JP2011145236A - Micro-fluid chip, and measuring method of specimen using the same - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a micro-fluid chip capable of dispensing easily a small amount of liquid. <P>SOLUTION: The micro-fluid chip 100 includes: a substrate 10 having a first surface 11 and a second surface 12 opposed to the first surface 11, and provided with a through hole region 13 on which a through hole 15 for connecting the first surface 11 to the second surface 12 is formed and a well region 14 which is adjacent to the through hole region 13, and on which wells 16 having each opening are formed on the first surface 11 side; a cover laid on the first surface 11 side, and having a fixed resin 21 fixed on the substrate 10 so as to enclose the through hole region 13 and the well region 14 in a plan view; and a container mounting mechanism 17 having a structure for communicating a container 200 with the through hole 15, which is a mechanism used for mounting the container 200 on the second surface 12 side. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a microfluidic chip and a specimen measurement method using the same.

  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. Microfluidic chips are sometimes referred to by names such as Micro Total Analytical System (Micro TAS) or Lab-on-a-chip.

  The microfluidic chip generally includes 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, chemicals, and the like (Patent Document 1).

  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, while the conventional microfluidic chip can reduce the amount of sample, the operation of injecting (dispensing) the small amount of sample into a plurality of reaction containers called wells is complicated. For example, there is a method of performing a dispensing operation on a microfluidic chip with a pipette or the like. However, since the dispensing operation is performed manually, it takes a long working time.

  One of the objects according to some embodiments of the present invention is to provide a microfluidic chip capable of easily dispensing a small amount of sample.

  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 first surface and a second surface opposite to the first surface, and in a plan view, a through hole region in which a through hole connecting the first surface and the second surface is formed, and adjacent to the through hole region And a substrate provided with a well region in which a well having an opening on the first surface side is formed,
A cover laid on the first surface side and having a fixing region fixed to the substrate so as to surround the through-hole region and the well region in plan view;
A mechanism used when mounting a container on the second surface side, the container mounting mechanism having a structure for communicating the container and the through hole;
Have

  According to the microfluidic chip of this application example, the container and the through-hole of the substrate can be communicated with each other by a container mounting mechanism used when mounting a container containing a sample (specimen), for example. There is no need to transfer the sample to the sample, and the sample can be more easily accommodated in the microfluidic chip. Further, when dispensing the sample into the well, the sample accommodated in the container can be easily introduced into the well through the gap between the substrate and the cover, for example, by the action of centrifugal force. Therefore, it is not necessary to perform a dispensing operation with a pipette or the like, and the dispensing can be performed more easily.

  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 substrate may have, on the second surface side, a groove extending in a direction separating the through-hole region and the well region between the through-hole region and the well region in a plan view.

  According to the microfluidic chip of this application example, it is easy to bend the microfluidic chip between the through-hole region and the well region, with the groove extending in the direction separating the through-hole region and the well region as a fold. . Thereby, the direction in which the through hole and the well are inclined can be adjusted. Therefore, for example, when a centrifugal force or the like is applied to the microfluidic chip, the direction in which the through hole and the well are inclined can be adjusted, and the sample can be introduced into the well more efficiently. Moreover, it is easy to divide the microfluidic chip between the through-hole region and the well region with a cutting tool or the like. Thereby, the microfluidic chip of this application example can be easily placed on a flat surface and easy to handle by cutting the through-hole region and the well region after introducing the sample into the well.

[Application Example 3]
In application example 1 or application example 2,
The area of the outline of the well in plan view is larger than the area of the outline of the opening in plan view,
In plan view, the outline of the opening may be in contact with the outline of the well on the side close to the through-hole region in the outline of the well.

  According to the microfluidic chip of this application example, the area of the well in plan view is larger than the area of the well opening, that is, the well opening is narrowed. For this reason, the sample introduced into the well is unlikely to flow backward and contamination is unlikely to occur. Further, when a reagent or the like that acts on the sample is arranged in the well, it is possible to suppress the reagent from diffusing outside the well. Thereby, the reliability of inspection or the like can be further increased. Furthermore, in the microfluidic chip of this application example, the well opening is disposed on the side close to the through-hole region side. As a result, when centrifugal force or the like is applied to the microfluidic chip, the sample is easily introduced from the opening of the well, and the sample introduced into the well is less likely to flow out of the opening. Can be introduced.

[Application Example 4]
In application example 3,
The well may include a first well portion extending in a thickness direction of the substrate, and a second well portion continuing to the first well portion and extending in a direction parallel to the first surface.

  According to the microfluidic chip of this application example, since the well has the first well portion and the second well portion, the cross-sectional shape of the well has a shape similar to the alphabet “L”. Therefore, when a reagent or the like that acts on the liquid is disposed in the well, it is possible to further suppress the reagent from diffusing outside the well. Moreover, the outflow of the liquid in a well can be reduced more.

[Application Example 5]
In any one of Application Examples 1 to 4,
Of the inner wall of the well, a reagent may be applied to the inner wall of the well whose normal direction is parallel to the thickness direction of the substrate.

  According to the microfluidic chip of this application example, a reagent can act on a sample in a well. Further, in the microfluidic chip of this application example, since the position where the reagent is arranged is the inner wall of the well inner wall whose normal is parallel to the thickness direction of the substrate, the reagent is used when the microfluidic chip is manufactured. Easy to place. The normal of the inner wall refers to a straight line perpendicular to the inner wall.

[Application Example 6]
In any one of Application Examples 1 to 5,
A container mounted on the container mounting mechanism may be further provided.

  According to the microfluidic chip of this application example, the container for storing the sample is provided. Therefore, after preparing a sample in a container, the sample according to a required quantity can be supplied to a well, without performing dispensing operation with a pipette etc.

[Application Example 7]
One aspect of a specimen measurement method using the microfluidic chip according to the present invention is as follows.
The microfluidic chip is:
A through hole region having a first surface and a second surface opposite to the first surface and having a through hole connecting the first surface and the second surface in plan view and adjacent to the through hole region And a substrate provided with a well region in which a well having an opening is formed on the first surface side;
A cover laid on the first surface side and having a fixing region fixed to the substrate so as to surround the through-hole region and the well region in plan view;
A mechanism used when mounting the container on the second surface side, the container mounting mechanism having a structure for communicating the container and the through hole,
Attaching the container containing the specimen to the microfluidic chip;
Applying an inertial force to the container and the microfluidic chip to fill the well with the specimen;
Fixing the substrate and the cover at least around the opening;
Reacting the specimen filled in the well; and
Observing the specimen in which the reaction has been performed;
Have

  According to the sample measurement method of this application example, for example, a container containing a sample (sample) can be attached to the microfluidic chip so that the container and the through hole of the substrate can communicate with each other. There is no need to transfer the sample, and the sample can be easily accommodated in the microfluidic chip. Further, by applying an inertial force (for example, centrifugal force) to the container and the microfluidic chip, the sample accommodated in the container can be easily filled in the well through the gap between the substrate and the cover. Further, by fixing the substrate and the cover at least around the opening of the well, the sample filled in the well can be prevented from flowing out of the well. Therefore, the sample can be reliably dispensed in a short time, and the measurement efficiency of the sample can be improved.

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. 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. The schematic diagram of the cross section of the principal part of the microfluidic chip | tip 100 of embodiment. The schematic diagram of the cross section of the microfluidic chip | tip 100 and the container 200 of embodiment. The schematic diagram of the cross section of the microfluidic chip | tip 100 and the container 200 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. 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 schematic diagram of the cross section of the principal part of the board | substrate 10 of embodiment. The top view which shows typically the principal part of the board | substrate 10 of embodiment. The schematic diagram of the cross section of the principal part of the board | substrate 10 of embodiment. The figure which shows typically an example of the usage method of the microfluidic chip of embodiment.

  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.

  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. The shape of the substrate 10 is plate-like, and the height of the well region 14 on the first surface 11 of the substrate 10 and the height of the surface of the substrate surrounding the well region 14 coincide with the height of the opening surface of the well opening 16. If it does not specifically limit. 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.

1.2. Through-hole region and well region As shown in FIG. 1, the substrate 10 is provided with a through-hole region 13 and a well region 14 in plan view. The through-hole region 13 and the well region 14 are provided adjacent to each other. The through-hole region 13 and the well region 14 may be provided adjacent to each other. A plurality of through-hole regions 13 and well regions 14 may be provided. The through-hole region 13 and the well region 14 are provided inside a fixing region 21 of the cover 20 described later in plan view. The positions where the through-hole region 13 and the well region 14 are provided in the substrate 10 are 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 through-hole region 13 toward the well region 14. There is no particular limitation as long as it can be done. The planar shapes of the through-hole region 13 and the well region 14 are not particularly limited, and may be a rectangle, a circle, or the like.

1.3. Cover FIG. 3 is a plan view schematically showing the microfluidic chip 100 of the present embodiment. FIG. 4 is a schematic view of a cross section of the microfluidic chip 100 of the present embodiment. 4 corresponds to a cross section taken along line BB in FIG.

  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. 3, 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, a gap is formed between the substrate 10 and the cover 20. It has a degree of flexibility or elasticity. In addition, the cover 20 can close an opening 16a of a well 16 (described later) provided in the substrate 10 while being in contact with the substrate 10, thereby making 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 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 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. In such a method, since only pressurization is performed, no heat is generated when fixing, the temperature increase of the microfluidic chip 100 can be suppressed, and the influence of heat on the liquid or the like can be suppressed. can do. As a specific example of the cover 20 having such a surface 20a, trade name: LightCycler 480 Sealing Foil, model name: 04 729 757 001, manufactured by Roche Diagnostics, trade name: polyolefin microplate sealing tape, model name : 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 fixing region 21 is formed by the ultrasonic welding method, since the bonding force between the substrate 10 and the cover 20 due to the welding is relatively strong, liquid leakage from the well region 14 can be more reliably prevented. .

  The cover 20 has a fixing region 21 fixed to the substrate 10. The fixing region 21 is disposed so as to surround the through-hole region 13 and the well region 14 of the substrate 10 in plan view. That is, the through-hole region 13 and the well region 14 are disposed inside the fixing region 21 when viewed in a plan view. The 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 centrifugal force is applied to the microfluidic chip 100, the surface 20a and the first surface 11 can be separated from the region other than the fixing region 21 of the laid cover 20. In the 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 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 formed inside. For example, a bag-like structure may be formed. Accordingly, the through-hole region 13 and the well region 14 can be communicated with each other on the first surface 11 side of the substrate 10 between the substrate 10 and the cover 20, and the liquid can be circulated between the two regions.

  As already described, the inertial force is applied to the microfluidic chip 100 from the through hole region 13 side toward the well region 14 side. Therefore, the 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 fixing region 21 due to inertial force. Further, the fixing region 21 can be continuous in an annular shape so as to surround both the through-hole 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 fixing region 21 when the liquid is introduced into the gap between the cover 20 and the substrate 10. In addition, the adhering area | region 21 can have a part which is not continuous in the through-hole area | region 13 side. When such a portion is provided, the discontinuous portion can function as a vent hole when an internal pressure is generated in the gap between the cover 20 and the substrate 10. Further, when the fixing region 21 is continuous in an annular shape so as to surround the through-hole region 13 and the well region 14 of the substrate 10 in a plan view, it is illustrated on the cover 20 at an appropriate position on the through-hole region 13 side. By providing the hole, the hole can function as a vent hole when an internal pressure is generated in the gap between the cover 20 and the substrate 10.

  The fixing region 21 can be formed, for example, by applying an adhesive in the shape of the fixing region 21 on the first surface 11 of the substrate 10 and laying the cover 20. Further, for example, when the fixing region 21 has a property of exerting an adhesive force by applying pressure without exerting an adhesive force when the surface 20a of the cover 20 does not press the cover 20 against the substrate 10. A jig or the like corresponding to the shape of the fixing region 21 is prepared, and the cover 20 can be formed by applying pressure to the substrate 10 with the jig. Further, for example, the fixing region 21 can be formed by ultrasonic welding using a jig having a shape corresponding to the shape of the fixing region 21.

1.4. Through hole A through hole 15 that connects the first surface 11 and the second surface 12 is formed in the through hole region 13 of the substrate 10. A plurality of through holes 15 may be formed in the through hole region 13. The planar shape of the through hole 15 can be a circle, a rectangle, or the like. When the planar shape of the through-hole 15 is circular, for example, a general-purpose tube (such as a micro tube or a PCR tube) can be easily mounted so as to communicate with the through-hole 15. The through-hole 15 can circulate a liquid such as a sample in both directions on the first surface 11 side and the second surface 12 side. For example, the through-hole 15 can circulate the contents of the container 200 described later in both directions on the first surface 11 side and the second surface 12 side.

1.5. In the well region 14 of the well substrate 10, a well 16 having an opening 16a on the first surface 11 side of the substrate 10 is formed (see FIGS. 1 to 4). FIG. 5 is a schematic diagram showing an enlarged cross section near the well 16 of the microfluidic chip 100.

  A plurality of wells 16 can be formed in the well region 14. 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 therein. In addition, a liquid reaction such as a specimen can be performed in the well 16. For example, the contents of a container 200 to be described later can be introduced and held in the well 16, and the function as a reaction container can be achieved.

  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 the present embodiment, a case will be described in which the well 16 has a cylindrical shape (a circular shape in a plan view and a rectangular shape in a sectional view) as shown in FIGS. Other shapes that can be taken by the well 16 will be further described in the section of modifications.

  In the well 16, a reagent 30 for reaction or inspection can be arranged 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 16 a of the well 16 can be blocked by the cover 20. 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.

1.6. Container Mounting Mechanism The substrate 10 has a container mounting mechanism 17 for connecting the container 200 to the second surface 12 side. FIG. 6 is a schematic cross-sectional view of an example of the microfluidic chip 100 in which the container 200 is connected to the container mounting mechanism 17. FIG. 7 is a schematic cross-sectional view of another example of the microfluidic chip 100 in which the container 200 is connected to the container mounting mechanism 17.

  The container 200 is provided so that the contents can be moved to the through-hole 15 by the container mounting mechanism 17. That is, the container 200 is provided by the container mounting mechanism 17 so that the space inside the container 200 communicates with the space inside the through hole 15. The container 200 is not limited in any way, and examples thereof include general-purpose tubes (microtubes, PCR tubes, etc.). In addition, when the container 200 is a thing of the aspect used exclusively with respect to the microfluidic chip | tip 100, the microfluidic chip | tip 100 may also be called the microfluidic chip | tip 100 including the container 200. FIG.

  The container mounting mechanism 17 communicates the space inside the container 200 and the space inside the through hole 15 so that the contents of the container 200 do not leak to the outside from the connection portion between the container 200 and the through hole 15 (substrate 10). It has a structure to make it. The container mounting mechanism 17 has, for example, a structure that connects the substrate 10 and the container 200, and can fix the container 200 to the substrate 10. The container mounting mechanism 17 has any structure as long as it has a structure that allows the space inside the container 200 and the space inside the through hole 15 to communicate with each other and fix the container 200 to the substrate 10. Good. For example, in the example illustrated in FIG. 6, the container mounting mechanism 17 has a structure in which the container 200 is fitted (a structure in which the container 200 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 200 so that the container 200 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 200. In the example shown in FIG. 7, the container mounting mechanism 17 has a shape of a screw receiver (internal thread) into which the container 200 can be screwed. In this example, the container 200 is fixed by being screwed (as a male screw) into the container mounting mechanism 17 formed in the through hole 15 from the second surface 12 side of the substrate 10. In addition, although not shown, the container mounting mechanism 17 may be a holding spring (elastic body) that holds the container 200 against the substrate 10 or a hook (claw) that hooks a part of the container 200. There may be.

  Thus, the container mounting mechanism 17 can have an arbitrary structure according to the shape of the container 200 to be used. Then, the container 200 is fixed to the second surface 12 side of the substrate 10 by the container mounting mechanism 17 so that the contents can be supplied to the through hole 15.

1.7. Other Configurations The substrate 10 of the microfluidic chip 100 can have grooves 18. FIG. 8 is a plan view schematically showing the microfluidic chip 100 having the substrate 10 in which the grooves 18 are formed. FIG. 9 is a schematic cross-sectional view of the microfluidic chip 100 having the substrate 10 in which the grooves 18 are formed. FIG. 9 corresponds to a cross section taken along line CC in FIG.

  The groove 18 is provided on the second surface 12 side of the substrate 10. The inner wall of the groove 18 is not connected to the first surface 11. Therefore, the thickness of the substrate 10 is reduced at the portion of the substrate 10 where the groove 18 is formed. The groove 18 has a shape extending in a direction separating the through hole region 13 and the well region 14 between the through hole region 13 and the well region 14 in plan view. The cross section in the longitudinal direction (extending direction) of the groove 18 has a cross-sectional shape of an alphabet “V” in the illustrated example. The cross-sectional shape in the longitudinal direction of the groove 18 may be an alphabet “U” shape or other shapes. In the illustrated example, one groove 18 is provided, but a plurality of grooves 18 may be provided. Furthermore, although not shown, when a plurality of grooves 18 are provided, the grooves 18 may be aligned in the longitudinal direction and aligned in a line in the longitudinal direction (approximately in the form of a broken line in plan view) The grooves 18 may be aligned in the direction crossing the longitudinal direction (for example, double lines in a plan view) with the longitudinal direction aligned.

  One function of the groove 18 is to facilitate the separation of the microfluidic chip 100 between the through-hole region 13 and the well region 14. For example, when the microfluidic chip 100 is separated between the through-hole region 13 and the well region 14, when this is performed with a cutting tool such as scissors, cutting is facilitated if the groove 18 is formed. One of the functions of the groove 18 is to serve as a guide when the microfluidic chip 100 is bent with the second surface 12 of the substrate 10 facing outward. This facilitates bending the microfluidic chip 100 between the through-hole region 13 and the well region 14 with the second surface 12 of the substrate 10 facing outside. Such effects and the like based on the function of the groove 18 will be described again after the usage method of the microfluidic chip 100 is described.

1.8. 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. 10 is a diagram schematically illustrating a state in which an inertial force is applied in a state where the container 200 containing the specimen 210 is connected to the microfluidic chip 100. The state of the specimen 210 is liquid. 10 shows a state where an inertial force is applied to the microfluidic chip 100 and the container 200, and FIG. 6 shows a state where no inertial force is applied.

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 fixing region 21 of the cover 20 is assumed to be continuous in an annular shape so as to surround both the through-hole 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 outer shapes of the substrate 10 and the cover 20 are rectangular, and the long sides are about 7 . A 5 cm short side is illustrated as about 2.5 cm.

  First, the specimen 210 containing the target nucleic acid is prepared in the container 200. 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, nucleotides, and a coenzyme such as MgCl). 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 container 200 is mounted on the container mounting mechanism 17 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.

  Next, an inertial force from the through hole region 13 side to the well region 14 side is applied to the microfluidic chip 100 and the container 200. In this embodiment, the inertia force is generated by a centrifuge. The rotation axis R of the centrifuge is arranged on the side opposite to the well region 14 when viewed from the through hole region 13 of the microfluidic chip 100. When an inertial force (centrifugal force) is generated in the microfluidic chip 100 and the container 200 by operating the centrifuge, the specimen 210 in the container 200 moves away from the rotation axis R as shown in FIG. The acceleration G (arrow in the figure) is received in the direction toward the region 14. 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. Note that the size (thickness) of the gap at this time is not particularly limited, but FIG. 10 shows an example in which the size of the gap is very small. Then, the specimen 210 is introduced into the well 16 from the opening 16 a of the well 16 formed in the well region 14 of the substrate 10 by being replaced with air in the well 16 by inertia. 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. 10, the 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 the direction perpendicular to the rotation axis R of the centrifuge. Although 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 container 200 can move from the container 200 to the well 16 by centrifugal force. Such an arrangement can be set as appropriate depending on the shape of the container 200 and the arrangement of the wells 16 of the substrate 10.

  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, in a state where the sample 210 is not present in the gap between the substrate 10 and the cover 20, the well 16 is sealed by pressing the cover 20 against the substrate 10, and the sample 210 is sealed between the substrate 10 and the cover 20. In this state, the well 16 can be sealed while pushing the excess specimen 210 back to the through-hole region 13 side. Thereby, all the surfaces which the board | substrate 10 and the cover 20 contact can be adhere | attached.

  FIG. 11 is a diagram schematically showing how the well 16 of the microfluidic chip 100 is sealed by the roller 300. In the example of FIG. 11, the state in which the well 16 is sealed by the roller 300 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 container 200 is shown.

  In this way, 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, if necessary, the container 200 is separated from the microfluidic chip 100, and if necessary, the through-hole region 13 and the well region 14 are separated and introduced into a temperature control device such as a thermal cycler. The 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, so that the target nucleic acid can be quantified (real-time PCR). Yes, 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.

1.9. Modification 1.9.1. Deformation of Well Shape In the above section “1.5. 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 a schematic diagram showing an enlarged cross section of the substrate 10 having the well 46 according to the modification. FIG. 13 is an enlarged plan view schematically showing the substrate 10 having the modified well 46. FIG. 14 is a schematic diagram showing an enlarged cross section of the substrate 10 having the well 46 of another modification. A cross section along line DD in FIG. 13 corresponds to FIG. 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.

  As shown in FIG. 12, the modified well 46 includes a first well portion 46b and a second well portion 46c.

  The first well portion 46 b is a cavity extending in the thickness direction of the substrate 10. The first well portion 46 b is connected to the opening 46 a of the well 46. The first well portion 46 b can have any shape as long as it extends in the thickness direction of the substrate 10. For example, the first well portion 46b extends along the thickness direction of the substrate 10 in the example shown in FIG. 12, and extends in a direction having an inclination in the thickness direction in the example shown in FIG. The depth of the first well portion 46b, that is, the length in the thickness direction of the substrate 10 is not particularly limited as long as the thickness of the substrate 10 and the second well portion 46c described later can be formed. The first well portion 46b has a cylindrical shape in the examples of FIGS. 12 and 13, and in the example of FIG. 14 has a shape such that the column is inclined, but is not limited thereto, A prism, a truncated cone and a truncated pyramid, a shape in which they are inclined, or a shape obtained by combining these shapes can be used.

  The second well portion 46 c is a cavity that is continuous with the first well portion 46 b and extends in a direction parallel to the first surface 11 of the substrate 10. The direction in which the second well 46c extends is along the direction from the through hole region 13 side of the substrate 10 toward the well region 14 side. The position at which the second well portion 46c is connected to the first well portion 46b is not particularly limited, and is arbitrary as long as the second well portion 46c is not connected to the opening 46a. There is no particular limitation on the size of the second well portion 46c, that is, the length of the second well portion 46c. Further, the contour of the second well portion 46c in a plan view viewed from a direction perpendicular to the first surface 11 of the substrate 10 is not particularly limited. In the example of FIGS. 12 and 13, the second well portion 46 c is connected to the portion farthest from the opening 46 a of the first well portion 46 b and is a horizontal hole having an oval outline in plan view. That is, in the example of FIGS. 12 and 13, the cross-sectional shape of the well 46 has a shape similar to the alphabet “L”. In the example of FIGS. 12 and 13, the outline of the opening 46 a in the outline of the well 46 is in contact with the outline of the well 46 on the side close to the through-hole region 13 in plan view.

  Further, the second well portion 46 c can have an arbitrary shape as long as it extends in a direction parallel to the first surface 11 of the substrate 10. Although not shown, for example, the second well portion 46 c may extend in a direction having an inclination with respect to a direction parallel to the first surface 11 of the substrate 10.

  A method for forming the well 46 having the first well portion 46b and the second well portion 46c on the substrate 10 is not particularly limited. For example, as shown in FIGS. 12 and 14, the substrate 10 is divided in the thickness direction. After the substrate 10a, the substrate 10b, and the substrate 10c are formed in advance, the substrate 10a, the substrate 10b, and the substrate 10c can be stacked. That is, in the example of FIG. 12, the shape that becomes the first well portion 46b is formed on the substrate 10a, the shape that becomes the second well portion 46c is formed on the substrate 10b, the substrate 10c is prepared, and these are laminated. By fixing, the substrate 10 having the well 46 is produced. In this example, the substrate 10c can be omitted, the second well 46c having a bottom surface can be formed on the substrate 10b, and the substrate 10a and the substrate 10b can be stacked to form the substrate 10. Alternatively, the substrate 10a and the substrate 10b may be integrated to form a well shape, and the substrate 10c may be stacked thereon.

  The modified well 46 has the following characteristics because it includes the first well portion 46b and the second well portion 46c as described above.

  As described above, the area of the outline of the well 46 in plan view is larger than the area of the outline of the opening 46a in plan view. Therefore, the shape of the well 46 is narrowed at the opening 46a. Thereby, when a liquid is introduced into the well 46, the liquid is unlikely to flow backward. That is, it is difficult for the liquid introduced into the well 46 to be discharged from the well 46 to the outside again. In addition, when a reagent or the like that acts on the liquid is disposed in the well 46, it is possible to prevent the reagent from diffusing outside the well 46. In other words, since such a well 46 has a large depth (path length from the opening 46a), the well 46 has high independence. For example, the outflow of liquid in the well 46 can be suppressed. Thereby, when using the microfluidic chip | tip 100 for a test | inspection etc., the reliability of a test | inspection etc. can be improved more.

  In addition, since the second well portion 46 c extends from the first well portion 46 b to the well region 14 side from the through hole region 13 side of the substrate 10, the opening 46 a of the well 46 has an outline of the well 46. Of these, the substrate 10 is disposed on the side close to the through-hole region 13. Therefore, when a centrifugal force or the like is applied to the microfluidic chip 100, the liquid can be more efficiently introduced into the second well portion 46c of the well 46. Furthermore, when the liquid path between the opening 46a of the well 46 and the position farthest from the opening 46a of the well 46 is long and a reagent or the like that acts on the liquid is disposed in the well 46, the reagent is not contained in the well 46 Diffusion outside 46 can be further suppressed.

  Furthermore, when the contour of the opening 46a in the contour of the well 46 is in contact with the contour of the well 46 on the side close to the through-hole region 13 in plan view, the opening 46a of the well 46 has the through-hole. It will be arranged at the end near the region 13. Thereby, when a centrifugal force or the like is applied to the microfluidic chip 100, the liquid can be introduced into the well 46 more efficiently.

1.9.2. Modification of Reagent Arrangement In the above section “1.5. Well”, it has been stated that the reagent 30 for reaction or inspection can be arranged in the well 16 in advance. Similarly, in the well 46 described in the section “1.9.1. Deformation of well shape”, the reagent 30 for reaction or inspection can be arranged in the well 46 in advance. .

  Even when the reagent 30 is arranged in the well 46 according to the modified example, the position in the well 46 where the reagent 30 is arranged is farther from the opening 46a and the liquid introduced with the reagent is located outside the well 46. This is preferable because the possibility of outflow is reduced. That is, as shown in FIGS. 12 to 14, the reagent 30a, the reagent 30b, and the like are arranged near the tip of the second well portion 46c (a position far from the first well portion 46b), so that the reagent is placed in the well 46. It is possible to prevent the liquid from flowing out of the well 46 together with the introduced liquid.

  Further, the substrate 10 having the well 46 as shown in FIGS. 12 to 14 can be formed by stacking a plurality of substrates as described above. Therefore, in order to arrange the reagent in the well 46, a method of applying the ink to the inner wall surface of the well 46 by an ink jet method can be adopted. That is, for example, when the reagent is arranged at the position of reference numeral 30b, the substrate 10a is formed with a shape that becomes the first well portion 46b, the substrate 10b is formed with the shape that becomes the second well portion 46c, and the substrate 10a and After laminating and fixing the substrate 10b, a reagent is applied to the position of the reference numeral 30b from the lower side of the substrate 10b (the side of the substrate 10b opposite to the surface on which the substrate 10a is laminated) by a liquid ejecting head or the like. Finally, the reagent 30b can be disposed in the well 46 by stacking the substrate 10c. Alternatively, for example, when the reagent is arranged at the position of reference numeral 30a, after forming the substrate 10c, forming the shape that becomes the second well portion 46c on the substrate 10b, and laminating and fixing the substrate 10c and the substrate 10b (The substrate 10c and the substrate 10b may be integrated) From the upper side of the substrate 10b (the side of the substrate 10b opposite to the surface on which the substrate 10c is laminated) by a liquid ejecting head or the like, the reference numeral 30a Apply the reagent to the position. Then, the reagent 30a may be arranged in the well 46 by forming a shape to be the first well portion 46b on the substrate 10a and finally laminating the substrate 10a on the substrate 10b. In these cases, the inner wall of the well 46 to which the reagent can be applied is an inner wall whose normal is parallel to the thickness direction of the substrate 10. In this way, the reagent can be easily placed in the well 46 when the microfluidic chip 100 is manufactured.

  In FIGS. 12 to 14, the reagent 30a and the reagent 30b are both shown for illustration of the position where the reagent is placed. However, the reagent is placed at one of the positions. May be.

  Furthermore, in the example of FIGS. 12 and 13, when the reagent 30b is arranged, the position where the reagent 30b is arranged can suppress the outflow of the reagent 30b to the outside. The further away from the opening 46a, the better. Furthermore, it is more preferable that the sum of the depth of the first well portion 46 b and the distance between the first well portion 46 b and the reagent 30 b in plan view is larger than the diameter of the opening 46 a of the well 46. By adopting such an arrangement, the outflow of the reagent 30b to the outside can be further suppressed.

1.9.3. FIG. 15 is a diagram schematically showing a modified example of the usage method of the microfluidic chip 100. The microfluidic chip 100 can be bent between the through-hole region 13 and the well region 14 of the substrate 10 if the material of the substrate 10 and the cover 20 is appropriately selected. Such bending is described in “1.7. Other Configurations”. When the substrate 10 has the groove 18, it is easy to bend the second surface 12 along the groove 18. Stated.

  FIG. 15 is an example in which the substrate 10 has a groove 18, and the microfluidic chip 100 is bent between the through hole region 13 and the well region 14 of the substrate 10 with the second surface 12 facing outside. When the microfluidic chip 100 is bent in this way, the following new effects can be achieved in the step of applying an inertial force in the method of using the microfluidic chip.

  In the above embodiment (FIG. 10), when an inertial force from the through hole region 13 side to the well region 14 side is applied to the microfluidic chip 100 and the container 200, the direction of the acceleration G caused by the centrifugal force (in FIG. 10, The container 200 and the well 16 were both inclined at a right angle with respect to (indicated by an arrow G). In this case, the effect of the present invention can be achieved without any problem by adjusting the elasticity of the cover 20 or the applied centrifugal force. However, when the microfluidic chip 100 is bent, the specimen is more reliably placed in the well 16. 210 can be introduced.

  As shown in FIG. 15, when the microfluidic chip 100 is bent and installed in a centrifuge, the container 200 and wells are directed against the direction of acceleration G (indicated by arrow G in FIG. 15) generated by centrifugal force. 16 can be arranged so as to be inclined in different directions. In this example, the mouth of the container 200 is inclined toward the direction along the direction of the acceleration G, and the opening 16a of the well 16 is inclined toward the direction opposite to the direction of the acceleration. Therefore, while the centrifugal force is applied, the container 200 easily discharges the content (specimen 210) (easy to pour out), and the well 16 does not easily discharge the content (specimen 210 and the reagent 30) (acceptance). Cheap). Thereby, the liquid (specimen 210) can be introduced into the well 16 more efficiently.

  The angle at which the microfluidic chip 100 is bent is not particularly limited as long as the above effects can be obtained. Further, the centrifuge capable of applying a centrifugal force to the bent microfluidic chip 100 is not particularly limited. FIG. 15 shows a part of an example of a centrifuge rotor 400 capable of such a modification.

1.10. Operational Effects, etc. The microfluidic chip 100 of the present embodiment has at least the following operational effects. According to the microfluidic chip 100 of this embodiment, a small amount of liquid can be accurately dispensed. For example, when a liquid is manually dispensed using a pipette, a minute amount of liquid is difficult to dispense. Is possible. That is, according to the microfluidic chip 100 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 100 of the present embodiment, since the flow path communicating with the well is not formed, the well can be arranged at a 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 100 of this embodiment can be improved. Furthermore, the microfluidic chip 100 of the present embodiment can significantly reduce the man-hours and costs for dispensing work. In addition, according to the microfluidic chip 100 of the present embodiment, the dispensing process can be greatly reduced, and there is no need 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 100 is used for PCR has been described as an example. However, the use of the microfluidic chip 100 of the present embodiment is not limited, and examples thereof include viruses, bacteria, and proteins. It can also be used for inspection of low molecular weight to high molecular weight compounds, cells, particles, colloids, for example, allergic substances 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 100 has been described. However, the reagent 30 may not be arranged in the well depending on the examination contents.

2. Sample Measuring Method A method for measuring the sample 210 using the above-described microfluidic chip 100 and its usage will be described. As an example of a method for measuring the specimen 210 according to the present embodiment, a process of mounting the container 200 containing the specimen 210 on the microfluidic chip 100, and applying an inertial force to the container 200 and the microfluidic chip 100, the specimen A step of filling the well with 210, 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 state in which the specimen 210 exists in the well And the step of observing.

  For example, as described above, the step of attaching the container 200 containing the specimen 210 to the microfluidic chip 100 is performed by injecting the specimen 210 into the container 200 with a micropipette or the like and using the container mounting mechanism 17 of the microfluidic chip 100. This can be done by mounting the container 200.

  The step of applying the inertial force to the container 200 and 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. This step may be performed by bending the microfluidic chip 100.

  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 300. 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. Further, when the substrate 10 is formed by stacking a plurality of substrates as in the modification example, only the substrate 10c can be observed as a transparent material.

  According to the sample measuring method as described above, a predetermined amount of the sample 210 can be easily and accurately filled in the well. 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, 10a, 10b, 10c ... Board | substrate, 11 ... 1st surface, 12 ... 2nd surface, 13 ... Through-hole area | region, 14 ... Well area | region, 15 ... Through-hole, 16 ... Well, 16a ... Opening, 17 ... Container mounting Mechanism, 17a ... protrusion, 18 ... groove, 20 ... cover, 20a ... surface, 21 ... fixed area, 30, 30a, 30b ... reagent, 46 ... well, 46a ... opening, 46b ... first well portion, 46c ... second Well part, 100 ... microfluidic chip, 200 ... container, 210 ... specimen, 300 ... roller, 400 ... rotor, R ... rotation axis, G ... acceleration

Claims (7)

A first surface and a second surface opposite to the first surface, and in a plan view, a through hole region in which a through hole connecting the first surface and the second surface is formed, and adjacent to the through hole region And a substrate provided with a well region in which a well having an opening on the first surface side is formed,
A cover laid on the first surface side and having a fixing region fixed to the substrate so as to surround the through-hole region and the well region in plan view;
A mechanism used when mounting a container on the second surface side, the container mounting mechanism having a structure for communicating the container and the through hole;
A microfluidic chip. In claim 1,
The microfluidic chip, wherein the substrate has a groove extending in a direction separating the through hole region and the well region between the through hole region and the well region in a plan view on the second surface side. In claim 1 or claim 2,
The area of the outline of the well in plan view is larger than the area of the outline of the opening in plan view,
In plan view, the outline of the opening is in contact with the outline of the well on the side close to the through-hole region in the outline of the well. In claim 3,
The microfluidic chip, wherein the well includes a first well portion extending in a thickness direction of the substrate and a second well portion extending in a direction parallel to the first surface and continuing from the first well portion. In any one of Claims 1 thru | or 4,
A microfluidic chip in which a reagent is applied to the inner wall of the well whose normal line is parallel to the thickness direction of the substrate among the inner walls of the well. In any one of Claims 1 thru | or 5,
A microfluidic chip further comprising a container mounted on the container mounting mechanism. A method for measuring a specimen using a microfluidic chip,
The microfluidic chip is:
A first surface and a second surface opposite to the first surface, and in a plan view, a through hole region in which a through hole connecting the first surface and the second surface is formed, and adjacent to the through hole region And a substrate provided with a well region in which a well having an opening on the first surface side is formed,
A cover laid on the first surface side and having a fixing region fixed to the substrate so as to surround the through-hole region and the well region in plan view;
A mechanism used when mounting the container on the second surface side, the container mounting mechanism having a structure for communicating the container and the through hole,
Attaching the container containing the specimen to the microfluidic chip;
Applying an inertial force to the container and the microfluidic chip to fill the well with the specimen;
Fixing the substrate and the cover at least around the opening;
Reacting the specimen filled in the well; and
Observing the specimen in which the reaction has been performed;
A method for measuring a specimen.

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