JP4391790B2 - Chip usage and inspection chip - Google Patents

Description

  The present invention relates to a method of using a chip into which a sample containing a target component has been introduced and a test chip for testing the target component.

  Biochemistry that measures the concentration of enzymes and their products that are active in the liver, kidneys, pancreas, etc., in order to diagnose liver and biliary tract diseases and alcoholic liver disorders and observe the treatment Inspection is widely conducted. As an apparatus for performing such a biochemical test, a health care device capable of separating serum or plasma from collected blood and taking it out of the chip and quantifying the concentration, pH, etc. of the serum or plasma is disclosed ( Patent References 1 to 5).

In addition, a sample processing card is disclosed in which a sample is introduced into a sample measuring means via a capillary by centrifugal force, and a weighed sample is mixed with a reagent (see Patent Document 6). Similarly, a microanalyzer that can separate and accurately weigh biological samples using a centrifugal force is disclosed (see Patent Document 7).
JP 2001-258868 A JP 2002-267777 A JP 2003-102710 A JP 2003-83958 A JP 2003-83926 A U.S. Pat. No. 4,883,763 US Pat. No. 6,399,361

  However, in the health care devices described in Patent Documents 1 to 5, the target component can be separated from the collected sample, but the target component cannot be separated and weighed together in the same chip. Therefore, in order to separate and weigh, it is necessary to separate the target component with the separation device and then to take out the target component from the separation device and introduce it into the weighing device. For this reason, operations such as separation of target components and accurate weighing are not performed in series in the same chip, which is complicated. In the sample processing card described in Patent Document 6, the target component can be weighed, but the target component cannot be separated and weighed from the sample in a lump. Furthermore, in the microanalyzer described in Patent Document 7, the WAX valve provided at a predetermined position must be removed in order to prevent the centrifuged fluid from flowing out, and the fluid flows out after removing the valve. To weigh. Therefore, it is necessary to provide a WAX valve in the microanalyzer described in Patent Document 7. Further, since it is necessary to apply heat such as infrared rays in order to remove the WAX bulb, temperature control is necessary and complicated. Furthermore, when the WAX valve is melted and dissolved and mixed with the sample, the target component is contaminated, and accurate weighing and quantification of the target component cannot be performed.

Therefore, an object of the present invention is to provide a method of using a chip that can be separated and weighed easily and collectively in a chip into which a sample containing a target component has been introduced.
Moreover, it aims at providing the test | inspection chip which can perform separation and weighing collectively and simply.

In order to solve the above problems, the first invention of the present application has a reagent reservoir for holding a reagent, a mixing section connected to the reagent reservoir and the weighing tube, and a chip into which a sample containing a target component is introduced. A method of use, wherein a separation step of centrifuging the target component from the sample by rotating the chip around a predetermined rotation axis, and a target component introduction step of introducing the target component into a weighing tube having a predetermined volume A weighing step in which the tip is rotated again around the rotating shaft, and the target component introduced into the weighing tube is weighed out, and the tip is rotated around the rotating shaft to move the tip from the reagent reservoir. A reagent introduction step for introducing the reagent into the mixing section;
There is provided a method of using a chip including a mixing step of rotating the chip around the rotation axis, introducing the target component weighed and collected in the weighing step into the mixing unit, and mixing with the reagent .

  In the separation step, the target component is separated from the sample by rotating the chip around a predetermined rotation axis. Next, in the target component introduction step, the separated target component is introduced into the weighing tube. Further, in the weighing step, the target component that fills the weighing tube is taken out from the weighing tube and weighed by rotating the tip around the same rotation axis again. Therefore, by using the above usage method, the target components in the sample can be separated and weighed easily and collectively. At this time, the target component introduced into the weighing tube is weighed and taken out by the volume of the weighing tube, so that the target component can be accurately weighed. Moreover, since the rotation axis of the chip in the separation step and the weighing step is the same, the chip is easily handled in the separation of the target component and the weighing step. Furthermore, since there is one rotating shaft, the chip design is easy and the chip can be miniaturized.

  The reagent is introduced into the mixing unit by rotation about the same rotation axis as the separation step and the weighing step. In addition, the separated and weighed target components are introduced into the mixing unit by rotation about the same rotation axis as the separation step and the weighing step, and mixed with the reagent. By using the above usage method, separation of the target component in the sample, weighing, and mixing with the reagent can be performed all at once. At this time, since the target component is accurately weighed, a mixed substance having a desired mixing ratio between the reagent and the target component can be obtained. In addition, since the rotation axes of the chips in the separation step, the reagent introduction step, and the mixing step are the same, it is easy to handle the chips when performing the above steps continuously. Thus, when there is only one rotating shaft, the design of the chip is easy, and the chip can be miniaturized. Further, it is possible to reduce the size of a device such as a rotating device on which the chip is placed.

Furthermore, since the target component is not taken out of the chip in the steps from when the sample is introduced and mixed with the reagent, contamination of the target component can be reduced.
Here, it is preferable that the method further includes a light irradiation step of irradiating the mixed material of the target component and the reagent with light, and a quantitative step of taking out the light after passing through the mixed material and quantifying the target component. By using this method of use, separation of target components in a sample, weighing, mixing with a reagent, and quantitative determination can be performed all at once. In addition, since the target component is accurately weighed, the target component can be accurately quantified using a mixed material having a desired mixing ratio between the reagent and the target component. Furthermore, since the target component is not taken out of the chip, contamination of the target component can be reduced and the target component can be accurately quantified.

A third invention of the present application provides the method of using a chip according to the second invention of the present application, wherein the mixing step is simultaneous with the weighing step.
By performing the mixing step and the weighing step at the same rotation, the target component introduced into the weighing tube can be weighed while being taken out and simultaneously introduced into the mixing unit. Therefore, handling of the chip is easy. Moreover, the mixed substance in which the sample and the target component are mixed can be obtained quickly.

A fourth invention of the present application provides the method for using a chip according to the second invention of the present application, wherein the reagent introduction step is simultaneous with a separation step, a weighing step, or a mixing step.
The introduction of the reagent into the mixing unit is performed when the chip is rotated in the separation step, the weighing step, or the mixing step. Therefore, a mixed substance can be obtained quickly.
A fifth invention of the present application is an inspection chip having at least one quantification unit for quantifying a target component in a sample, wherein the quantification unit rotates the inspection chip around a predetermined rotation axis, thereby A centrifuge tube for centrifuging the target component, a weighing tube connected to one end of the centrifuge tube for weighing the target component by rotation about the rotation axis, and the weighing tube A transfer means for transferring the target component from the centrifuge tube to the weighing tube; at least one reagent reservoir for storing a reagent; the reagent reservoir and the weighing tube; Mixing in which the target component introduced from the weighing tube by re-rotation about the axis is mixed with the reagent introduced from the reagent reservoir by rotation about the rotation axis And a light detection path that is connected to the mixing unit and allows the mixed substance in which the reagent and the target component are mixed to pass therethrough, and a light inlet that is connected to the light detection path and introduces light into the light detection path And an optical outlet connected to the light detection path for extracting light after passing through the light detection path.

  The sample is introduced into the centrifuge tube, the chip is rotated around a predetermined rotation axis, and the target component is centrifuged from the sample in the centrifuge tube. Next, the separated target component is transferred to a weighing tube by transfer means. The target component in the weighing tube is introduced into the mixing unit from the weighing tube by re-rotation around the same rotation axis as that of the centrifuge tube, and mixed with the reagent. Here, the reagent is introduced from the reagent reservoir into the mixing unit by rotation about a predetermined rotation axis. The mixed substance mixed is introduced into the light detection path, and the target component is quantified by detecting the light that has passed through the light detection path. By using the above-described inspection chip, separation of target components in a sample, weighing, mixing with a reagent, and quantification can be performed all at once. At this time, since the target component is accurately weighed, a mixed substance having a desired mixing ratio between the reagent and the target component can be obtained, so that the target component can be accurately quantified. Moreover, since it can carry out collectively as mentioned above, the contamination of a target component can be reduced and it can quantify accurately. Furthermore, since the rotation axis of rotation for performing centrifugation in the centrifuge tube, introduction of the reagent into the mixing unit, and mixing of the reagent and the target component in the mixing unit is the same, the design of the test chip is easy, Further, the inspection chip can be reduced in size. Furthermore, since the structure which can isolate | separate and weigh a target component is simple, manufacture of a test | inspection chip is easy. In addition, it is possible to reduce the size of devices such as a rotating device on which the inspection chip is placed.

Here, when the inspection chip is connected to the centrifuge tube and further includes a collection needle for collecting the sample, the collection needle is connected to the inspection chip, so that the collected sample is separated, weighed, and quantified. Can be performed collectively and simply. Therefore, it is preferable because contamination of the sample can be reduced and accurate quantification can be performed.
In addition, a reagent reservoir is connected to the mixing part via a connection part, and the connection part has a rotation axis so that an acute angle formed by the connection part with a radial direction of a circle around the rotation axis is smaller than 90 degrees. It is preferable that the reagent reservoir is provided closer to the rotating shaft than the connection portion. By forming the reagent reservoir and the connecting portion as described above, the force in the direction of the mixing portion is exerted on the reagent reservoir and the reagent in the connecting portion by rotation around the rotation axis. It can be well introduced into the mixing section.

A sixth invention of the present application provides the inspection chip according to the fifth invention, further comprising a capillary valve having a flow path width larger than that of the weighing tube between the weighing tube and the mixing portion.
The capillary valve provided between the weighing tube and the mixing unit has a flow channel width larger than the flow channel width of the weighing tube. Therefore, when the target component is introduced from the centrifuge tube into the weighing tube by the transfer means, the capillary valve can prevent the target component from flowing out from the weighing tube to the mixing unit. Moreover, it can prevent that the target component moved to the mixing part from the weighing tube flows back into the weighing tube due to capillary action. Therefore, the target component accurately weighed can be introduced into the mixing unit.

  The seventh invention of the present application is the fifth invention, wherein the weighing tube is generally U-shaped, the weighing tube opens to the rotating shaft side, and one end of the U-shape of the weighing tube is connected to the centrifuge tube. The other end is connected to the transfer means, the bottom of the U-shape is connected to the mixing unit, and the target component is mixed from the bottom of the weighing tube by the rotation about the rotation axis. Provide inspection chip to move to.

  When the generally U-shaped weighing tube is arranged and connected to the rotation axis as described above, the target component can be efficiently moved to the mixing unit when rotated about the rotation axis. That is, the direction from the opening of the U-shaped weighing tube to the bottom is substantially the same as the centrifugal force when rotated about the rotation axis. At this time, when rotating around the rotation axis, the centrifugal force acts most on the bottom of the weighing tube, so that the target component can be efficiently moved from the weighing tube to the mixing unit.

  Here, the weighing tube is formed symmetrically with respect to the line, the transfer means has substantially the same shape as the centrifuge tube, and the centrifuge tube and the transfer means are formed symmetrically with respect to the weighing tube. Preferably. Since the transfer means, the centrifuge tube, and the transfer pipe are formed in line symmetry, the target component can be easily and accurately moved from the centrifuge tube to the weighing tube by the transfer means. Therefore, the target component can be accurately weighed.

An eighth invention of the present application provides the inspection chip according to the fifth invention, wherein the transfer means is connected to a pump. By sucking the target component in the centrifuge tube using the pump, the target component can be moved from the centrifuge tube to the weighing tube.
The ninth invention of the present application is the fifth invention, further comprising: a holding portion that is provided at the bottom of the centrifuge tube and into which a component other than the target component in the sample is introduced by rotation about the rotation axis. Provide inspection chip including.

By providing the holding part at the bottom of the centrifuge tube, components other than the target component can be introduced into the holding part during the centrifugation around the rotation axis. By holding components other than the target component in the holding unit, the target component can be separated efficiently.
The tenth invention of the present application is the fifth invention according to the fifth invention, wherein the rotary shaft is provided with an inlet for taking in a sample to be introduced into two or more quantification units, and a circumference of the same circle centered on the rotary shaft. A plurality of partition plates provided at intervals along the partition plate, wherein the partition plates are arranged on the outer peripheral side in the radial direction of a circle centered on the rotation axis of the sample taken from the intake port. Providing test chips that hinder progress.

  The sample taken into the intake port on the rotating shaft tends to travel to the outer peripheral side in the radial direction by rotation about the rotating shaft. Although the progress of the sample toward the outer peripheral side in the radial direction is hindered by the partition plate, the sample proceeds toward the outer peripheral side in the radial direction through the plurality of partition plates. At this time, the sample proceeds to the outer peripheral side in the radial direction so as to be dispersed through the partition plates by rotation about the rotation axis. Therefore, the sample introduced into the inspection chip can be dispersed and introduced into a plurality of quantitative units.

  In the present invention, the target component is separated from the sample by rotating the chip around a predetermined rotation axis. Next, the separated target component is introduced into a weighing tube in the chip, and the target substance is taken out from the weighing tube and weighed by rotating the chip around the same rotation axis again. Therefore, separation and weighing of the target component in the sample can be performed collectively and simply. At this time, the target component introduced into the weighing tube is weighed and taken out by the volume of the weighing tube, so that the target component can be accurately weighed. Moreover, since the rotation axis of the chip in separation and weighing is the same, the chip can be easily handled in separation and weighing of the target component. Furthermore, since there is one rotating shaft, the chip design is easy and the chip can be miniaturized.

<Basic configuration>
1A and 1B are perspective views of an inspection chip according to the present invention, FIG. 2 is a plan view of FIG. 1A, and FIG. 3 is a schematic view of an apparatus on which the inspection chip is placed.
(1) Configuration of Inspection Chip The inspection chip 1 has a first substrate 3 and a second substrate 5 which are plate-like substrates. The first substrate 3 is formed with an inlet 7a, a transfer means 13a, and an outlet 15a. Further, the second substrate 5 has an inlet 7b corresponding to the inlet 7a, a centrifuge tube 9, a weighing tube 11, a transfer means 13b corresponding to the transfer means 13a, and an outlet 15b corresponding to the outlet 15a. Is formed. The inspection chip 1 is placed on the device 20 shown in FIG. The apparatus 20 includes a rotating table 21 on which the inspection chip 1 is mounted and rotated, and a predetermined rotation shaft 23 that rotates the inspection chip 1 mounted on the rotating table 21. The apparatus 20 includes, for example, a centrifuge control unit 25, a pump control unit 27, a temperature control unit 29, a current amplification unit 31, and the like. From the sample 40 introduced into the inspection chip 1 by the control of each unit. The target component 41 is separated and weighed.

  The sample 40 to be inspected is taken into the inspection chip 1 into the intake ports (7a, 7b) 7 of the inspection chip 1. The centrifuge tube 9 is connected to the intake port 7, and the sample 40 is introduced into the centrifuge tube 9 from the intake port 7. The centrifuge tube 9 is generally U-shaped, with one open end connected to the intake 7 and the other open end connected to a weighing tube 11 having a predetermined volume. Further, the centrifuge tube 9 is placed so that the U-shaped opening is substantially directed to the rotating shaft 23 side. And when the test | inspection chip 1 rotates centering | focusing on the predetermined rotating shaft 23, the target component 41 is centrifuged from the introduce | transduced sample 40. FIG. At this time, it is preferable that the U-shaped opening is positioned on the rotating shaft 23 side, and the U-shaped bottom is positioned on the radially outer peripheral side of a circle centering on the predetermined rotating shaft 23. When the centrifuge tube is arranged in this manner, the target component 41 can be efficiently centrifuged when the centrifuge tube is rotated about the rotation shaft 23. In other words, the direction from the opening of the U-shaped centrifuge tube 9 to the bottom and the centrifugal force when rotated about the rotation shaft 23 are substantially the same. Therefore, when rotating around the rotation shaft 23, the centrifugal force works most on the bottom of the centrifuge tube 9. Therefore, the non-target component 43 other than the target component 41 can be efficiently moved from the sample 40 to the bottom of the centrifuge tube 9, and the target component 41 can be separated. The weighing tube 11 connected to the centrifuge tube 9 is further connected to a transfer means (13a, 13b) 13 and an outlet (15a, 15b) 15. The weighing tube 11 has a desired volume for weighing the target component 41, and the target component 41 centrifuged from the sample 40 is introduced into the weighing tube 11. The transfer means 13 introduces the target component 41 centrifuged in the centrifuge tube 9 from the centrifuge tube 9 to the weighing tube 11. For example, the transfer means 13 is connected to a pump, and the target component 41 in the centrifuge tube 9 is transferred to the weighing tube 11 by applying a suction pressure to the centrifuge tube 9 and the weighing tube 11 by the pump. The take-out port 15 connected to the weighing tube 11 is located on the outer peripheral side in the radial direction of the circle centered on the predetermined rotation shaft 23 from the weighing tube 11, and the inspection chip 1 is centered on the predetermined rotation shaft 23. When rotating, the target component 41 is taken out from the weighing tube 11. At this time, in particular, the weighing tube 11 has a substantially U-shape or V-shape, and when the outlet 15 is connected to the bottom of the U-shape or V-shape, the target component 41 is efficiently utilized using centrifugal force. This is preferable because it can be introduced into the outlet 15. However, the centrifuge tube 9 is not limited to a U-shape, and may be formed to have, for example, a cup-shaped sample reservoir 9 'as shown in FIG. 1B, for example. At this time, in the sample 40 introduced into the sample reservoir 9 ′, the non-target component 43 in the sample 40 is accumulated at the bottom of the sample reservoir 9 ′ by rotating around the rotation shaft 23. Then, the target component 41 in the supernatant of the sample reservoir 9 ′ is introduced into the weighing tube 11 by the transfer means 13 and weighed in the same manner as described above.

Here, the rotating shaft 23 separates the target component 41 in the centrifuge tube 9 by rotation about the rotating shaft 23, and removes the target component 41 in the weighing tube 11 by rotating again about the rotating shaft 23. What is necessary is just to be formed so that it may introduce into the extraction port 15 from the weighing tube 11. Therefore, the predetermined rotating shaft 23 may be located outside the inspection chip 1 as shown in FIG. 3 or may be located inside the inspection chip 1.
(2) Method of using test chip Next, an example of a method of using the test chip 1 when the target component 41 is separated and weighed will be described with reference to FIGS.

The sample 40 containing the target component 41 is introduced into the centrifuge tube 9 (U-shaped tube indicated by the solid line in FIG. 4) from the intake 7 in the inspection chip 1 in advance, and the inspection chip 1 is fixed to the apparatus 20. Then, the target component 41 is separated and weighed as follows.
Step 1: The test chip 1 is rotated around a predetermined rotation shaft 23, and the target component 41 is centrifuged from the sample 40 introduced into the centrifuge tube 9 (see FIG. 5). At this time, centrifugal force acts on the U-shaped centrifuge tube 9 from the opening of the centrifuge tube 9 toward the bottom by rotation about the predetermined rotation shaft 23. Therefore, the non-target component 43 other than the target component 41 in the sample 40 moves to the bottom of the centrifuge tube 9, and the target component 41 is separated from the sample 40.

Step 2: Next, the centrifuged target component 41 is introduced from the centrifuge tube 9 into the weighing tube 11 (portion indicated by the solid line in FIG. 6) so as to fill the weighing tube 11 (see FIG. 6). The target component 41 is introduced into the weighing tube 11 by applying a pressure for sucking the target component 41 to the weighing tube 11 or the centrifuge tube 9 by the transfer means 13 connected to the weighing tube 11.
Step 3: Further, the test chip 1 is rotated again about the same rotation axis 23 as that in the centrifugation in Step 1 above, and the target component 41 introduced into the weighing tube 11 is removed from the outlet 15 (shown by the solid line in FIG. 7). (See FIG. 7). At this time, centrifugal force acts on the weighing tube 11 in the direction from the weighing tube 11 to the take-out port 15 by rotation about the predetermined rotation shaft 23. Accordingly, the target component 41 moves to the outlet 15. Here, the weighing tube 11 has a desired volume, and the target component 41 that satisfies the volume of the weighing tube 11 is taken out to the take-out port 15, so that the target component 41 in which a desired amount is weighed is obtained. be able to. Specifically, the target component 41 located between the connection portion AA ′ between the centrifuge tube 9 and the weighing tube 11 and the connection portion BB ′ between the weighing tube 11 and the transfer means 13 in FIG. It is introduced into the outlet 15 by force. The target component located in the other part can be accurately weighed by being introduced into the centrifuge tube 9 or the transfer means 13.
(3) Inspection Chip Manufacturing Method The above inspection chip 1 can be produced by an imprint method or an injection molding method. Depending on the method of manufacturing the substrate, PET (polyethylene terephthalate), Si, Si oxide film, quartz, glass, PDMS (polydimethylsiloxane), PMMA (polymethyl methacrylate), PC (polycarbonate), PP ( Polypropylene), PS (polystyrene), PVC (polyvinyl chloride), polysiloxane, allyl ester resin, cycloolefin polymer, Si rubber, and the like can be used.
(4) Effect By handling the inspection chip 1 into which the sample 40 is introduced as described above, the target component 41 is separated in step 1, and the target component 41 is weighed in steps 2 and 3. Therefore, the separation and weighing of the target component 41 can be performed collectively in the inspection chip 1 and can be easily performed. Moreover, since the target component 41 introduced into the weighing tube 11 is weighed and taken out by the volume of the weighing tube 11, the target component 41 can be accurately weighed. Furthermore, since it is not necessary to apply heat or the like for separation / weighing, the sample 40 is not affected by additives or heat. Therefore, contamination of the sample 40 can be reduced and the target component 41 included in the sample 40 can be accurately weighed. Further, since the rotating shafts 23 of Step 1 and Step 3 for separating and weighing the target component 41 are the same, it is easy to handle the inspection chip 1 in the separation and weighing of the target component 41. Further, since the single rotating shaft 23 is provided, the inspection chip 1 can be easily designed, and the inspection chip 1 can be downsized and the entire apparatus 20 on which the inspection chip 1 is placed can be reduced. For example, when the rotation axis 23 in the separation of the target component 41 and the weighing is different, the mounting position of the test chip 1 to the apparatus 20, the arrangement position of the centrifuge tube 9 and the weighing pipe 11 in the test chip 1, and the like. It is necessary to consider the relationship with the plurality of rotating shafts 23, which is complicated.

  Further, the inspection chip 1 is easy to manufacture because it has a simple configuration that can separate and weigh the target component 41 without providing a valve that needs to be removed during the separation and weighing. In addition, as shown in FIG. 1, it is preferable to have a two-dimensional direction extending along the radial direction of a circle centered on the rotation shaft 23. When the inspection chip 1 is such a plate-like substrate, the centrifuge tube 9, the weighing tube 11 and the like can be easily produced in the inspection chip 1 by using the above-described injection molding method or imprint method. . Moreover, the test | inspection chip 1 can be easily produced by producing the centrifuge tube 9, the weighing tube 11, etc. on one board | substrate, and bonding another board | substrate together. Furthermore, the inspection chip 1 can be reduced in thickness and size.

8A, when the sampling needle 50 and the syringe 51 are provided in the inspection chip 1, the sample 40 can be sampled, separated, and weighed in a lump. Therefore, the trouble of introducing the sample 40 collected by another means into the inspection chip 1 can be saved, and contamination of the sample 40 when introduced into the inspection chip 1 can be reduced. Furthermore, since it is possible to collect blood directly from the vein by the collection needle 50, it is possible to accurately weigh almost pure target components. When blood is collected from a vein, whole blood can be introduced into the test chip 1 by blood pressure without providing the syringe 51. Further, the sampling needle 50 and the syringe 51 may be removed when the inspection chip 1 is attached to the apparatus 20. Further, as shown in FIG. 8B, a syringe 52 may be provided instead of the syringe 51, and the sample 40 may be collected by the syringe 52.
<First embodiment>
9 is a plan view of the inspection chip according to the first embodiment of the present invention, FIGS. 10A and 10B are enlarged views of the main part of FIG. 9, and FIG. 11A is an enlarged view of the intake port. FIG. 11B is an enlarged view of the mixer section, FIG. 12 is a cross-sectional view of the intake port, and FIG. 13 is a schematic view of an apparatus on which the inspection chip is placed.
(1) Overall Configuration of Inspection Chip and Device The inspection chip 100 of the first embodiment is fixed to fix the sample inlet 105, the partition plate 110, and the inspection chip 100 including the target component to the apparatus 300 described later. It has the fixed part 200 into which the hole 115 and the sample are introduced. The intake port 105 is provided at the center so as to correspond to the rotation center of the inspection chip 100. The fixed amount unit 200 is evenly arranged at four places around the intake port 105. Further, as shown in FIG. 10A, the quantification unit 200 includes a centrifuge tube 201, a holding unit 203, a weighing tube 205, an air hole 206, a transfer means 207, a first connection tube 209, a capillary valve 211, and a second connection. The tube 215 includes a primary mixing unit 217, reagent reservoirs 219 a and 219 b for storing reagents, a secondary mixing unit 220, a light detection path 230, a light inlet 233, a light outlet 235, and an outlet 240. Here, the weighing tube 205, the first connection tube 209, the capillary valve 211, the second connection tube 215, and the primary mixing unit 217 in the quantification unit 200 are arranged on the radially outer peripheral side of a circle centering on the rotation shaft 310 described later. Are connected in order. Further, as shown in FIG. 12, a sampling needle 250 for sampling a sample is connected to the intake port 105 and connected to a spring 255. In FIG. 9, four determination units 200 are provided, but the number is not limited to four.

This inspection chip 100 is fixed to the apparatus 300 shown in FIG. The apparatus 300 fixes the inspection chip 100 to the apparatus 300, a rotating table 301 for rotating, a predetermined rotating shaft 310 for rotating the inspection chip 100 mounted on the rotating table 301, and a control for controlling the apparatus 300. A section 320, a pump section 333, a support section 331 for supporting the turntable 301, an optical fiber 332, a xenon lamp 335, and the like. Here, the rotation shaft 310 is provided so as to correspond to the intake port 105 of the inspection chip 100. The turntable 301 has a fixing portion 340 that fits into the fixing hole 115 of the inspection chip 100. The fixing table 301 further includes a transfer unit 207, reagent reservoirs 219 a and 219 b, and a connection pipe 343 that connects the outlet 240 and the pump unit 333 of the apparatus 300. The pump unit 333 performs pressure supply to the transfer unit 207, reagent supply to the reagent reservoirs 219 and 217, discharge of the solution from the outlet 340, and the like. The control unit 320 includes, for example, a centrifuge control unit 321, a pump control unit 323, a temperature control unit 325, a light control unit 327, a current amplification unit 329, and the like, and controls each unit of the apparatus 300. The turntable 301 is made of a metal having high thermal conductivity such as Al. The support portion 331 that supports the turntable 301 is made of a Peltier element thermocouple or the like, and can control the temperature of the test chip 100.
(2) Configuration of Each Part of Inspection Chip Next, the configuration of each part of the inspection chip will be described in detail.
(2-1) Intake Port The intake port 105 has a partition plate 110 shown in FIG. 11A and a sampling needle 250 with a spring 255 shown in FIG. In addition, the intake port 105 is provided so as to correspond to a predetermined rotation shaft 310 provided in the apparatus 300, and the sample 500 to be inspected is taken into the inspection chip 100.
(2-2) Sampling Needle Sampling of the sample 500 to the intake port 105 is performed as follows. Here, FIG. 12 is a cross-sectional view of the inside of the intake port 105, and the sampling needle 250 and the spring 255 are built in the intake port 105. Except when the sample 500 is collected, the spring 255 is contracted so that the collection needle 250 is built into the intake port 105 as shown in FIG. When the sample 500 is collected, the spring 255 is extended as shown in FIG. 12B, the collection needle 250 protrudes from the intake port 105, and the sample 500 is collected from the collection needle 250. When the sample 500 is collected by the collection needle 250 in this way, the trouble of introducing the collected sample into the inspection chip 100 can be saved. Further, contamination of the sample 500 when introduced into the inspection chip 100 can be reduced.
(2-3) Partition plate The partition plate 110 is provided at intervals along the circumference of the same circle centered on a predetermined rotation shaft 310 as shown in FIG. Here, the rotation shaft 310 is attached to the central portion of the intake port 105. The partition plate 110 prevents the sample 500 from traveling to the outer peripheral side in the radial direction of the circle having the rotation axis 310 as the center by rotation about the predetermined rotation axis 310. The progression of the sample 500 toward the outer peripheral side in the radial direction is hindered by the partition plate 110, but the sample 500 proceeds toward the outer peripheral side in the radial direction through the plurality of partition plates 110. At this time, the sample 500 advances to the outer peripheral side in the radial direction so as to be dispersed through the partition plates 110 by rotation about the rotation shaft 310. Therefore, the sample 500 introduced into the inspection chip 100 can be dispersed and introduced into the plurality of centrifuge tubes 201. Therefore, introduction of the sample 500 into the unidirectional centrifuge tube 201 can be reduced, and a bias in the amount of the sample 500 to be introduced can be reduced.
(2-4) Centrifugal tube and holding unit The centrifugal tube 201 is connected to the intake port 105, and the sample 500 is introduced into the centrifugal tube 201 from the intake port 105 via the partition plate 110. The centrifuge tube 201 is generally U-shaped, and one open end is connected to the intake port 105, and the other open end is connected to a weighing tube 205 having a predetermined volume. A holding part 203 is provided at the bottom of the U-shape of the centrifuge tube 201. Further, the U-shaped opening of the centrifuge tube 201 is located on the rotating shaft 310 side, and the U-shaped bottom is located on the radially outer peripheral side of a circle centered on the predetermined rotating shaft 310. When the centrifuge tube 201 is arranged in this way, the target component 510 can be efficiently centrifuged when rotated about the rotation shaft 310. That is, the direction from the opening to the bottom of the U-shaped centrifuge tube 201 is substantially the same as the centrifugal force when rotated about the rotation shaft 310. Therefore, when the rotating shaft 310 is rotated as a center, the centrifugal force works most on the bottom of the centrifuge tube 201. Therefore, the target component 510 can be separated by efficiently moving the non-target component 520 other than the target component 510 from the sample 500 to the bottom of the centrifuge tube 201. Further, since the holding unit 203 is provided at the bottom of the centrifuge tube 201, the non-target component 520 that has moved to the U-shaped bottom by centrifugation can be introduced into the holding unit 203. Thus, by introducing the non-target component 520 into the holding unit 203, the target component 510 can be efficiently separated.

Further, it is preferable that the width of the connecting portion between the holding unit 203 and the centrifuge tube 201 is formed to be larger than the width of the centrifuge tube 201. When the width of the connection portion is formed large, when the sample 500 and the non-target component 520 are introduced into the holding unit 203, the air existing in the holding unit 203 is efficiently transferred from the holding unit 203 to the centrifuge tube 201. I can miss well.
Here, the centrifuge tube 201 is not limited to a U-shape, and may be formed to have, for example, a cup shape as shown in FIG.
(2-5) Weighing tube, transfer unit, air hole The weighing tube 205 is connected to the centrifuge tube 201, the transfer unit 207, and the first connection tube 209. The target component 510 centrifuged by the centrifuge tube 201 is introduced by the transfer means 207 so as to fill the weighing tube 205. The target component 510 is introduced into the weighing tube 205 by, for example, applying a pressure for sucking the target component 510 from the pump unit 333 of the apparatus 300 connected to the transfer unit 207 to the weighing tube 205 or the centrifuge tube 201. At this time, the weighing tube 205 has a desired volume for weighing the target component 510, and the target component 510 can be accurately weighed by taking out the target component 510 in the weighing tube 205. Here, in particular, the weighing tube 205 is generally U-shaped or V-shaped. When the first connection tube 209 is connected to the bottom of the U-shape or V-shape, the weighing tube 205 is efficiently targeted using centrifugal force. The component 510 is preferable because it can be introduced into the primary mixing section 217. Further, when the air hole 206 is provided in the connecting portion (see FIG. 10A) of the weighing tube 205 to the centrifuge tube 201 and the transfer means 207, the target component 510 is easily separated in the weighing tube 205. Therefore, it is preferable.

Further, as shown in FIG. 14, the transfer means 207 is formed in substantially the same shape as the U-shaped centrifuge tube 201, and the centrifuge tube 201 and the transfer means 207 are symmetrical with respect to the weighing tube 205. Preferably it is formed. If it forms as shown in FIG. 14, the movement of the target component 510 from the centrifuge tube 201 to the weighing tube 205 by the transfer means 207 can be performed easily and accurately. In addition, the target component 510 can be easily and accurately weighed in the weighing tube 205. When the line is symmetrical in this way, the distance between the rotating shaft 310 and each part of the weighing tube 205, the centrifuge tube 201 and the transfer means 207, for example, the distance L from the rotating shaft 310 to the left and right vertices of the weighing tube 205 in FIG. 'And L''are equal. Therefore, since the centrifugal force at the left and right vertices of the weighing tube 205 is equal, the target component 510 introduced into the weighing tube 205 at the left and right vertices of the weighing tube 205 is easily transferred to the left and right centrifuge tubes 201 and the transfer means 207. And can be separated evenly. Therefore, the target component 510 can be accurately weighed.
(2-6) Reagent reservoir, primary mixing unit, capillary valve The capillary valve 211 is connected to the weighing tube 205 via the first connection tube 209. The primary mixing unit 217 is connected to the capillary valve 211 via the second connection pipe 215. The reagent reservoirs 219a and 219b are connected to the primary mixing unit 217.
(2-6-1) Primary Mixing Unit The target component 510 in the weighing tube 205 is transferred from the weighing tube 205 to the primary mixing unit 217 by re-rotation around the same rotation axis 310 as the rotation of the centrifuge tube 201. Introduce. Here, the primary mixing unit 217 is located on the outer peripheral side in the radial direction of the circle centering on the predetermined rotation shaft 310 with respect to the weighing tube 205, and the inspection chip 100 is rotated about the predetermined rotation shaft 310. In this case, the target component 510 is efficiently introduced from the weighing tube 205 into the primary mixing unit 217.
(2-6-2) Reagent reservoir The reagent reservoir (219a, 219b) 219 is connected to the primary mixing unit 217, and the reagent 550 is stored therein. The reagent 550 in the reagent reservoir 219 is introduced into the primary mixing unit 217 by rotation about a predetermined rotation shaft 310. The introduction of the reagent 550 from the reagent reservoir 219 to the primary mixing unit 217 is performed by rotating at the time of introducing the sample 500 from the intake port 105 to the centrifuge tube 201, rotating at the time of centrifugation, or performing primary mixing from the weighing tube 205. It is preferable to carry out simultaneously with the introduction of the target component 510 to the part 217 and the rotation during mixing, since the process can be simplified and speeded up. Here, the number of reagent reservoirs 219 is not necessarily one, and a plurality of reagent reservoirs 219 can be provided according to the inspection item.

  The reagent reservoir 219 is preferably designed as shown in FIG. The reagent reservoir 219 a includes a reagent reservoir body 219 a ′, a reagent reservoir body 219 a ′, and a connecting portion 219 a ″ that connects the primary mixing portion 217. At this time, it is preferable that the design is made so that the acute angle θ formed by the connecting portion 219a ″ and the radial direction of the circle centered on the rotation shaft 310 is positioned on the rotation shaft 310 side so as to be smaller than 90 degrees. The reagent reservoir main body 219a 'is provided on the rotating shaft 310 side with respect to the connecting portion 219a' '. By designing in this way, a force in the direction of the primary mixing part 217 acts on the reagent 550 in the reagent reservoir main body 219a ′ and the connection part 219a ″ by the rotation about the rotation shaft 310, so that the reagent 550 is efficiently used. It can be well introduced into the primary mixing section 217.

In the reagent reservoir 219, the reagent 550 can be placed in the capsule as follows. FIG. 16A is a plan view showing a state in which the reagent enclosed in the capsule is placed in the reagent reservoir, and FIGS. 16B and 16C are schematic views showing how the reagent flows out from the reagent reservoir. is there.
In the reagent reservoir 219 portion of the test chip 100, a space 605 for placing the capsule 600 in which the reagent 550 is enclosed, a reagent introduction unit 607 for introducing the reagent 550 into the primary mixing unit 217, a lid 610, and A suction port 630 for applying pressure to the lid 610 is provided. In addition, a protrusion 609 is provided at a position facing the reagent 550 in the test chip 100 forming the space 605. In addition, a lid 610 that covers the reagent reservoir 619 is provided in the upper portion of the space 605. The lid 610 has an extruding part 615 at a position facing the protrusion 609. When the pressure in the direction of pushing the capsule 600 is not applied to the lid 610, the capsule 600 is not pierced by the protrusion 609 as shown in FIG. On the other hand, for example, when the force for sucking the air between the lid 610 and the test chip 100 works through the suction port 630 and pressure in the capsule 600 direction is applied to the reagent reservoir 619, the protrusion 609 is pushed by the pusher 615. It is. Then, as shown in FIG. 16C, the protrusion 609 breaks through the capsule 600 and causes the reagent 550 to flow out of the capsule 600. The reagent 550 that has flowed out is introduced into the primary mixing unit 217 from the reagent introduction unit 607 connected to the primary mixing unit 217. With such a configuration, the reagent 550 can be held in the capsule 600, so that contact between the reagent 550 and the outside can be avoided. Therefore, it is possible to prevent pH change due to dissolution of carbon dioxide in the air and degradation of enzymes and pigments due to light. The capsule 600 may be broken by pressing the lid 610 from the outside of the substrate. The material of the capsule 600 is preferably an aluminum / plastic composite.
(2-6-3) Capillary valve The capillary valve 211 is connected to the weighing pipe 205 via the first connection pipe 209 so that the flow path width is larger than that of the weighing pipe 205 and / or the first connection pipe 209. Is formed. Here, the flow path width is a length in a direction substantially along the circumference of a circle centered on the rotation shaft 310, and is a length indicated by L in FIG. When the target component 510 is introduced into the weighing tube 205, the capillary valve 211 prevents the target component 510 from being introduced from the weighing tube 205 into the first connection tube 209 and the primary mixing unit 217. This is because the target component 510 in the weighing tube 205 is in contact with the capillary valve 211 having a larger flow path width than the weighing tube 205, thereby reducing the surface area of the target component 510 and keeping the free energy small. In addition, the capillary valve 211 prevents the target component 510 introduced from the weighing tube 205 into the primary mixing unit 217 from flowing back into the weighing tube 205 due to capillary action. Therefore, the target component accurately weighed can be introduced into the primary mixing unit 217. It is effective to provide a capillary valve when the contact angle between the target component 510 and the flow path wall or the substrate of each part that the target component 510 contacts is smaller than 90 degrees.
(2-7) Secondary Mixing Unit The secondary mixing unit 220 is connected to the primary mixing unit 217, and further mixes the mixture material 560 in which the target component 510 and the reagent 550 are mixed in the primary mixing unit 217. To do. The secondary mixing unit 220 includes a plurality of connected mixer units 220a. The mixer unit 220a is configured, for example, as shown in FIG. The mixer unit 220 a has an H-shaped wall 225, and a microchannel 227 is formed so as to surround the H-shaped wall 225. Such a fine microchannel 227 can increase the integration rate of the secondary mixing unit 220 and reduce the area of the quantitative unit 200.
(2-8) Photodetection Path, Light Entrance, Light Exit, and Takeout The mixed substance 560 in which the reagent 550 and the target component 510 are mixed in the secondary mixing unit 220 is introduced into the light detection path 230. Light is introduced into the light detection path 230 from the light entrance 233, and light after passing through the light detection path 230 is taken out from the light exit 235. Then, the target component 510 is quantified by measuring the amount of transmitted light. The light detection path 230 is preferably coated with a material having a high light reflectance such as Al. The light entrance 233 and the light exit 235 are optical waveguides, and as these materials, materials having a refractive index higher than that of the upper and lower substrates and easily collecting light are used. In addition, when performing ultraviolet light measurement, a material having a higher ultraviolet light transmittance than the upper and lower substrates is used. The light inlet 233 and the light outlet 235 are created by forming the upper and lower substrates on the upper and lower substrates, for example, after forming the portions other than the optical waveguide of the light inlet 233 and the light outlet 235, and then molding the upper and lower substrates by injection molding.
(3) Method for Using Test Chip Next, an example of a method for using the test chip 100 when quantifying the target component 510 from the sample 500 will be described with reference to FIGS.

  Step 1: First, as shown in FIG. 17, the inspection chip 100 is fixed to the turntable 301 so that the rotation shaft 310 of the apparatus 300 and the intake port 105 substantially coincide with each other. Then, the sample 500 is collected using the collection needle 250 with the spring 255. As the sample 500, for example, blood is collected from a finger as shown in FIG. Next, the sample 500 is quantified as follows.

  Step 2: The inspection chip 100 is rotated around a predetermined rotation axis 310, the collected sample 500 is moved to the outer peripheral side in the radial direction so as to be dispersed by the partition plate 110, and centrifuged as shown in FIG. It introduces into the separation tube 201. At this time, the sample 500 introduced into the intake port 105 behaves as indicated by an arrow shown in FIG. 19B and is introduced into the centrifuge tube 201 provided in four directions.

Step 3: The reagent 550 is introduced from the reagent reservoir 219 to the primary mixing unit 217 by rotating the test chip 100 about a predetermined rotation shaft 310 (see FIG. 20).
Step 4: The target component 510 is centrifuged from the sample 500 in the centrifuge tube 201 by rotating the test chip 100 around a predetermined rotation shaft 310 (see FIG. 21). At this time, centrifugal force acts on the U-shaped centrifuge tube 201 from the opening of the centrifuge tube 201 toward the bottom by rotation about the predetermined rotation shaft 310. Therefore, non-target components 520 other than the target component 510 in the sample 500 move to the bottom of the centrifuge tube 201 and are introduced into the holding unit 203. In this way, the target component 510 is separated from the sample 500.

  Step 5: Next, the target component 510 centrifuged in the centrifuge tube 201 is introduced from the centrifuge tube 201 into the weighing tube 205 so as to fill the weighing tube 205 (see FIG. 22). The target component 510 is introduced into the weighing tube 205 by the transfer means 207. For example, the pump unit 333 of the apparatus 300 is connected to the transfer unit 207, and a pressure for sucking the target component 510 is applied to the weighing tube 205 and the centrifuge tube 201. As a result, the target component in the centrifuge tube 201 is sucked into the weighing tube 205. At this time, the weighing pipe 205 and the capillary valve 211 are connected via the first connection pipe 209, and the flow path width of the capillary valve is formed to be larger than that of the first connection pipe 209 and / or the weighing pipe 205. Has been. Therefore, introduction of the target component 510 in the weighing pipe 205 from the weighing pipe 205 to the capillary valve 211 via the first connection pipe 209 is prevented. This is because the target component 510 in the weighing tube 205 attempts to reduce the surface area of the target component 510 and keep the free energy small.

  Step 6: Further, the test chip 100 is rotated again around the same rotation shaft 310 as that in the centrifugation in Step 4, and the target component 510 in the weighing tube 205 is introduced into the primary mixing unit 217 (see FIG. 23). . Further, in the primary mixing unit 217 by rotation about the rotation shaft 310, the target component 510 and the reagent 550 are mixed to obtain a mixed substance 560 (see FIG. 24).

  Here, the weighing tube 205 has a desired volume, and a desired amount of the target component 510 can be obtained by taking it out from the weighing tube 205 to the primary mixing unit 217. Specifically, the target component 510 located between the connection portion CC ′ between the centrifuge tube 201 and the weighing tube 205 in FIG. 23 and the connection portion DD ′ between the weighing tube 205 and the transfer means 207 includes: It is introduced into the primary mixing unit 217 by centrifugal force. Since the target component 510 located in the other part is introduced into the centrifuge tube 201 and the transfer means 207, the target component 510 accurately weighed can be obtained.

If the introduction of the target component 510 from the weighing tube 205 into the primary mixing unit 217 and the mixing of the target component 510 and the reagent 550 in the primary mixing unit 217 are performed at the same rotation, the test chip 100 is handled. This is preferable because it is easy and the mixed material 560 can be obtained quickly.
Step 7: The mixture material 560 in which the target component 510 and the reagent 550 are mixed in the primary mixing unit 217 is introduced into the secondary mixing unit 220 and further mixed (see FIG. 25).

Step 8: The mixed substance 560 is introduced into the light detection path 230 (see FIG. 26).
Step 9: When the bubble 570 exists in the light detection path 230 filled with the mixed substance 560, the inspection chip 100 is rotated again by the rotation around the predetermined rotation axis 310, and the light detection path 230 is removed from the light detection path 230. The bubble 570 is removed (see FIG. 27).
In order to remove bubbles more effectively, an air hole may be provided in the light detection path 230 as shown in FIG. In FIG. 28A, an air hole 247 is provided on the rotating shaft 310 side of the light detection path 230. Here, when the bubbles 570 existing in the mixed substance 560 introduced into the light detection path 230 around the rotation axis 310 are removed, the air holes 247 are closest to the rotation axis 310 in the light detection path 230. Provide in position. That is, the air hole 247 is provided at a position where the distance L3 from the position in the light detection path 230 where the air hole 247 is provided is smaller than the distances L2 and L4 from the position where the light detection path 230 is located to the rotation shaft 310. . When the air hole 247 is provided in this way, when the test chip 100 is rotated around the rotation shaft 310, the bubbles 570 are removed from the air hole 247 as shown in FIG. On the other hand, when the air hole 247 is not provided, a space 575 is generated between the light detection path 230 and the mixed material 560 by rotation about the rotation shaft 310. Therefore, it is difficult to remove the bubbles 570 due to the space 575 or the bubbles 570 are mixed into the mixed material 560.

Step 10: Light is introduced into the light detection path 230 from the light entrance 233, and light after passing through the light detection path 230 is taken out from the light exit 235. The target component 510 is quantified by measuring the amount of transmitted light.
The step of introducing the reagent in step 3 includes the step of introducing the target component 510 into the centrifuge tube 201 from the inlet 105 in step 2, the step of separating the target component 510 in the centrifuge tube 201 in step 4, and the step. 6 may be introduced simultaneously with the introduction of the target component 510 into the primary mixing unit 217. By introducing the reagents simultaneously, the mixed substance can be obtained quickly.
(4) Effect By handling the inspection chip 100 into which the sample 500 is introduced as described above, the target component 510 in the sample 500 can be separated, weighed, mixed with the reagent, and quantified in a batch. Since the target component 510 can be quantified without being taken out of the inspection chip 100, contamination of the target component 510 can be reduced and accurately quantified. At this time, the target component 510 introduced into the weighing tube 205 is weighed and taken out by the volume of the weighing tube 205, so that the target component 510 can be accurately weighed. Therefore, it is possible to obtain a mixed material 560 in which the reagent 550 and the target component 510 have a desired mixing ratio. Furthermore, since it is not necessary to apply heat or the like during separation, weighing, mixing with a reagent, and determination, the sample 500 is not affected by heat or the like. Therefore, contamination of the sample 500 can be reduced and the target component 510 contained in the sample 500 can be accurately weighed. Further, the rotation axis 310 of rotation for performing centrifugation in the centrifuge tube 201, introduction of the reagent into the primary mixing unit 217, mixing of the reagent 550 and the target component 510 in the primary mixing unit 217, and the like is the same. Therefore, the test chip 100 can be easily designed, and the test chip 100 can be downsized.
<Second Embodiment>
FIG. 29 is a plan view of a test chip according to the second embodiment of the present invention, and FIG. 30 is an enlarged view of a main part of FIG. The weighing tube shown in FIG. 30 of the second embodiment and an adjustment tube to be described later are different from the first embodiment except for the other configurations, and the same reference numerals represent the same components.

  The test chip 100 includes an intake port 105, a centrifuge tube 201, a holding unit 203, a weighing tube 650, an air hole 655, and a quantitative unit 640. The first connection pipe 209, the capillary valve 211, the second connection pipe 215, the primary mixing section 217, the reagent reservoirs 219a and 219b in which the reagent is stored, and the secondary mixing provided in the quantification section 200 of the first embodiment. The same components as the unit 220, the light detection path 230, the light inlet 233, the light outlet 235, and the outlet 240 are included in the quantitative unit 640 shown in FIG. As shown in FIG. 30, the intake port 105 is connected to one open end of a substantially U-shaped centrifuge tube 201. A weighing tube 650 and an adjustment tube 660 are connected to the other open end.

  The weighing tube 650 is composed of a series of V-shaped V-shaped tubes. The V-shaped bottom portion of the weighing tube 650 is located on the radially outer peripheral side of a circle centering on a predetermined rotation shaft 310 corresponding to the intake port 105. A first connection pipe 209 following the capillary valve 211 is connected to the bottom of the V shape. Each V-shape is substantially the same in shape or according to each inspection item, and is formed such that the distance L1 from the center 310 'of the rotating shaft 310 to the bottom of the V-shape is the same.

  The adjustment tube 660 is connected to the adjustment tube 660 and the centrifuge tube 201, and is connected to the connection portion 651 shown in the α portion of FIG. Part 653. The connection portion 651 of the α portion is located closer to the rotating shaft 310 than the reservoir portion 653. At this time, if the α portion of the adjustment tube 690 is formed in an inverted U shape opposite to the U shape of the centrifuge tube 201 and the β portion is formed in a U shape, the sample 500 and the centrifuge tube 201 in the adjustment tube 660 are formed. Separation from the sample 500 is easy and preferable.

  Here, before the target component 510 is centrifuged, the sample 500 is first introduced into the centrifuge tube 201 and the adjustment tube 660, and the centrifuge tube 201 is filled with the sample 500. That is, as shown in FIG. 31A, the centrifuge tube 201 is filled with the sample 500, and the sample 500 is introduced into the connection portion 651 and the reservoir portion 653 of the adjustment tube 660. When rotating around the rotation shaft 360 in this state, the sample 500 is separated with the connection portion 651 as a boundary. Specifically, the sample 500 on the side of the centrifuge tube 201 (the centrifuge tube 201 from E-E ′ in FIG. 31A) from the connection portion 651 is introduced into the centrifuge tube 201 and centrifuged. On the other hand, the sample 500 on the adjustment tube 660 side (the adjustment tube 660 from E-E ′ in FIG. 31A) from the connection portion 651 is centrifuged in the reservoir 653. Thus, by centrifuging with the centrifuge tube 201, a substantially constant amount of the target component 510 can be obtained from a constant amount of sample filling the centrifuge tube 201. When the sample 500 introduced into the test chip 100 is blood and the target component 510 is plasma, it is preferable to design the centrifuge tube 201 and the holding unit 203 as follows in order to obtain a certain amount of plasma. Since the ratio of blood cells in the blood is about 30 to 40%, the ratio of the volume of the holding unit 203 to the centrifuge tube 201 is centrifuge when the total volume of the centrifuge tube 201 and the holding unit 203 is 100%. The tube 201 is designed so that the holding portion 203 = 50%: 50%. It is preferable that the volume ratio is centrifuge tube 201: holding portion 203 = 60%: 40% because only blood cell components are introduced into holding portion 203, so that plasma can be centrifuged without waste. For example, if the volume of the holding unit 203 is 50% or more, a lot of plasma in the blood is introduced into the holding unit 203, so that the plasma component is wasted. On the other hand, when the volume of the holding unit 203 is 40% or less, blood cell components overflow from the holding unit 203, and thus it is difficult to separate plasma components.

  Next, the target component 510 centrifuged by the centrifuge tube 201 is introduced by the transfer means 207 so as to fill the continuous V-shaped weighing tube 650. Then, the target component 510 in the weighing tube 650 is introduced into the primary mixing unit 217 through the first connection tube 209 and the capillary valve 211 by rotation around the rotation shaft 310. Thus, the centrifuge tube 201 can be made into one by making the weighing tube 650 into the continuous V shape. Therefore, the weighing portion and the quantitative portion 640 can be densely provided in the inspection chip 1. Therefore, the inspection chip 100 having a plurality of quantification units can be reduced in size.

  Furthermore, it is preferable to provide an air hole 655 in the V-shaped peak portion of the weighing tube 650 located on the rotating shaft 310 side. When the air hole 655 is provided in the V-shaped peak portion, the target component 510 that fills the weighing tube 650 as shown in FIG. 32A due to air flowing in from the peak portion is shown in FIG. Can be easily separated and introduced into the primary mixing section 217 via the first connection pipe 209. Moreover, it is preferable that the first connecting pipe 209 has a hydrophobic surface in the pipe. If the hydrophobic treatment is performed, it is possible to further prevent the target component 510 from flowing into the first connecting pipe 209 due to capillary action. Here, in the case where Al is used in the step of performing the hydrophobic treatment, the hydrophobic treatment can be performed in the same step as the Al sputtering in the light detection path.

Here, the centrifuge tube 201 is not limited to a U-shape, and may be formed to have a cup shape, for example, as shown in FIG. Further, the adjustment tube 650 can be provided on the left side of the centrifuge tube 201 in FIG.
<Third Embodiment>
FIG. 34 is a plan view of an inspection chip according to the third embodiment of the present invention, and FIG. 35 is an enlarged view of the main part of FIG. The third embodiment is largely the same as the first embodiment except that the shape of the weighing tube and the installation position of the capillary valve are different, and the same reference numerals denote the same components.

  The weighing tube 205 of the third embodiment is V-shaped. Further, the transfer means 207 is formed in substantially the same shape as the U-shaped centrifuge tube 201, and the centrifuge tube 201 and the transfer means 207 are formed symmetrically about the weighing tube 205. Further, between the weighing tube 205 and the primary mixing unit 217, a capillary valve 211a adjacent to the weighing tube 205, a capillary valve 211b adjacent to the primary mixing unit 217, the primary mixing unit 217 and the secondary mixing unit are provided. And a capillary valve 211d provided between the secondary mixing unit 220 and the light detection path 230.

In the third embodiment, the weighing tube 205 is V-shaped, and the centrifuge tube 201 and the transfer means 207 are formed symmetrically with respect to the weighing tube 205, so that they are introduced into the weighing tube 205. The target component 510 can be accurately weighed and introduced into the primary mixing unit 217. The capillary valve 211a prevents the target component 510 introduced into the weighing tube 205 from leaking out of the weighing tube 205. The capillary valve 211b prevents the target component 510 introduced into the primary mixing unit 217 from flowing back from the primary mixing unit 217 to the weighing tube 205. The capillary valve 211c mainly suppresses the natural introduction of the mixed substance 560 from the primary mixing unit 217 to the secondary mixing unit 220 having a channel width or volume smaller than that of the primary mixing unit 217. That is, the capillary valve 211c can prevent the reagent 550 and the target component 510 from being introduced into the secondary mixing unit 220 in a state where they are hardly mixed. For example, only the reagent 550 or the target component 510 is prevented from being introduced into the secondary mixing unit 220. Then, the mixed material 560 in which the reagent 550 and the target component 510 are mixed is introduced into the secondary mixing unit 220 from the primary mixing unit 217 by further centrifugation or by suction by a pump means, and further mixing is performed. I do. The capillary valve 211d prevents the mixed material 560 introduced into the light detection path 230 from flowing back from the light detection path 230 to the secondary mixing unit 220 having a flow path width or volume smaller than that of the light detection path 230.
<Experimental example 1>
In Experimental Example 1, an experiment was conducted to prove the usefulness of a capillary valve using an inspection chip having a capillary valve. FIG. 36 is a photograph showing the main part of the inspection chip 1 used in Experimental Example 1. The test chip 1 shown in Experimental Example 1 includes a V-shaped weighing pipe 700, a first connection pipe 705, a leakage prevention capillary valve 710, a second connection pipe 715, a backflow prevention capillary valve 720, and a liquid reservoir 730. ing. These are connected in order toward the outer peripheral side in the radial direction of a circle centered on the rotation axis. The flow channel width of the weighing tube 700 is 200 μm, the volume of the broken line portion γ of the weighing tube 700 is 0.33 μl, the V-shaped angle θ of the weighing tube 700 is 60 degrees, and the flow channel width of the leakage preventing capillary valve 710 is 1000 μm, The channel width of the backflow prevention capillary valve 720 is 1000 μm, the channel width of the liquid reservoir 730 is 4000 μm, and the channel depth of all channels is 200 μm. A standard serum colored with ink was introduced into the weighing tube 700. Then, the position of the standard serum after a rotation time of 30 seconds was observed at a rotation radius of 1.3 cm and a rotation number of 3000 rpm.
<Comparative Example 1>
In Comparative Example 1, an experiment similar to that in Experimental Example 1 was performed using a test chip in which the leakage preventing capillary valve 710 and the backflow preventing capillary valve 720 of Experimental Example 1 were not provided.

  The results of Experimental Example 1 are shown in FIGS. The results of Comparative Example 1 are shown in FIGS. 37 and 39 are compared, in the test chip used in Experimental Example 1, the standard serum in the weighing tube 700 fills the weighing tube 700. That is, the leakage prevention capillary valve 710 prevents the standard serum from leaking from the weighing tube 700 (see FIG. 37). On the other hand, in the test chip used in Comparative Example 1, the leakage prevention capillary valve 710 is not provided, the standard serum leaks from the weighing tube 700, and the weighing tube 700 is not completely filled (FIG. 39). reference). In the test chip used in Experimental Example 1, the standard serum introduced into the liquid reservoir 730 is introduced and held in the liquid reservoir 730 (see FIG. 38). On the other hand, in the test chip used in Comparative Example 1, the backflow prevention capillary valve 720 is not provided, and the standard serum flows backward from the liquid reservoir 730 (see FIG. 40).

  Therefore, it has been demonstrated that the provision of the capillary valve can prevent the leakage of the solution in the weighing tube and the backflow of the solution introduced into the liquid reservoir. Since the capillary valve in Experimental Example 1 effectively prevents solution leakage and backflow, it is preferable to design the capillary valve and the primary mixing unit 217 integrally as shown in FIG.

  If this invention is used, isolation | separation and weighing of the target component in a sample can be performed collectively and simply. At this time, the target component introduced into the weighing tube is accurately weighed. Further, since the rotation axis of the chip in the separation and weighing is the same, the separation and weighing of the target component is easy, and the chip can be miniaturized. Therefore, it can be used for a portable inspection chip.

(A) The perspective view of the test | inspection chip based on this invention. (B) The perspective view of another test | inspection chip based on this invention. The top view of Fig.1 (a). Schematic of an apparatus on which an inspection chip is placed. An example (1) of the usage method of a test | inspection chip. An example (2) of the usage method of a test | inspection chip. An example (3) of the usage method of a test | inspection chip. An example (4) of the usage method of a test | inspection chip. (A) Inspection tip with sampling needle and syringe. (B) Inspection chip with a dropper. The top view of the test | inspection chip which concerns on the example of 1st Embodiment of this invention. (A) The enlarged view of a fixed_quantity | quantitative_assay part. (B) An example of a centrifuge tube. (A) Enlarged view of the inlet. (B) The enlarged view of a mixer part. (A) Sectional drawing of the intake in an unused state. (B) Sectional drawing of the intake port in use condition. Schematic of an apparatus on which an inspection chip is placed. Explanatory drawing of the principal part formed in line symmetry. Explanatory drawing of the inclination of a reagent reservoir. (A) A state in which the reagent enclosed in the capsule is placed in the reagent reservoir. (B) The schematic diagram (1) which shows a mode that a reagent flows out from a reagent reservoir. (C) The schematic diagram (2) which shows a mode that a reagent flows out from a reagent reservoir. An example (1) of the usage method of the test | inspection chip 100. FIG. An example (2) of the usage method of the test | inspection chip 100. FIG. (A) An example (3-1) of using the test chip 100. (B) An example of usage of the inspection chip 100 (3-2). An example (4) of using the test chip 100. An example (5) of how to use the test chip 100. An example (6) of the usage method of the test | inspection chip 100. FIG. An example (7) of using the test chip 100. An example (8) of using the test chip 100. An example (9) of using the test chip 100. An example (10) of how to use the inspection chip 100. An example (11) of using the test chip 100. (A) Explanatory drawing (1) of the air hole position of a photon detection path. (B) Explanatory drawing (2) of the air hole position of a photon detection path. The top view of the test | inspection chip which concerns on 2nd Embodiment of this invention. The enlarged view of the principal part of FIG. (A) Explanatory drawing of an adjustment pipe. (B) Explanatory drawing of an adjustment pipe. (A) Explanatory drawing (1) of the behavior of the target component in a weighing tube. (B) Explanatory drawing (2) of the behavior of the target component in a weighing tube. The top view of the test | inspection chip which concerns on the example of 3rd Embodiment of this invention. The top view of the test | inspection chip which concerns on the example of 3rd Embodiment of this invention. The enlarged view of the principal part of FIG. The photograph which shows the principal part of the test | inspection chip 1 used in Experimental example 1. FIG. Result (1) of Experimental Example 1. Result (2) of Experimental Example 1. Result (1) of Comparative Example 1. Result (2) of Comparative Example 1.

Explanation of symbols

1, 100: Inspection chip
7, 105: Inlet
9, 201: Centrifuge tube
11, 205, 650, 700: Weighing tube
13, 207: Transfer means
15, 240: Exit
20, 300: apparatus
23, 310: Rotating shaft
40, 500: Sample
41, 510: target component
50, 250: sampling needle
110: Partition plate
200: Quantitative part
203: Holding unit
209: First connecting pipe
211: Capillary valve
215: Second connecting pipe
217: Primary mixing section
219: Reagent reservoir
220: Secondary mixing section
230: Photodetection path
233: Light entrance
235: Light exit
247, 655: Air hole
301: Turntable
550: Reagent
560: mixture material
570: Bubble
660: Adjustment pipe
710: Capillary valve for leak prevention
720: Backflow prevention capillary valve

Claims (9)

A method for using a chip having a reagent reservoir for holding a reagent, a mixing section connected to the reagent reservoir and the weighing tube, and into which a sample containing a target component is introduced,
A separation step of centrifuge the target component from the sample by rotating the chip around a predetermined rotation axis;
A target component introduction step of introducing the target component into a weighing tube having a predetermined volume;
A weighing step of rotating the tip again around the rotation axis and weighing the target component introduced into the weighing tube;
A reagent introduction step of introducing the reagent from the reagent reservoir into the mixing unit by rotating the chip about the rotation axis;
A mixing step of rotating the chip around the rotation axis, introducing the target component weighed and collected in the weighing step into the mixing unit, and mixing with the reagent;
How to use the chip including. The method according to claim 1 , wherein the mixing step is simultaneous with the weighing step. The method of using a chip according to claim 1 , wherein the reagent introduction step is simultaneous with a separation step, a weighing step, or a mixing step. A test chip having at least one quantification unit for quantifying a target component in a sample,
The quantitative unit is
A centrifuge tube for centrifuging the target component from the sample by rotating the inspection chip around a predetermined rotation axis;
A weighing tube connected to one end of the centrifuge tube and weighing the target component by rotation about the rotation axis;
A transfer means connected to the weighing tube for transferring the target component from the centrifuge tube to the weighing tube;
At least one reagent reservoir in which reagents are stored;
The target component is connected to the reagent reservoir and the weighing tube, and the target component introduced from the weighing tube by re-rotation about the rotation axis is rotated by the rotation of the target component about the rotation axis. A mixing section that is mixed with a reagent introduced from a reagent reservoir; and a light detection path that is connected to the mixing section and allows a mixed substance in which the reagent and the target component are mixed to pass therethrough;
A light inlet connected to the light detection path for introducing light into the light detection path;
An inspection chip having an optical outlet connected to the optical detection path and for extracting light after passing through the optical detection path. The inspection chip according to claim 4 , further comprising a capillary valve having a flow path width larger than that of the weighing tube between the weighing tube and the mixing unit. The weighing tube is generally U-shaped, the weighing tube opens to the rotating shaft side, one end of the U-shape of the weighing tube is connected to the centrifuge tube, and the other end is the transfer means The inspection component according to claim 4 , wherein a U-shaped bottom portion is connected to the mixing portion, and the target component is moved from the bottom portion of the weighing tube to the mixing portion by rotation about the rotation axis. Chip. The inspection chip according to claim 4 , wherein the transfer means is connected to a pump. The inspection chip according to claim 4 , further comprising a holding portion that is provided at a bottom portion of the centrifuge tube and into which a component other than the target component in the sample is introduced by rotation around the rotation axis. An inlet for taking in a sample to be introduced into two or more of the quantification units, provided on the rotating shaft;
A plurality of partition plates provided at intervals along the circumference of the same circle around the rotation axis;
The inspection chip according to claim 4 , wherein the partition plate prevents a sample taken in from the take-in port from proceeding to a radially outer peripheral side of a circle having the rotation axis as a center.

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