JP2011163946A - Microfluid chip - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microfluid chip that facilitates dispensing a small number of samples. <P>SOLUTION: The microfluid chip 100 includes a substrate 10 that has a first surface 11 and a second surface 12 facing the first surface 11, has a reservoir region 13 having a reservoir 15 and a well region 14 adjacent to the reservoir region 13 in the plane view, and has a well 16 having an opening 16a on the first surface 11 side in the well region 14, and a cover having a first fixed region 21 that is laid on the first surface 11 side of the substrate 10 and is fixed to the substrate 10 so as to surround the reservoir region 13 and well region 14 in the plane view and a second fixed region 22 that has a discontinuous part on the reservoir region 13 side and is fixed to the substrate 10 along the contour of the opening 16a. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a microfluidic chip.

  A method of performing chemical analysis, chemical synthesis, or bio-related analysis using a microfluidic chip in which a liquid flow path is provided on a glass substrate or the like has attracted attention. The microfluidic chip may be referred to by a name such as Micro Total Analytical System (Micro TAS) or Lab-on-a-chip.

  Some microfluidic chips generally include a plurality of minute reaction containers called wells, and each of the plurality of reaction containers can perform different reactions. The microfluidic chip has the advantage that the amount of sample and reagent can be greatly reduced compared to conventional analyzers, analytical instruments, reaction vessels, etc., and waste generated by operation can be reduced. There is. Therefore, it is expected to be used in a wide range of fields such as medical diagnosis, on-site analysis of the environment and food, production of pharmaceuticals and chemicals, and the like. For example, Patent Document 1 describes a micro card (chip) in which a reservoir and a well are connected by a flow path called a channel, and a sample introduced into the reservoir is dispensed into the well.

  Since the microfluidic chip requires a small amount of reagent, the cost of various tests can be reduced, and the required amount of sample (analyte) can be small, so the reaction time can be greatly shortened. . In particular, when a microfluidic chip is applied to medical examinations and the like, there is an advantage that, for example, the burden on the patient can be reduced because the required amount of specimen such as blood is small.

JP 2006-509199 A

  However, in a microfluidic chip in which a channel is provided between a reservoir and a well, as in the chip of Patent Document 1, for example, when dispensing a sample, a plurality of neighboring wells pass through the channel. In some cases, the sample was mixed to cause contamination.

  One object of some embodiments of the present invention is to provide a microfluidic chip that is less prone to sample contamination during dispensing.

  SUMMARY An advantage of some aspects of the invention is to solve at least a part of the problems described above, and the invention can be implemented as the following aspects or application examples.

[Application Example 1]
One aspect of the microfluidic chip according to the present invention is:
A reservoir region having a first surface and a second surface opposite to the first surface, a reservoir region in which a reservoir is formed, and a well region adjacent to the reservoir region in plan view, and the first region in the well region A substrate in which a well having an opening on the surface side is formed;
A first fixing region which is laid on the first surface side of the substrate and fixed to the substrate so as to surround the reservoir region and the well region in a plan view; and a discontinuous portion on the reservoir region side. A cover having a second fixing region fixed to the substrate along the contour of the opening;
including.

  According to the microfluidic chip of this application example, when the sample is dispensed into the well, the sample introduced into the well is less likely to flow backward, and the sample in the well is less likely to be mixed with the sample in the neighboring well. be able to. That is, contamination can be suppressed.

  In the present invention, “laying” means a state in which a cover is laid on the first surface of the substrate, and “adhesion” means a state in which the cover is laid and fixed on the first surface of the substrate. Point to. Therefore, the state in which the cover is laid and has the fixing region is a state in which there is a portion where it is easy to form a gap between the substrate and the cover, and a portion where the substrate and the cover are difficult to separate. The part where the substrate and the cover are difficult to separate is expressed as a fixed region.

  In addition, in the present invention, “in a plan view” or “when seen in a plan view” refers to a case when viewed from a direction orthogonal to the first surface of the substrate.

[Application Example 2]
In application example 1,
The well may have a protruding portion in which a contour of the opening protrudes toward the reservoir region in a plan view.

  According to the microfluidic chip of this application example, the introduction of the sample can be facilitated by the shape of the protruding portion.

[Application Example 3]
In application example 2,
You may have two or more said protrusion parts.

  According to the microfluidic chip of this application example, it is possible to more efficiently replace the gas in the well with the sample to be introduced.

[Application Example 4]
In any one of Application Examples 1 to 3,
The total length of the second fixing regions in the outline of the opening may be 50% or more of the length of the outline of the opening.

  According to the microfluidic chip of this application example, it is possible to more reliably prevent the sample in the well from being mixed with the sample in the neighboring well.

[Application Example 5]
In any one of Application Examples 1 to 4,
The length of the longest continuous part of the second fixed region in the outline of the opening may be 50% or more of the length of the outline of the opening.

  According to the microfluidic chip of this application example, it is possible to more reliably prevent the sample in the well from being mixed with the sample in the neighboring well.

[Application Example 6]
In any one of Application Examples 1 to 3,
When the opening is partitioned by a straight line connecting both ends of the longest continuous portion of the second fixing region, an area of the partitioning portion of the opening surrounded by the straight line and the second fixing region is an area of the opening. It can occupy 50% or more.

  According to the microfluidic chip of this application example, it is possible to more reliably prevent the sample in the well from being mixed with the sample in the neighboring well.

The top view which shows typically the board | substrate 10 of embodiment. The schematic diagram of the cross section of the board | substrate 10 of embodiment. The top view which expands and shows typically the well 16 vicinity of embodiment. Sectional drawing which expands and shows typically the well 16 vicinity of embodiment. FIG. 2 is a plan view schematically showing the microfluidic chip 100 of the embodiment. FIG. 3 is a schematic cross-sectional view of the microfluidic chip 100 of the embodiment. FIG. 2 is a plan view schematically showing the microfluidic chip 100 of the embodiment. The figure which shows typically an example of the usage method of the microfluidic chip of embodiment. The figure which shows typically an example of the usage method of the microfluidic chip of embodiment. The figure which shows typically an example of the usage method of the microfluidic chip of embodiment. The figure which shows typically the microfluidic chip | tip of a reference example. The top view which shows typically the well 46 of the microfluidic chip | tip of a modification. The schematic diagram of the cross section of the well 46 vicinity of the microfluidic chip | tip of a modification. The schematic diagram of the cross section of the well 46 vicinity of the microfluidic chip | tip of a modification. The schematic diagram of the cross section of the well 46 vicinity of the microfluidic chip | tip of a modification. The schematic diagram of the cross section of the well 46 vicinity of the microfluidic chip | tip of a modification. The top view which shows typically the well 46 of the microfluidic chip | tip of a modification. The schematic diagram of the cross section of the microfluidic chip | tip 200 of a modification. The schematic diagram of the cross section of the microfluidic chip | tip 300 of a modification.

  Preferred embodiments of the present invention will be described below with reference to the drawings. In addition, the following embodiment demonstrates an example of this invention. Therefore, this invention is not limited to the following embodiment, The various modifications implemented in the range which does not change a summary are also included. Note that not all the configurations described in the following embodiments are essential constituent requirements of the present invention.

1. Microfluidic Chip The microfluidic chip 100 of this embodiment includes a substrate 10 and a cover 20.

1.1. Substrate FIG. 1 is a plan view schematically showing a substrate 10 of the microfluidic chip 100 of the present embodiment. FIG. 2 is a schematic diagram of a cross section of the substrate 10 of the microfluidic chip 100 of the present embodiment. FIG. 2 corresponds to a cross section taken along line AA of FIG.

1.1.1. Substrate The substrate 10 is a plate-like member that becomes the base of the microfluidic chip 100. As shown in FIG. 2, the substrate 10 has a first surface 11 and a second surface 12 that have a front-back relationship (opposite each other). The shape of the board | substrate 10 will not be specifically limited if it is flat form. The thickness of the substrate 10 (the distance between the first surface 11 and the second surface 12) is not particularly limited, but is preferably 0.5 mm or more and 5 mm or less in terms of ease of handling and resistance to breakage. The planar outer shape of the substrate 10 is not particularly limited, and may be a rectangle, a circle, or the like. In the present embodiment, an example in which the planar shape of the substrate 10 is a rectangle is shown.

  The material of the substrate 10 is not particularly limited, and examples thereof include inorganic materials (for example, single crystal silicon, Pyrex (registered trademark) glass) and organic materials (for example, resins such as polycarbonate and polypropylene), and these composite materials. It may be. When using the microfluidic chip 100 as a reaction vessel (reaction chip) for PCR (Polymerase Chain Reaction), the substrate 10 is made of a material with low spontaneous fluorescence. It is desirable to be formed by. Examples of such a material having a small spontaneous fluorescence include polycarbonate and polypropylene. When the microfluidic chip 100 is used for PCR, the substrate 10 is preferably made of a material that can withstand heating in PCR.

  Further, the substrate 10 may be made of carbon black, graphite, titanium black, aniline black, Ru, Mn, Ni, Cr, Fe, Co or Cu oxide, Si, Ti, Ta, Zr or Cr carbide. A black substance such as can be blended. By blending such a black material into the material of the substrate 10, the spontaneous fluorescence of the resin or the like can be further suppressed. In addition, when the microfluidic chip 100 is used for an application (for example, real-time PCR or the like) in which a well 16 or the like described later is observed from the outside of the microfluidic chip 100, the material of the substrate 10 is made transparent as necessary. Can be. In addition, when the microfluidic chip 100 is used as a PCR reaction chip, the material of the substrate 10 is preferably a material that hardly adsorbs nucleic acids and proteins and does not inhibit enzyme reactions such as polymerase.

  In the case where the substrate 10 is formed of an inorganic material, it can be formed and processed by performing dry etching using a photolithography method. Further, when the substrate 10 is formed using a resin as a main component, it can be molded and processed by a method such as mold molding, injection molding, or hot embossing.

1.1.2. Reservoir Region and Well Region As shown in FIG. 1, the substrate 10 is provided with a reservoir region 13 and a well region 14 in plan view. The reservoir region 13 and the well region 14 are provided adjacent to each other. The reservoir region 13 and the well region 14 may be provided adjacent to each other. A plurality of reservoir regions 13 and well regions 14 may be provided. The reservoir region 13 and the well region 14 are provided inside a first fixing region 21 of the cover 20 described later in plan view. The position where the reservoir region 13 and the well region 14 are provided on the substrate 10 is such that when an inertial force such as a centrifugal force is applied to the substrate 10, the inertial force acts in a direction from the reservoir region 13 toward the well region 14. However, there is no particular limitation as far as possible. The planar shape of the reservoir region 13 and the well region 14 is not particularly limited, and may be a rectangle, a circle, or the like.

1.1.3. Reservoir A reservoir 15 is formed in the reservoir region 13 of the substrate 10. The reservoir 15 is a space for storing a sample (liquid such as a specimen) at least before an inertial force is applied when the microfluidic chip 100 is used. The reservoir 15 is formed with an opening in the first surface 11 of the substrate 10. The planar shape of the reservoir 15 is not limited, and may be a circle, a rectangle, or the like. For example, the volume of the reservoir 15 is preferably equal to or larger than the total volume of the wells 16.

  In the example of FIGS. 1 and 2, the reservoir 15 is formed in a hollow shape that opens to the first surface 11 side, but is not limited thereto, and may penetrate to the second surface 12 side. When using the microfluidic chip 100, a sample is supplied to the reservoir 15 from the outside. In the example of FIGS. 1 and 2, for example, the sample may be supplied before the cover 20 is attached. After the cover 20 is mounted, a hole may be formed in a region corresponding to the reservoir 15 of the cover 20 and supplied from the hole, as in a modified example described later. Only the region corresponding to the above may be peeled off and supplied. Other forms that can be taken by the reservoir 15 will also be described in the section of modifications.

1.1.4. A well 16 having an opening 16a on the first surface 11 side of the substrate 10 is formed in the well region 14 of the well substrate 10 (see FIGS. 1 and 2). FIG. 3 is an enlarged plan view showing the vicinity of the well 16 of the microfluidic chip 100. FIG. 4 is a schematic diagram showing an enlarged cross section near the well 16 of the microfluidic chip 100. A cross section taken along line BB in FIG. 3 corresponds to FIG.

  A plurality of wells 16 can be formed in the well region 14. Arrangement of the wells 16 when forming a plurality in the well region 14 is arbitrary as long as the function is not impaired. The well 16 has a container shape having an opening 16 a on the first surface 11 side of the substrate 10. The well 16 can hold a liquid such as a specimen (sample) therein. In addition, a liquid reaction such as a specimen can be performed in the well 16. For example, the contents of the reservoir 15 can be introduced and held in the well 16 and can function as a reaction container.

  The shape of the well 16 is not particularly limited as long as it is a container shape, and can take various forms. For example, the shape of the well 16 may be any of a cylinder, a prism, a truncated cone and a truncated pyramid, a shape in which they are inclined, and a combination of these. In addition, the shape of the well 16 may have a larger outline than the opening 16a in a plan view. Examples of such a shape include a first well extending from the opening 16a to the substrate 10 in the thickness direction, and It may be an alphabetic “L” shape or the like having a second well connected to the first well inside the substrate 10 and extending in a direction opposite to the reservoir region 13. In the present embodiment, the case where the well 16 has a cylindrical shape (a shape in a plan view and a shape in a sectional view) as shown in FIGS. 1 to 4 will be described. Other shapes that can be taken by the well 16 will be further described in the section of modifications.

  As shown in FIG. 4, a reagent 30 for reaction or inspection can be placed in the well 16 in advance. The state of the reagent 30 to be arranged is preferably solid or liquid. For example, when the microfluidic chip 100 is used as a PCR chip, the reagent 30 includes a primer (nucleic acid) for amplifying the target nucleic acid and a fluorescent reagent for measuring the amount of amplification product (for example, SYBR GREEN ™). ), And if necessary, other nucleic acids can be arranged. When a reagent 30 having a high stability with respect to drying, such as the above-described example, is used as the reagent 30, the reagent 30 may be dried in the well 16. For example, as shown in FIG. You may arrange | position in the state apply | coated to the wall surface and dried. As a method for applying the reagent 30 to the inner wall surface of the well 16 as described above, for example, a method of applying the reagent 30 by a liquid ejecting head or the like used for ink jet printing can be used.

  When the reagent 30 is disposed in the well 16, the same reagent 30 may be disposed in the plurality of wells 16, or different reagents 30 may be disposed in the well 16. In the case where a plurality of wells 16 are formed, how the reagent 30 is arranged for each well 16 can be arbitrarily designed according to the desired reaction and inspection mode.

  The position where the reagent 30 is arranged in the well 16 is not particularly limited, but the farther from the opening 16a, the smaller the possibility that the reagent 30 will flow out of the well 16 together with the introduced liquid. More preferred.

  The opening 16a of the well 16 can be blocked by a cover 20 described later. Thereby, the well 16 can be an independent sealed container. By thus sealing the wells 16 independently, mixing with the contents of other wells 16 and contamination can be suppressed. The well 16 can be an independent sealed container when the cover 20 is laid and fixed around the opening 16 a of the well 16.

1.2. Cover FIG. 5 is a plan view schematically showing the microfluidic chip 100 of the present embodiment. FIG. 6 is a schematic view of a cross section of the microfluidic chip 100 of the present embodiment. 6 corresponds to a cross section taken along the line CC of FIG.

1.2.1. Cover The cover 20 is laid on the first surface 11 side of the substrate 10. In this specification, the term “laying” is used in the meaning including the case where the cover 20 and the first surface 11 of the substrate 10 are fixed (fixed) and the case where they are not fixed. The cover 20 has a film or sheet shape. Although the thickness of the cover 20 is not specifically limited, For example, it can be 0.01 mm or more and 5 mm or less. The planar outer shape of the cover 20 may or may not coincide with the planar outer shape of the substrate 10. In the example of FIG. 5, the planar outer shape of the cover 20 matches the planar outer shape of the substrate 10. The cover 20 is in contact with the substrate 10 in a static state, but when an inertial force such as a centrifugal force is applied to the microfluidic chip 100, the substrate 10 and the cover 20 are in a region other than the fixing region described later. Flexible or elastic enough to form a gap between the two. Moreover, the cover 20 can close the opening 16a of the well 16 provided in the board | substrate 10 in the state which contact | connected the board | substrate 10, and can make the well 16 an independent space (container). When the cover 20 is pressed against the substrate 10, it is preferable that the cover 20 is not easily bent in the vicinity of the opening 16 a of the well 16 and has elasticity that does not cause a large change in the volume of the well 16. The degree of elasticity of the cover 20 is appropriately designed by selecting a material in consideration of the thickness of the cover 20, the size of the opening 16 a of the well 16, and the pressure when contacting the substrate 10. Can.

  Examples of the material of the cover 20 include those having elasticity that can be bent by an inertial force such as centrifugal force. For example, organic materials (resins such as polycarbonate and polypropylene and various rubbers), organic materials, A composite material of an inorganic material can be given. When the microfluidic chip 100 is used for an application involving fluorescence measurement, such as when the microfluidic chip 100 is used as a PCR reaction chip, the cover 20 is desirably formed of a material having a small spontaneous fluorescence. Examples of such a material having a small spontaneous fluorescence include polycarbonate and polypropylene. When the microfluidic chip 100 is used as a PCR reaction chip, the cover 20 is preferably made of a material that has little nucleic acid or protein adsorption and does not inhibit an enzyme reaction such as polymerase. In addition, when the microfluidic chip 100 is used as a PCR chip, the cover 20 is preferably made of a material that can withstand heating in PCR.

  The material of the cover 20 is carbon black, graphite, titanium black, aniline black, or an oxide of Ru, Mn, Ni, Cr, Fe, Co or Cu, Si, Ti, Ta, as with the substrate 10. A black substance such as Zr or Cr carbide can be blended. By blending such a black substance into the material of the cover 20, the spontaneous fluorescence of the resin or the like can be further suppressed. Further, when the microfluidic chip 100 is used for the purpose of observing the well 16 described later from the outside of the microfluidic chip 100, the material of the cover 20 can be made transparent as necessary. The cover 20 can be molded and processed by a method such as film molding, sheet molding, injection molding, or press molding.

  The cover 20 is laid so that one surface 20 a faces the substrate 10. That is, the surface 20 a is provided so as to face the first surface 11 of the substrate 10. The surface 20a of the cover 20 may have adhesiveness. When the surface 20a has adhesiveness, it is preferable that the surface 20a and the first surface 11 have such an adhesive force that can be peeled off when an inertial force such as centrifugal force is applied to the microfluidic chip 100. However, this is not limited to this after the application of an inertial force such as centrifugal force to the microfluidic chip 100, and the adhesive force is sufficient to adhere (fix) the surface 20a and the first surface 11 so that they are not easily separated. May be.

  The surface 20a of the cover 20 may not have an adhesive force. In such a case, in order to fix the surface 20a to the first surface 11, an adhesive can be used or heat fusion can be performed. Further, the surface 20a may have a property of exhibiting an adhesive force by pressurization without exhibiting an adhesive force in a state where the cover 20 is not pressed against the substrate 10. An example of such a surface 20a is a porous surface. With such a method, when the substrate 10 and the cover 20 are fixed, only pressure is applied, so that no heat is generated, and an increase in the temperature of the microfluidic chip 100 can be suppressed. The influence of the heat to give can be suppressed. Specific examples of the cover 20 having such a surface 20a include a trade name: LightCycler 480 Sealing Foil, a model name: 04 729 757 001, manufactured by Roche Diagnostics, a trade name: a polyolefin microplate sealing tape : 9793-3M, trade name: amplification tape 96, model name: 232702, manufactured by Nunc, etc.

  Furthermore, the surface 20a may have a potential adhesion. That is, the surface 20a does not have an adhesive force before an inertial force such as a centrifugal force is applied to the microfluidic chip 100, and an energy ray (for example, an ultraviolet ray) at a desired time after the inertial force is applied. , An electron beam, etc.) may be used to exert adhesive force.

  Further, as a method for fixing the surface 20a and the first surface 11 of the substrate 10, an ultrasonic welding method may be used. For example, a jig having a shape corresponding to the shape of the region to be fixed may be pressed against the portion of the cover 20 to be fixed to the substrate 10 and ultrasonically vibrated to weld (fix) the cover 20 and the substrate 10. Good. In this case, since the substrate 10 and the cover 20 are welded (fixed) by ultrasonic irradiation, for example, when the liquid is a biochemical sample, heating of the sample can be suppressed. Damage to the can be reduced. In addition, for example, it is possible to suppress a decrease in the activity of the reagent contained in the well. Furthermore, when the first fixing region 21 is formed by the ultrasonic welding method, the bonding force between the substrate 10 and the cover 20 due to the welding is relatively strong, so that liquid leakage from the well region 14 can be more reliably prevented. Can do. The first fixing region 21 and the second fixing region 22 described below can be formed by the method described above.

1.2.2. First fixing area The cover 20 has a first fixing area 21 and a second fixing area 22 fixed to the substrate 10.

  The first fixing region 21 is disposed so as to surround the reservoir region 13 and the well region 14 of the substrate 10 in plan view. That is, the reservoir region 13 and the well region 14 are disposed inside the first fixing region 21 as viewed in a plan view. The first fixing region 21 is a region in which the laid cover 20 and the substrate 10 are not easily separated even when an inertial force such as a centrifugal force is applied to the microfluidic chip 100. When an inertial force such as a centrifugal force is applied to the microfluidic chip 100, the surface 20a and the first surface 11 are peeled off in regions other than the first fixing region 21 and the second fixing region 22 of the laid cover 20. can do. In the first fixing region 21, for example, the cover 20 and the substrate 10 may be welded or bonded with an adhesive, an adhesive, or the like in accordance with the property of the surface 20 a of the cover 20.

  As one of the functions of the first fixing region 21, when an inertial force such as a centrifugal force is applied to the microfluidic chip 100 to form a gap between the cover 20 and the substrate 10, the gap is set inside. Forming a bag (pocket) -like structure to be formed. Accordingly, the reservoir region 13 and the well region 14 can be communicated between the substrate 10 and the cover 20 on the first surface 11 side of the substrate 10, and the sample (liquid) can be circulated between the two regions.

  As already described, the inertial force is applied to the microfluidic chip 100 from the reservoir region 13 side toward the well region 14 side. Therefore, the first fixing region 21 is continuously provided at least at a position surrounding the well region 14. Therefore, when the liquid is introduced into the gap between the cover 20 and the substrate 10, the liquid is prevented from leaking from the well region 14 side to the outside of the first fixing region 21 due to inertial force. Further, the first fixing region 21 can be continuous in an annular shape so as to surround both the reservoir region 13 and the well region 14 of the substrate 10 in a plan view. In this way, it is possible to prevent the liquid from leaking outside the first fixing region 21 when the liquid is introduced into the gap between the cover 20 and the substrate 10. The first fixing region 21 may have a non-continuous portion on the reservoir region 13 side.

  The first fixing region 21 can be formed, for example, by applying an adhesive in the shape of the first fixing region 21 on the first surface 11 of the substrate 10 and laying the cover 20. In addition, for example, the first fixing region 21 has a property that the surface 20a of the cover 20 does not exert an adhesive force in a state where the cover 20 is not pressed against the substrate 10 and exhibits an adhesive force by pressurization. Alternatively, a jig or the like corresponding to the shape of the first fixing region 21 is prepared, and the cover 20 can be formed by applying a pressing force to the substrate 10 with the jig. Further, for example, the first fixing region 21 can be formed by ultrasonic welding using a jig having a shape corresponding to the shape of the first fixing region 21.

1.2.3. Second Fixed Region The second fixed region 22 is provided along the outline of the opening 16a of the well 16 in plan view. The second fixing region 22 is provided with a discontinuous portion 22a on the reservoir region 13 side in a plan view. That is, the well 16 is disposed in contact with the inside of the second fixing region 22 in plan view. The second fixing region 22 is a region in which the cover 20 and the substrate 10 are not easily separated even when an inertial force such as a centrifugal force is applied to the microfluidic chip 100. Therefore, when an inertial force such as centrifugal force is applied to the microfluidic chip 100, the surface 20a and the first surface 11 can be separated at the discontinuous portion 22a on the reservoir region 13 side. In the second fixing region 22, according to the property of the surface 20a of the cover 20, for example, the cover 20 and the substrate 10 may be welded or may be bonded with an adhesive, an adhesive, or the like.

  One of the functions of the second fixing region 22 is that when an inertial force such as centrifugal force is applied to the microfluidic chip 100 to form a gap between the cover 20 and the substrate 10, In the discontinuous portion 22a, the gap and the inside of the well 16 are communicated. Further, as one of the functions of the second fixing region 22, an inertial force such as centrifugal force is applied to the microfluidic chip 100, and a sample (liquid) is introduced into the well 16 (the gas in the well 16 is the sample). And a bag-like structure that makes it difficult for the excess sample to leak out in the direction opposite to the reservoir region 13 of the well 16. This function is manifested when the second fixing region 22 is continuously provided at a position opposite to the reservoir region 13 side at least in the outline of the opening 16a of the well 16. Thereby, when the liquid is introduced into the gap between the cover 20 and the substrate 10, an excessive amount of liquid with respect to the volume of the well 16 is separated from the reservoir region 12 of the second fixing region 22 due to inertial force. It is difficult to leak in the opposite direction.

  In addition, as one of the functions of the second fixing region 22, a gap formed between the cover 20 and the substrate 10 when an inertial force such as centrifugal force is applied to the microfluidic chip 100 and the inside of the well 16 are used. The path of movement of the sample (liquid) is narrowed. Since the second fixing region 22 has such a shape, the sample once introduced into the well 16 can be prevented from leaking out of the well 16 again. As a result, it is possible to enhance the effect of preventing mixing between the sample introduced into the well and the sample introduced into the adjacent well, and contamination due to the mixing.

  The second fixing region 22 can take various shapes. The second fixing region 22 shown in FIG. 3 shows some examples of the shape, and each example is provided along the contour of the opening 16a of the well 16 and is not formed on the reservoir region 13 side. A continuous portion 22a is provided. In the example of FIG. 3, the second fixing region 22 having a different shape is formed for each well 16, but the second fixing region 22 may have the same shape in the plurality of wells 16.

  The second fixing regions 22 shown in FIG. 3 are all formed for wells that do not have protrusions, which will be described later. However, the matter described below is that the wells have protrusions. Can also be applied.

  As shown in FIG. 3, the second fixing region 22 may have any shape as long as it has a discontinuous portion on the reservoir region 13 side. However, the total length in the contour of the opening 16a of the well 16 in the second fixing region 22 is 50% or more of the contour length of the opening 16a, or the second fixing region 22 in the contour of the opening 16a. The length of the longest continuous portion is preferably 50% or more of the contour length of the opening 16a. Thereby, mixing of the contents of the adjacent wells 16 and contamination, which are one of the effects of the microfluidic chip 100 of the present embodiment, can be more reliably suppressed. The second fixing region 22 illustrated in FIG. 3A is formed only on the opposite side of the reservoir region 13. In this example, the total length of the contour of the opening 16a of the well 16 in the second fixing region 22 is 50% of the length of the contour of the opening 16a. The second fixing region 22 illustrated in FIG. 3B is formed on the entire opposite side of the reservoir region 13 and a part on the reservoir region 13 side. In this example, the length of the longest continuous portion of the second fixing region 22 in the outline of the opening 16a is 50% or more of the length of the outline of the opening 16a.

  The shape of the second fixing region 22 is such that when the opening 16 a is partitioned by a straight line connecting both ends of the longest continuous portion of the second fixing region 22, the opening 16 a surrounded by the straight line and the second fixing region 22 is formed. The area Sp of the partition portion may occupy 50% or more of the area of the opening 16a. Thereby, mixing of the contents of the adjacent wells 16 and contamination, which are one of the effects of the microfluidic chip 100 of the present embodiment, can be more reliably suppressed. In the example of FIG. 3B, when the opening 16 a is defined by a straight line connecting both ends of the longest continuous portion of the second fixing region 22, the partition of the opening 16 surrounded by the straight line and the second fixing region 22. The area Sp of the portion occupies 50% or more of the area of the opening 16a.

  If a plurality of discontinuous portions 22a are formed in the well 16 as in the example of FIG. 3B, for example, the number of paths for introducing a sample increases and the width of each path can be reduced. it can. Thereby, for example, by disposing the discontinuous portions 22 a as appropriate, the path for introducing the sample 210 into the well 16 and the path for discharging the gas from the well 16 can be separated. This makes it possible to more efficiently introduce the sample into the well 16 when using the microfluidic chip.

  Regarding the size of the second fixing region 22, the second fixing regions 22 formed around the adjacent wells 16 are continuous with each other in a range where the first fixing region 21 and the second fixing region 22 are not continuous. Also good. FIG. 7 schematically shows a state in which the second fixing regions 22 of adjacent wells 16 are continuous in the microfluidic chip 100. Even if such a second fixing region 22 is formed, the sample by centrifugal force is applied to the well 16 located on the opposite side of the reservoir region 13 of the continuous second fixing region 22 as shown by the arrow in the figure. Therefore, a sample can be introduced without hindrance.

  The second fixing region 22 can be formed, for example, by applying an adhesive in the shape of the second fixing region 22 on the first surface 11 of the substrate 10 and laying the cover 20. Further, for example, the second fixing region 22 has a property in which the surface 20a of the cover 20 does not exert an adhesive force in a state where the cover 20 is not pressed against the substrate 10 and exhibits an adhesive force by pressurization. Alternatively, a jig or the like corresponding to the shape of the second fixing region 22 is prepared, and the cover 20 can be formed by applying a pressing force to the substrate 10 with the jig. Further, for example, the second fixing region 22 can also be formed by ultrasonic welding using a jig having a shape corresponding to the shape of the second fixing region 22.

1.3. Method of Using Microfluidic Chip The microfluidic chip 100 described above can be used for a wide range of applications. Hereinafter, the method of using the microfluidic chip 100 as an example of a chip for PCR will be described. To do.

  FIG. 8 is a diagram schematically illustrating a state in which an inertial force is applied in a state where the specimen 210 is placed in the reservoir 15 of the microfluidic chip 100. The state of the specimen 210 is liquid. FIG. 6 schematically shows a state in which the inertial force is not applied in a state where the specimen 210 is placed in the reservoir 15 of the microfluidic chip 100.

  In the following example, it is assumed that the surface 20a of the cover 20 has a property of exerting an adhesive force by pressurization without exhibiting an adhesive force in a state where the cover 20 is not pressed against the substrate 10. . In the following example, the first fixing region 21 of the cover 20 is assumed to be continuous in an annular shape so as to surround both the reservoir region 13 and the well region 14 of the substrate 10 in plan view. In the following example, the cover 20 is formed of a transparent material. In this example, the well 16 has a cylindrical shape, and the bottom surface has a diameter of about 1 mm and a depth of about 0.5 mm. Further, in this example, the planar external shapes of the substrate 10 and the cover 20 are rectangular, and the long side is about 7.5 cm and the short side is about 2.5 cm.

  First, a specimen 210 containing a target nucleic acid is prepared. Examples of the PCR specimen 210 include an aqueous solution containing a target nucleic acid, primer DNA, and a PCR master mix (for example, containing a polymerase, a nucleotide, and a coenzyme such as magnesium chloride). In the sample 210, examples of the target nucleic acid to be measured include DNA extracted from blood, urine, saliva, spinal fluid, etc., or cDNA reverse-transcribed from RNA. The amount of the specimen 210 is appropriately determined according to the entire volume of the well 16, and is preferably the same as or larger than the total volume of the plurality of wells 16, for example. More preferably, it is larger than the total volume of the plurality of wells 16 in that 210 can be filled.

  Next, the specimen 210 is accommodated in the reservoir 15 of the microfluidic chip 100 (see FIG. 6). At this time, the specimen 210 is not introduced into the well 16 of the microfluidic chip 100, and the reagent 30 and the like are arranged in each well 16. Examples of the reagent 30 and the like include primer DNA and fluorescent probe DNA. In addition, the method of putting the specimen 210 in the reservoir 15 is not particularly limited, and as described above, for example, it may be performed before the cover 20 is attached, or after the cover 20 is attached, A hole may be formed in an area corresponding to the reservoir 15 of the 20 and supplied from the hole, and the cover 20 may be supplied after being peeled only in an area corresponding to the reservoir 15.

  Next, an inertial force is applied to the microfluidic chip 100 from the reservoir region 13 side toward the well region 14 side. In this embodiment, the inertial force is applied by a centrifuge. The rotation axis R of the centrifuge is disposed on the side opposite to the well region 14 when viewed from the reservoir region 13 of the microfluidic chip 100. When an inertial force (centrifugal force) is generated in the microfluidic chip 100 by operating the centrifuge, the specimen 210 in the reservoir 15 moves away from the rotation axis R, that is, in the well region 14 as shown in FIG. Acceleration G (arrow in the figure) is received in the direction of heading. In a state where acceleration is applied, a gap is generated between the substrate 10 and the cover 20, and the specimen 210 is transferred in the gap toward the well region 14. The size (thickness) of the gap at this time is not particularly limited, but FIG. 8 shows an example in which the size of the gap is very small. Then, the specimen 210 is replaced with the air in the well 16 by inertia force from the opening 16 a of the well 16 formed in the well region 14 of the substrate 10 through the discontinuous portion of the second fixing region 22. To be introduced. The specimen introduced into the well 16 is mixed with the reagent 30 or the like previously arranged in the well 16.

  In the example of FIG. 8, a state in which the plane of the microfluidic chip 100 (for example, the first surface 11 of the substrate 10) is arranged and rotated along a direction perpendicular to the rotation axis R of the centrifuge. As shown, the arrangement of the microfluidic chip 100 with respect to the rotation axis R of the centrifuge is arbitrary as long as the specimen 210 inside the reservoir 15 can move toward the well 16 by centrifugal force. Such an arrangement can be appropriately set depending on the arrangement of the wells 16 of the substrate 10 or the like.

  Next, the centrifuge is stopped and the application of inertia force is stopped. In this state, the specimen 210 is filled in the well 16. Further, for example, when the volume of the specimen 210 is larger than the total volume of the well 16, the specimen 210 exists in the well 16 and in the gap between the substrate 10 and the cover 20. In any case, by pressing the cover 20 against the substrate 10 from the outside of the cover 20 with a jig such as a roller, a squeegee or a blade, the cover 20 is brought into close contact with the microfluidic chip 100 and the well 16 is sealed. The specimen 210 having an accurate volume can be enclosed in the well 16. That is, when the specimen 210 is not present in the gap between the substrate 10 and the cover 20, the discontinuous portion 22 a is pressed by pressing the cover 20 against the substrate 10, the well 16 is sealed, and the specimen 210 is sealed. In the state where it exists in the gap between the substrate 10 and the cover 20, the well 16 can be sealed while pushing the excess specimen 210 back toward the reservoir region 13. Thus, the well 16 can be a sealed reaction container having a predetermined volume.

  FIG. 9 is a diagram schematically showing how the well 16 of the microfluidic chip 100 is sealed by the roller 500. In the example of FIG. 9, a state in which the well 16 is sealed by the roller 500 while returning the excessive specimen 210 existing in the gap between the substrate 10 and the cover 20 from the through hole 15 to the reservoir 15 is shown. FIG. 10 is a plan view schematically showing a state where the well 16 of the microfluidic chip 100 is sealed.

  FIG. 10 is a plan view schematically showing the microfluidic chip 100 in which the well 16 is sealed. As shown in FIG. 10, after the cover 20 is pressed by the roller 500, the contact surface between the substrate 10 and the cover 20 is at least around the well 16 (in the illustrated example, the entire surface where the substrate 10 and the cover 20 contact each other). ).

  As described above, as shown in FIG. 10, an accurate volume of the specimen 210 can be introduced into the well 16 of the microfluidic chip 100. The same applies to the case where a plurality of wells 16 are formed in the microfluidic chip 100. The plurality of wells 16 become independent sealed spaces as a result of the above operation, and a desired amount of the specimen 210 is precisely dispensed. can do.

  Thereafter, the microfluidic chip 100 is introduced into a temperature control device such as a thermal cycler, and the PCR reaction is performed in the well 16. As an example of the temperature control of the PCR reaction, there is a method of repeating a three-stage temperature change of 55 ° C., 74 ° C., and 95 ° C. with a period of several minutes. By such a temperature cycle, the target nucleic acid can be amplified twice per cycle. Furthermore, since the cover 20 is made of a transparent material here, the inside of the well 16 can be observed from the outside of the cover 20 through the cover 20 as necessary. Therefore, the target nucleic acid can be quantified (real-time PCR), and the progress of the reaction can be confirmed even during the PCR reaction. In addition, the microfluidic chip 100 can be used to analyze various nucleic acids (DNA, RNA) using the principle of PCR, such as gene mutation such as SNP and DNA methylation.

  Another effect of the second fixing region 22 is that the gap between the substrate 10 and the cover 20 does not become too large when the microfluidic chip 100 is used. FIG. 11 shows a microfluidic chip of a reference example that does not have the second fixing region. For example, as shown in FIG. 11, if the inertial force applied when the microfluidic chip 100 is used is too large, the gap between the substrate 10 and the cover 20 becomes too large near the end opposite to the reservoir region 13. There is. In the microfluidic chip 100 according to the present embodiment, since the second fixing region 22 is formed, the gap between the substrate 10 and the cover 20 can be prevented from becoming too large at least in the vicinity of the well 16. Therefore, for example, the waste of the sample can be kept small.

1.4. Modification 1.4.1. Deformation of Well Shape In the above section “1.1.4. Well”, it was described that the well 16 can be variously deformed. Hereinafter, a well 46 that is a modified embodiment of the well 16 will be described.

  FIG. 12 is an enlarged plan view schematically showing the vicinity of the well 46 of the modification. FIGS. 13 to 16 are plan views schematically showing an enlarged cross section of a well 46 according to a modification. 13 to 16 correspond to cross sections taken along the lines DD, EE, FF, and GG of FIG. 12, respectively. In the following modification, the same reference numerals are given to the same configurations as those described in the above-described embodiment, and detailed description thereof is omitted. In the configurations described in the following modification examples, configurations indicated by the same reference numerals as those in the above-described embodiment have the same function.

  The well 46 may have a protrusion 47 having an opening 47a on the first surface 11 side of the substrate 10 as shown in FIG. In other words, the well 46 may have a protruding portion 47 formed so that the outline of the opening 46a protrudes toward the reservoir region 13 in a plan view. In the example of FIG. 12, the well 46 has one protrusion 47, and the contour of the opening 47 a of the protrusion 47 is continuous with the contour of the well 46 and from the contour of the well 46 toward the reservoir region 13. It protrudes.

  The protrusion 47 is a space that communicates with the well 46 in the substrate 10. The protrusion 47 has an opening 47 a on the first surface 11 side of the substrate 10, and is integrated with the well 46 to have a container shape. The shape of the protrusion 47 is not particularly limited. For example, the shape of the protrusion 47 may be any of a cylinder, a prism, a truncated cone and a truncated pyramid, a shape in which they are inclined, and a combination of these. Although the magnitude | size of the protrusion part 47 is not specifically limited, It is preferable to have a volume smaller than the well 46 used as a main container. The depth of the protrusion 47 is not particularly limited, and may be the same as or shallower than the depth of the well 46. Furthermore, the depth of the protrusion 47 may not be uniform, and the bottom surface may be inclined.

  The position where the protrusion 47 is formed is preferably on the reservoir region 13 side of the well 46. By forming the protruding portion 47 on the reservoir region 13 side of the well 46, the sample is easily introduced into the well 46 through the protruding portion 47 when the microfluidic chip 100 is used. When the protrusion 47 is formed in the well 46, the flow rate of the sample flowing through the protrusion 46 can be adjusted according to the shape of the protrusion 46.

  In addition, when the protrusion 47 is formed, it is possible to facilitate the formation of the second fixing region 22. That is, in the above-mentioned section “1.2.3. Second fixing region”, the second fixing region 22 is prepared by, for example, preparing a jig corresponding to the shape of the second fixing region 22. As described above, the cover 20 can be formed by applying a pressing force to the substrate 10. When the protrusion 47 is formed, the shape of the jig at this time can be simplified. When the protrusion 47 is not provided, it is necessary to form a shape corresponding to the discontinuous portion of the second fixing region 22 in the jig, but when the protrusion 47 is provided, the well 46 is formed. Even if a larger cylindrical or columnar jig is used, the discontinuous portion of the second fixing region 22 can be formed.

  FIGS. 13 to 16 show a state in which an inertial force is applied to the microfluidic chip, and a sample 210 is drawn in any of the drawings. In the example of FIGS. 13 to 16, a gap is formed between the substrate 10 and the cover 20 in a region other than the second fixing region 22, and the sample is in the gap, the well 46, and the protrusion 47. 210 shows the existence of 210. As shown in FIG. 13, since the gap and the protruding portion 47 communicate with each other, the sample 210 can be introduced into the well 46 through the protruding portion 47.

  In addition, a plurality of protrusions 47 can be formed. FIG. 17 is a plan view schematically showing an enlarged well 46 having two protrusions 47. Also in the example of FIG. 17, the outline of the opening 47 a of the protrusion 47 is continuous with the outline of the well 46. The protrusion 47 is a space communicating with the well 46 in the substrate 10 protruding from the outline of the well 46, has an opening 47 a on the first surface 11 side of the substrate 10, and is integrated with the well 46. It has a container shape.

  When a plurality of protrusions 47 are formed in the well 46, for example, it becomes easy to form a plurality of discontinuous portions of the second fixing region 22. When a plurality of protrusions 47 are formed in the well 46, the number of sample channels increases, and the width of each channel can be reduced. Thereby, for example, by appropriately arranging the projecting portions 47, the path for introducing the sample 210 into the well 46 and the path for discharging the gas from the well 46 can be separated. This makes it possible to more efficiently introduce the sample into the well 46 when using the microfluidic chip.

  In any of the above examples, the opening 47 a of the protrusion 47 can be blocked by the cover 20 together with the opening 46 a of the well 46. Thereby, the well 46 and the protrusion part 47 connected to this can become one independent sealed container. As described above, the well 46 can be prevented from being mixed and contaminated with the contents of the other well 46 by sealing the well 46 independently.

1.4.2. Through-hole The microfluidic chip of this embodiment can include a mechanism for supplying a sample (liquid such as a specimen) to the reservoir 15 from the outside. Examples of such a mechanism include a through-hole 15a that allows the reservoir 15 to communicate with the outside, and a through-hole 15a in which a general-purpose tube (such as a microtube or a PCR tube) is directly attached to the reservoir 15.

  18 and 19 are diagrams schematically showing a cross section of a microfluidic chip having a through hole 15a.

  In the reservoir region 13 of the substrate 10 of the microfluidic chip 200 shown in FIG. 18, a reservoir 15 for accommodating the sample 210 is formed, and a through hole 15a that communicates the reservoir 15 with the outside is formed. . As shown in FIG. 18, the microfluidic chip 200 uses a pipette P without special processing on the substrate 10 and the cover 20, and the sample 210 is externally stored in the reservoir 15 through the through hole 15a. Can be introduced. And if necessary, the opening of the through-hole 15a can be closed using a seal 50 as shown in the figure. According to the microfluidic chip 200 of such a modification, the workability when the sample is introduced into the reservoir 15 can be improved.

  Further, in the microfluidic chip 300 shown in FIG. 19, a container mounting mechanism 17 capable of connecting a microtube or the like to the reservoir 15 for storing a sample is formed. The container mounting mechanism 17 can connect the container 400 to the second surface 12 side of the substrate 10 to allow the inside of the container 400 to communicate with the reservoir 15 to form one closed space.

  Examples of the container 400 include general-purpose tubes (microtubes, PCR tubes, etc.). The container mounting mechanism 17 has, for example, a structure that connects the substrate 10 and the container 400, and can fix the container 400 to the substrate 10. For example, in the example shown in FIG. 19, the container mounting mechanism 17 has a structure in which the container 400 is fitted (a structure in which the container 400 can serve as a cap). In this example, a cylindrical container mounting mechanism 17 is formed on the second surface 12 side around the through-hole 15, and the container mounting mechanism 17 is inserted into the inner surface of the container 400 so that the container 400 is fixed. Has been. Further, as in the illustrated example, the container mounting mechanism 17 may have a structure having an auxiliary function such as a protrusion 17 a for making it difficult to remove the container 400.

  According to the microfluidic chip 300 of the modified example, the workability when the sample is introduced into the reservoir 15 can be further improved. That is, by preparing the sample 210 in the container 400 in advance, it is not necessary to transfer the sample from the container 400 to the microfluidic chip. Therefore, workability is improved and the sample can be introduced into the reservoir 15 without waste.

1.5. Effects and the like The microfluidic chip of the present embodiment has at least the following effects. The microfluidic chip according to this embodiment includes a cover having a discontinuous portion on the reservoir region 13 side and having a second fixing region fixed to the substrate so as to follow the outline of the well opening. For this reason, the path for flowing the liquid through the well is narrowed, and the liquid once introduced into the well is unlikely to flow backward. Therefore, contamination such as mixing of samples is suppressed.

  Further, according to the microfluidic chip of the present embodiment, a small amount of liquid can be accurately dispensed. For example, a minute amount of liquid that is difficult to dispense manually by using a pipette. Note is possible. That is, according to the microfluidic chip of this embodiment, it is possible to easily introduce the liquid into the well in an amount very close to the design amount. Further, according to the microfluidic chip of the present embodiment, since the flow path communicating with the well is not formed, the well can be arranged with high density and the amount of liquid to be used can be suppressed to a very small amount. Therefore, the efficiency and reliability when performing inspection, reaction, etc. using the microfluidic chip of this embodiment can be improved. Furthermore, the microfluidic chip of the present embodiment can greatly reduce the man-hours and costs for dispensing work. In addition, according to the microfluidic chip of the present embodiment, the dispensing process can be greatly reduced, and for example, it is not necessary to use expensive equipment such as an automatic dispensing device. Can be dispensed.

  In the present embodiment, a part of the case where the microfluidic chip is used for PCR has been described as an example. However, the use of the microfluidic chip of the present embodiment is not limited, and examples thereof include viruses, bacteria, proteins, low It can also be used for inspection of molecules to polymer compounds, cells, particles, colloids, for example, allergens such as pollen, poisonous substances, harmful substances, and environmental pollutants. In the present embodiment, the case where the reagent 30 is arranged in the well of the microfluidic chip has been described. However, the reagent 30 may not be arranged in the well depending on the examination contents.

2. Specimen Measurement Method A specimen 210 measurement method using the above-described microfluidic chip and its usage will be described. As an example of a method for measuring the specimen 210 according to the present embodiment, a step of introducing the specimen 210 into the reservoir 15 of the microfluidic chip 100, and an inertial force is applied to the microfluidic chip 100 to fill the specimen 210 into the well. A step of fixing the substrate 10 and the cover 20 at least around the opening of the well; a step of reacting the specimen 210 in the well; and a step of observing the specimen 210 in a state of being present in the well; Can be mentioned.

  First, the specimen 210 is introduced into the reservoir 15 of the microfluidic chip 100. For example, as described above, this step can be performed by injecting the specimen 210 into the reservoir 15 with a micropipette or the like and sealing the reservoir 15 with a seal 50 or the like as necessary.

  Next, the step of applying the inertial force to the microfluidic chip 100 and filling the specimen 210 into the well can be performed by applying the inertial force (centrifugal force) using a centrifuge, for example, as described above.

  The step of fixing the substrate 10 and the cover 20 at least around the opening 16a of the well 16 is, for example, as described above, in which the surface 20a of the cover 20 exhibits an adhesive force by pressure, and the cover 20 is mounted on the substrate by the roller 500. 10 can be performed.

  The step of reacting the specimen 210 in the well 16 can be performed by introducing the microfluidic chip 100 into the thermal cycler after the above step, for example, as described above.

  For example, as described above, the step of observing the specimen 210 in the well 16 is performed by observing the inside of the well 16 from the outside using at least one of the substrate 10 and the cover 20 of the microfluidic chip 100 as a transparent material. It can be carried out.

  According to the sample measuring method as described above, a predetermined amount of the sample 210 can be easily and accurately filled in a well, and mixing between adjacent wells can be suppressed to be small. In addition, foreign matter can be prevented from being mixed into the specimen 210, and the specimen 210 can be accurately and reliably dispensed at low cost, so that the measurement accuracy of the specimen 210 can be improved.

  The embodiments described above and the modified embodiments can be arbitrarily combined with a plurality of forms. Thereby, combined embodiment can have an effect which each embodiment has, or a synergistic effect.

  The present invention is not limited to the above-described embodiments, and various modifications can be made. For example, the present invention includes substantially the same configuration (for example, a configuration having the same function, method, and result, or a configuration having the same purpose and effect) as the configuration described in the embodiment. In addition, the invention includes a configuration in which a non-essential part of the configuration described in the embodiment is replaced. In addition, the present invention includes a configuration that achieves the same effect as the configuration described in the embodiment or a configuration that can achieve the same object. In addition, the invention includes a configuration in which a known technique is added to the configuration described in the embodiment.

DESCRIPTION OF SYMBOLS 10 ... Board | substrate, 11 ... 1st surface, 12 ... 2nd surface, 13 ... Reservoir area | region, 14 ... Well area | region, 15 ... Reservoir, 15a ... Through-hole, 16 ... Well, 16a ... Opening, 17 ... Container mounting mechanism, 17a ... Projection, 20 ... Cover, 20a ... Surface, 21 ... First fixing region, 22 ... Second fixing region, 22a ... Discontinuous portion, 30 ... Reagent, 46 ... Well, 46a ... Opening, 47 ... Protrusion, 47a ... Opening 50 ... Seal 100,200,300 ... Microfluidic chip 400 ... Container 210 ... Sample 500 ... Roller R ... Rotating shaft G ... Acceleration P ... Pipette

Claims (6)

A reservoir region having a first surface and a second surface opposite to the first surface, a reservoir region in which a reservoir is formed, and a well region adjacent to the reservoir region in plan view, and the first region in the well region A substrate in which a well having an opening on the surface side is formed;
A first fixing region which is laid on the first surface side of the substrate and fixed to the substrate so as to surround the reservoir region and the well region in a plan view; and a discontinuous portion on the reservoir region side. A cover having a second fixing region fixed to the substrate along the contour of the opening;
Including a microfluidic chip. In claim 1,
The well has a microfluidic chip having a projecting portion in which an outline of the opening projects toward the reservoir region in a plan view. In claim 2,
A microfluidic chip having a plurality of the protrusions. In any one of Claims 1 to 3,
The microfluidic chip, wherein the total length of the second fixing region in the outline of the opening is 50% or more of the length of the outline of the opening. In any one of Claims 1 thru | or 4,
The length of the longest continuous part of the second fixed region in the outline of the opening is 50% or more of the length of the outline of the opening. In any one of Claims 1 to 3,
When the opening is partitioned by a straight line connecting both ends of the longest continuous portion of the second fixing region, an area of the partitioning portion of the opening surrounded by the straight line and the second fixing region is an area of the opening. Microfluidic chip occupying 50% or more.