JP2009281869A - Microchip - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microchip capable of separately mixing a plurality of kinds of liquid reagents with a specimen and further enhanced in the precision of inspection and analysis in the microchip more than before. <P>SOLUTION: A fluid circuit has a first mixing part for mixing the specimen with a first reaction liquid to produce a primary mixed liquid, a second mixing part for mixing the primary mixed liquid with a second reaction liquid to produce a secondary mixed liquid, and a third mixing part for mixing the secondary mixed liquid with a third reaction liquid to produce a tertiary mixed liquid. The first mixing part includes a residue separating part for separating the residue at least in one of the first, second and third mixed liquids, and the residue separating part is constituted so that a residue is separated by applying a centrifugal force to the microchip in a first direction and only a supernatant liquid is allowed to flow out of the residue separating part by applying the centrifugal force to the microchip in a second direction. <P>COPYRIGHT: (C)2010,JPO&amp;INPIT

Description

  The present invention relates to a microchip useful as a micro-TAS (Micro Total Analysis System) suitably used for biochemical tests such as DNA, proteins, cells, immunity and blood, chemical synthesis, and environmental analysis. Specifically, the present invention relates to a microchip that performs inspection and analysis by optical measurement.

  In recent years, the importance of detecting, detecting or quantifying biological substances such as DNA (Deoxyribo Nucleic Acid), enzymes, antigens, antibodies, proteins, viruses and cells, and chemical substances in fields such as medicine, health, food, and drug discovery There have been proposed various biochips and microchemical chips (hereinafter collectively referred to as microchips) that can be easily measured. Microchips can perform a series of experiments and analysis operations performed in the laboratory within a chip of several centimeters to 10 centimeters and a thickness of several millimeters to several centimeters. However, it has many advantages such as low cost, high reaction speed, high-throughput testing, and the ability to obtain test results immediately at the sample collection site.

  The microchip has a fluid circuit therein, and the fluid circuit holds, for example, a liquid reagent for mixing or reacting with a specimen (for example, blood) or holding a liquid reagent for processing the specimen. Appropriately connect these units to the holding unit, the measuring unit for measuring the sample and the liquid reagent, the mixing unit for mixing the sample and the liquid reagent, and the detection unit for analyzing and / or inspecting the mixed liquid. It is mainly composed of a fine fluid circuit (for example, a width of about several hundred μm). The microchip is typically used by being mounted on a device (centrifuge) capable of applying a centrifugal force thereto. By applying a centrifugal force in an appropriate direction to the microchip, the sample and liquid reagent can be weighed, mixed, introduced into the detection unit, and the like. Inspecting / analyzing the liquid mixture introduced into the detection unit (for example, detecting a specific component in the liquid mixture), for example, irradiating the detection unit containing the liquid mixture with detection light and measuring the transmittance (For example, refer to Japanese Patent Laid-Open No. 2006-300741 (Patent Document 1)).

At present, various patterns of fluid circuits have been developed for the microchip according to the type of specimen and the measurement substance in the specimen.
JP 2006-300741 A

  As described above, various patterns of microchips have been developed according to the measurement substance in the specimen.

  An object of the present invention is to provide a microchip capable of separately mixing a plurality of types of liquid reagents with the sample and further improving the accuracy of inspection and analysis in the microchip.

  In order to solve the above problems, the present inventor has determined that the type and number of liquid reagents to be mixed with a specimen, the order in which the specimen and a plurality of kinds of liquid reagents are mixed, and the specimen and the liquid reagent are mixed. In this way, attention was focused on removing residues (solid content) unnecessary for analysis and the like on the microchip.

  The present inventors diligently studied a pattern in which a plurality of types of liquid reagents and specimens as described above are mixed and a residue unnecessary for analysis or the like on a microchip is removed. In addition, the microchip fluid circuit includes a pattern for mixing a plurality of types of liquid reagents and specimens and a residue separation unit, thereby further improving the accuracy of inspection and analysis in the microchip. I found out that I can do it. That is, the present invention is as follows.

  In the present invention, at least a first substrate having a groove provided on the substrate surface and a second substrate are bonded together, and the groove and the first substrate side surface of the second substrate are formed. A microchip having a fluid circuit therein, the microchip having a sample introduction port for introducing a sample into the fluid circuit, which penetrates from one surface to the fluid circuit. A first mixing section for mixing the first reaction liquid to produce a primary mixed liquid, and a second mixing section for mixing the primary mixed liquid and the second reaction liquid to produce a secondary mixed liquid. A first mixing unit, a second mixing unit, and a third mixing unit; and a third mixing unit for mixing the secondary mixed solution and the third reaction solution to produce a tertiary mixed solution. At least one of the mixing parts in at least one of the primary mixed solution, the secondary mixed solution and the tertiary mixed solution A residue separator for separating the residue, and the residue separator separates the residue by applying centrifugal force to the microchip in the first direction, and applies centrifugal force in the second direction. The present invention relates to a microchip that allows only the supernatant liquid to flow out from the residue separator.

  In the microchip of the present invention, the internal angle θ formed by the first direction and the second direction is preferably 30 to 60 degrees (where θ is an internal angle of less than 90 degrees).

  In the microchip of the present invention, the residue separation portion is formed of a substantially V-shaped wall surface having an opening and a squeezed tail portion, and one side of the substantially V-shaped wall surface extends in the first direction. When a centrifugal force is applied, the length is such that at least one supernatant and residue of the primary mixed solution, the secondary mixed solution, and the tertiary mixed solution do not flow out from the residue separation unit, and is centrifuged in the second direction. It is preferable to have a length that allows only the supernatant liquid to flow out of the residue separator by applying force.

  Moreover, it is preferable that the microchip of the present invention further includes a measuring unit for measuring only the supernatant liquid flowing out from the residue separating unit.

  In this specification, a liquid reagent is also referred to as a reaction solution.

  The present invention can provide a microchip in which a plurality of types of liquid reagents can be separately mixed with the specimen, and the accuracy of inspection and analysis in the microchip is further improved as compared with the conventional technique.

  In one preferred embodiment, the microchip of the present invention is formed by bonding a first substrate having a groove on the substrate surface and a second substrate. Such a microchip has a hollow portion formed by a groove provided on the surface of the first substrate and a first substrate side surface (bonding surface of the second substrate) of the second substrate. A fluid circuit is provided.

  In another preferred embodiment, the microchip of the present invention is formed by bonding a third substrate, a first substrate having grooves provided on both surfaces of the substrate, and a second substrate in this order. . Such a microchip composed of three substrates is formed from a first substrate side surface (bonding surface of the third substrate) of the third substrate and a groove provided on the third substrate side surface of the first substrate. 1st fluid circuit comprised, 1st board | substrate side surface (bonding surface of 2nd board | substrate) in a 2nd board | substrate, and the groove | channel provided in the 2nd board | substrate side surface in 1st board | substrate A second fluid circuit and a two-layer fluid circuit. Here, two layers means that fluid circuits are provided at two different positions in the thickness direction of the microchip. The 1st fluid circuit and the 2nd fluid circuit may be connected by one or two or more penetration holes penetrated in the thickness direction formed in the 1st substrate.

  The material of each substrate constituting the microchip of the present invention is not particularly limited. For example, polyethylene terephthalate (PET), polybutylene terephthalate (PBT), polymethyl methacrylate (PMMA), polycarbonate (PC), polystyrene (PS) , Polypropylene (PP), polyethylene (PE), polyethylene naphthalate (PEN), polyarylate resin (PAR), acrylonitrile / butadiene / styrene resin (ABS), vinyl chloride resin (PVC), polymethylpentene resin (PMP), Organic materials such as polybutadiene resin (PBD), biodegradable polymer (BP), cycloolefin polymer (COP), and polydimethylsiloxane (PDMS); inorganic materials such as silicon, glass, and quartz can be used.

  The method for forming the grooves constituting the fluid circuit on the surface of the first substrate is not particularly limited, and examples thereof include an injection molding method using a mold having a transfer structure, an imprint method, and the like. In the case of forming a substrate using an inorganic material, an etching method or the like can be used.

  In the microchip of the present invention, the fluid circuit (the first fluid circuit and the second fluid circuit when the fluid circuit having two layers is provided) is suitable for the fluid (particularly, liquid) in the fluid circuit. In order to be able to perform various processes, various parts are provided at appropriate positions in the fluid circuit, and these parts are appropriately connected through fine flow paths.

  In the microchip of the present invention, the fluid circuit includes, as one of the above-described parts, a centrifuge for separating a sample introduced into the microchip into a target component and a non-target component by centrifugation. Here, “specimen” in this specification means a substance (for example, blood) to be examined and analyzed that is introduced into the fluid circuit. In the present specification, the “target component” means a specific component in a specimen that is prepared in a microchip and that constitutes a sample to be used for examination / analysis. Is a specific component in the sample to be mixed or reacted with the liquid reagent previously held in the container. When the specimen is blood, for example, the non-target component can include a plasma component, and the target component can include a blood cell component.

  The microchip of the present invention has a sample introduction port, and the sample is introduced into the fluid circuit through the sample introduction port. The sample introduction port can be configured as a through-hole penetrating from one surface of the microchip to the fluid circuit. Specifically, when the microchip includes a first substrate and a second substrate each having a groove on the substrate surface, the sample introduction port includes a through-hole penetrating the first substrate in the thickness direction. can do. When the microchip is formed by laminating the third substrate, the first substrate having grooves provided on both surfaces of the substrate, and the second substrate in this order, the sample introduction port is connected to the third substrate. The through hole penetrating through the substrate (or the first substrate) in the thickness direction can be used. The centrifuge unit is connected to the sample introduction port via a flow path, and the sample injected from the sample introduction port can be introduced into the centrifuge unit.

  In the present invention, the fluid circuit may include a part other than the centrifugal separator, such as a liquid reagent holding part for holding a liquid reagent, a target component measuring part for measuring a target component, A liquid reagent metering unit for measuring a liquid reagent, a mixing unit for mixing the target component and the liquid reagent, and an inspection / analysis of the obtained liquid mixture (sample to be used for the inspection / analysis described above) (for example, And a detection unit for detecting or quantifying a specific component in the mixed solution. The microchip of the present invention may have all of these exemplified portions, or may not have any one or more. Moreover, you may have site | parts other than these illustrated site | parts.

  The “liquid reagent” is a substance (reagent) for mixing or reacting with the target component, and is usually built in the liquid reagent holding part of the fluid circuit in advance before using the microchip. The liquid mixture finally obtained by mixing the target component and the liquid reagent is not particularly limited. For example, light that is transmitted through irradiation of light to a portion (for example, a detection unit) in which the liquid mixture is accommodated. It is used for optical measurement such as a method for detecting the intensity (transmittance) of the light and inspected and analyzed.

  Fluid processing in the fluid circuit such as extraction of the target component from the specimen, measurement of the target component and / or liquid reagent, mixing of the target component and liquid reagent, introduction of the obtained liquid mixture into the detection unit, etc. This can be performed by sequentially applying a centrifugal force in an appropriate direction to the microchip. Application of centrifugal force to the microchip is typically performed by placing the microchip on a device (centrifuge) that can apply centrifugal force thereto. Hereinafter, the microchip of the present invention will be described in more detail with reference to embodiments.

  Hereinafter, a preferred embodiment of the present invention will be described with reference to the drawings. In the following drawings, the same or corresponding parts are denoted by the same reference numerals, and description thereof will not be repeated. In addition, dimensional relationships such as length, size, and width in the drawings are changed as appropriate for clarity and simplification of the drawings, and do not represent actual dimensions.

  FIG. 1A is a schematic top view showing a preferred example of the microchip of the present invention. FIG. 1B is a schematic top view in which a part of FIG. 1A is enlarged.

  Hereinafter, description will be given based on FIG. 1A and FIG. First, the structure of the microchip 100 of this embodiment will be described. The microchip 100 of the present embodiment is manufactured by bonding a second substrate to a groove forming side surface of a first substrate having a groove forming a fluid circuit on the surface and a through hole penetrating in the thickness direction. Yes. FIG. 1A is a top view showing the first substrate side surface of the microchip 100. Actually, the grooves constituting the fluid circuit are formed on the surface of the first substrate opposite to the surface shown in FIG. 1 (bonding surface with the second substrate). The groove pattern is shown as a solid line so that it can be clearly understood.

  In the present embodiment, the microchip 100 has a longitudinal direction of 48 to 52 mm and a lateral direction of 38 to 42 mm. Further, the microchip 100 is held in a disk-shaped centrifuge having a diameter of 45 to 50 cm and applied with a centrifugal force.

  The microchip 100 according to the present embodiment has a sample introduction port 30 for introducing a sample into the fluid circuit that penetrates from one surface to the fluid circuit. Then, the fluid circuit mixes the first mixing unit 41 for producing the primary mixed liquid by mixing the specimen and the first reaction liquid, the first mixed liquid and the second reaction liquid, A second mixing unit 42 for manufacturing the secondary mixed solution and a third mixing unit 43 for mixing the secondary mixed solution and the third reaction solution to manufacture a tertiary mixed solution are provided.

  Moreover, in this embodiment, the 1st mixing part 41 is provided with the residue separation part 1 for isolate | separating the residue in this primary liquid mixture. However, in the present invention, the residue separating unit 1 may be included in at least one of the first mixing unit 41, the second mixing unit 42, and the third mixing unit 43. It is only necessary to be provided for separating a residue in at least one of the secondary mixed liquid and the tertiary mixed liquid.

  And in the residue separation part 1, the residue in this primary liquid mixture can be isolate | separated by applying a centrifugal force to the microchip 100 in the 1st direction. In the present embodiment, the first direction is the right direction with respect to FIG. 1A and the longitudinal direction of the microchip 100. Then, by applying a centrifugal force to the microchip 100 in the second direction, only the supernatant liquid in the primary mixed liquid can be caused to flow out from the residue separating unit 1. In the present embodiment, the supernatant liquid moves to the measuring unit 40, and the weighed amount of the supernatant liquid moves to the second mixing unit 42.

  The microchip 100 sufficiently mixes the specimen with each of the three types of the first reaction liquid, the second reaction liquid, and the third reaction liquid, and is detrimental to the inspection and analysis in the microchip 100. It incorporates a structure that can remove residues.

  As shown in FIG. 1B, the residue separating portion 1 is formed of a substantially V-shaped wall surface including an opening and a swelled tail. The substantially V-shaped wall surface is formed of a first side 2 on the substantially V-shaped wall surface and a second side 3 on the substantially V-shaped wall surface.

  The first side 2 in the substantially V-shape has a length that prevents the supernatant and residue of the primary mixed solution from flowing out of the residue separating unit 1 when centrifugal force is applied in the first direction, and It has a length that allows only the supernatant liquid to flow out of the residue separator 1 by applying centrifugal force in the second direction.

  In the present embodiment, the left direction toward FIG. 1B is the first direction, and the direction 45 degrees obliquely above the first direction (upper left direction) is the second direction. At this time, θ is preferably 30 to 60 degrees, and particularly preferably 45 to 60 degrees. This is because when the centrifugal force is applied in the first direction, the residue and the supernatant can be further away. Further, in the present embodiment, it is necessary to appropriately set the liquid level of the supernatant liquid of the primary mixed solution to the left of the line that descends perpendicularly to the first direction from the first side 2 in the substantially V shape. There is.

  In the present embodiment, it is preferable that the second side 3 in the substantially V shape is refracted inward toward the first mixing portion 41. Specifically, in this embodiment, θa is preferably 90 to 180 degrees, and particularly preferably 120 to 150 degrees. This is because when the centrifugal force is applied in the first direction, the residue can be more easily moved in the application direction.

  In the present embodiment, the first direction is the left direction of the microchip as described above. However, in the present invention, the first direction is not particularly limited. However, the interior angle θ formed by the first direction for removing the residue of the primary mixed solution in the residue separating unit 1 and the second direction for allowing only the supernatant liquid in the primary mixed solution to flow out of the residue separating unit 1 is formed. Is preferably 30 to 60 degrees, particularly preferably 40 to 50 degrees (however, θ is an internal angle of less than 90 degrees). This is because a part of the residue is prevented from flowing out to the measuring unit 40.

  2 to 15 are schematic views showing each step of the operation in the microchip 100 shown in FIG. Hereinafter, a description will be given with reference to FIGS. In addition, in FIG. 11 and FIG. 12, the operation | movement in the residue separation part in this embodiment is demonstrated. The white arrows in FIGS. 2 to 15 indicate the direction of the centrifugal force received by the central portion of the microchip 100, and the directions will be described below from the left to the right and up and down in the drawings.

  In this embodiment, blood (whole blood) is described as an example of the sample. However, in the present invention, the sample is not particularly limited, and is composed of two or more types of solutions having different specific gravities. It can be applied as appropriate.

  First, as shown in FIG. 2, when a centrifugal force is applied to the microchip 100 in the left direction as viewed in FIG. 2, the blood 10 is introduced into the sample introduction unit 31. In addition, the first holding part 32 holds the first reaction liquid 11, the second holding part 34 holds the second reaction liquid 12, and the third holding part 33 holds the second reaction liquid 12. Holds the third reaction solution 13.

  Next, as shown in FIG. 3, when centrifugal force is applied to the microchip 100 in the downward direction toward FIG. 3, the blood 10 is introduced into the centrifugal separator 35, and the blood cell component 10 b and plasma with different specific gravity are further introduced. Separated into component 10a.

  Next, as shown in FIG. 4, when a centrifugal force is applied to the microchip 100 in the left direction toward FIG. 4, the plasma component 10 a flows out from the centrifugal separator 35. Then, only the blood cell component 10b is held in the centrifugal separator 35.

  Next, as shown in FIG. 5, by applying centrifugal force to the microchip 100 in the downward direction toward FIG. 5, the blood cell component 10b, the first reaction liquid 11, the second reaction liquid 12, and the first The liquid level of the reaction liquid 13 is adjusted. This is done for the reason that the plasma component 10a is introduced into the waste liquid space and the blood cell component 10b is sent without any remaining liquid in the next step.

  Next, as shown in FIG. 6, by applying a centrifugal force to the microchip 100 in the right direction toward FIG. 6, the blood cell component 10 b moves from the centrifuge unit 35 to the measuring unit 50, and the measuring unit At 50, only a predetermined amount is retained. Further, the first reaction liquid 11 moves from the first holding unit 32 to the liquid reagent measuring unit 36, and only a predetermined amount is held in the liquid reagent measuring unit 36. Then, the second reaction liquid 12 moves from the second holding unit 34 to the preliminary chamber 51. Further, the third reaction liquid 13 moves from the third holding part 33 to the preliminary chamber 52. These preliminary chambers are portions that temporarily hold the second reaction liquid 12 and the third reaction liquid 13 before being measured by the liquid reagent measuring section.

  Next, as shown in FIG. 7, by applying centrifugal force to the microchip 100 in the downward direction toward FIG. 7, the weighed blood cell component 10b and the weighed first reaction liquid 11 are The primary mixed liquid 14 is manufactured by mixing. Then, the third reaction solution 13 moves from the preliminary chamber 52 to the liquid reagent measuring unit 37, and only a predetermined amount is held in the liquid reagent measuring unit 37.

  Next, as illustrated in FIG. 8, the primary mixed solution 14 is sufficiently mixed in the first mixing unit 41 by applying a centrifugal force to the microchip 100 in the right direction toward FIG. 8. In addition, the third reaction solution 13 that has been reduced in weight by the liquid reagent measuring unit 37 moves to the third mixing unit 43 and is held there.

  Next, as shown in FIG. 9, by applying a centrifugal force to the microchip 100 in the left direction toward FIG. 9, the second reaction solution 12 moves from the preliminary chamber 51 to the liquid reagent measuring unit 39. Only a predetermined amount is held in the liquid reagent measuring unit 39. Further, the primary mixed solution 14 is more sufficiently mixed in the first mixing unit 41. In addition, the blood cell component 10b that is not held in the measuring unit 50 moves to the error sensing unit 60, and the measuring unit 50 can check whether or not the predetermined amount of blood cell component 10b has been measured.

  Next, as shown in FIG. 10, the primary mixed solution 14 is further sufficiently mixed in the first mixing unit 41 by applying a centrifugal force to the microchip 100 in a downward direction toward FIG. 10. Further, in the lower left of the microchip 100, the waste liquid 15 accumulates.

  Next, as shown in FIG. 11, by applying a centrifugal force in the left direction toward FIG. 11, the residue 14 a of the primary mixture 14 and the supernatant 14 b of the primary mixture 14 in the residue separator 1. And separated. At this time, the residue 14a of the primary mixed liquid 14 that is a solid content is retained on the swelled tail side of the substantially V-shaped wall surface that forms the residue separation unit 1. Since one side of the substantially V-shaped wall has such a length that the supernatant liquid 14b of the primary mixed liquid does not flow out of the residue separating section 1, the supernatant liquid is added to the measuring section 40 described later in this stage. It can suppress that 14b leaks out.

  Next, as shown in FIG. 12, only the supernatant 14 b of the primary mixed solution 14 is moved to the measuring unit 40 by applying a centrifugal force obliquely upward 45 degrees toward the left in FIG. 12. At this time, one side of the substantially V-shaped wall has a length that prevents the supernatant liquid 14b of the primary mixed solution from flowing out of the residue separating unit 1, and therefore the supernatant is added to the measuring unit 40 described later in this stage. The liquid 14b does not leak out. Moreover, it is preferable that the other side of the substantially V-shaped wall surface is not linear but is refracted inward to the first mixing portion 41 side. This is because only the supernatant liquid 14b can be smoothly moved to the measuring unit 40 by being refracted inward.

  Next, as shown in FIG. 13, by applying a centrifugal force in the left direction toward FIG.

  Next, as shown in FIG. 14, by applying a centrifugal force in the downward direction toward FIG. 14, the supernatant liquid 14 b moves to the second mixing unit 42, and the second reaction liquid 12 and The secondary mixed liquid 16 is manufactured by mixing. Further, the waste liquid 17 is accumulated in the lower left of the microchip 100.

  Finally, as shown in FIG. 15, by applying a centrifugal force in the right direction toward FIG. 15, the secondary mixed solution 16 moves to the third holding unit 43 and the third reaction solution 13. And the tertiary mixture 20 is produced.

  The embodiment disclosed this time should be considered as illustrative in all points and not restrictive. The scope of the present invention is defined by the terms of the claims, rather than the description above, and is intended to include any modifications within the scope and meaning equivalent to the terms of the claims.

(A) is a schematic top view which shows a preferable example of the microchip of this invention, (b) is the typical top view which expanded a part in (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a). It is a schematic diagram which shows the process of the operation | movement in the microchip shown to Fig.1 (a).

Explanation of symbols

  DESCRIPTION OF SYMBOLS 1 Residue isolation | separation part, 1st edge | side in a substantially V-shaped wall surface, 3rd edge | side in a substantially V-shaped wall surface, 10 blood, 10a plasma component, 10b blood cell component, 11 1st reaction liquid, 12 Second reaction liquid, 13 Third reaction liquid, 14 Primary mixed liquid, 14a Residue, 14b Supernatant liquid, 15, 17 Waste liquid, 16 Secondary mixed liquid, 20 Tertiary mixed liquid, 30 Sample inlet, 31 Sample introduced Part, 32 first holding part, 33 third holding part, 34 second holding part, 35 centrifuge part, 36, 37, 39 liquid reagent measuring part, 40, 50 measuring part, 41 first mixing part , 42 2nd mixing part, 43 3rd mixing part, 51,52 preliminary chamber, 60 error sensing part, 100 microchip.

Claims (4)

A fluid formed by bonding at least a first substrate having a groove provided on the substrate surface and a second substrate, and formed by the groove and the first substrate side surface of the second substrate. A microchip having a circuit therein,
The microchip has a sample introduction port for introducing a sample into the fluid circuit, penetrating from the one surface to the fluid circuit.
The fluid circuit is:
A first mixing unit for mixing the specimen and the first reaction solution to produce a primary mixture;
A second mixing unit for mixing the primary mixed solution and the second reaction solution to produce a secondary mixed solution;
A third mixing unit for mixing the secondary mixed solution and the third reaction solution to produce a tertiary mixed solution;
At least one of the first mixing unit, the second mixing unit, and the third mixing unit separates a residue in at least one of the primary mixed solution, the secondary mixed solution, and the tertiary mixed solution. And a residue separator for
The residue separation unit separates the residue by applying a centrifugal force to the microchip in a first direction,
A microchip in which only a supernatant liquid is caused to flow out of the residue separation unit by applying a centrifugal force in a second direction.   2. The microchip according to claim 1, wherein an inner angle θ formed by the first direction and the second direction is 30 to 60 degrees (where θ is an inner angle of less than 90 degrees). The residue separator is
Formed with a substantially V-shaped wall with an opening and a squeezed tail,
When the centrifugal force is applied in the first direction, one side of the substantially V-shaped wall surface is at least one of the supernatant of the primary mixed solution, the secondary mixed solution, and the tertiary mixed solution, and the Having a length that does not allow the residue to flow out of the residue separator,
And the microchip of Claim 1 or 2 which has the length which flows out only a supernatant liquid from the said residue separation part by applying a centrifugal force in the said 2nd direction.   The microchip according to claim 1, further comprising a measuring unit for measuring only the supernatant liquid that has flowed out of the residue separation unit.

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