JP2009109467A - Microchip - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a microchip, capable of easily and rapidly inspecting and analyzing an inspection/analysis target stored in a plurality of cuvettes for optical measurement that the microchip possesses. <P>SOLUTION: The microchip has a fluid circuit inside, where a first substrate having a groove formed on a substrate surface and a plurality of through-holes penetrating in a thickness direction are bonded to one or not less than two second substrates. The microchip has not less than two cuvettes for optical measurement, including not less than two through-holes in the plurality of through-holes, and the substrate surface of the second substrate, and not less than two through-holes for composing not less than two cuvettes for optical measurement are arranged on a concentric circumference on the first substrate surface. <P>COPYRIGHT: (C)2009,JPO&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 including an optical measurement cuvette for performing inspection / analysis and the like 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. Each unit such as a holding unit, a measuring unit for measuring the sample and the liquid reagent, a mixing unit for mixing the sample and the liquid reagent, an optical measurement cuvette (detection unit) for analyzing and / or inspecting the mixed solution, It is mainly composed of fine flow paths (for example, a width of about several hundred μm) that appropriately connect these parts. 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 specimen and liquid reagent can be weighed, mixed, and introduced into the optical measurement cuvette. For inspection / analysis of the liquid mixture introduced into the optical measurement cuvette (for example, detection of a specific component in the liquid mixture), for example, the optical measurement cuvette containing the liquid mixture is irradiated with detection light and its transmittance It can be performed by measuring the like.

  Here, a microchip, particularly a microchip capable of performing a plurality of items of inspection / analysis on one type of specimen introduced into the microchip (in this case, the microchip has a plurality of optical measurement cuvettes. In order to make the best use of the above-mentioned merit of the microchip, a simple and quick detection operation is required.

  In addition, although the prior art about this invention was demonstrated based on the general technical information which the applicant acquired, the information which should be disclosed as prior art literature information before filing in the range which an applicant memorize | stores. Applicant does not have.

  The present invention has been made to solve the above-described problems, and has as its object to test and analyze objects (for example, the above-described specimen and liquid reagent contained in a plurality of optical measurement cuvettes included in a microchip. The present invention is to provide a microchip that can be easily and rapidly inspected and analyzed for (mixed liquid).

  That is, according to the present invention, a first substrate having a groove provided on the substrate surface and a plurality of through holes penetrating in the thickness direction is bonded to one or more second substrates. A microchip having a fluid circuit, wherein the microchip comprises two or more through holes and a substrate surface of the second substrate among the plurality of through holes. A microchip having a cuvette and having two or more through holes constituting the two or more optical measurement cuvettes arranged on the circumference of the same circle on the first substrate surface is provided.

  In the microchip of the present invention, the fluid circuit is connected to the liquid reagent holding unit for storing the liquid reagent, one or more measuring units for measuring the liquid reagent or the sample, and the measuring unit. And one or more overflow liquid storage units for storing the liquid reagent or specimen overflowing from the liquid. In this case, it is preferable that the overflow liquid container is disposed on the circumference where the two or more through holes are disposed on the surface of the first substrate.

  The fluid circuit includes one or more liquid reagent measuring units for measuring a liquid reagent and one or more sample measuring units for measuring a sample, and the liquid reagent overflowing from the measuring unit during measurement or You may have a some overflow liquid accommodating part for accommodating a test substance.

  In addition, the microchip of the present invention is formed by bonding a second substrate to both surfaces of a first substrate having grooves provided on both surfaces of the substrate and a plurality of through holes penetrating in the thickness direction. It is preferable that the microchip has a fluid circuit.

  Here, the second substrate is preferably a transparent substrate. The first substrate is preferably an opaque substrate, and more preferably a black substrate.

  According to the present invention, a plurality of optical measurements can be easily and rapidly performed in a microchip having a plurality of optical measurement cuvettes and capable of performing inspection and analysis of a plurality of items for one kind of specimen.

  Hereinafter, the present invention will be described in detail with reference to embodiments. 1A and 1B are outline views showing an example of a microchip according to the present invention. FIG. 1A is a top view, FIG. 1B is a side view, and FIG. 1C is a bottom view. A microchip 100 shown in FIG. 1 is formed by bonding two second substrates 102 and 103, which are transparent substrates, to both surfaces of a first substrate 101, which is a black substrate (see FIG. 1B). . The vertical and horizontal lengths of these substrates are not particularly limited, but in this embodiment, the horizontal (A in FIG. 1) is approximately 62 mm × vertical (B in FIG. 1) is approximately 30 mm. In the present embodiment, the thicknesses of the second substrate 102, the first substrate 101, and the second substrate 103 (C, D, and E in FIG. 1, respectively) are about 1.6 mm, about 9 mm, and about 1 respectively. .6 mm. However, the vertical and horizontal lengths and thicknesses of the substrate are not limited to these.

  Into the second substrate 102, liquid reagent introduction ports 110 (total of 11 in the present embodiment) and specimens (for example, whole blood etc.) penetrating in the thickness direction are introduced into the microchip fluid circuit. A mouth 120 is formed. The liquid reagent is incorporated in the liquid reagent holding part of the fluid circuit in advance before actual use of the microchip (examination / analysis of the specimen), and is a reagent for mixing or reacting with the specimen or processing the specimen. . Usually, after injecting a liquid reagent from the liquid reagent introduction port 110, the microchip is sealed with the liquid reagent introduction port 110 with a sealing label or the like for actual use.

  The first substrate 101 is formed with grooves formed on both surfaces thereof and through-holes penetrating in the thickness direction, and by attaching the second substrates 102 and 103 to this, two layers are formed inside the microchip. The fluid circuit is formed. Here, two layers means that fluid circuits are provided at two different positions in the thickness direction of the microchip. These two fluid circuits are communicated by a through hole formed in the first substrate 101.

  Among the plurality of through holes formed in the first substrate 101, the through holes 311, 312, 313, 314, 315, and 316 (6 in total) shown in FIG. Together with the surfaces of the second substrate 102 and 103 substrate for sealing the optical cuvette (also referred to as a detection unit in this specification) 161, 162, 163, 164, 165, 166. In the present embodiment, the diameter of the opening on the second substrate 102 side of the through holes 141 to 146 is about 1.5 mm, and the diameter of the opening on the second substrate 103 side is about 1 mm. It is not something. By irradiating the optical measurement cuvette with detection light in a substantially vertical direction from the lower side (or upper side) of the microchip to the surface of the microchip, and measuring the transmittance and the like, the optical measurement cuvette is measured. Inspection / analysis (for example, detection of a specific component in the liquid mixture) of an inspection / analysis object (for example, a liquid mixture of a specimen and a liquid reagent) accommodated in the inside is performed. Note that the sample mixed with the liquid reagent may be the sample itself or a specific component separated from the sample inside or outside the microchip. In the present specification, the specimen includes both meanings.

  Here, the microchip of the present embodiment is characterized in that the through holes 311 to 316 constituting the optical measurement cuvette are arranged on the circumference 150 of the same circle on the surface of the first substrate 101 ( FIG. 1 (a)). With this configuration, it is possible to easily and quickly perform inspection / analysis of each inspection / analysis target introduced into the six optical measurement cuvettes (detection units).

  As described above, in the microchip having the fluid circuit inside, such as the microchip of the present invention, the measurement of the specimen and the liquid reagent, the mixing of the specimen and the liquid reagent, and the specimen and the liquid reagent in the fluid circuit. A series of operations such as movement of the mixed solution to each part can be performed by applying a centrifugal force in an appropriate direction to the microchip. Application of centrifugal force to the microchip is performed using, for example, a centrifuge having a microchip mounting portion for mounting the microchip.

  A centrifugal device that applies centrifugal force to a microchip has, for example, a circular stage that can freely rotate (rotate to revolve the microchip) around a centrifugal center, and is provided on the surface of the stage or on the stage. The microchip mounting portion can be provided on the surface of a circular stage for rotating the microchip. The configuration of the microchip mounting portion is not particularly limited, and can be, for example, a groove for fitting a microchip having substantially the same shape as the outer shape of the microchip, or a fixed wall that supports the mounted microchip. It can also consist of. FIG. 2 is a schematic perspective view showing a state in which a microchip is mounted on a centrifuge provided with a microchip mounting portion composed of a fixed wall for supporting the microchip. As shown in FIG. 2, the microchip mounting portion 200 can be provided with a fixture 203 using, for example, a leaf spring or a spring, for fixing the position of the mounted microchip. The microchip placed on the circular stage 201 of the centrifuge and placed along the fixing walls 202a and 202b for supporting the microchip is pressed by the fixture 203, and the circular stage 201 is moved in this state. Centrifugal force is applied by rotating.

  After a predetermined centrifugation operation is performed, a light source (not shown) located below the circular stage 201 is irradiated with detection light 204 toward the optical measurement cuvette of the microchip, and its transmittance is measured. Thus, the inspection / analysis of the inspection / analysis object in the optical measurement cuvette is performed. At this time, in the microchip of this embodiment having a plurality of optical measurement cuvettes, it is necessary to irradiate each optical measurement cuvette with the detection light, but in this embodiment, these optical measurement cuvettes are used. Since the cuvettes are arranged on the circumference of the same circle, the detection light 204 is emitted from a fixed light source, and the circular stage 201 is rotated so that each optical measurement cuvette is rotated by the light of the detection light 204. By arranging them on the axis in order, inspection and analysis can be performed easily and quickly. The detection light may be irradiated from the upper surface side of the microchip.

  Here, the circle in the “same circle” is preferably a circle centered on the centrifugal center for imparting centrifugal force to the microchip. More specifically, the microchip is usually placed on the circular stage of the centrifuge having a rotatable circular stage as described above, and centrifugal force is applied to the microchip. The circle centered on the centrifugal center for applying centrifugal force can also be referred to as a circle centered on the rotational center of the circular stage.

  Next, the configuration of the fluid circuit included in the microchip of the present embodiment will be described more specifically.

  FIG. 3 is a perspective view showing a fluid circuit formed on the first substrate 101, and FIG. 3A is a surface of the second substrate 102 side (hereinafter simply referred to as “upper side”). FIG. 3B is a perspective view showing the fluid circuit formed on the surface of the second substrate 103 side (hereinafter sometimes simply referred to as “lower side”). is there. As shown in FIG. 3, each portion of the first substrate 101 is subjected to fluid treatment of a specimen, a liquid reagent, or a mixed solution thereof by a groove formed on the surface and a through-hole penetrating in the thickness direction. And a fine channel that appropriately connects these portions.

  4 and 5 are a top view and a bottom view of the first substrate 101, respectively. FIG. 4 shows the upper fluid circuit of the first substrate 101, and FIG. 5 shows the lower fluid circuit. In FIG. 5, the lower fluid circuit of the first substrate is shown in a horizontally inverted state so that the correspondence with the upper fluid circuit shown in FIG. 4 can be clearly understood. The microchip 100 of the present embodiment is a multi-item chip that can perform six items of inspection / analysis for one specimen, and the fluid circuit includes six items so that six items of inspection / analysis can be performed. It is divided into sections (sections 1 to 6 in FIG. 4) (however, they are connected to each other in the specimen measuring section installation area (lower fluid circuit upper area)). Each section is provided with one or two liquid reagent holders containing liquid reagents (liquid reagent holders 301a, 301b, 302a, 302b, 303a, 303b, 304a, 304b, 305a in FIG. 4). , 305b and 306a in total 11). The sample introduced from the sample introduction port 120 in FIG. 1 is distributed to each section after the blood cell component is separated and removed, and when it is weighed, one or two kinds of liquid in each section weighed separately. It is mixed with the reagent and introduced into the through holes 311, 312, 313, 314, 315 and 316 constituting the optical measurement cuvette, respectively. As described above, the liquid mixture introduced into each optical measurement cuvette (detection unit) irradiates light to the optical measurement cuvette from a direction substantially perpendicular to the microchip surface, and measures the transmitted light. It is subjected to optical measurement, and a specific component in the mixed solution is detected. In these series of processes, by applying a centrifugal force in an appropriate direction to the microchip, the liquid reagent, the specimen, or a mixture thereof is supplied to each part in the two-layer fluid circuit provided in each section. This is done by moving them in an appropriate order.

  In each of the above sections, in the lower fluid circuit, there are sample metering units (six units 401, 402, 403, 404, 405, and 406 in FIG. 5) for metering the sample and liquid reagent metering for metering the liquid reagent. (A total of 11 liquid reagent measuring units 411a, 411b, 412a, 412b, 413a, 413b, 414a, 414b, 415a, 415b and 416a in FIG. 5) are provided. Each sample measurement part is connected in series by the flow path.

  Here, in the microchip of the present embodiment, as shown in FIG. 4, from the overflow sample storage unit 330 for storing the sample overflowing from the sample measurement unit during measurement and from the liquid reagent measurement unit during measurement. Overflow reagent storage portions 331a, 331b, 332a, 332b, 333a, 333b, 334a, 334b, 335a, 335b, and 336a are provided for storing overflowing liquid reagents. The overflow specimen storage part 330 is connected to the specimen measurement part 406 via the flow path 16a (see FIG. 5), the through hole 26a penetrating in the thickness direction, and the flow path 16b (see FIG. 4). In addition, each overflow reagent storage unit is connected to a corresponding liquid reagent metering unit via a flow path and a through hole. For example, in the section 1, the liquid reagent measuring unit 411a for measuring the liquid reagent stored in the liquid reagent holding unit 301a and the overflow reagent storage unit 331a for storing the overflowing liquid reagent include the flow path 11a ( 5) and a through-hole 21a penetrating in the thickness direction and a flow path 11b (see FIG. 4). The same applies to other overflow reagent storage units.

  As described above, the microchip includes the overflow specimen storage section and the overflow reagent storage section (hereinafter collectively referred to as the overflow liquid storage section), thereby detecting the presence or absence of the overflow liquid in the overflow liquid storage section. Easily confirm whether the sample or liquid reagent is reliably transferred to the sample measuring unit or liquid reagent measuring unit by centrifugation and whether the sample measuring unit or liquid reagent measuring unit is filled with the sample or liquid reagent Can do. That is, if it is detected that the overflow liquid is present in the overflow liquid storage unit, it is guaranteed that the sample or the liquid reagent is accurately measured in the sample measurement unit or the liquid reagent measurement unit. As a result, the reliability of the examination / analysis of the specimen is improved, and if the measurement abnormality is confirmed, it is possible to determine that the obtained examination / analysis data is not adopted. For example, the sample or liquid reagent is not introduced into the sample measuring unit or the liquid reagent measuring unit due to a malfunction of the apparatus; evaporating of the liquid reagent, insufficient sample introduction amount due to misuse by the user, or substrate during microchip manufacturing. Examples include cases where the amount of specimen or liquid reagent to be weighed has not been weighed due to poor bonding or the like.

  Here, the method for detecting whether or not the overflowing specimen or liquid reagent is present in the overflow liquid storage part is not particularly limited, but for example, a second transparent substrate is used for the overflow liquid storage part. A method of irradiating light from the side of the second substrate 102 and measuring the intensity of the reflected light can be preferably used. The light to be used is not particularly limited, and may be, for example, monochromatic light (for example, laser light) having a wavelength of about 400 to 1000 nm, or may be mixed light such as white light. The intensity of reflected light can be measured using, for example, a commercially available reflection sensor.

  In the method of detecting the presence or absence of the overflow liquid by measuring the reflected light intensity, basically, the second liquid is introduced into the overflow liquid storage section before the overflow liquid is introduced into the overflow liquid storage section. The reflected light intensity obtained by irradiating light from the substrate 102 side and the second substrate 102 with respect to the overflow liquid storage unit after the sample or liquid reagent is introduced into the sample measuring unit or the liquid reagent measuring unit. The ratio of the reflected light intensity obtained by irradiating light from the side is obtained, and the presence or absence of the overflow liquid is detected from the intensity ratio. That is, when the ratio (reflected light intensity after introduction / reflected light intensity before introduction) is smaller than 1 (when the reflected light intensity after introduction is smaller), the overflow liquid is present in the overflow liquid container. To be judged. However, if the manufacturing fluctuation between microchips is small and the reflected light intensity before introducing the overflow liquid can be regarded as almost constant between the microchips, the measurement of the reflected light intensity before introducing the overflow liquid should be omitted. Is possible.

  Note that the method for detecting whether or not there is a liquid by measuring the reflected light intensity is not limited to the overflow liquid container, but can be applied to other parts of the microchip. For example, by irradiating light to the liquid reagent holding part before actual use of the microchip and measuring the reflected light intensity, it can be confirmed whether the liquid reagent is present in the liquid reagent holding part, It is possible to detect an abnormality that the liquid reagent is not stored in the liquid reagent holding part due to outflow or evaporation due to impact or the like during transportation. In addition, the sample measuring unit, the liquid reagent measuring unit, and the mixing unit where the sample and the liquid reagent are mixed are irradiated with light, and the reflected light intensity is measured, so that the sample measuring unit and the mixing unit can be reliably tested. Since it can be confirmed that the liquid reagent or the mixed solution thereof is present, it is possible to ensure that the predetermined processing is performed by applying the centrifugal force.

  As described above, the microchip 100 of the present embodiment has a total of 11 overflow reagent storage portions and one overflow sample storage portion corresponding to each liquid reagent, all of which are the second substrate. It is preferably formed in the fluid circuit on the 102 side (upper fluid circuit) (see FIG. 4). By forming all overflow liquid storage parts in one fluid circuit, there is no need to turn the microchip over when measuring reflected light intensity, and it is possible to detect the presence or absence of overflow liquid in all storage parts easily and quickly. it can. Further, these overflow liquid storage portions are more preferably arranged on the circumference of the same circle in one fluid circuit formed on the surface of the first substrate 101, and constitute the optical measurement cuvette. It is particularly preferable to arrange on the circumference where the through holes 311 to 316 are arranged (see FIG. 4). By arranging the optical measurement cuvette and the overflow liquid container on the circumference of the same circle, the detection light from the fixed transmitted light measurement light source and reflected light measurement light source (which may be the same light source) In addition, by rotating the circular stage and arranging each optical measurement cuvette and each overflow liquid storage part in order on the optical axis of the detection light, inspection and analysis and overflow can be performed easily and quickly. The presence or absence of liquid can be detected.

  Next, an example of fluid processing using the microchip 100 of the present embodiment will be described with reference to FIGS. 6 to 12 show the state of the liquid (specimen, liquid reagent, and mixed liquid thereof) and the lower surface (second substrate 103) of the upper surface (surface on the second substrate 102 side) of the first substrate 101 in each step of fluid processing. It is a figure which shows the state of the liquid of a side surface. (A) in each figure is a figure which shows the state of the liquid on the 1st board | substrate 101 upper surface, (b) is a figure which shows the state of the liquid on the 1st board | substrate 101 lower surface. 6 (b) to 12 (b), as in FIG. 5, the left and right are reversed so that the correspondence with the upper fluid circuit shown in FIGS. 6 (a) to 12 (a) can be clearly understood. The lower fluid circuit of the 1st board | substrate is shown in the made state. In the following description, only the fluid processing in the fluid circuit of section 1 will be mainly described, but the same processing is performed in the other sections, and this is clearly understood by referring to the drawings. can do. Furthermore, in the following, a case where the specimen is whole blood (hereinafter, a plasma component separated from whole blood may also be referred to as a specimen) will be described as an example, but the type of specimen is limited to this. It is not a thing.

(1) Blood Cell Separation, Liquid Reagent Metering Step First, in this step, the microchip in the state shown in FIGS. 4 and 5 is directed downward in FIG. 6 (hereinafter simply referred to as downward. The same applies to FIGS. The same applies to the other directions.) A centrifugal force is applied. Thereby, the whole blood introduced from the specimen introduction port of the second substrate 102 moves to the lower fluid circuit through the through hole 20a and is introduced into the blood cell separation unit 420 (see FIG. 6B). . The whole blood 600 introduced into the blood cell separation unit 420 is centrifuged by the blood cell separation unit 420 by a downward centrifugal force and separated into a plasma component (upper layer) and a blood cell component (lower layer). Whole blood overflowing from the blood cell separator 420 moves to the upper fluid circuit through the through hole 20b and is stored in the waste liquid reservoir 430 (see FIG. 6A). Further, by applying a downward centrifugal force, the liquid reagents in the liquid reagent holding portions 301a and 301b reach the liquid reagent measuring portions 411a and 411b through the through holes 21b and 21c, respectively, and are weighed (FIG. 6B). reference). The liquid reagent overflowing from the measuring unit passes through the through holes 21a and 21d, and is stored in the overflow reagent storage units 331a and 331b in the upper surface side fluid circuit (see FIG. 6A).

(2) Specimen weighing step Next, a leftward centrifugal force is applied. As a result, the plasma component separated in the blood cell separation unit 420 is introduced into the sample measurement unit 401 (at the same time, also introduced into the sample measurement units 402, 403, 404 and 405, 406) and measured (FIG. 7 ( b)). The plasma component overflowing from the measuring portion is moved into the upper fluid circuit through the through hole 26a (see FIG. 7A). The liquid reagent remains in the mixing portion 441a of the lower fluid circuit.

(3) First mixing step Next, a downward centrifugal force is applied. As a result, the measured liquid reagent (the liquid reagent held in the liquid reagent holding unit 301a) and the plasma component measured by the sample measuring unit 401 are mixed in the liquid reagent measuring unit 411a (first). First step of the mixing process, see FIG. 8B). Next, by applying a leftward centrifugal force, the mixed solution is further mixed with the liquid reagent remaining in the mixing unit 441a (see the second step of the first mixing step, see FIG. 9B). These first step and second step are performed a plurality of times as necessary to ensure mixing. Finally, a state similar to the state shown in FIG. 9 is obtained.

(4) Second mixing step Next, an upward centrifugal force is applied. Thereby, the mixed liquid in the mixing unit 441a reaches the mixing unit 441b through the through hole 21e, and the other measured liquid reagent (the liquid reagent held in the liquid reagent holding unit 301b) is also Through the through hole 21e, it reaches the mixing section 441b and is mixed (see the second step of the second mixing step, see FIG. 10A). Next, by applying a rightward centrifugal force, as shown in FIG. 11A, the mixed solution moves in the mixing unit 441b and the mixing is promoted (second step of the second mixing step, FIG. 11 (a)). Further, the liquid reagent is accommodated in the overflow reagent accommodating portion 332b by the rightward centrifugal force (see FIG. 11A). These first step and second step are performed a plurality of times as necessary to ensure mixing. Finally, a state similar to the state shown in FIG. 11 is obtained.

(5) Detection part introduction process Finally, downward centrifugal force is applied. As a result, the mixed solution is introduced into the through hole 311 constituting the optical measurement cuvette (detection unit) (refer to FIGS. 12A and 12B for other mixed solutions as well). The overflow reagent storage units 331a and 331b and the overflow sample storage unit 330 are in a state where a liquid reagent or a sample (plasma component) is stored. The same applies to other overflow reagent storage units. Through the above-described steps, each optical measurement cuvette is filled with the liquid mixture to be inspected and analyzed, and each overflow reagent storage unit and the overflow sample storage unit are filled with the overflow solution. In such a state, according to the above-described method, each of the transmitted light measurement light source and the reflected light measurement light source (which may be the same light source) is irradiated with detection light, and the circular stage is rotated. The optical measurement cuvette and each overflow liquid storage unit are sequentially arranged on the optical axis of the detection light, thereby performing inspection / analysis and detection of the presence or absence of the overflow liquid. It should be noted that the presence or absence of the sample or liquid reagent does not necessarily have to be performed at this stage, but it is in this state that the sample or liquid reagent can be stored in all the overflow sample storage units and overflow reagent storage units. In order to simplify the operation, it is preferable to confirm the presence or absence of the sample or liquid reagent after the optical measurement cuvette introduction step.

  As described above, the microchip of the present invention and the method of using the microchip have been described by taking the microchip having a two-layer fluid circuit as an example. The microchip formed by bonding together the 1st board | substrate with which the groove | channel and through-hole which comprise a fluid circuit were formed, and the 1st 2nd board | substrate which is a transparent substrate may be sufficient.

  Further, the number of optical measurement cuvettes included in the microchip of the present invention is not particularly limited as long as it has at least two optical measurement cuvettes. Further, in the present invention, the structure of the fluid circuit is not limited to that shown in the above embodiment, and various structures can be adopted according to the processing to be performed on the specimen. The fluid circuit is not necessarily required to have the overflow liquid storage unit, and may have at least a plurality of optical measurement cuvettes.

  In the case where the microchip of the present invention has a two-layer fluid circuit, referring to the above embodiment, the second substrates 102 and 103 do not necessarily need to be transparent substrates, but constitute at least an optical measurement cuvette. The surface region to be processed needs to be transparent so that the transmitted light of the incident light can be measured. Further, as a method of bonding the first substrate 101 and the second substrates 102 and 103, when using a welding method in which light is applied to the bonding surface of the substrate and the surface is melted, the bonding is used. The first substrate 101 is preferably an opaque substrate (preferably a black substrate) and the second substrates 102 and 103 are preferably transparent substrates so that the absorbed light can be absorbed more efficiently. Thus, the first substrate 101 and the second substrate 102, 103 are bonded by irradiating light from the second substrate 102, 103 side and melting the bonding surface of the first substrate 101. Can be easily performed.

  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.

It is an outline view showing an example of a microchip concerning the present invention. It is a schematic perspective view which shows the state which mounted the microchip in the centrifuge provided with a microchip mounting part. It is a perspective view which shows an example of the fluid circuit formed in the 1st board | substrate of the microchip based on this invention. It is a top view which shows an example of the 1st board | substrate of the microchip which concerns on this invention. It is a bottom view which shows an example of the 1st board | substrate of the microchip which concerns on this invention. It is a figure which shows the state of the liquid of the upper surface of the 1st board | substrate in the blood cell separation and a liquid reagent measurement process, and the state of the liquid of a lower surface. It is a figure which shows the state of the liquid of the upper surface of the 1st board | substrate in a sample measurement process, and the state of the liquid of a lower surface. It is a figure which shows the state of the liquid of the upper surface of the 1st board | substrate in the 1st step of a 1st mixing process, and the state of the liquid of a lower surface. It is a figure which shows the state of the liquid of the upper surface of the 1st board | substrate in the 2nd step of a 1st mixing process, and the state of the liquid of a lower surface. It is a figure which shows the state of the liquid of the upper surface of the 1st board | substrate in the 2nd mixing process 1st step, and the state of the liquid of a lower surface. It is a figure which shows the state of the liquid of the upper surface of the 1st board | substrate in the 2nd step of a 2nd mixing process, and the state of the liquid of a lower surface. It is a figure which shows the state of the liquid of the upper surface of the 1st board | substrate in the detection part introduction | transduction process, and the state of the liquid of a lower surface.

Explanation of symbols

  100 Microchip, 101 First substrate, 102, 103 Second substrate, 110 Liquid reagent inlet, 120 Sample inlet, 150 circumference, 161, 162, 163, 164, 165, 166 Optical measurement cuvette, 200 Microchip mounting part, 201 circular stage, 202a, 202b fixing wall, 203 fixing tool, 204 detection light, 301a, 301b, 302a, 302b, 303a, 303b, 304a, 304b, 305a, 305b, 306a liquid reagent holding part, 311, 312, 313, 314, 315, 316 Through hole constituting the cuvette for optical measurement, 330 overflow specimen storage part, 331a, 331b, 332a, 332b, 333a, 333b, 334a, 334b, 335a, 335b, 336a overflow reagent Containment section, 01, 402, 403, 404, 405, 406 Sample weighing unit, 411a, 411b, 412a, 412b, 413a, 413b, 414a, 414b, 415a, 415b, 416a Liquid reagent weighing unit, 420 Blood cell separation unit, 430 Waste liquid reservoir, 441a, 441b mixing section, 11a, 11b, 16a, 16b flow path, 20a, 20b, 21a, 21b, 21c, 21d, 21e, 26a through hole, 600 whole blood.

Claims (7)

A microchip having a fluid circuit therein, which is formed by laminating a first substrate having a groove provided on the substrate surface and a plurality of through holes penetrating in the thickness direction, and one or more second substrates. Because
The microchip has two or more optical measurement cuvettes constituted by any two or more through holes and the substrate surface of the second substrate among the plurality of through holes,
The two or more through-holes constituting the two or more optical measurement cuvettes are microchips arranged on the circumference of the same circle on the surface of the first substrate. The fluid circuit is:
A liquid reagent holding unit for storing the liquid reagent;
One or more measuring units for measuring the liquid reagent or specimen;
One or more overflow liquid storage units connected to the measurement unit, for storing the liquid reagent or the sample overflowing from the measurement unit during measurement,
2. The microchip according to claim 1, wherein the overflow liquid storage portion is disposed on a circumference where the two or more through-holes are disposed on the surface of the first substrate.   The fluid circuit includes one or more liquid reagent measuring units for measuring the liquid reagent and one or more sample measuring units for measuring a sample, and the liquid reagent overflows from the measuring unit during measurement. The microchip according to claim 2, further comprising a plurality of overflow liquid storage units for storing the specimen.   2. A fluid circuit having two layers inside, wherein a second substrate is bonded to both surfaces of a first substrate having grooves provided on both surfaces of the substrate and a plurality of through holes penetrating in the thickness direction. 4. The microchip according to any one of 3.   The microchip according to claim 1, wherein the second substrate is a transparent substrate.   The microchip according to claim 1, wherein the first substrate is an opaque substrate.   The microchip according to claim 6, wherein the first substrate is a black substrate.

Images