WO2011122216A1

WO2011122216A1 - Microchip

Info

Publication number: WO2011122216A1
Application number: PCT/JP2011/054730
Authority: WO
Inventors: 直紀清水
Original assignee: コニカミノルタオプト株式会社
Priority date: 2010-03-31
Filing date: 2011-03-02
Publication date: 2011-10-06

Abstract

In order to provide a microchip that can easily and effectively perform temperature adjustment of a temperature-adjusted region, and that can sufficiently support a heat cycle operation or the like wherein a heating/cooling step is repeated in short cycles, disclosed is a microchip (100)—provided with: a first substrate (1) to which a microchannel (11) that introduces a sample and/or a reagent to the inside is formed on at least one side; and a second substrate (2) that is layered/bonded to the first substrate (1), contacting the surface to which the microchannel (11) has been formed—wherein a receiving section (13), which interconnects with the microchannel (11) and performs temperature adjustment of the sample and/or reagent, is provided to at least one of the first substrate (1) and the second substrate (2); and a temperature adjustment channel (14) that does not interconnect with the microchannel (11) and that is for passing a temperature adjustment medium is formed at the vicinity of the receiving section (13).

Description

Microchip

The present invention relates to a microchip used for chemical analysis.

In recent years, as a device for analyzing samples in the field of medical and environmental measurement, μ-TAS (Micro-Total Analysis System), which aims to perform from sample pretreatment to measurement / detection on a chip, Various microchips such as a lab-on-a-chip and a sample analysis system using the microchips have attracted attention.

In such a microchip, it may be necessary to locally adjust the temperature such as heating / cooling for pretreatment of the specimen.

For example, when performing DNA analysis, a sample containing DNA is set on a microchip and reacted with a reagent or the like on the microchip for analysis. However, since the amount of target DNA present in the sample is usually very small, this method is used. When used, it is necessary to amplify and cultivate a specific type of DNA (target DNA) using a PCR (polymerase chain reaction) method or the like as a pretreatment for analysis in order to improve analysis sensitivity.

In this PCR method, a solution in which DNA (target DNA) to be amplified, DNA synthase (DNA polymerase) and a large amount of primers (oligonucleotide) are mixed in advance is prepared as a sample, and heating / cooling of this sample is repeated. This technique amplifies DNA.

In the PCR method, a solution containing double-stranded DNA is denatured into single-stranded DNA by heating at a high temperature (eg, about 94 degrees), and then the solution that has become single-stranded DNA is cooled to, for example, about 60 degrees. I will do it. Thereby, a primer couple | bonds with a part of long single stranded DNA (annealing). In this state, when the primer is not separated and heated to a temperature suitable for the activity of the DNA polymerase (eg, about 72 ° C.), DNA complementary to the single-stranded portion is synthesized starting from the portion to which the primer is bound. .

In the PCR method, by performing a heat cycle operation in which such heating / cooling steps are repeated in a short cycle, DNA synthesis can be repeated and target DNA can be amplified and cultured.

In order to perform such a heat cycle operation on the specimen, conventionally, for example, as shown in FIG. 12, heating means 91 such as a heater using electrothermal conversion elements such as Peltier elements above and below the microchip 9, respectively. And a cooling unit 92 having a water cooling type (that is, for example, a water pipe is disposed at a contact portion with the microchip) or an air cooling type (that is, a cooling fan is disposed at a contact portion with the microchip, for example). It is common.

In such a chip unit, local heating or local cooling can be appropriately performed by switching ON / OFF of the heating unit 91 and the cooling unit 92.

Note that if a temperature change due to heating / cooling is transmitted to a region other than the region where temperature adjustment is necessary, for example, it may adversely affect a sample, a reagent, etc. that are easily deteriorated due to the temperature change, and the accuracy of inspection / analysis may be reduced.

Therefore, it has also been proposed to provide heat insulation means at the boundary between different types of temperature regions (see, for example, Patent Document 1).

JP 2007-120399 A

However, if heating / cooling heating means and cooling means are provided outside the microchip, the heat generated by the heating means and the temperature drop due to the cooling means are sufficiently transmitted to the area where the temperature inside the microchip needs to be adjusted. There is a problem that it is not possible to sufficiently cope with a heat cycle operation or the like in which the heating / cooling process is repeated for a short period, for example.

In particular, microchips are widely molded from resin because of their ease of processing and the like, but since resin has low thermal conductivity, this problem is significant in the case of microchips by resin molding. It becomes.

Therefore, in order to sufficiently transmit the heat generated by the heating means provided outside the microchip and the temperature decrease caused by the cooling means to the inside of the microchip, the material layer of the microchip in the region where temperature adjustment is required is as thin as possible. It is necessary to.

However, providing a thin local portion on the microchip may be unevenly shaped in terms of molding, and requires an area that requires temperature adjustment (that is, a specimen accommodating section (chamber section for accommodating a specimen, for example). As the volume of)) increases, molding becomes more difficult, and depending on the thickness and area, molding may not be possible. Therefore, it has been difficult to provide a microchip capable of efficiently adjusting the temperature.

Therefore, the present invention has been made in view of the circumstances as described above, and it is possible to easily and effectively adjust the temperature in the temperature adjustment region, and to perform a heat cycle operation in which the heating / cooling process is repeated in a short cycle. It is an object of the present invention to provide a microchip that can sufficiently cope with the above.

In order to solve the above problems, according to the present invention,
A first substrate having a test flow channel for introducing a sample and / or a reagent into at least one side thereof, and a surface on which the test flow channel is formed, and is laminated on the first substrate. In a microchip comprising a second substrate to be joined,
At least one of the first substrate and the second substrate includes a temperature adjustment region that communicates with the test flow channel and adjusts the temperature of the specimen and / or reagent,
There is provided a microchip characterized in that a temperature adjusting channel for allowing a temperature adjusting medium to pass therethrough is formed in the vicinity of the temperature adjusting region so as not to communicate with the inspection channel.

According to the present invention, the temperature adjustment flow path that does not communicate with the inspection flow path is formed in the vicinity of the temperature adjustment area that requires temperature adjustment. Therefore, the temperature adjustment in the temperature adjustment area is easily and effectively performed. Therefore, it is possible to provide a microchip that can sufficiently cope with a heat cycle operation in which the heating / cooling process is repeated in a short cycle.

It is a top view which shows schematic structure of the microchip in the 1st Embodiment of this invention. FIG. 2 is a cross-sectional view taken along line AA in FIG. It is the expanded sectional view which expanded the B section of FIG. It is sectional drawing which shows schematic structure of the modification of the microchip in 1st Embodiment shown in FIG. It is a top view which shows schematic structure of the microchip in the 2nd Embodiment of this invention. FIG. 6 is a cross-sectional view taken along the line CC in FIG. 5. FIG. 6 is a cross-sectional view taken along the line DD in FIG. 5. It is sectional drawing which shows schematic structure of the modification of the microchip in 2nd Embodiment shown in FIG. It is sectional drawing which shows schematic structure of the modification of the microchip in 2nd Embodiment shown in FIG. It is a top view which shows schematic structure of the microchip in the 3rd Embodiment of this invention. It is sectional drawing which follows the EE line | wire of FIG. It is sectional drawing which shows schematic structure of an example of the conventional microchip.

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

A microchip (100) according to the following embodiment uses a microfabrication technique to form a fine channel or circuit on one surface of a resin substrate, and allows nucleic acid, protein, blood, etc. A micro-analysis chip that performs chemical reaction, separation, and analysis of a liquid sample, or an apparatus called a μ-TAS (Micro-Total Analysis System), and its practical application is in progress. Here, the microchip is described as being made of resin, but the material is not particularly limited, and materials such as glass can also be used. However, considering moldability, it is preferably made of resin.

As an advantage of such a microchip, it is conceivable to realize an inexpensive system that can be carried in a small space because the amount of sample or reagent used or the amount of discharged waste liquid is reduced.
[First Embodiment]
First, a microchip according to a first embodiment of the present invention will be described with reference to FIGS.

FIG. 1 is a plan view of the microchip 100 according to the first embodiment, and FIG. 2 is a cross-sectional view taken along the line AA in FIG.

As shown in FIG. 1, the microchip 100 has a substantially rectangular shape in plan view, and includes a rectangular first substrate 1 (the front side in FIG. 1 and a lower side in FIG. 2) and a second substrate 2 ( 1 is provided on the back side of the paper in FIG. 1, and the upper side in FIG.

The outer shape of the microchip 100 may be any shape that is easy to handle and analyze, and is preferably a square or a rectangle. As an example, the size may be 10 to 200 mm square. Further, the size may be 10 to 100 mm square.

The first substrate 1 is, for example, a resin plate member, and the second substrate 2 is, for example, a resin film. In addition, the 2nd board | substrate 2 is not limited to a film, A sheet-like (plate-shaped) member may be sufficient.

Note that the material for forming the first substrate 1 and the second substrate 2 is not particularly limited, but it is preferable to use a resin in consideration of moldability.

The resin for forming the first substrate 1 and the second substrate 2 has good moldability (transferability, releasability), high transparency, low self-fluorescence for ultraviolet light and visible light, etc. Are preferable conditions, and for example, a thermoplastic resin can be applied.

Examples of the thermoplastic resin include polycarbonate, polymethyl methacrylate, polystyrene, polyacrylonitrile, polyvinyl chloride, polyethylene terephthalate, nylon 6, nylon 66, polyvinyl acetate, polyvinylidene chloride, polypropylene, polyisoprene, polyethylene, polydimethyl. It is preferable to use siloxane, cyclic polyolefin or the like. It is particularly preferable to use polymethyl methacrylate and cyclic polyolefin.

Note that the same material may be used for the first substrate 1 and the second substrate 2, or different materials may be used.

The thickness of the first substrate 1 is not particularly limited, but is preferably 0.2 to 5 mm, more preferably 0.5 to 2 mm in consideration of moldability.

The thickness of the second substrate 2 is not particularly limited, but is preferably 30 μm to 300 μm, and more preferably 40 to 150 μm, for example.

The microchip 100 includes a plurality of microchannels 11 that are test channels for introducing an analysis sample or reagent as a specimen into the microchip 100 and temperature adjustment of the analysis sample or reagent in communication with the microchannel 11. The accommodating part 13 which is a temperature adjustment area | region which performs is formed.

In the present embodiment, the fine flow path 11 is a groove formed on the surface of the first substrate 1, and the accommodating portion 13 is a recess formed on the surface of the first substrate 1.

The second substrate 2 is laminated and bonded to the first substrate 1 in contact with the surface on which the fine flow path 11 and the accommodating portion 13 are formed. Thereby, the fine flow path 11 and the accommodating part 13 are sealed, and the second substrate 2 functions as a lid member (cover).

In the present embodiment, the grooves constituting the microchannel 11 and the recesses constituting the accommodating portion 13 are provided on the first substrate 1 side, but these are the second substrate 2 side which is a lid member. These grooves and recesses may be formed in both the first substrate 1 and the second substrate 2. Moreover, the microchannel 11 etc. may be formed by forming the groove part and recessed part corresponding to each of the 1st board | substrate 1 and the 2nd board | substrate 2, and superimposing both.

The shape of the microchannel 11 is within the range of 10 to 200 μm in both width and depth in consideration of the fact that the amount of analysis sample and reagent used can be reduced, and the fabrication accuracy of molds, transferability and mold release properties are taken into consideration. Although it is preferable that it is the value of, it does not specifically limit.

The width and depth of the fine channel 11 may be determined according to the use of the microchip 100 or the like.

In addition, the cross-sectional shape of the fine channel 11 may be a rectangular shape or a curved surface shape.

The shape, size and the like of the accommodating portion 13 are not particularly limited, and may be a square shape or the like in addition to the circular shape as shown in FIG.

Further, the cross-sectional shape of the accommodating portion 13 may be a rectangular shape or a curved surface shape.

In this embodiment, the microchip 100 is used for, for example, DNA analysis using a PCR (polymerase chain reaction) method, and in this case, DNA to be amplified (target DNA), DNA synthesis A solution in which an enzyme (DNA polymerase) and a large amount of primers (oligonucleotide) are mixed in advance is accommodated in the accommodating portion 13 which is a temperature adjustment region as an analysis sample (specimen).

In the following, a case where DNA analysis using the PCR method is performed will be described as an example, and the case where the solution is stored in the storage unit 13 and temperature adjustment is performed will be described. It is not limited. The storage unit 13 stores various analysis samples (specimens), reagents, and the like that require temperature adjustment according to the purpose of inspection and analysis.

The microchip 100 is formed with a plurality of wells 12 communicating with the fine flow path 11.

The well 12 is a hole that opens on either side of the microchip 100 to inject and discharge the analysis sample and the reagent. Although not shown in FIG. 1 and the like, the accommodating portion 13 is also formed with a well communicating therewith.

As shown in FIGS. 1 and 2, the surface of the first substrate 1 and in the vicinity of the accommodating portion 13 that is the temperature adjustment region do not communicate with the fine channel 11 that is the inspection channel, and the temperature A temperature adjusting flow path 14 for passing the adjusting medium is formed so as to surround the accommodating portion 13. Specifically, when viewed from a direction perpendicular to the flow path forming surface of the first substrate 1, the accommodating portion is located at a position surrounding the accommodating portion except for the vicinity of the connecting portion between the accommodating portion 13 and the fine channel 11. A temperature adjusting flow path 14 is formed along the periphery of 13.

The temperature adjusting flow path 14 is a groove formed on the surface of the first substrate 1 and is sealed by laminating the second substrate 2 on the first substrate 1.

The width and depth of the temperature adjusting flow path 14 are not particularly limited, and may be determined according to the type of the temperature adjusting medium. The width and depth of the temperature adjusting channel 14 may be the same as the width and depth of the fine channel 11 or may be larger and deeper than this.

Further, the shape and the like of the temperature adjusting flow path 14 are not particularly limited, and the cross-sectional shape of the temperature adjusting flow path 14 may be rectangular or curved.

The temperature adjusting medium passed through the temperature adjusting flow path 14 is a heating medium for heating the temperature adjusting area or a cooling medium for cooling the temperature adjusting area.

The type of the temperature adjusting medium is not particularly limited. For example, hot water can be used as the heating medium, and cold water can be used as the cooling medium.

The temperature adjusting medium may be either liquid or gas. Further, a gas may be used as a temperature adjusting medium during heating, and a liquid may be used during cooling.

The well 15 which is a hole for injecting and discharging the temperature adjusting medium is formed in the temperature adjusting flow path 14. In the present embodiment, the temperature adjusting flow path 14 bends in the vicinity of the connecting portion between the housing portion 13 and the fine flow path 11 and extends away from this portion, and the end portion of the temperature adjusting flow path 14 is the first substrate 1. A well 15 is formed near the side.

The well 15 is connected to a pump or the like (not shown) so that the temperature adjusting medium is injected into and discharged from the temperature adjusting channel 14.

In this embodiment, for example, as temperature adjustment related information, the elapsed time from the start of injection of the temperature adjustment medium, the temperature of the storage unit 13 or the analysis sample (sample) stored therein, the temperature adjustment medium As information acquisition means for acquiring temperature and the like, time measuring means (timer) for measuring time, temperature sensor for measuring temperature (not shown) and the like are provided, and temperature adjustment related information acquired by these is provided. Based on this, injection / discharge of the temperature adjusting medium to / from the temperature adjusting flow path 14 by the pump is controlled.

Note that the information acquisition means is not limited to those exemplified here. Further, the temperature adjustment related information acquired by the information acquisition means is not limited to the information shown here, but other information may be acquired, for example, only the elapsed time may be acquired. You may make it acquire only some of them.

For example, when DNA analysis using the PCR method is performed, in order to improve analysis sensitivity, DNA contained in the analysis sample is amplified and cultured as a pretreatment for the test.

Specifically, first, the analysis sample accommodated in the accommodation unit 13 is heated at a high temperature of, for example, about 94 degrees to denature double-stranded DNA contained in the analysis sample into single-stranded DNA. Thereafter, the solution containing the single-stranded DNA is rapidly cooled to about 60 degrees, for example. Thereby, a primer couple | bonds with a part of long single stranded DNA (annealing). Further, in this state, the analysis sample is heated for a certain period of time to a temperature suitable for the activity of the DNA polymerase (for example, about 72 ° C.) without separation of the primers. As a result, DNA complementary to the single-stranded portion is synthesized starting from the portion to which the primer is bound.

In the present embodiment, under such time and temperature control, injection / discharge of the temperature adjustment medium to / from the temperature adjustment flow path 14 is controlled to perform a heat cycle operation in which the heating / cooling process is repeated in a short cycle. Thus, DNA synthesis can be repeated and target DNA can be amplified and cultured.

FIG. 3 is an enlarged view of a portion B surrounded by an alternate long and short dash line in FIG.

The inner surface of the temperature adjusting flow path 14 is subjected to a permeation preventing process capable of preventing the temperature adjusting medium from penetrating into the first substrate 1 and the second substrate 2.

In the present embodiment, a coating process such as a fluorine coating process is performed as the permeation preventing process, and the coating film 141 is formed on the entire inner surface of the temperature adjusting flow path 14 as shown in FIG.

As the permeation prevention process, for example, a peening process is performed on a mold when the first substrate 1 and the second substrate 2 are molded, or a fluorine coating process is performed on the first substrate 1 and the second substrate 2. A water repellent treatment may be performed by performing a film treatment or a coating treatment, or a hydrophilic treatment may be performed by performing plasma treatment or corona discharge treatment on the surface of the first substrate 1 or the second substrate 2, for example. It is done.

The permeation prevention treatment may be at least one of water repellency treatment such as peening treatment or fluorine coating treatment shown here, and hydrophilic treatment such as plasma treatment or corona discharge treatment, or a combination thereof. However, the present invention is not limited to those exemplified here. That is, it may be a combination of two or more peening treatments or fluorine coating treatments as a water repellent treatment, or may be a combination of two or more plasma treatments or corona discharge treatments that are hydrophilic treatments. A combination of two or more of the water-repellent treatment and one of the hydrophilic treatments.

Whether water repellent treatment or hydrophilic treatment is performed as the permeation prevention treatment is appropriately selected depending on the type of temperature adjusting medium used, the first substrate, the material forming the second substrate 2, and the like. To do.

Further, the permeation prevention treatment is not limited as long as it can prevent the temperature adjusting medium from permeating into the first substrate 1 and / or the second substrate 2, and is performed by a method other than the water repellent treatment and the hydrophilic treatment. May be.

Next, the operation of this embodiment will be described.

First, when forming the microchip 100, the first substrate 1 and the second substrate 2 are formed by injection molding of a resin or the like.

At this time, on one surface of the first substrate 1, a groove portion constituting the fine flow path 11, a groove portion constituting the temperature adjusting flow path 14, and an accommodating portion 13 communicating with the fine flow path 11 are formed. A recess is formed. Further, a well 12 that communicates with the fine flow path 11 and penetrates the first substrate 1 in the thickness direction of the substrate and a temperature adjustment flow path 14 and penetrates the first substrate 1 in the thickness direction of the substrate. 15. A well (not shown) communicating with the accommodating portion 13 is formed.

Next, the second substrate 2 is laminated and bonded to the first substrate 1 so as to be in contact with the surface of the first substrate 1 on which the fine flow path 11 and the like are formed.

Thereby, the groove part which comprises the fine flow path 11, the groove part which comprises the flow path 14 for temperature control, and the recessed part which comprises the accommodating part 13 connected with the fine flow path 11 are sealed by the 2nd board | substrate 2, The microchip 100 is completed.

When performing analysis / analysis of an analysis sample (specimen) using the microchip 100, an analysis sample or the like that requires temperature adjustment is stored in the storage unit 13. When the analysis sample or the like is to be heated, a heating medium such as hot water is passed through the temperature adjusting channel 14 as a temperature adjusting medium. When the analysis sample or the like is to be cooled, a cooling medium such as cold water is passed through the temperature adjusting channel 14 as a temperature adjusting medium.

For example, when DNA analysis is performed by the PCR method, first, the accommodating portion 13 and the analysis sample accommodated therein are heated to about 94 ° C. through a heating medium such as hot water through the temperature adjusting channel 14. Thereafter, the storage unit 13 and the analysis sample stored in the storage unit 13 are passed through the cooling medium such as cold water through the temperature adjustment channel 14 to about 60 degrees, and the temperature adjustment channel 14 is again supplied with hot water or the like. Heat to about 72 degrees through the heating medium. Thereby, the DNA in the analysis sample can be amplified.

Then, after repeating such heating / cooling steps to amplify the DNA to a desired amount, a reagent or the like is injected into the fine channel 11 and reacted with the analysis sample to perform DNA analysis.

As described above, according to the present embodiment, by passing a heating medium or a cooling medium as a temperature adjustment medium through the temperature adjustment flow path 14, the storage portion 13 and the analysis sample (specimen) stored therein ) And the like can be performed.

In this way, both heating / cooling can be performed using only one temperature adjusting flow path 14 by changing the temperature adjusting medium passed therethrough. The temperature can be adjusted freely.

Further, since the temperature adjusting flow path 14 is provided in the vicinity of the accommodating portion 13 so as to surround the accommodating portion 13, it is not affected by the thickness of the substrate of the microchip 100, the thermal conductivity of the material, or the like. 13 and the analysis sample (specimen) accommodated therein can be effectively heated / cooled. For this reason, it can fully respond to heat cycle operation etc. which repeat a heating / cooling process with a short cycle.

In the present embodiment, the groove part constituting the fine flow path 11, the groove part constituting the temperature adjusting flow path 14, and the concave part constituting the accommodating part 13 communicating with the fine flow path 11 are all the first substrate. Although the case where it is formed on one surface of 1 is taken as an example, these are not limited to the case where it is provided on the first substrate 1.

For example, as shown in FIG. 4, the concave portion constituting the accommodating portion 13 is formed in the first substrate 1, and the groove portion constituting the temperature adjusting flow path 31 is in contact with the first substrate 1 in the second substrate 3. It may be provided in a distributed manner such as being formed on the surface.

Further, in the present embodiment, the case where both the first substrate 1 and the second substrate 2 are formed of a resin has been described as an example, but one or both are formed of a material other than a resin such as glass. May be.

For a substrate having a complicated structure, it is preferable to form it with a resin because it is easier to process. However, for example, when the second substrate 2 is a simple plate-like member to be bonded to the first substrate 1, the first The two substrates 2 may be formed of glass.
[Second Embodiment]
Next, a second embodiment of the microchip according to the present invention will be described with reference to FIGS. In addition, since this embodiment is different from the first embodiment in the configuration of the temperature adjusting flow path, the following description will particularly focus on differences from the first embodiment.

5 is a plan view of the microchip in the present embodiment, FIG. 6 is a cross-sectional view taken along the line CC in FIG. 5, and FIG. 7 is a cross-sectional view taken along the line DD in FIG. is there.

As shown in FIG. 5 to FIG. 7, the microchip 200 in this embodiment includes a first substrate 1 and

second substrates

4 and 5 bonded to the front and back of the first substrate 1.

On the surface of the first substrate 1 (the upper surface in FIGS. 6 and 7), there are formed a recess that constitutes the accommodating portion 13 and a groove that constitutes the microchannel 11 that communicates therewith.

Further, a groove portion constituting the temperature adjusting channel 16 is formed on the back surface (the lower surface in FIGS. 6 and 7) of the first substrate 1.

The concave portion constituting the accommodating portion 13 and the groove portion constituting the fine flow path 11 are sealed by the second substrate 4 being laminated and bonded to the surface of the first substrate 1, and the temperature adjusting flow path 16 is defined. The groove portion to be configured is sealed by the second substrate 5 being laminated and bonded to the back surface of the first substrate 1.

As shown in FIG. 7 and the like, the temperature adjusting flow path 16 is in the stacking direction of the first substrate 1 and the

second substrates

4 and 5 with respect to all or part of the accommodating portion 13 that is the temperature adjusting region. It arrange | positions so that it may overlap below.

In addition, the position where the temperature adjusting flow path 16 is provided is not limited to this, and may be disposed above the accommodating portion 13.

In the present embodiment, the temperature adjusting flow path 16 has a crank shape which is bent back a plurality of times, and is provided corresponding to almost the entire area of the accommodating portion 13.

The temperature adjusting flow path 16 overlaps with the whole or a part of the accommodating portion 13 that is a temperature adjusting region so as to overlap above or below in the stacking direction of the first substrate and the

second substrates

4 and 5. The range in which the temperature adjusting flow path 16 is provided is not limited to the one corresponding to substantially the entire area of the accommodating portion 13.

Further, the microchip 200 is provided with a well 17 that communicates with the temperature adjusting flow path 16.

Since other configurations are substantially the same as those described in the first embodiment, the same members are denoted by the same reference numerals and description thereof is omitted.

Next, the operation of this embodiment will be described.

First, when the microchip 200 is formed, the first substrate 1 and the

second substrates

4 and 5 are formed by injection molding of a resin or the like.

At this time, on the surface of the first substrate 1, a groove part constituting the fine flow path 11 and a concave part constituting the accommodating part 13 are formed. In addition, on the back surface of the first substrate 1, a groove portion constituting the temperature adjusting flow path 16 is formed. Further, a well 12 that communicates with the fine channel 11 and penetrates the first substrate 1 in the thickness direction of the substrate and a temperature adjustment channel 16 and penetrates the first substrate 1 in the thickness direction of the substrate. 17. A well (not shown) communicating with the accommodating portion 13 is formed.

Next, the second substrate 4 is laminated and bonded to the first substrate 1 so as to be in contact with the surface of the first substrate 1 on which the fine flow path 11 and the like are formed. Further, the second substrate 5 is laminated and bonded to the first substrate 1 so as to be in contact with the back surface of the first substrate 1 on which the temperature adjusting flow path 16 is formed.

Thereby, the groove part constituting the fine flow path 11 and the concave part constituting the accommodating part 13 communicating with the fine flow path 11 are sealed by the second substrate 4, and the groove part constituting the temperature adjusting flow path 16 is The microchip 200 is completed by being sealed by the second substrate 5.

The method of testing / analyzing an analysis sample (specimen) using the microchip 200 is the same as that in the first embodiment, and a description thereof will be omitted.

As described above, according to the present embodiment, by passing the heating medium or the cooling medium as the temperature adjustment medium through the temperature adjustment flow path 16, the storage portion 13 and the analysis sample (specimen) stored therein ) And the like can be performed.

As described above, both the heating / cooling can be performed only by changing the temperature adjusting medium passed through the single temperature adjusting flow path 16, and therefore, with a simple configuration, the analysis sample (specimen), etc. The temperature can be adjusted freely.

Further, since the temperature adjusting flow path 16 is provided in the vicinity of the accommodating portion 13 and below the accommodating portion 13 so as to correspond to substantially the entire area of the accommodating portion 13, the substrate of the microchip 200 is provided. It is possible to effectively heat / cool the accommodating portion 13 and the analysis sample (specimen) accommodated in the accommodating portion 13 without being influenced by the thickness, the thermal conductivity of the material, or the like. For this reason, it can fully cope with a heat cycle operation or the like in which the heating / cooling process is repeated in a short cycle.

In the present embodiment, the groove part constituting the fine flow path 11, the groove part constituting the temperature adjusting flow path 16, and the concave part constituting the accommodating part 13 communicating with the fine flow path 11 are all the first substrate. Although the case where it is formed on the front surface and the back surface of 1 is taken as an example, all of these may not be provided on the first substrate 1.

For example, as shown in FIGS. 8 and 9, the microchip has a three-layer structure in which a first substrate 1, a second substrate 4, and a third substrate 6 are laminated, and a fine structure is formed on the surface of the first substrate 1. A third substrate is formed on the surface side of the first substrate 1 via the second substrate 4 by forming a groove portion constituting the flow channel 11 and a concave portion constituting the accommodating portion 13 communicating with the fine flow channel 11. 6 may be formed on the lower side surface (the lower surface in FIGS. 8 and 9), and in this case, a well 62 communicating with the temperature adjustment channel 61 is provided. The third substrate 6 is formed so as to penetrate therethrough.

In this case, the temperature adjusting flow path 61 is provided in the vicinity of the accommodating portion 13 and above the accommodating portion 13 so as to correspond to the almost entire area of the accommodating portion 13. As in the case of being provided below 13, heating / cooling of the accommodating portion 13 and the analysis sample (specimen) accommodated therein is effectively performed without being affected by the thickness or material of the microchip substrate. Can do. For this reason, it becomes possible to fully cope with a heat cycle operation or the like in which the heating / cooling process is repeated in a short cycle.
[Third Embodiment]
Next, a third embodiment of the microchip according to the present invention will be described with reference to FIGS. In addition, since this embodiment is different from the first embodiment and the second embodiment in the configuration of the temperature adjusting flow path, the first embodiment and the second embodiment will be particularly described below. Different points will be described.

FIG. 10 is a plan view of the microchip in the present embodiment, and FIG. 11 is a cross-sectional view taken along the line EE in FIG.

As shown in FIGS. 10 and 11, the microchip 300 in the present embodiment includes a first substrate 1 and

second substrates

4 and 5 bonded to the front and back of the first substrate 1.

On the surface of the first substrate 1 (the upper surface in FIG. 11), there are formed a concave portion that constitutes the accommodating portion 13 and a groove portion that constitutes the fine channel 11 that communicates therewith.

Further, a groove portion constituting the temperature adjusting flow path 18 is formed on the back surface (the lower surface in FIG. 11) of the first substrate 1.

The concave portion constituting the accommodating portion 13 and the groove portion constituting the fine channel 11 are sealed by laminating the second substrate 4 on the surface of the first substrate 1 to constitute the temperature adjusting channel 18. The groove is sealed by laminating the second substrate 5 on the back surface of the first substrate 1.

As shown in FIG. 11, the temperature adjusting flow path 18 is lower in the stacking direction of the first substrate 1 and the

second substrates

4, 5 than the whole or a part of the accommodating portion 13 that is the temperature adjusting region. It is arranged to overlap.

It should be noted that the position where the temperature adjusting flow path 18 is provided is not limited to this, and the temperature adjusting flow path 18 may be disposed above the accommodating portion 13.

In the present embodiment, the temperature adjusting flow path 18 has a crank shape which is bent back a plurality of times, and is provided corresponding to almost the entire area of the accommodating portion 13.

The temperature adjusting flow path 18 overlaps with the whole or a part of the accommodating portion 13 that is a temperature adjusting region above or below in the stacking direction of the first substrate 1 and the

second substrates

4 and 5. The range in which the temperature adjusting flow path 18 is provided is not limited to that corresponding to almost the entire region of the accommodating portion 13.

In addition, the microchip 300 is provided with a well 19 that communicates with the temperature adjusting flow path 18.

Further, in the microchip 300, a gap portion 20 is disposed as a heat insulating means between the temperature adjusting flow path 18 and the fine flow path 11 as the inspection flow path. The gap 20 is a groove that separates the temperature adjusting flow path 18 and the fine flow path 11. Since air has a low thermal conductivity of about 0.03 W / m · k, a high heat insulating effect can be expected by providing the air gap 20 as an air layer.

Note that the width, depth, shape, and the like of the gap 20 are not particularly limited. It is preferable that the gap portion 20 is provided as large as possible within the range of constraints such as layout because a heat insulating effect can be obtained.

Next, the operation of this embodiment will be described.

First, when the microchip 300 is formed, the first substrate 1 and the

second substrates

4 and 5 are formed by injection molding of a resin or the like.

At this time, on the surface of the first substrate 1, a groove portion constituting the fine flow path 11, a recess portion constituting the accommodating portion 13, and a gap portion 20 are formed. In addition, a groove portion constituting the temperature adjusting flow path 18 is formed on the back surface of the first substrate 1. Further, a well 12 that communicates with the fine channel 11 and penetrates the first substrate 1 in the thickness direction of the substrate and a temperature adjustment channel 18 and penetrates the first substrate 1 in the thickness direction of the substrate. 19, a well (not shown) communicating with the accommodating portion 13 is formed.

Next, the second substrate 4 is laminated and bonded to the first substrate 1 so as to be in contact with the surface of the first substrate 1 on which the fine flow path 11 and the like are formed. Further, the second substrate 5 is laminated and bonded to the first substrate 1 so as to be in contact with the back surface of the first substrate 1 on which the temperature adjusting flow path 18 is formed.

Thereby, the groove part which comprises the fine flow path 11, the recessed part which comprises the accommodating part 13 connected with the fine flow path 11, and the space | gap part 20 are sealed by the 2nd board | substrate 4, and the flow path 18 for temperature adjustment is comprised. The groove portion to be sealed is sealed by the second substrate 5 to complete the microchip 300.

The inspection / analysis method for the analysis sample (specimen) using the microchip 300 is the same as that in the first embodiment, and thus the description thereof is omitted.

As described above, according to the present embodiment, by passing the heating medium or the cooling medium as the temperature adjusting medium through the temperature adjusting flow path 18, the accommodating portion 13 and the analysis sample (specimen) accommodated therein are included. ) And the like can be performed.

In this way, both the heating / cooling can be performed only by changing the temperature adjusting medium passed through the single temperature adjusting flow path 18, so that the analysis sample (specimen) or the like can be made with a simple configuration. The temperature can be adjusted freely.

In the present embodiment, since the gap 20 as the heat insulating means is disposed between the temperature adjustment flow path 18 and the fine flow path 11, the temperature adjustment flow path 18 is accommodated through the temperature adjustment medium. When the part 13 and the analysis sample (sample) accommodated therein are heated / cooled, this temperature change is not transmitted to the surrounding fine channel 11 or the like. As a result, even when a reagent or the like that is altered by heating / cooling is used for inspection or analysis, the reagent or the like is not affected by the temperature adjustment in the temperature adjustment flow path 18 and performs accurate inspection and analysis. be able to.

In this embodiment, the case where the heat insulating means is the gap portion 20 that separates the temperature adjusting flow path 18 and the fine flow path 11 has been described as an example, but the heat insulating means is not limited thereto. For example, a heat insulating flow path may be formed by providing a groove portion that separates the temperature adjusting flow path 18 and the fine flow path 11 and enclosing a low thermal conductivity material in the groove section.

In this case, the low thermal conductivity material may be a material having a thermal conductivity lower than that of the resin forming the substrate.

Preferably, an epoxy resin having a thermal conductivity lower than 0.3 W / m · k (thermal conductivity 0.21 W / m · k) or the like can be applied.

Further, a heat insulating layer made of a low heat conductive material may be integrally disposed between the temperature adjusting flow path 18 and the fine flow path 11 in the first substrate or the second substrate.

Note that the heat insulating means shown in the third embodiment is arranged between the temperature adjustment flow path and the inspection flow path in the first embodiment and the second embodiment. It may be.

Of course, the present invention is not limited to the above embodiment, and can be modified as appropriate.

DESCRIPTION OF SYMBOLS 1 1st board | substrate 2 2nd board | substrate 11 Micro flow path 13 Storage part 14 Temperature control flow path 20 Cavity part 100 Microchip 141 Coating

Claims

A first substrate having a test flow channel for introducing a sample and / or a reagent into at least one side thereof, and a surface on which the test flow channel is formed, and is laminated on the first substrate. In a microchip comprising a second substrate to be joined,
At least one of the first substrate and the second substrate includes a temperature adjustment region that communicates with the test flow channel and adjusts the temperature of the specimen and / or reagent,
A microchip, characterized in that a temperature adjusting channel for allowing a temperature adjusting medium to pass therethrough is formed in the vicinity of the temperature adjusting region.
2. The microchip according to claim 1, wherein the temperature adjustment flow path is disposed so as to surround the temperature adjustment region.
The temperature adjusting flow path is disposed so as to overlap above or below in the stacking direction of the first substrate and the second substrate with respect to all or part of the temperature adjusting region. The microchip according to claim 1.
The portion of the temperature adjusting flow path that overlaps in the stacking direction of the first substrate and the second substrate with respect to the temperature adjusting region has a shape that is folded back multiple times. 2. The microchip according to 1.
The said temperature adjustment medium is a heating medium which heats the said temperature adjustment area | region, or a cooling medium which cools the said temperature adjustment area | region, The Claim 1 characterized by the above-mentioned. Microchip.
The microchip according to any one of claims 1 to 5, wherein the temperature adjusting medium is a liquid or a gas.
The microchip according to any one of claims 1 to 6, wherein at least one of the first substrate and the second substrate is formed of a resin.
The inner surface of the temperature adjusting flow path is subjected to a permeation preventing process capable of preventing permeation of the temperature adjusting medium into the first substrate and / or the second substrate. The microchip according to any one of claims 1 to 7.
The microchip according to claim 8, wherein the permeation prevention treatment is at least one of water repellent treatment and hydrophilic treatment, or a combination thereof.
10. The microchip according to claim 9, wherein the water repellent treatment is at least one of a peening treatment and a fluorine coating treatment.
10. The microchip according to claim 9, wherein the hydrophilic treatment is at least one of plasma treatment and corona discharge treatment.
The microchip according to any one of claims 1 to 11, wherein a heat insulating means is disposed between the temperature adjusting flow path and the inspection flow path.
The heat insulating means includes a gap that separates the temperature adjusting flow path and the inspection flow path, and a heat insulating flow path in which a low heat conductive material is sealed in a groove that separates the temperature adjusting flow path and the inspection flow path. And at least one of the heat insulating layers in which a low thermal conductive material is integrally disposed between the temperature adjusting flow path and the inspection flow path in the first substrate or the second substrate. The microchip according to claim 12.