Fluid supply device, microfluidic chip, test kit, and separation and recovery kit

Abstract

This fluid supply device 10 comprises: a storage part 2 for storing a fluid; and a drainage passage 3 for draining the fluid stored in the storage part 2. The storage part 2 comprises: a bottom surface 21; a ceiling surface 22 facing the bottom surface 21; and a partition wall 23 surrounding the bottom surface 21 and the ceiling surface 22 and separating the bottom surface 21 and the ceiling surface 22. The ceiling surface 22 is formed of a flexible material. At least one among the ceiling surface 22 and the bottom surface 21 is coated with an adhesive 10Ab for maintaining the contact between the bottom surface 21 and the ceiling surface 22 when the bottom surface 21 and the ceiling surface 22 come into contact with each other.

Description

Fluid supply device, microfluidic chip, test kit, and separation and recovery kit

[0001] The present invention relates to a fluid supply device for supplying a fluid, and in particular to a microfluidic chip provided with the same.

[0002] ELISA has traditionally been used to detect biomolecules (biomarkers). Currently, samples collected from subjects are sent to specialized testing institutions, where they are analyzed using ELISA. However, there is a time lag between the results being known and the subject receiving the report. Therefore, there is a need for an analytical method that can analyze the sample immediately upon receiving it from the subject and return the results immediately. Furthermore, if the subject could easily perform the analysis themselves, testing would be possible at any time, which would be even more desirable.

[0003] In general, ELISA requires sequential flow of multiple types of solutions, such as a sample, a washing solution, and a reaction substrate solution, into an observation area. In contrast, Non-Patent Document 1 discloses a microfluidic chip that allows subjects to perform ELISA analysis themselves. Specifically, multiple types of solutions are sealed in thin tubes within the microfluidic chip, and an absorbent pad is connected to the tubes. This initiates the movement of the solutions by negative pressure due to capillary action, allowing multiple types of solutions to be automatically and continuously flowed in sequence.

[0004] Parandakh et al, “3D-printed capillaric ELISA-on-a-chip with aliquoting”, Lab on a Chip, Issue 6, 2023, Internet <URL: https: / / pubs.rsc.org / en / content / articlelanding / 2023 / lc / d2lc00878e> Kim et al, “Large dropscale femtoliterlet array for digital counting of single biomolecules”, Lab on a Chip, Issue 12, 2012, 4986-4991Peoples et al, “Microfluidic Capillary System for Immunoaffinity Separations of C-Reactive Protein in Human Serum and Cerebrospinal Fluid”, Anal.Chem., 2008, 80, 3853-3858

[0005] However, with the technology of Non-Patent Document 1, once the absorbent pad is connected, the movement of the solutions cannot be stopped. Therefore, in order to ensure that each solution reacts in the observation area, it is necessary to prepare more solutions than the minimum necessary, which results in the problem of solution waste.

[0006] The present invention has been made in view of the above problems, and an object of the present invention is to provide a fluid supply device that can easily control the supply of fluid.

[0007] In order to solve the above problems, the present invention includes the following aspects. Item 1. A fluid supply device comprising: a storage section for storing a fluid; and an outflow path for outflowing the fluid stored in the storage section, wherein the storage section comprises: a bottom surface, a ceiling surface facing the bottom surface, and a partition wall surrounding the bottom surface and the ceiling surface to separate the bottom surface and the ceiling surface, wherein the ceiling surface is formed of a flexible material, and at least one of the ceiling surface and the bottom surface is coated with an adhesive for maintaining contact between the bottom surface and the ceiling surface when they come into contact. Item 2. The fluid supply device according to Item 1, wherein the shape of the storage section in a plan view is rectangular. Item 3. The fluid supply device according to Item 1 or 2, comprising a plurality of the storage sections. Item 4. The fluid supply device according to Item 3, wherein the plurality of storage sections are arranged in a row. Item 5. The fluid supply device according to Item 4, wherein the plurality of storage sections are arranged along the longitudinal direction of each storage section. Item 6. Item 7. A microfluidic chip comprising the fluid supply device according to any one of Items 1 to 5. Item 7. The microfluidic chip according to Item 6, further comprising: a connection path connected to each of the outlet paths of the plurality of storage units; and a contact object that sequentially brings each of the fluids stored in the storage units into contact, wherein the fluid that has passed through the connection path is supplied to a region where the contact object is provided. Item 8. A test kit comprising the microfluidic chip according to Item 7; and a hand roller having a roller part for pressing the storage units to cause the fluid to flow out of the storage units. Item 9. A separation and recovery kit comprising the microfluidic chip according to Item 7; and a hand roller having a roller part for pressing the storage units to cause the fluid to flow out of the storage units.

[0008] According to the present invention, it is possible to provide a fluid supply device that can reduce the waste of fluid.

[0009] 1 is a schematic diagram showing the configuration of a test kit according to Embodiment 1. FIG. 2 is a partial cross-sectional view of a microfluidic chip according to Embodiment 1. FIGS. 1(a) to 1(c) are plan views of an upper layer, a middle layer, and a lower layer, respectively, of a microfluidic chip according to Embodiment 1. FIG. 3 is a flowchart showing a procedure for using a test kit according to Embodiment 1. FIG. 4 is a plan view showing one state of a microfluidic chip according to Embodiment 1. FIGS. 1(a) and 1(b) are partial cross-sectional and plan views, respectively, showing one state of a microfluidic chip according to Embodiment 1. FIGS. 1(a) and 1(b) are partial cross-sectional and plan views, respectively, showing one state of a microfluidic chip according to Embodiment 1. FIG. 5 is a plan view showing one state of a microfluidic chip according to Embodiment 1. FIG. 6 is a plan view showing one state of a microfluidic chip according to Embodiment 1. FIG. 7 is a schematic diagram showing the configuration of a test kit according to Embodiment 2. FIGS. 1(a) to 1(c) are plan views of an upper layer, a middle layer, and a lower layer, respectively, of a microfluidic chip according to Embodiment 2. FIG. 8 is a flowchart showing a procedure for using a test kit according to Embodiment 2. FIG. 9 is a plan view showing one state of a microfluidic chip according to Embodiment 2. FIG. 10 is a plan view showing one state of a microfluidic chip according to Embodiment 2. FIG. 1 is a plan view showing one state of a microfluidic chip according to embodiment 2. FIG. 2 is a plan view showing one state of a microfluidic chip according to embodiment 2. FIG. 3 is a plan view showing one state of a microfluidic chip according to embodiment 2. FIG. 4 is a schematic view showing the configuration of a test kit according to embodiment 3. (a) to (c) are plan views of an upper layer, a middle layer, and a lower layer of a microfluidic chip according to embodiment 3, respectively. FIG. 5 is a flowchart showing the procedure for using a test kit according to embodiment 3. FIG. 6 is a plan view showing one state of a microfluidic chip according to embodiment 3. FIG. 7 is a plan view showing one state of a microfluidic chip according to embodiment 3. FIG. 8 is a plan view showing one state of a microfluidic chip according to embodiment 3. FIG. 9 is a plan view showing one state of a microfluidic chip according to embodiment 3. FIG. 10 is a plan view showing one state of a microfluidic chip according to embodiment 3. FIG. 11 is a plan view showing one state of a microfluidic chip according to embodiment 3. FIG. 12 is a plan view showing one state of a microfluidic chip according to embodiment 3. (a) and (b) are a partial plan view and a partial cross-sectional view, respectively, of a microfluidic chip according to a modified example.

[0010] Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. Note that the present invention is not limited to the following embodiments, and various modifications are possible without departing from the spirit of the present invention.

[0011] [Embodiment 1] (Overall Configuration) Fig. 1 is a schematic diagram showing the configuration of a test kit 1 according to embodiment 1 of the present invention. The test kit 1 mainly comprises a microfluidic chip 10, a dropper 20, and a hand roller 30. In this embodiment, the test kit 1 is used to detect PSA (prostate specific antigen), a prostate cancer marker protein, by digital ELISA assay.

[0012] The microfluidic chip 10 has a rectangular, flat plate shape in plan view, and mainly includes a storage section 2, an outflow channel 3, a connection channel 4, a capture / observation section 5, a waste liquid channel 6, a waste liquid storage section 7, inlet ports 8a-8c, and air holes 9a-9d. The storage section 2 and the outflow channel 3 constitute a fluid supply device as defined in the claims.

[0013] The storage unit 2 is a space for storing a fluid and functions as a reservoir for supplying the fluid to the capturing and observation unit 5. The shape of the storage unit 2 in a plan view is not particularly limited, but a rectangular shape is preferable. Furthermore, the width of each storage unit 2 is preferably slightly smaller than the width of the roller portion 31 of the hand roller 30, as will be described later.

[0014] The microfluidic chip 10 includes three storage units 2, specifically, a first storage unit 2a, a second storage unit 2b, and a third storage unit 2c. The first storage unit 2a stores a sample (blood, saliva, urine, swab of an affected area, etc.) collected from a subject, the second storage unit 2b stores a cleaning solution, and the third storage unit 2c stores a reaction substrate solution. The arrangement of the first storage unit 2a, the second storage unit 2b, and the third storage unit 2c in a plan view is not particularly limited, but it is preferable that they are arranged in a row. Hereinafter, when the first storage unit 2a, the second storage unit 2b, and the third storage unit 2c are not to be distinguished from one another, they will be simply referred to as storage units 2.

[0015] 2 is a partial cross-sectional view of the microfluidic chip 10. The storage section 2 has a bottom surface 21, a ceiling surface 22, and a partition wall 23. As will be described later, the microfluidic chip 10 has a three-layer structure consisting of an upper layer 10A, a middle layer 10B, and a lower layer 10C, with a portion of the lower layer 10C forming the bottom surface 21. The ceiling surface 22 faces the bottom surface 21, and a portion of the upper layer 10A forms the ceiling surface 22. The partition wall 23 is provided to surround the bottom surface 21 and the ceiling surface 22, separating them. A portion of the middle layer 10B forms the partition wall 23.

[0016] 1 is a channel for discharging the fluid stored in the storage unit 2. The microfluidic chip 10 has three outflow channels 3, specifically, an outflow channel 3a connected to the first storage unit 2a, an outflow channel 3b connected to the second storage unit 2b, and an outflow channel 3c connected to the third storage unit 2c. Each outflow channel 3 includes a water-repellent treated region W.

[0017] The connecting channel 4 is a channel connected to the outflow channels 3a, 3b, and 3c and extending to the capturing and observation unit 5. That is, the fluid that has passed through the outflow channels 3a, 3b, and 3c is supplied to the capturing and observation unit 5 through the connecting channel 4.

[0018] The capture / observation section 5 is a space for capturing a specimen, reacting it with a reaction substrate liquid, and observing the reaction state. The capture / observation section 5 is provided with an antibody-binding microchamber (contact target) M. In this embodiment, the capture / observation section 5 is arranged in a row together with the first storage section 2a, the second storage section 2b, and the third storage section 2c.

[0019] The waste liquid path 6 is a channel connected to the capturing and observation unit 5 and extending to the waste liquid reservoir 7. That is, the fluid flowing out from the capturing and observation unit 5 is supplied to the waste liquid reservoir 7 through the waste liquid path 6.

[0020] The waste liquid reservoir 7 is a space for storing the fluid that flows out from the capture / observation unit 5 as waste liquid.

[0021] The inlets 8a, 8b, and 8c are holes for injecting fluid into the first storage section 2a, the second storage section 2b, and the third storage section 2c, respectively. The air holes 9a, 9b, and 9c are holes for venting air to the outside when injecting fluid into the first storage section 2a, the second storage section 2b, and the third storage section 2c, respectively. The air hole 9d is a hole for venting air to the outside when waste liquid flows into the waste liquid storage section 7.

[0022] The dropper 20 is a tool for injecting fluid from the injection ports 8a, 8b, and 8c.

[0023] As will be described later, the hand roller 30 is used to press the roller part 31 against the upper side of the reservoir 2 to cause the fluid stored in the reservoir 2 to flow out.

[0024] The microfluidic chip 10 has a three-layer structure: Fig. 3(a) is a plan view of the upper layer 10A of the microfluidic chip 10, Fig. 3(b) is a plan view of the middle layer 10B of the microfluidic chip 10, and Fig. 3(c) is a plan view of the lower layer 10C of the microfluidic chip 10.

[0025] The upper layer 10A can be composed of a single-sided adhesive film. The upper layer 10A constitutes the ceiling surface 22 shown in FIG. 2, and the base material 10Aa of the upper layer 10A is formed of a flexible material. An adhesive 10Ab is formed on one side of the base material 10Aa. The adhesive 10Ab is preferably water-insoluble. In this embodiment, Scotch (registered trademark) Silent Tape 145RN (3M Corporation) is used as the upper layer 10A. The upper layer 10A has injection ports 8a-8c and air holes 9a-9d formed therein. These holes can be made, for example, with a biopsy trephine (Kai Industries Co., Ltd.).

[0026] The middle layer 10B can be made of a single-sided adhesive film or a double-sided adhesive film. Holes and slits are formed in the middle layer 10B to form the storage section 2, outflow channel 3, connection channel 4, capture / observation section 5, waste channel 6, and waste liquid storage section 7 shown in FIG. 1 . The area surrounding the hole for forming the storage section 2 in the middle layer 10B forms the partition wall 23 shown in FIG. 2 . In this embodiment, Kapton (registered trademark) double-sided tape 760H #25 (Teraoka Seisakusho Co., Ltd.) cut with a seal cutter CM300 (Brother Industries, Ltd.) is used as the middle layer 10B.

[0027] The lower layer 10C can be composed of a chip substrate. The lower layer 10C constitutes the bottom surface 21 shown in FIG. 2. The lower layer 10C also has an antibody-binding microchamber M and a water-repellent region W formed therein. The material of the lower layer 10C is preferably transparent for optical detection, and may be glass or hard plastic (polystyrene, cycloolefin polymer, PMMA, etc.). In this embodiment, a polystyrene flat slide is used as the lower layer 10C.

[0028] The antibody-binding microchambers M can also be formed by a method of microfabricating fluororesin using photolithography (Non-Patent Document 2) or by injection molding of plastic using a microfabricated mold. The size and shape of the antibody-binding microchambers M may be, for example, a large number of cylindrical depressions, each 3 μm in diameter and 3 μm deep, formed at a pitch of 7 μm. A capture antibody that specifically binds to the test substance is bound to the antibody-binding microchambers M in advance.

[0029] The water-repellent treated area W can be formed using, for example, A-PAP Pen (Daido Sangyo Co., Ltd.).

[0030] The microfluidic chip 10 is fabricated by bonding together the upper layer 10A, the middle layer 10B, and the lower layer 10C.

[0031] (Procedure for Using the Test Kit) FIG. 4 is a flowchart showing the procedure for using the test kit 1.

[0032] In step S1, a cleaning solution and a substrate solution are injected into the second storage section 2b and the third storage section 2c, respectively. The cleaning solution can be a 100 mM potassium phosphate buffer solution containing 0.1% (w / v) Tween 20, pH 7.5. The substrate solution can be a 100 mM phosphate buffer solution containing 1 mM fluorescein di-β-galactopyranoside (FDG) and 0.02% (w / v) Tween 20, pH 7.5. After the cleaning solution and the substrate solution are injected, the injection ports 8b and 8c and the air holes 9b and 9c are sealed. Because the water-repellent regions W are formed in the outflow channels 3b and 3c shown in FIG. 1, the cleaning solution and the substrate solution do not flow out of the second storage section 2b and the third storage section 2c.

[0033] Step S1 may be performed by the subject or the examiner, or may be performed when the microfluidic chip 10 is fabricated.

[0034] In step S2, a test reaction solution containing a sample collected from a subject is injected into the first storage section 2a. The sample may be blood, serum, interstitial fluid, urine, or an extract thereof. In this embodiment, the sample is mixed with an appropriate amount of a detection antibody solution and allowed to stand for a certain period of time to allow the detection antibody and PSA to fully bind to each other, resulting in a test reaction solution. The detection antibody is a monoclonal antibody capable of specifically binding to the test target substance, to which a detection enzyme is bound. This can be prepared using the method described in Non-Patent Document 2. The detection enzyme may be, for example, beta-galactosidase. The detection antibody solution may be, for example, a 100 mM, pH 7.5 phosphate buffer solution containing 4 pM of detection antibody and 0.5% (w / v) bovine serum albumin. The mixture ratio of the sample to the detection antibody solution may be, for example, 1:10. After injecting 50-100 μL of the test reaction solution, the injection port 8a and air hole 9a are sealed with a seal. Here, since the water-repellent region W is formed in the outflow channel 3a shown in FIG. 1, the test reaction liquid does not flow out from the first storage section 2a.

[0035] 5 is a plan view showing the state of the microfluidic chip 10 after step S2 is completed. For convenience, the upper layer 10A is omitted from the illustration in FIG. 5. The upper layer 10A is also omitted from the illustration in FIGS. 6(b), 7(b), and 8 to 10. Furthermore, to distinguish between the fluids stored in the first storage section 2a, the second storage section 2b, and the third storage section 2c, the fluids are color-coded for convenience, but the colors differ from the actual colors of the fluids.

[0036] The processes from step S2 onwards are performed by the subject or the examiner.

[0037] In step S3, the roller part 31 of the hand roller 30 is rotated while being pressed against the first storage section 2a. Specifically, as shown in FIGS. 6(a) and 6(b), the roller part 31 of the hand roller 30 is pressed against a portion of the upper layer 10A of the microfluidic chip 10 that corresponds to one end of the first storage section 2a. Then, the support 32 is moved leftward in the figure, and the roller part 31 is rotated while being pressed. This brings the bottom surface 21 and the ceiling surface 22 of the first storage section 2a into contact with each other.

[0038] Here, an adhesive 10Ab is applied to the ceiling surface 22 to maintain contact between the bottom surface 21 and the ceiling surface 22 when they come into contact. Therefore, as shown in Figure 7(a), the bottom surface 21 and the ceiling surface 22 remain attached to each other even after the roller unit 31 has passed, and the volume of the first storage unit 2a decreases as the roller unit 31 moves. Therefore, the test reaction liquid stored in the first storage unit 2a is gradually pushed out of the first storage unit 2a as the roller unit 31 moves, passes over the water-repellent region W, and flows out of the outflow channel 3a.

[0039] As a result, as shown in FIG. 7( b), the test reaction liquid flowing out of the outlet channel 3 a is supplied to the capture and observation section 5 through the connecting channel 4. As described above, the bottom surface 21 and the ceiling surface 22 of the first storage section 2 a remain bonded together, preventing backflow of the test reaction liquid. Furthermore, once the test reaction liquid reaches the capture and observation section 5, the movement of the roller unit 31 can be stopped as needed, allowing the test reaction liquid to remain in the capture and observation section 5 for a desired period of time. This allows the test reaction liquid to be sufficiently brought into contact with the antibody-binding microchambers M, ensuring that the specimen (a complex of a PSA molecule and a detection antibody) is reliably captured in the antibody-binding microchambers M. The adhesive may be applied only to the bottom surface 21, or to both the bottom surface 21 and the ceiling surface 22.

[0040] In step S4 shown in Fig. 4, the roller unit 31 is rotated while being pressed against the second storage unit 2b in the same manner as in step S3. This causes the washing solution stored in the second storage unit 2b to flow out from the outlet channel 3b. As a result, as shown in Fig. 8, the washing solution flowing out from the outlet channel 3b is supplied to the capture and observation unit 5 through the connection channel 4. As in step S3, the movement of the roller unit 31 can be interrupted as necessary when the washing solution reaches the capture and observation unit 5, allowing the washing solution to remain in the capture and observation unit 5 for a desired period of time. This allows the antibody-binding microchambers M to be properly washed.

[0041] In the first storage section 2a, the bottom surface 21 and the ceiling surface 22 are bonded together, so that the cleaning liquid does not flow into the first storage section 2a.

[0042] In step S5 shown in Fig. 4, the roller unit 31 is rotated while being pressed against the third storage unit 2c in the same manner as in steps S3 and S4. This causes the reaction substrate liquid stored in the third storage unit 2c to flow out from the outlet channel 3c. As a result, as shown in Fig. 9, the reaction substrate liquid flowing out from the outlet channel 3c is supplied to the capture / observation unit 5 through the connection channel 4. If a complex of a PSA molecule and a detection antibody is present in the antibody-binding microchamber M, a fluorescent substance is produced by the β-galactosidase enzyme bound to the detection antibody and accumulates in the antibody-binding microchamber M.

[0043] In steps S3 to S5, the first storage section 2a, the second storage section 2b, and the third storage section 2c are arranged in a row along the longitudinal direction of each storage section 2, so that the first storage section 2a, the second storage section 2b, and the third storage section 2c can be pressed sequentially by simply moving the roller section 31 in one direction.

[0044] In step S6, the roller unit 31 is rotated while being pressed against the capture and observation unit 5. This causes the reaction substrate liquid stored in the capture and observation unit 5 to flow out into the waste liquid channel 6, and the bottom and ceiling surfaces of the capture and observation unit 5 are pressed together. As a result, as shown in Figure 10, the inflow of fluid into the antibody-binding microchamber M is prevented, and the antibody-binding microchamber M becomes isolated from other fluids.

[0045] In step S7, the detection antibody is allowed to react with the reaction substrate after a certain time has passed, and the capture / observation unit 5 is observed by irradiating it with fluorescent light. If a fluorescent substance has accumulated in the antibody-bound microchambers M, the number of microchambers with increased fluorescence can be counted using a microscope or other method, and the concentration of the detection antibody can be determined by comparing this with a calibration curve.

[0046] (Summary) As described above, in this embodiment, the ceiling surface 22 of the storage unit 2 is formed from a flexible material, and an adhesive is applied to the ceiling surface 22. Then, as in steps S3 to S5, the roller unit 31 is pressed against the storage unit 2 and rotated, causing the fluid to flow out of the storage unit 2. At this time, the bottom surface 21 and the ceiling surface 22 of the storage unit 2 are bonded together by the adhesive, preventing backflow of the fluid. Furthermore, the movement of the fluid can be stopped by stopping the movement of the roller unit 31. Therefore, the supply of fluid from the storage unit 2 can be easily controlled.

[0047] Furthermore, in this embodiment, the fluid can be maintained in a state where it has reached the desired position, allowing the fluid (test reaction solution, washing solution, reaction substrate solution) to be in contact with the contact object (antibody-binding microchamber M) for a sufficient time. Therefore, it is only necessary to prepare an amount of fluid to be stored in each storage section 2 that can reach the area where the contact object is located (capture / observation section 5), thereby reducing fluid waste. Furthermore, the speed of the fluid can be controlled by controlling the movement speed of the roller section 31.

[0048] [Embodiment 2] (Overall Configuration) Figure 11 is a schematic diagram showing the configuration of a test kit 1' according to embodiment 2 of the present invention. The test kit 1' mainly comprises a microfluidic chip 10', a dropper 20, and a hand roller 30. In this embodiment, the test kit 1' is used to detect PSA by the ELISA method. Note that components having the same functions as those in embodiment 1 above are designated by the same reference numerals, and their description will be omitted.

[0049] The microfluidic chip 10' has a rectangular, flat plate shape in a plan view, and mainly includes three storage sections 2 (first storage section 2a, second storage section 2b, third storage section 2c), three outflow channels 3 (outflow channels 3a, 3b, 3c), a connection channel 4, a capture / observation section 5', a waste liquid channel 6, a waste liquid storage section 7, injection ports 8a to 8c, and air holes 9a to 9d.

[0050] The structures of the reservoir 2, outflow channel 3, and waste fluid storage section 7 are the same as those in the microfluidic chip 10 shown in FIG. 1 . Meanwhile, in the microfluidic chip 10′, a portion of the connection channel 4 forms a water-repellent section 4a having a rectangular shape in a planar view, and a portion of the waste fluid channel 6 forms a water-repellent section 6a having a rectangular shape in a planar view. Water-repellent regions W are formed on the bottom surfaces of the water-repellent sections 4a and 6a, sandwiching the flow path. Furthermore, similar to the reservoir 2 and the capture / observation section 5′, an adhesive is applied to the ceiling surface. Furthermore, the capture / observation section 5′ is coated with a capture antibody T, which specifically forms an image of the test substance, as a contact object. The binding method may be, for example, nonspecific adsorption due to hydrophobic interaction between the antibody molecules and the substrate surface. Furthermore, in embodiment 1, the first storage section 2a, the second storage section 2b, the third storage section 2c, and the capture / observation section 5 are arranged in a row, but in this embodiment, the first storage section 2a, the second storage section 2b, the third storage section 2c, the water-repellent section 4a, and the water-repellent section 6a are arranged in a row.

[0051] 12(a) to 12(c) are plan views of the upper layer 10A', middle layer 10B', and lower layer 10C' of the microfluidic chip 10', respectively. The materials of the upper layer 10A', middle layer 10B', and lower layer 10C' are the same as those of the upper layer 10A, middle layer 10B, and lower layer 10C shown in FIG.

[0052] (Procedure for Using the Test Kit) FIG. 13 is a flowchart showing the procedure for using the test kit 1'.

[0053] In step S11, a cleaning solution and a substrate reaction solution are poured into the second storage section 2b and the third storage section 2c, respectively. The cleaning solution and the substrate reaction solution are the same as those in embodiment 1. After the cleaning solution and the substrate reaction solution are poured, the injection ports 8b and 8c and the air holes 9b and 9c are sealed.

[0054] In step S12, an assay reaction liquid containing a specimen collected from a subject is injected into the first storage section 2a. The assay reaction liquid is the same as that in embodiment 1. After the assay reaction liquid is injected, the injection port 8a and the air hole 9a are sealed.

[0055] Fig. 14 is a plan view showing the state of the microfluidic chip 10' after step S12 is completed. For convenience, the upper layer 10A' is not shown in Fig. 14. The upper layer 10A' is also not shown in Figs. 15 to 18.

[0056] In step S13, the roller part 31 of the hand roller 30 is rotated while being pressed against the first storage section 2a. This brings the bottom surface 21 and the ceiling surface 22 of the first storage section 2a into contact with each other, causing the test reaction liquid to flow out of the outflow channel 3a. As a result, as shown in FIG. 15, the test reaction liquid flowing out of the outflow channel 3a is supplied to the capturing and observation section 5' through the connecting channel 4.

[0057] In step S14, roller unit 31 is rotated while being pressed against second storage unit 2b. This causes the cleaning liquid stored in second storage unit 2b to flow out from outlet channel 3b. As a result, as shown in FIG. 16, the cleaning liquid flowing out from outlet channel 3b is supplied to capture / observation unit 5' through connection channel 4.

[0058] In step S15, the roller unit 31 is rotated while being pressed against the third storage unit 2c. This causes the substrate reaction liquid stored in the third storage unit 2c to flow out from the outlet channel 3c. As a result, as shown in FIG. 17, the substrate reaction liquid flowing out from the outlet channel 3c is supplied to the capture / observation unit 5' through the connecting channel 4.

[0059] In step S16, roller unit 31 is rotated while being pressed against water-repellent sections 4a and 6a. This causes the bottom and ceiling surfaces of water-repellent sections 4a and 6a to be pressed together. As a result, as shown in Figure 18, the substrate reaction liquid is prevented from flowing out of capture / observation section 5' and fluids are prevented from flowing into capture / observation section 5', and capture / observation section 5' becomes isolated from other fluids.

[0060] In step S17, the detection antibody is allowed to react with the reaction substrate after a certain time has passed, and the capture / observation unit 5' is illuminated with fluorescent light and observed. If a fluorescent substance has accumulated in the capture antibody T, the number of microchambers where fluorescence has increased can be counted using a microscope or the like, and the concentration of the detection antibody can be determined by comparing this with a calibration curve.

[0061] [Embodiment 3] (Overall configuration) Figure 19 is a schematic diagram showing the configuration of a separation and recovery kit 1" according to embodiment 3 of the present invention. The separation and recovery kit 1" mainly comprises a microfluidic chip 10", a dropper 20, and a hand roller 30. The separation and recovery kit 1" is used to separate and recover a target substance from a sample containing many impurities, and in this embodiment, it is particularly used to separate and recover CEP (C-reactive protein), an inflammatory marker protein, from a sample such as blood or cerebrospinal fluid, which is a solution containing many impurities. Note that components having the same functions as those in the above-mentioned embodiments 1 and 2 are given the same reference numerals, and their description will be omitted.

[0062] The microfluidic chip 10" has a rectangular, flat plate shape in a plan view, and mainly includes three storage sections 2 (first storage section 2a, second storage section 2b, third storage section 2c), three outflow channels 3 (outflow channels 3a, 3b, 3c), a connection channel 4, a capture section 5", an extraction channel 5a, a waste liquid channel 6, a waste liquid storage section 7, injection ports 8a to 8c, air holes 9a to 9d, and a recovery port 9e.

[0063] The structures of the storage section 2, outflow channel 3, waste channel 6, and waste liquid storage section 7 are the same as those in the microfluidic chip 10 shown in Figure 1. On the other hand, in the microfluidic chip 10'', a capture section 5'' is provided instead of the capture / observation section 5, and the width of the capture section 5'' is approximately the same as that of the connection channel 4 and the waste liquid channel 6. A capture substance T' is formed on the entire bottom surface of the capture section 5'', and an extraction channel 5a and a waste liquid channel 6 are connected downstream of the capture section 5''. A water-repellent region W is formed in part of the extraction channel 5a, and the extraction channel 5a extends to the recovery port 9e. A water-repellent region W is also formed in part of the waste liquid channel 6.

[0064] 20(a) to 20(c) are plan views of an upper layer 10A", a middle layer 10B", and a lower layer 10C" of a microfluidic chip 10", respectively. The materials of the upper layer 10A", the middle layer 10B", and the lower layer 10C" are the same as those of the upper layer 10A, the middle layer 10B", and the lower layer 10C shown in FIG. 3, respectively.

[0065] The capture substance T' in the lower layer 10C" is an antibody that captures the target substance, CRP, bound to the surface of the lower layer 10C". The binding method may be nonspecific adsorption or the like. Furthermore, to increase the capture efficiency, a treatment may be performed such as forming a structure (pillars or the like) on the lower layer 10C" that increases the surface area of ​​the capture part 5".

[0066] (Procedure for Using Separation and Recovery Kit) FIG. 21 is a flowchart showing the procedure for using the separation and recovery kit 1 ″.

[0067] In step S21, a cleaning solution and an elution solution are injected into the second storage section 2b and the third storage section 2c, respectively. The cleaning solution may be Dulbecco's PBS buffer solution, which is commonly used in cell culture, etc. The elution solution may be a glycine hydrochloride buffer solution with a pH of 1.8, as described in Non-Patent Document 3. After the injection of the cleaning solution and the elution solution, the injection ports 8b and 8c and the air holes 9b and 9c are sealed with seals.

[0068] In step S22, a sample liquid such as blood or cerebrospinal fluid collected from a subject is diluted as necessary and injected into the first storage section 2a. After the sample liquid is injected, the injection port 8a and the air hole 9a are sealed. Note that in this embodiment, since detection is not the purpose, there is no need to mix the sample liquid with a detection antibody.

[0069] FIG. 22 is a plan view showing the state of the microfluidic chip 10" after step S22 is completed. For convenience, the upper layer 10A" is not shown in FIG. 22. Note that the upper layer 10A" is also not shown in FIGS. 23 to 27.

[0070] In step S23, roller part 31 of hand roller 30 is rotated while being pressed against first storage part 2a. This brings bottom surface 21 and ceiling surface 22 of first storage part 2a into contact, causing the sample liquid to flow out of outlet channel 3a. As a result, as shown in FIG. 23, the sample liquid flowing out of outlet channel 3a is supplied to capture part 5" through connection channel 4, and CRP in the sample liquid is captured by capture substance T'.

[0071] In step S24, roller unit 31 is rotated while being pressed against second storage unit 2b. This causes the cleaning solution stored in second storage unit 2b to flow out from outlet channel 3b. As a result, as shown in FIG. 24, the cleaning solution flowing out from outlet channel 3b is supplied to capture unit 5" through connection channel 4, and removes components of the sample liquid other than CRP from capture unit 5".

[0072] In step S25, the roller unit 31 is rotated while being pressed against the waste liquid path 6. This causes the bottom surface and the ceiling surface to be pressed together in the portion of the waste liquid path 6 where the water-repellent treated area W is formed. As a result, the waste liquid path 6 is blocked as shown in FIG.

[0073] In step S26, the roller unit 31 is rotated while being pressed against the third storage unit 2c. This causes the eluate stored in the third storage unit 2c to flow out from the outlet channel 3c. As a result, as shown in FIG. 26, the eluate flowing out from the outlet channel 3c is supplied to the capture unit 5" through the connecting channel 4, and CRP is eluted from the capture unit 5". Because the waste liquid channel 6 is blocked, the eluate containing CRP passes through the capture unit 5" and then flows toward the recovery port 9e shown in FIG. 19.

[0074] In step S27, the eluate is collected from the collection port 9e. As shown in FIG. 25, before the roller unit 31 reaches the third storage unit 2c, cleaning liquid remains in the capture unit 5" and its upper and lower flow paths. Therefore, cleaning liquid flows out from the collection port 9e until the roller unit 31 reaches partway through the third storage unit 2c, and then the eluate flows out as the roller unit 31 is moved further. Therefore, it is preferable to temporarily suspend the movement of the roller unit 31 once the outflow of cleaning liquid has finished, prepare to collect the eluate, and then resume the movement of the roller unit 31.

[0075] (Additional Notes) Although the embodiments of the present invention have been described above, the present invention is not limited to the above-described embodiments, and various modifications are possible without departing from the spirit of the present invention. For example, forms obtained by appropriately combining the technical means disclosed in the embodiments also fall within the technical scope of the present invention.

[0076] In the above embodiment, the fluid is caused to flow out of the reservoir 2 by pressing the roller 31 against the reservoir 2, but the reservoir 2 may be pressed with a finger or a spatula instead of the roller 31.

[0077] In the above embodiment, the fluid is injected into the storage unit 2 using the dropper 20, but as shown in Fig. 28, a pack P containing the fluid may be placed in the storage unit 2. It is preferable to process a part of the pack P so that it can easily burst when the roller unit 31 presses the storage unit 2, so that the fluid can easily flow out of the pack P.

[0078] The present invention can be applied not only to microfluidic chips but also to any device that supplies fluid.

[0079] DESCRIPTION OF SYMBOLS 1 Test kit 1' Test kit 1" Separation and recovery kit 2 Storage section 2a First storage section 2b Second storage section 2c Third storage section 3 Outlet channel 3a Outlet channel 3b Outlet channel 3c Outlet channel 4 Connection channel 4a Water-repellent section 5 Capture and observation section 5' Capture and observation section 5" Capture section 5a Extraction channel 6 Waste channel 6a Water-repellent section 7 Waste liquid storage section 8a Inlet 8b Inlet 8c Inlet 9a Air hole 9b Air hole 9c Air hole 9d Air hole 9e Recovery port 10 Microfluidic chip (fluid supply device) 10' Microfluidic chip (fluid supply device) 10" Microfluidic chip (fluid supply device) 10A Upper layer 10A' Upper layer 10A" Upper layer 10Aa Substrate 10Ab Adhesive 10B Middle layer 10B' Middle layer 10B" Middle layer 10C Lower layer 10C' Lower layer 10C" Lower layer 20 Dropper 21 Bottom surface 22 Ceiling surface 23 Partition wall 30 Hand roller 31 Roller portion 32 Support M Antibody-binding microchamber (contact object) T Capture antibody (contact object) T' Capture substance (contact object) P Pack W Water-repellent treated area

Claims

1. A fluid supply device comprising: a storage section for storing a fluid; and an outlet path for discharging the fluid stored in the storage section, wherein the storage section comprises: a bottom surface, a ceiling surface opposite the bottom surface, and a partition wall surrounding the bottom surface and the ceiling surface to separate the bottom surface and the ceiling surface, wherein the ceiling surface is formed from a flexible material, and at least one of the ceiling surface and the bottom surface is coated with an adhesive to maintain contact between the bottom surface and the ceiling surface when they come into contact.

2. The fluid supply device according to claim 1, wherein the storage portion has a rectangular shape in a plan view.

3. The fluid delivery device of claim 1, comprising a plurality of said reservoirs.

4. The fluid delivery device of claim 3, wherein a plurality of said reservoirs are arranged in a row.

5. The fluid supply device of claim 4, wherein a plurality of said reservoirs are arranged along the longitudinal direction of each reservoir.

6. A microfluidic chip equipped with a fluid supply device according to any one of claims 1 to 5.

7. The microfluidic chip according to claim 6, further comprising: a connection path connected to each of the outlet paths of the plurality of storage sections; and a contact object that sequentially contacts each of the fluids stored in the storage sections, wherein the fluid that has passed through the connection path is supplied to an area where the contact object is provided.

8. A test kit comprising: the microfluidic chip according to claim 7; and a hand roller having a roller portion for pressing the reservoir to cause the fluid to flow out of the reservoir.

9. A separation and recovery kit comprising: the microfluidic chip according to claim 7; and a hand roller having a roller portion for pressing the reservoir to cause the fluid to flow out of the reservoir.

Images