of thermosetting resin,
A hole for inserting the tube into the mold, and the tube and the microchip can be integrally formed by inserting the tube into the hole during the microchip molding. Type.

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