JPWO2009034819A1 - Microchip manufacturing method, microchip, and vacuum bonding apparatus - Google Patents

Abstract

In a microchip manufacturing method in which at least two substrates are laminated, a positioning step of inserting a reference hole of a lower substrate into a positioning pin and then inserting a reference hole of an upper substrate into the positioning pin with an adhesive layer interposed therebetween, and positioning A method for manufacturing a microchip, comprising: a bonding step of bonding substrates subjected to the steps; and a pressing step of pressing and pressing the uppermost substrate.

Description

  The present invention relates to a microchip manufacturing method, a microchip, and a vacuum sticking apparatus.

  In recent years, by making full use of micromachine technology and ultrafine processing technology, devices and means (for example, pumps, valves, flow paths, sensors, etc.) for performing chemical analysis, chemical synthesis, etc. are miniaturized and integrated on one chip. Such a system has been developed (see, for example, Patent Document 1). This is also called μ-TAS (Micro Total Analysis System), bioreactor, lab-on-chip, biochip, medical examination, diagnostic field, environmental measurement field, agricultural production Its application is expected in the manufacturing field. In reality, as seen in genetic testing, automated, faster, and simplified microanalysis systems are costly and necessary when complex processes, skilled techniques, and equipment operations are required. The benefits of enabling time-and-location analysis as well as sample size and time are enormous.

  In various types of analysis and inspection, these analysis chips (hereinafter referred to as “microchips” in which a microchannel is provided in the chip and performs various reactions in the microchannel) are used. . For example, a working fluid is supplied into a microchip composed of at least two substrates, and a reagent and a sample that are stored in advance are merged, and the detection unit formed in the downstream area of the merging unit uses an optical detection unit. A micro integrated analysis system that automatically inspects an inspection object is disclosed (for example, Patent Document 2).

  Since such a microchip encloses a reagent or the like in a flow channel space formed by bonding a flow channel substrate in which a groove forming a flow channel is processed and a coated substrate that covers the groove, and injects a working fluid. If there is a gap between the substrates, liquid leakage will occur and the inspection will not be performed correctly.

As a method of closely attaching two substrates of a microchip, a method of joining a flow path substrate made of an elastic material having a self-sealing property with patterned flow paths and a glass substrate (for example, Patent Document 3) is disclosed. In addition, a method of joining two thin plates made of a semiconductor such as metal or silicon (for example, Patent Document 4) is disclosed.
JP 2004-28589 A JP 2006-275734 A JP 2004-108285 A Japanese Patent Laid-Open No. 2006-18785

  In the micro total system as disclosed in Patent Document 2, since the microchip is disposable every time, it is desirable to manufacture the microchip with a resin that is inexpensive and easy to manufacture. Generally, the flow path substrate is processed by injection molding a resin substrate using a mold to form a groove for forming a flow path or a hole for injecting a fluid.

  However, generally, when a resin substrate is injection-molded, sink marks and burrs may occur in grooves and holes, and warpage may occur in the substrate. When such a resin substrate is bonded, a gap is generated, and the remaining air may interfere with liquid feeding in the flow path. In particular, when the microchip is heated to promote the reaction, the influence of the remaining air is great. Patent Documents 2, 3, and 4 do not disclose a method for solving these problems when bonding resin substrates.

  This invention is made | formed in view of the said subject, Comprising: It aims at providing the manufacturing method of a microchip, a microchip, and a vacuum sticking apparatus with a high yield and easy production.

  The object of the present invention can be achieved by the following constitution.

1.
In a method of manufacturing a microchip in which at least two substrates are laminated,
A positioning step of inserting the reference hole of the upper substrate into the positioning pin after inserting the reference hole of the lower substrate into the positioning pin;
A bonding step in which the substrate subjected to the positioning step is bonded under vacuum with an adhesive layer interposed therebetween;
A pressure-bonding step of pressing and pressing the uppermost substrate;
A method for producing a microchip, comprising:

2.
2. The method of manufacturing a microchip according to 1, wherein at least one of the substrates is formed by injection molding a resin material.

3.
In the laminating step, the inside of the hermetic chamber on which at least two substrates are placed is evacuated, and an elastic sheet disposed in parallel with the upper substrate on the upper surface of the hermetic chamber is placed inside the hermetic chamber. 3. The method for producing a microchip according to 1 or 2, wherein the upper substrate and the lower substrate are brought into close contact with each other.

4).
4. The method of manufacturing a microchip according to any one of claims 1 to 3, wherein the crimping step is performed under vacuum.

5).
5. The method of manufacturing a microchip according to any one of claims 1 to 4, wherein in the pressing step, a roller is moved and pressed onto the elastic sheet in close contact with the substrate.

6).
In a microchip formed by laminating at least two substrates,
A microchip comprising a reference hole serving as a reference for stacking on the substrate.

7).
In a vacuum applicator for laminating at least two substrates,
An airtight chamber for placing the substrate;
A positioning pin provided in the hermetic chamber for inserting a reference hole of the substrate;
An elastic sheet disposed in parallel with the substrate on the upper surface of the hermetic chamber;
A vacuum pump for evacuating the inside of the hermetic chamber;
And inserting the substrates into the positioning pins in the stacking order, and then evacuating the inside of the hermetic chamber by the vacuum pump to draw the elastic sheet into the hermetic chamber so that at least two of the substrates are A vacuum bonding apparatus, wherein the substrates are bonded together.

  According to the present invention, after aligning the reference hole of the flow path substrate in which the groove-shaped flow path is formed on one side and the reference hole of the coated substrate, the laminated microchip is bonded by pressure bonding under vacuum. Can be easily produced.

It is sectional drawing of the microchip 1 in embodiment of this invention. It is a top view for demonstrating an example of the flow-path board | substrate 2 and the coating substrate 4 of the microchip 1 in this invention. It is sectional drawing of the sticking apparatus 90 when performing a positioning process. 4 is a plan view for explaining positioning of a flow path substrate 2 and a coated substrate 4. FIG. It is sectional drawing of the sticking apparatus 90 when performing a bonding process. It is sectional drawing of the sticking apparatus 90 when performing a crimping | compression-bonding process. It is an external view of the reaction detection apparatus 80 using the microchip 1 of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 Microchip 2 Flow path board | substrate 4 Coated board | substrate 6 Flow path 8 Waste liquid storage part 11 Communication hole 13 Reagent storage part 17 Sample receiving part 18 Reaction part 19 Detection part 53 Elastic sheet 55, 56 Positioning pin 57 Inner lid 59 Lower stand 61 Vacuum pump 63, 64, 65 Valve 80 Reaction detection device 90 Pasting device 82 Storage body 83 Insertion port 84 Display unit 85 Memory card slot 86 Print output port 87 Operation panel 88 Input / output terminal 200, 201 Reference hole 400, 401 Reference hole

  Hereinafter, the present invention will be described in detail. The microchip of the present invention performs a reaction between a sample and a reagent for various inspections, chemical analysis, chemical synthesis, sample processing / separation, etc., in a fine channel or structure provided in a plate-shaped chip. Is what you do.

  Examples of the use of the microchip of the present invention include biological substance inspection / analysis by gene amplification reaction, antigen-antibody reaction, etc., inspection / analysis of other chemical substances, chemical synthesis of target compounds by organic synthesis, drug efficacy screening, Includes chemical extraction and metal complex formation / separation.

  Hereinafter, embodiments of the present invention will be described with reference to the drawings.

  FIG. 1 is a cross-sectional view of a microchip 1 in an embodiment of the present invention.

  The vertical and horizontal sizes of the entire chip of the microchip 1 are typically several tens of mm, and the height is typically several millimeters although it depends on the application.

  The microchip 1 has a two-layer structure including a flow path substrate 2 in which a flow path 6 is formed on the inner surface and a covering substrate 4 that covers the flow path 6. The flow path 6 provided on the flow path substrate 2 includes a fine flow path having a flow width of 200 μm or less and a coarse flow path having a flow width exceeding 200 μm.

  Here, the “flow path width” represents the horizontal width when the cross section perpendicular to the flow direction is a rectangle, and the average value of the horizontal width when the cross section is another shape similar to a rectangle. To express. The height of the channel is appropriately set according to the purpose regardless of the narrow channel and the wider channel, and is, for example, 10 μm to 1000 μm.

  Reference numeral 78 denotes a driving liquid injection port. The driving liquid injected from the driving liquid injection port 78 sends a reagent or the like accommodated in a part of the flow path 6 through the communicating flow path 6. On the other hand, the sample injected from the sample injection port 79 flows into the flow path 6 and performs a predetermined reaction with a reagent or the like injected from a reagent injection hole (not shown), and the waste liquid after the reaction is stored in the waste liquid storage unit 8. The

  The air communication hole 11 is provided to allow air in the flow path 6 to escape when liquid such as driving liquid is injected.

  In the microchip 1 of the present invention, the flow path substrate 2 and the coated substrate 4 are produced by injection molding, and the flow path substrate 2 and the coated substrate 4 are overlapped to form a flow path. Various resin materials can be used as the resin material of the flow path substrate 2 in which the flow path grooves are formed, and examples thereof include polystyrene, polyethylene, polypropylene, polyethylene terephthalate, polymethyl methacrylate, and polycarbonate. The coated substrate 4 is made of an elastic material such as transparent or translucent silicon rubber, for example, and the coated substrate 4 previously coated with a silicon-based adhesive material or the like is overlapped and bonded to the flow path substrate 2. The thickness of the coated substrate 4 is about 100 μm.

  FIG. 2 is a plan view for explaining an example of the flow path substrate 2 and the covering substrate 4 of the microchip 1 according to the present invention. 2A shows the substrate pattern of the coated substrate 4, and FIG. 2B shows the substrate pattern of the flow path substrate 2.

  As shown in FIG. 2A, the coated substrate 4 is a sheet-like substrate in which only the communication holes 11 and the reference holes 400 and 401 are formed, and the reference holes 400 and 401 and the communication holes 11 are formed using a mold. To do.

  The flow path substrate 2 is a substrate made of a resin material in which the groove pattern and the reference holes 200 and 201 shown in FIG. 2B are formed, and is manufactured by injection molding using a mold. Polypropylene or the like is used as the resin material.

  The reference holes 400 and 401 and the reference holes 200 and 201 are used for positioning in the process of bonding the coated substrate 4 and the flow path substrate 2 described later. The hole diameters of the reference holes 400 and 401 are smaller than the hole diameters of the reference holes 200 and 201. Further, the reference hole 401 and the reference hole 201 are long holes in order to absorb errors at the time of manufacturing the substrate.

  The microchip 1 of the present embodiment is used for, for example, a gene amplification reaction. As shown in FIG. 2B, a flow path containing two types of reagents is provided on one surface of the flow path substrate 2. Three reagent storage portions 13 are provided.

  A recess 211 is provided on the upstream side of the reagent storage unit 13 of the flow path substrate 2 shown in FIG. When the flow path substrate 2 and the covering substrate 4 are bonded together, the recess 211 communicates with the communication hole 11a provided in the communication hole substrate 4 shown in FIG. The communication hole 11a is a reagent injection port. After the flow path substrate 2 and the covering substrate 4 are bonded together, the reagent is injected from the communication hole 11a to close the communication hole 11a.

  The communication hole 11b is also a reagent injection port. Similarly, after injecting a reagent from the communication hole 11b into the sample receiving portion 17, the communication hole 11b is closed. The other communication holes 11 shown in FIG. 2A are provided as air vent holes for releasing the air in the flow path when the driving liquid is injected into the microchip 1.

  The reagent stored in the reagent storage unit 13 of the flow path substrate 2 is injected with a driving liquid by a separate micropump (not shown) communicated with the driving liquid injection port 78, and is pushed out of the reagent storage unit 13 to be merged. 15, the mixed reagent is stored in the mixed reagent storage unit 16 on the downstream side.

  This mixed reagent merges downstream with the sample injected into the channel-shaped sample receiving portion 17. The mixed reagent and the sample are extruded and mixed downstream with the driving liquid by a micropump connected to a separate driving liquid inlet 78. The mixed solution of the mixed reagent and the sample is accommodated in the reaction unit 18 and the amplification reaction is started by heating.

  The liquid after the reaction is sent to the detection unit 19, and the target substance is detected by, for example, an optical detection method.

  A waste liquid reservoir 8 is provided on the most downstream side of the flow path in FIG. 2B, and waste liquid from the upstream flow path is sent to the waste liquid reservoir 8.

  As shown in the partial cross-sectional view of FIG. 1, in the microchip 1 of this embodiment, the flow path is formed by bonding the covering substrate 4 to the flow path substrate 2 in which the flow path grooves are formed. Next, the process of bonding the covering substrate 4 to the flow path substrate 2 will be described with reference to FIGS.

  3, 5, and 6 are cross-sectional views of a bonding device 90 used when the coated substrate 4 is bonded to the flow path substrate 2 in the embodiment of the present invention, and FIG. 4 is a positioning of the flow path substrate 2 and the coated substrate 4. It is a top view for demonstrating.

  In the present invention, the flow path substrate 2 and the cover substrate 4 are bonded together under vacuum so that air does not remain in the gap between the flow path substrate 2 and the cover substrate 4. FIG. 3 is a cross-sectional view of the sticking device 90 when performing the positioning step. FIG. 5 is a cross-sectional view of the sticking device 90 when performing the bonding step, and FIG. 6 is a cross-sectional view of the sticking device 90 when performing the crimping step. In addition, the same number is attached | subjected to the same functional element as the functional element demonstrated so far, and description is abbreviate | omitted.

  The structure of the sticking device 90 will be described with reference to FIG.

  The hermetic chamber B of the sticking device 90 is configured by sealing the lower base 59 with an inner lid 57 to which an elastic sheet 53 is attached. The inner lid 57 can be opened and closed. When the inner lid 57 is closed, a closed space is formed with the recess of the lower base 59. The elastic sheet 53 is a sheet made of an elastic body such as rubber, and is supported on four sides by an inner lid 57.

  In addition, an openable / closable outer lid 58 that covers the inner lid 57 is provided. When the outer lid 58 is closed, the space formed by the outer lid 58 and the inner lid 57 becomes a sealed space. A space sealed by the outer lid 58 and the inner lid 57 is referred to as an outer lid space A.

  The outer lid space A formed by the recesses of the inner lid 57 and the lower base 59 and the airtight chamber B formed by the outer lid 58 and the inner lid 57 are each sucked by the vacuum pump 61 communicating from the pipe 66 to be in a vacuum state. be able to. The pipe 66 communicates with the outer lid space A, the airtight chamber B, and the vacuum pump 61, and the path can be opened and closed by valves 63 and 64. A valve 65 attached to a hole provided in the outer lid 58 is a valve for introducing air into the outer lid space A.

[Positioning process]
Reference numeral 56 denotes a positioning pin, which positions the flow path substrate 2 and the covering substrate 4 by passing the reference hole 400 and the reference hole 200 through the positioning pin 56. The sticking device 90 is also provided with a positioning hole 55 for positioning the reference hole 401 and the reference hole 201. The tip of the positioning pin 56 has a small diameter so as to fit in the reference hole 400 as shown in FIG. Further, as shown in FIG. 3, at least a portion longer than the thickness of the flow path substrate 2 after the tip of the positioning pin 56 has a diameter that fits into the reference hole 200, and a lower portion has a diameter that the reference hole 200 does not enter. It has become. The positioning pin 56 is biased upward by a spring 67 in the drawing. The positioning pin 55 has the same shape as the positioning pin 56, and is fitted into the reference hole 401 and the reference hole 201 and is urged by a spring (not shown).

  FIG. 4 shows a state where the coated substrate 4 is intended to overlap the flow path substrate 2. When sticking is performed using the sticking device 90, first, the reference hole 200 of the flow path substrate 2 is inserted into the positioning pin 56 and the reference hole 201 is inserted into the positioning pin 55 and placed in the lower base 59. Next, the reference holes 400 and 401 of the coated substrate 4 are inserted into the positioning pins 56 and 55 so that the alignment can be easily performed even when the two substrates are transparent. Further, for example, the alignment of the recess 211 of the flow path substrate 2 illustrated in FIG. 4 and the communication hole 11a of the covering substrate 4 can be performed with high accuracy. The same applies to other communication holes and recesses, and by providing a reference hole in the flow path substrate 2 and the covering substrate 4, alignment can be performed with high accuracy.

  In FIG. 3, the reference hole 200 of the flow path substrate 2 is fitted to the positioning pin 56, and the positioning pin 55 is also inserted into the reference hole 201 (not shown) of the flow path substrate 2. Further, the reference hole 400 of the coated substrate 4 is inserted into the positioning pin 56, and the reference hole 401 (not shown) is also inserted into the positioning pin 55. The positioning pin 56 is urged upward by a spring 67 and the positioning pin 55 by a spring (not shown), and both the positioning pins 56 and 55 are movable in the vertical direction of the paper.

  In the state of FIG. 3, the coated substrate 4 is supported by the positioning pins 56 and 55, and a gap is left between the coated substrate 4 and the flow path substrate 2. An adhesive layer 5 to which an adhesive is applied in advance is formed on the surface of the coated substrate 4 that faces the flow path substrate 2.

[Lamination process]
First, the valves 63 and 64 are opened, the valve 65 is closed, and the air in the outer lid space A and the hermetic chamber B is sucked into the vacuum state by the vacuum pump 61.

  Next, after the valve 63 is closed, the valve 65 is opened and air is introduced into the outer lid space A. Then, as shown in FIG. 5, the elastic sheet 53 is drawn into the airtight chamber B in a vacuum state, and the coated substrate 4 supported by the positioning pins 56 and 55 is pushed in so that the coated substrate 4 flows through the adhesive layer 5. Adheres closely to the road substrate 2. If it does in this way, the elastic sheet 53 can be drawn in to the airtight chamber B side in a short time, and the coating substrate 4 can be bonded together.

  The outer lid space A does not necessarily need to be evacuated. The valve 63 is closed from the beginning, the valve 65 is opened, and only the airtight chamber B is sucked by the vacuum pump 61 so that the coated substrate 4 is bonded. be able to.

[Crimping process]
In the crimping step, the outer lid 58 is opened as shown in FIG. 6, and the upper portion of the elastic sheet 53 is opened. The hermetic chamber B remains in a vacuum state. The pressure roller 62 is lowered by the driving mechanism 69 in the downward direction on the paper surface and is brought into pressure contact with the elastic sheet 53. The drive mechanism 69 moves the pressure roller 62 in the left-right direction on the paper surface to press the coated substrate 4 and the flow path substrate 2 from above the elastic sheet 53 so that no floating occurs between the coated substrate 4 and the flow path substrate 2. To.

  When bonding is performed under vacuum in this way, even if sink marks or burrs are generated in the grooves or holes of the flow path substrate 2 or the substrate is warped, the gap is between the coated substrate 4 and the flow path substrate 2. The coated substrate 4 and the flow path substrate 2 can be bonded without leaving air. Further, the produced microchip 1 is not warped.

  FIG. 7 is an external view of a reaction detection device 80 using the microchip 1 of the present invention.

  The reaction detection device 80 is a device that automatically detects a reaction between a specimen previously injected into the microchip 1 and a reagent and displays the result on the display unit 84.

  The housing 82 of the reaction detection device 80 has an insertion port 83, and the microchip 1 is inserted into the insertion port 83 and set inside the housing 82. The insertion port 83 is sufficiently higher than the thickness of the microchip 1 so as not to contact the insertion port 83 when the microchip 1 is inserted. Reference numeral 85 denotes a memory card slot, 86 denotes a print output port, 87 denotes an operation panel, and 88 denotes an input / output terminal.

  The person in charge of inspection inserts the microchip 1 in the direction of the arrow in FIG. 7 and operates the operation panel 87 to start the inspection. Inside the reaction detector 80, the reaction in the microchip 1 is automatically inspected, and when the inspection is completed, the result is displayed on the display unit 84 constituted by a liquid crystal panel or the like. The inspection result can be output from the print output port 86 or stored in a memory card inserted into the memory card slot 85 by operating the operation panel 87. Further, data can be stored in the personal computer or the like from the external input / output terminal 88 using, for example, a LAN cable.

  As described above, according to the present invention, it is possible to provide a microchip manufacturing method, a microchip, and a vacuum sticking apparatus that are easy to produce with a high yield.

Claims (7)

In a method of manufacturing a microchip in which at least two substrates are laminated,
A positioning step of inserting the reference hole of the upper substrate into the positioning pin after inserting the reference hole of the lower substrate into the positioning pin;
A bonding step in which the substrate subjected to the positioning step is bonded under vacuum with an adhesive layer interposed therebetween;
A pressure-bonding step of pressing and pressing the uppermost substrate;
A method for producing a microchip, comprising: The method for manufacturing a microchip according to claim 1, wherein at least one of the substrates is formed by injection molding a resin material. In the laminating step, the inside of the hermetic chamber on which at least two substrates are placed is evacuated, and an elastic sheet disposed in parallel with the upper substrate on the upper surface of the hermetic chamber is placed inside the hermetic chamber. 3. The method of manufacturing a microchip according to claim 1, wherein the upper substrate and the lower substrate are brought into close contact with each other. The method for manufacturing a microchip according to any one of claims 1 to 3, wherein the crimping step is performed under vacuum. 5. The microchip manufacturing method according to claim 1, wherein in the pressure-bonding step, pressure is applied by moving a roller on the elastic sheet that is in close contact with the substrate. 6. Method. In a microchip formed by laminating at least two substrates,
A microchip comprising a reference hole serving as a reference for stacking on the substrate. In a vacuum applicator for laminating at least two substrates,
An airtight chamber for placing the substrate;
A positioning pin provided in the hermetic chamber for inserting a reference hole of the substrate;
An elastic sheet disposed in parallel with the substrate on the upper surface of the hermetic chamber;
A vacuum pump for evacuating the inside of the hermetic chamber;
And inserting the substrates into the positioning pins in the stacking order, and then evacuating the inside of the hermetic chamber by the vacuum pump to draw the elastic sheet into the hermetic chamber so that at least two of the substrates are A vacuum bonding apparatus, wherein the substrates are bonded together.