WO2017115733A1 - 試料分析用基板、試料分析装置、試料分析システムおよび試料分析システム用プログラム - Google Patents
試料分析用基板、試料分析装置、試料分析システムおよび試料分析システム用プログラム Download PDFInfo
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Definitions
- the present application relates to a sample analysis substrate, a sample analysis device, a sample analysis system, and a sample analysis system program.
- Patent Document 1 uses a disk-shaped sample analysis substrate on which a channel, a chamber, and the like are formed, and rotates the sample analysis substrate to transfer, distribute, and mix the components in the sample solution. The technology which performs analysis etc. of this is disclosed.
- Analysis of specific components in a sample includes an analysis method using complicated reaction steps using enzyme reaction, immune reaction, and the like. There has been a demand for a technique capable of performing an analysis method through such a complicated reaction step in a sample analysis substrate.
- Non-limiting exemplary embodiments of the present application provide a sample analysis substrate, a sample analysis apparatus, a sample analysis system, and a sample analysis that can be applied to an analysis method in which components in a specimen are analyzed through more complicated reaction steps. Provide system programs.
- a sample analysis substrate that transfers liquid by the rotational motion of the present disclosure includes a substrate having a rotation axis, and a first holding chamber that is located in the substrate and has a first space for holding the first liquid.
- a reaction chamber located in the substrate and having a space for holding a liquid sample containing a specimen, and a first flow path located in the substrate and having a first opening and a second opening. The first opening and the second opening are located in the substrate, the first flow path connected to the first holding chamber and the reaction chamber, respectively, and the ligand is fixed to the liquid sample containing the specimen and the surface.
- a main chamber having a space for holding the magnetic particles formed, and a second flow path located in the substrate and having a third opening and a fourth opening, wherein the third opening and the fourth opening Open Are each provided with a second flow path connected to the reaction chamber and the main chamber, and a magnet storage chamber located in the substrate and capable of storing a magnet, wherein the first opening has the second opening.
- the magnet storage chamber When the second opening is positioned closer to the rotation axis than the third opening, and the magnet storage chamber has a magnet stored in the magnet storage chamber.
- the magnetic particles in the main chamber can be captured by the magnet in the main chamber.
- the sample analysis substrate According to the sample analysis substrate, the sample analysis apparatus, the sample analysis system, and the sample analysis system program according to one aspect of the present application, it is possible to cope with an analysis method in which components in a specimen are analyzed through complicated reaction steps.
- FIG. 1 is an example of a schematic diagram illustrating a sandwich immunoassay method using magnetic particles.
- FIG. 2A is a schematic diagram illustrating an example of a configuration of the sample analysis system of the embodiment.
- FIG. 2B is a schematic diagram illustrating an example of a configuration for detecting the origin of the sample analysis substrate in the sample analysis system.
- FIG. 3A is an exploded perspective view showing an example of a sample analysis substrate.
- FIG. 3B is a plan view showing an example of a sample analysis substrate.
- FIG. 3C is a plan view showing a configuration relating to the transfer of the reaction liquid in the sample analysis substrate shown in FIG. 3A.
- FIG. 3A is an example of a schematic diagram illustrating a sandwich immunoassay method using magnetic particles.
- FIG. 2A is a schematic diagram illustrating an example of a configuration of the sample analysis system of the embodiment.
- FIG. 2B is a schematic diagram illustrating an example of a configuration for detecting the origin of the sample analysis substrate in the
- FIG. 3D is a plan view showing a configuration relating to a siphon structure of a main chamber and a third flow path in the sample analysis substrate shown in FIG. 3A.
- FIG. 3E is a perspective view showing an example of a method for holding a magnet of the sample analysis substrate.
- FIG. 3F is a plan view showing a configuration related to the transfer of the cleaning liquid in the sample analysis substrate shown in FIG. 3A.
- FIG. 3G is an enlarged plan view showing a part of the structure of the first flow path of the sample analysis substrate.
- FIG. 3H is a plan view showing a configuration related to the transfer of the substrate solution in the sample analysis substrate shown in FIG. 3A.
- FIG. 4 is a flowchart showing an example of the operation of the sample analysis system.
- FIG. 4 is a flowchart showing an example of the operation of the sample analysis system.
- FIG. 5 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 6 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 7 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 8 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 9 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 10 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 11 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 12 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 13 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 14 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 15 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 16 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 17 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 18 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 19 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 20 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 21 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 22 is a diagram schematically illustrating an example of the stop angle of the sample analysis substrate and the position of the liquid during the operation of the sample analysis system.
- FIG. 23A is a plan view showing another example of the sample analysis substrate.
- FIG. 23B is a plan view showing another example of the sample analysis substrate.
- FIG. 24A is a plan view showing another example of the sample analysis substrate.
- FIG. 24B is a plan view showing another example of the sample analysis substrate.
- FIG. 24C is a plan view showing another example of the sample analysis substrate.
- FIG. 25A is a plan view showing another example of the sample analysis substrate.
- FIG. 25B is a plan view showing another example of the sample analysis substrate.
- FIG. 25C is a plan view showing another example of the sample analysis substrate.
- FIG. 26 is a plan view showing another example of the sample analysis substrate.
- FIG. 27 is a plan view showing another example of the sample analysis substrate.
- FIG. 28A is a plan view showing another example of the reaction chamber.
- FIG. 28B is a plan view showing another example of the reaction chamber.
- FIG. 28C is a plan view showing another example of the reaction chamber.
- FIG. 28D is a plan view showing another example of the reaction chamber.
- FIG. 28E is a plan view showing another example of the reaction chamber.
- a binding reaction between an analyte as an analysis target and a ligand that specifically binds to the analyte may be used.
- analysis methods include immunoassay methods and genetic diagnosis methods.
- immunoassay methods include competitive methods and non-competitive methods (sandwich immunoassay methods).
- An example of a gene diagnosis method is a gene detection method by hybridization.
- magnetic particles sometimes referred to as “magnetic beads”, “magnetic particles” or “magnetic beads”.
- a sandwich immunoassay method using magnetic particles will be specifically described.
- an antigen-antibody reaction between a primary antibody 304 immobilized on the surface of a magnetic particle 302 (hereinafter referred to as “magnetic particle-immobilized antibody 305”) and an antigen 306 as a measurement object.
- the secondary antibody to which the labeling substance 307 is bound (hereinafter referred to as “labeled antibody 308”) and the antigen 306 are bound by an antigen-antibody reaction.
- labeled antibody 308 the secondary antibody to which the labeling substance 307 is bound
- a complex 310 in which the magnetic particle-immobilized antibody 305 and the labeled antibody 308 are bound to the antigen 306 is obtained.
- the signal based on the labeled substance 307 of the labeled antibody 308 bound to the complex 310 is detected, and the antigen concentration is measured according to the amount of the detected signal.
- the labeling substance 307 include enzymes (for example, peroxidase, alkaline phosphatase, luciferase, etc.), chemiluminescent substances, electrochemiluminescent substances, fluorescent substances, etc., and dyes corresponding to the respective labeling substances 307, Signals such as luminescence and fluorescence are detected.
- the sandwich immunoassay method using magnetic particles has been described as an example, but B / F separation is performed by immunoassay or hybridization using competitive or non-competitive methods regardless of the presence or absence of magnetic particles. Necessary for gene detection.
- magnetic particles for example, a ligand immobilized on a solid phase composed of a material such as polystyrene or polycarbonate by physical adsorption, a ligand immobilized on a solid phase by chemical bonding, a metal composed of gold, etc.
- SAM self-assembled monolayer
- the magnetic particles including the composite 310 In order to sufficiently perform the B / F separation, it is preferable to wash the magnetic particles including the composite 310 with a washing solution a plurality of times. Specifically, first, in the reaction solution containing the complex 310, the unreacted antigen 306, the labeled antibody 308, and the like, only the reaction solution is removed while the complex 310 containing the magnetic particles is captured by the magnet. . Thereafter, a cleaning liquid is added to clean the composite 310, and the cleaning liquid is removed. By repeating this washing a plurality of times, B / F separation in which unreacted substances and non-specifically adsorbed substances are sufficiently removed can be achieved.
- the complex 310 is reacted with the substrate solution, and a signal based on the labeling substance 307 is generated.
- the remaining reaction solution is transferred to the chamber holding the complex 310 and reacts with the substrate solution, thereby causing an erroneous signal. May be generated. Measurement errors due to such signals are suppressed, and more accurate signal measurement is required.
- a sample analysis substrate, a sample analysis device, a sample analysis system, and a program for the sample analysis system were conceived.
- a sample analysis substrate, a sample analysis device, a sample analysis system, and a sample analysis system program according to an aspect of the present application are as follows.
- a sample analysis substrate that transfers liquid by a rotational motion, A substrate having a rotation axis; A first holding chamber located in the substrate and having a first space for holding a first liquid; A reaction chamber located in the substrate and having a space for holding a liquid sample containing a specimen; A first flow path located in the substrate and having a first opening and a second opening, wherein the first opening and the second opening are connected to the first holding chamber and the reaction chamber, respectively.
- the sample analysis substrate according to Item 5 wherein the non-capillary space has a portion located closer to the rotation axis than the capillary space.
- the reaction chamber includes a first portion and a second portion;
- the substrate has a wall portion located between the first portion and the second portion of the reaction chamber;
- the wall portion forms a convex portion in a direction toward the rotation axis, In the first part and the second part, a part of the capillary space and the non-part on the side farther from an arc having a line segment connecting the rotation axis of the wall part closest to the rotation axis and the rotation axis.
- Each part of the capillary space is located, Item 6.
- a part of the capillary space that connects the first part and the second part is located in a part or all of the wall part located on the first part side, Item 8.
- a collection chamber located in the substrate and having a space;
- a third flow path located in the substrate and having a fifth opening and a sixth opening, wherein the fifth opening and the sixth opening are connected to the main chamber and the recovery chamber, respectively.
- [Item 10] 10 10. The sample analysis substrate according to any one of items 1 to 9, wherein the first channel is a non-capillary channel.
- the first chamber has an outermost peripheral side surface that is located farthest from the rotation axis, and an adjacent side surface adjacent to the outermost peripheral side surface, Forming a recess in the outermost peripheral side surface and the adjacent side surface; Item 11.
- the sample analysis substrate according to Item 10 wherein the first liquid is held by the recess when the sample analysis substrate is held at a predetermined angular position.
- the first channel is a capillary channel.
- Item 13 Item 13.
- Item 14 Item 13.
- the space of the first holding chamber includes a first portion and a second portion, and a connecting portion that is located between the first portion and the second portion and connects the first portion and the second portion.
- the substrate has a wall portion separating the first portion and the second portion of the space of the first holding chamber;
- the reaction chamber is located farther from the axis of rotation than the second portion of the first holding chamber;
- the connecting portion of the space of the first holding chamber is located closer to the rotating shaft than the wall portion of the substrate; 13.
- a fourth holding chamber located in the substrate and having a space for holding a second liquid; An eighth flow path for connecting the fourth holding chamber and the reaction chamber and transferring the second liquid;
- the first holding chamber has an outermost peripheral side surface farthest from the rotation shaft, an adjacent side surface adjacent to the outermost peripheral side surface, and a recess formed by the outermost peripheral side surface and the adjacent side surface.
- the fourth holding chamber has an outermost peripheral side surface farthest from the rotation axis, an adjacent side surface adjacent to the outermost peripheral side surface, and a recess formed by the outermost peripheral side surface and the adjacent side surface.
- the adjacent side surface of the first holding chamber and the adjacent side surface of the fourth holding chamber are non-parallel when viewed from a direction parallel to the rotation axis.
- Item 15. The sample analysis substrate according to any one of Items 1 to 14.
- [Item 17] The sample analysis substrate according to any one of items 1 to 16, and A motor for rotating the sample analysis substrate around the rotation axis;
- a rotation angle detection circuit for detecting a rotation angle of the rotation shaft of the motor; Based on the detection result of the rotation angle detection circuit, the drive circuit that controls the rotation angle of the motor when rotating and stopping, and the arithmetic unit, the memory, and the memory are configured to be executable by the arithmetic unit
- a sample analysis system comprising: The program is When the sample analysis substrate in which the first liquid and the liquid sample are introduced into the first holding chamber and the Seki reaction chamber is loaded in the
- a rotation angle detection circuit for detecting a rotation angle of the rotation shaft of the motor;
- the drive circuit that controls the rotation angle of the motor when rotating and stopping, and the arithmetic unit, the memory, and the memory are configured to be executable by the arithmetic unit
- the program is When the sample analysis substrate in which the first liquid and the liquid sample are introduced into the first holding chamber and the Seki reaction chamber is loaded in the sample analyzer, (A) The liquid sample in the reaction chamber is transferred to the main chamber by rotating the sample analysis substrate, (B) by rotating the sample analysis substrate, to transfer the first liquid in the first holding chamber to the reaction chamber after the step (a), (C) transferring the first liquid in the reaction chamber to the main chamber by rotating the sample analysis substrate; Sample analyzer.
- a sample analysis substrate that transfers liquid by a rotational motion, A substrate having a rotation axis; A first holding chamber located in the substrate and having a first space for holding a first liquid; A reaction chamber located in the substrate and having a space for holding a liquid sample containing the analyte; A first flow path located in the substrate and having a first opening and a second opening, wherein the first opening and the second opening are connected to the first holding chamber and the reaction chamber, respectively.
- a first flow path A main chamber having a space for holding a liquid sample containing the analyte and a magnetic particle having a ligand immobilized on the surface thereof, located in the substrate;
- a second flow path located in the substrate and having a third opening and a fourth opening, wherein the third opening and the fourth opening are connected to the reaction chamber and the main chamber, respectively.
- a magnet storage chamber located in the substrate and capable of storing magnets; With The first opening is located closer to the rotation axis than the second opening, The second opening is located closer to the rotation axis than the third opening, The magnet storage chamber is disposed at a position where, when a magnet is stored in the magnet storage chamber, the magnetic particles in the main chamber can be captured in the main chamber by the magnet.
- a liquid feeding method using (A) introducing a first liquid and a liquid sample into the first holding chamber and the reaction chamber, respectively; (B) transferring the liquid sample in the reaction chamber to the main chamber; (C) after the step (b), transferring the first liquid in the first holding chamber to the reaction chamber; (D) transferring the first liquid in the reaction chamber to the main chamber; How to use the sample analysis substrate.
- the sample analysis substrate, sample analysis apparatus, sample analysis system, and sample analysis system program according to the present embodiment measure signals with high accuracy even when liquid remains in the chamber that holds the reaction solution. Can do.
- the two or more liquids held in different chambers are transferred to other chambers by various rotations of the sample analysis substrate, it is possible to more reliably prevent the liquids from being sent at unnecessary timing.
- the liquid is a substrate solution and a cleaning liquid, but the liquid is not limited to the substrate solution and the cleaning liquid, and may be various liquids used for sample analysis.
- FIG. 2A is a schematic diagram showing the overall configuration of the sample analysis system 501.
- the sample analysis system 501 includes a sample analysis substrate 100 and a sample analysis device 200.
- the sample analyzer 200 includes a motor 201, an origin detector 203, a rotation angle detection circuit 204, a control circuit 205, a drive circuit 206, and an optical measurement unit 207.
- the motor 201 has a rotation axis A inclined from the gravity (vertical) direction G at an angle ⁇ greater than 0 ° and not more than 90 ° with respect to the turntable 201a and the gravity direction, and the sample analysis placed on the turntable 201a
- the substrate 100 is rotated around the rotation axis A. Since the rotation axis A is inclined, in addition to the centrifugal force due to the rotation, the movement due to the gravity can be used for the transfer of the liquid in the sample analysis substrate 100.
- the inclination angle of the rotation axis A with respect to the gravity direction G is preferably 5 ° or more, more preferably 10 ° or more and 45 ° or less, and further preferably 20 ° or more and 30 ° or less.
- the motor 201 may be, for example, a direct current motor, a brushless motor, an ultrasonic motor, or the like.
- the origin detector 203 detects the origin of the sample analysis substrate 100 attached to the motor 201.
- the origin detector 203 includes a light source 203a, a light receiving element 203b, and an origin detection circuit 203c, and is arranged such that the sample analysis substrate 100 is positioned between the light source 203a and the light receiving element 203b. Is done.
- the light source 203a is a light emitting diode
- the light receiving element 203b is a photodiode.
- the sample analysis substrate 100 has a marker 210 provided at a specific position.
- the marker 210 has a light shielding property of shielding at least a part of light emitted from the light source 203a.
- the area of the marker 210 has a low transmittance (for example, 10% or less), and the area other than the marker 210 has a high transmittance (for example, 60% or more).
- the light receiving element 203b When the sample analysis substrate 100 is rotated by the motor 201, the light receiving element 203b outputs a detection signal corresponding to the amount of incident light to the origin detection circuit 203c. Depending on the direction of rotation, the detection signal increases or decreases at the edges 210a and 210b of the marker 210. For example, when the sample analysis substrate 100 rotates clockwise as indicated by an arrow, the origin detection circuit 203c detects a decrease in the detected light amount and outputs it as an origin signal. In the present specification, the position of the edge 210a of the marker 210 is treated as the origin position of the sample analysis substrate 100 (the angular position serving as the reference of the sample analysis substrate 100).
- the position of a specific angle arbitrarily determined from the position of the edge 210a of the marker 210 may be determined as the origin.
- the marker 210 has a sector shape and the central angle thereof is smaller than the angle detection accuracy necessary for sample analysis, the marker 210 itself may be determined as the origin position.
- the origin position is used for the sample analyzer 200 to acquire information on the rotation angle of the sample analysis substrate 100.
- the origin detector 203 may have other configurations.
- the sample analysis substrate 100 may include an origin detection magnet, and the origin detector 203 may be a magnetic detection element that detects the magnetism of the magnet. Moreover, you may use the magnet for catching the magnetic particle mentioned later for origin detection.
- the origin detector 203 may not be provided.
- the rotation angle detection circuit 204 detects the angle of the rotation axis A of the motor 201.
- the rotation angle detection circuit 204 may be a rotary encoder attached to the rotation axis A.
- the rotation angle detection circuit 204 includes a hall element provided in the brushless motor and a detection circuit that receives an output signal of the hall element and outputs an angle of the rotation axis A. Also good.
- the drive circuit 206 rotates the motor 201. Specifically, based on a command from the control circuit 205, the sample analysis substrate 100 is rotated clockwise or counterclockwise. Further, based on the detection results of the rotation angle detection circuit 204 and the origin detector 203 and the command from the control circuit 205, the swing and rotation of the sample analysis substrate 100 are stopped.
- the optical measurement unit 207 detects a signal (for example, dye, luminescence, fluorescence, etc.) corresponding to the labeling substance 307 of the labeled antibody 308 bound to the complex 310 (FIG. 1) held on the sample analysis substrate 100.
- a signal for example, dye, luminescence, fluorescence, etc.
- the control circuit 205 includes a CPU provided in the sample analyzer 200, for example.
- the control circuit 205 executes a computer program read into a RAM (Random Access Memory; not shown), and sends instructions to other circuits according to the procedure of the computer program.
- Each circuit that receives the instruction operates as described in this specification to realize the function of each circuit.
- the command from the control circuit 205 is sent to the drive circuit 206, the rotation angle detection circuit 204, the optical measurement unit 207, etc., as shown in FIG. 2A, for example.
- the procedure of the computer program is shown by the flowchart in the accompanying drawings.
- the RAM into which the computer program is read in other words, the RAM that stores the computer program may be volatile or non-volatile.
- Volatile RAM is RAM that cannot store stored information unless power is supplied.
- dynamic random access memory DRAM
- the nonvolatile RAM is a RAM that can hold information without supplying power.
- magnetoresistive RAM (MRAM), resistance change memory (ReRAM), and ferroelectric memory (FeRAM) are examples of nonvolatile RAM. In the present embodiment, it is preferable to employ a nonvolatile RAM.
- Both volatile RAM and non-volatile RAM are non-transitory examples of computer-readable recording media.
- a magnetic recording medium such as a hard disk or an optical recording medium such as an optical disk is an example of a computer-readable recording medium that is not temporary. That is, the computer program according to the present disclosure can be recorded on various non-transitory computer-readable media other than a medium such as the atmosphere (temporary medium) that propagates the computer program as a radio wave signal.
- control circuit 205 is described as a separate component from the rotation angle detection circuit 204 and the origin detection circuit 203c of the origin detector 203.
- these may be realized by common hardware.
- a CPU computer
- a CPU provided in the sample analyzer 200 functions as a computer program that functions as the control circuit 205, a computer program that functions as the rotation angle detection circuit 204, and a computer program that functions as the origin detection circuit 203 c of the origin detector 203. May be executed serially or in parallel. Thereby, the CPU can be apparently operated as a different component.
- FIG. 3A is an exploded perspective view of the sample analysis substrate 100.
- the sample analysis substrate 100 includes a rotating shaft 110 and a plate-shaped substrate 100 ′ having a predetermined thickness in a direction parallel to the rotating shaft 110.
- the substrate 100 ′ of the sample analysis substrate 100 is composed of a base substrate 100a and a cover substrate 100b.
- the substrate 100 ′ of the sample analysis substrate 100 has a circular shape, but may have, for example, a polygonal shape, an elliptical shape, a sector shape, or the like.
- the substrate 100 ′ has two main surfaces 100c and 100d.
- the main surface 100c and the main surface 100d are parallel to each other, and the thickness of the substrate 100 ′ (the distance between the two main surfaces) defined by the distance between the main surface 100c and the main surface 100d is the substrate.
- the main surfaces 100c and 100d need not be parallel.
- a part of two main surfaces may be non-parallel or parallel, or may be totally non-parallel.
- FIG. 3B is a plan view of the base substrate 100a.
- the sample analysis substrate 100 includes a first holding chamber 101, a second holding chamber 102, a third holding chamber 103, a first storage chamber 104, and a second storage chamber, which are located in the substrate 100 ′, respectively.
- the shape of each chamber is not limited as long as it is not specifically mentioned below, and may have any shape.
- Each chamber generally has a space defined by upper and lower surfaces parallel to the two major surfaces 100c, 100d (FIG. 3A) of the substrate 100 ', and three or more side surfaces located therebetween. Two adjacent surfaces of the upper surface, the lower surface, and the side surface may not be separated by a clear ridge line.
- the shape of each chamber may be a flat sphere or a spheroid.
- the sample analysis substrate 100 further includes a first flow path 111, a second flow path 112, a third flow path 113, a fourth flow path 114, a fifth flow path 115, and a sixth flow, which are located in the substrate 100 ′, respectively. It has a channel 116 and a seventh channel 117.
- the first flow path 111 connects the first holding chamber 101 and the reaction chamber 106.
- the second flow path 112 connects the reaction chamber 106 and the main chamber 107.
- the third flow path 113 connects the main chamber 107 and the recovery chamber 108.
- the fourth flow path 114 connects the first storage chamber 104 and the first holding chamber 101.
- the fifth flow path 115 connects the second storage chamber 105 and the second holding chamber 102.
- the sixth flow path 116 connects the second holding chamber 102 and the third holding chamber 103.
- the seventh flow path 117 connects the third holding chamber 103 and the main chamber 107.
- the first holding chamber 101 connected to the first storage chamber 104 via the fourth flow path 114 is connected to the reaction chamber 106 instead of the main chamber 107 by the first flow path 111.
- the liquid transfer between the chambers through the flow path can be realized by various methods. For example, transfer by gravity and transfer by capillary force and centrifugal force by rotation can be used. Hereinafter, these two transfer methods will be described generally.
- the liquid can move in the flow path by gravity.
- the sample analysis substrate 100 is supported with the rotation axis 110 tilted with respect to the direction of gravity G in a range of greater than 0 degrees and less than 90 degrees.
- the transfer source chamber in which the liquid exists is arranged at a higher position than the transfer destination chamber. “High” means being higher in the direction of gravity G.
- the flow path capable of being transferred by gravity is not a capillary path described below.
- the flow path that can be transferred by gravity has, for example, a thickness of 1 mm or more.
- the channel may be a capillary channel.
- a “capillary channel” refers to a channel having a narrow cross section that can fill at least a portion of the liquid with capillary force due to capillary action.
- the liquid transfer through the capillary channel will be described by taking as an example a configuration having chambers A and B that are not capillary spaces, and a capillary channel that connects chamber A and chamber B.
- air holes are provided in the chamber A and the chamber B so as not to cause a pressure difference due to the movement of the liquid, and the pressure in the two chambers is set to the pressure with the external environment. Match.
- the liquid in the flow path is stationary due to the balance of capillary force, atmospheric pressure, and gravity, and no liquid is transferred from chamber A to chamber B. Further, by rotating the sample analysis substrate, the liquid is not transferred even when a centrifugal force less than the capillary force acts on the liquid in the channel.
- chamber B is arranged at a position farther than chamber A with respect to the rotation axis, and the sample analysis substrate is rotated so that a centrifugal force larger than the capillary force acts on the liquid in the flow path of the capillary channel. Then, the liquid in the chamber A can be transferred to the chamber B by centrifugal force.
- the flow channel has a thickness of 50 ⁇ m to 300 ⁇ m, for example.
- different thicknesses can be realized by changing the depths of the spaces provided in the base substrate 100a.
- the depth of the space provided in the base substrate 100a is made constant, and convex portions having different heights are provided at positions corresponding to the respective chambers and flow paths of the cover substrate 100b, so that the thicknesses of the respective flow paths and chambers are different. It may be allowed.
- part or all of the flow path may constitute a capillary space so that part or all of the chamber is reliably filled with the retained liquid.
- the thickness of the region serving as the capillary space is 50 ⁇ m to 300 ⁇ m as described above.
- the sample analysis substrate 100 having a diameter of 60 mm can be rotated in the range of 100 rpm to 8000 rpm.
- the rotation speed is determined according to the shape of each chamber and flow path, the physical properties of the liquid, the timing of liquid transfer and processing, and the like.
- Hydrophilic treatment may be applied to the inner surface of the flow path or chamber in which the capillary force works and the connection portion of the chamber to which the flow path is connected.
- Capillary force works greatly by hydrophilic treatment.
- the hydrophilic treatment is performed, for example, by applying a nonionic, cationic, anionic or zwitterionic surfactant on the inner surface, performing corona discharge treatment, or providing physical fine irregularities. (For example, refer to JP 2007-3361 A).
- hydrophilic treatment may be applied to these flow paths as well.
- Each of the first holding chamber 101, the second holding chamber 102, the third holding chamber 103, the first storage chamber 104, the second storage chamber 105, the reaction chamber 106, the main chamber 107, and the recovery chamber 108 has at least one air hole. 122 is provided. Thereby, the inside of each chamber is maintained at atmospheric pressure, and each flow path can be moved by the capillary phenomenon and siphon principle. Further, the first storage chamber 104, the second storage chamber 105, and the reaction chamber 106 may be provided with an opening 123 for injecting the cleaning solution and the substrate solution. The air hole 122 may also serve as the opening 123.
- Each of the first holding chamber 101, the second holding chamber 102, the third holding chamber 103, the first storage chamber 104, the second storage chamber 105, the reaction chamber 106, the main chamber 107, and the recovery chamber 108 includes a base substrate 100a.
- the upper and lower portions of each space are formed by covering the base substrate 100a with the cover substrate 100b. That is, these spaces are defined by the inner surface of the substrate 100 '.
- the first channel 111, the second channel 112, the third channel 113, the fourth channel 114, the fifth channel 115, the sixth channel 116, and the seventh channel 117 are also formed in the base substrate 100a.
- the base substrate 100a and the cover substrate 100b define an upper surface and a lower surface, respectively.
- the substrate 100 ′ can be made of a resin such as acrylic, polycarbonate, or polystyrene.
- Table 1 shows the substance or liquid introduced at the start of sample analysis, the first introduction chamber, and the order in which the introduced substance or liquid is introduced into the main chamber in the sample analysis substrate 100 of the present embodiment. Shows the combination.
- the combinations shown in Table 1 are only one example illustrated, and the substances and liquids to be introduced into the chamber and the order of introduction into the main chamber 107 are not limited to the substances and orders shown in Table 1.
- the magnetic particle-immobilized antibody 305, the specimen containing the antigen 306, and the labeled antibody 308 are introduced into the reaction chamber 106, and the complex 310 is generated in the reaction chamber 106.
- the magnetic particle-immobilized antibody 305 and the labeled antibody 308 are arranged in advance in the reaction chamber 106 as the drying reagent 125.
- a substrate solution is introduced into the second storage chamber 105.
- a cleaning liquid is introduced into the first storage chamber 104. As will be described in detail below, the cleaning liquid held in the first storage chamber 104 is introduced into the main chamber 107 via the reaction chamber 106.
- FIG. 3C to FIG. 3H for the sake of simplicity, the structure that is not related or not mentioned of the sample analysis substrate 100 is not shown.
- reaction chamber 106 As shown in FIG. 3C, a reaction chamber 106 is provided on the sample analysis substrate 100. As described with reference to FIG. 1, the reaction chamber 106 is a reaction field in which a magnetic particle-immobilized antibody 305, a specimen containing an antigen 306, and a labeled antibody 308 are reacted to form a complex 310. .
- the reaction chamber 106 includes a first portion 106q and a second portion 106r.
- the first portion 106q and the second portion 106r are generally arranged in the circumferential direction around the rotation axis 11.
- a wall portion 126 constituted by the inner surface of the substrate 100 ′ is located between the first portion 106 q and the second portion 106 r.
- the wall portion 126 has a convex shape on the rotating shaft 110 side, and divides the first portion 106q and the second portion 106r.
- the first portion 106q and the second portion 106r are connected to each other at a radial position connecting the rotation shaft 110 and the point 126p closest to the rotation shaft 110 of the wall portion 126.
- the first portion 106q includes a first region 106qf that is a non-capillary space and a second region 106qe that is a capillary space.
- the first region 106qf and the second region 106qe are connected adjacent to each other, and the mutual space is in communication.
- the second region 106qe is a narrow region along the wall portion 126 in the first portion 106q.
- the second region 106qe is in contact with the outermost peripheral side surface 106qa located farthest from the rotation shaft 110 among the side surfaces of the first portion 106q.
- the first region 106qf is a sufficiently large space that can hold the specimen. In the first portion, the first region 106qf is located closer to the rotation shaft 110 than the second region 106qe.
- the second portion 106r includes a first region 106rf that is a non-capillary space and a second region 106re that is a capillary space.
- the first region 106rf and the second region 106re are connected adjacent to each other, and the spaces are in communication with each other.
- the first region 106rf is located along a side surface that faces the outermost circumferential side surface 106ra that is farthest from the rotation shaft 110 among the side surfaces that define the second portion 106r. That is, the first area 106rf is located closer to the rotation shaft 110 than the second area 106re.
- the first region 106qf of the first portion 106q is connected to the first region 106rf of the second portion 106r, and the second region 106re of the second portion 106r is connected to the second region 106re of the second portion 106r.
- the first portion 106q and the second portion 106r are more than an arc 126ar having a radius that is a line segment connecting the point 126p closest to the rotation axis of the wall portion 126 and the rotation axis 110.
- Each has a portion located on the far side. Specifically, a part of the first region 106qf of the first part 106q and a part of the second region 106re of the second part 106r are located outside the arc 126ar.
- the non-capillary space and the capillary space have different thicknesses.
- the dried reagent 125 including the dried magnetic particle-immobilized antibody 305 and the labeled antibody 308 is held in advance in the second region 106re of the second portion 106r.
- the liquid containing the sample solution containing the antigen 306 is introduced into the first region 106qf of the first portion 106q of the reaction chamber 106 at the start of sample analysis.
- the second region 106qe is filled by capillary force, and further the second region 106re of the second portion is filled.
- the liquid and the drying reagent 125 come into contact with each other, and the magnetic particle-immobilized antibody 305 and the labeled antibody 308 of the drying reagent 125 are eluted or dispersed in the liquid.
- the antigen 306, the magnetic particle-immobilized antibody 305, and the labeled antibody 308 are mixed in the liquid to form the complex 310.
- the drying reagent 125 is preferably held in the capillary space (second region 106re) of the reaction chamber 106.
- the liquid in the non-capillary space in the reaction chamber moves to the capillary space due to the capillary force.
- the force acting on the liquid is smaller than the centrifugal force due to the rotation of the sample analysis substrate 100.
- all of the magnetic particle-immobilized antibody 305 in the drying reagent 125 does not move to the capillary space, but remains in the non-capillary space. Because there is a possibility.
- the introduction of the specimen solution containing the antigen 306, the magnetic particle-immobilized antibody 305, and the labeled antibody 308 into the reaction chamber 106 is not limited thereto.
- the sample analysis substrate 100 does not include the drying reagent 125, and at the start of sample analysis, the magnetic particle-immobilized antibody 305, the specimen containing the antigen 306, and the labeled antibody 308 are placed in the first region of the first portion 106q of the reaction chamber. 106qf may be introduced.
- the sample analysis substrate 100 includes, for example, a chamber for holding the magnetic particle-immobilized antibody 305, the specimen containing the antigen 306, and the labeled antibody 308, and a flow path (for example, the chamber and the reaction chamber 106 are connected).
- the specimen containing the antigen 306 and the labeled antibody 308 are weighed into the respective chambers, and the magnetic particle-immobilized antibody 305 and the specimen containing the antigen 306 and the labeled antibody 308 injected into each chamber are transferred to the reaction chamber 106 and the reaction chamber 106. They may be mixed in to form the composite 310.
- the solution containing the complex 310 in the reaction chamber 106 is transferred to the main chamber 107 via the second flow path 112.
- the second flow path 112 has an opening 112g and an opening 112h.
- the opening 112g of the second flow path 112 is the outermost peripheral side surface 106ra located on the side farthest from the rotating shaft 110 among the side surfaces defining the second region 106re of the second portion 106r of the reaction chamber 106, or the outermost peripheral side surface. It is preferable that the adjacent side surface adjacent to 106ra is provided at a position including a connection portion with the outermost peripheral side surface 106a.
- FIG. 3C shows an example in which the opening 112g is provided in a part of the outermost peripheral side surface 106a.
- the opening 112h of the second flow path 112 is located farther from the rotation shaft 110 than the opening 112g.
- the opening 112h is connected to the side surface of the main chamber 107 as described below.
- the solution containing the complex 310 in the reaction chamber 106 is subjected to centrifugal force. It is transferred to the main chamber 107 via the second flow path 112.
- the second channel 112 may be a capillary channel or a channel that can be transferred by gravity.
- the main chamber 107 is a place where B / F separation of the solution containing the complex 310 is performed.
- the sample analysis substrate 100 includes a magnet storage chamber 120 located in the substrate 100 ′ and a magnet 121 disposed in the magnet storage chamber 120.
- the magnet storage chamber 120 is located in the vicinity of the space of the main chamber 107 in the sample analysis substrate 100. More specifically, the magnet storage chamber 120 is disposed in the vicinity of the outermost peripheral side surface 107 a located farthest from the rotation shaft 110 among the plurality of side surfaces of the main chamber 107. However, the magnet storage chamber 120 in the sample analysis substrate 100 may be disposed at a position close to the upper surface or the lower surface other than the outermost peripheral side surface 107 a of the main chamber 107. That is, the position is not particularly limited as long as the magnetic particles can be captured on the wall surface of the main chamber 107 by the magnet 121 arranged in the magnet storage chamber 120.
- the magnet 121 may be configured to be removable according to B / F separation, or may be non-detachably attached to the substrate 100 ′ from the magnet storage chamber 120, or provided on the sample analyzer 200 side. It may be configured.
- the substrate 100 ′ may include a concave magnet storage chamber 120 having an opening 120 a on the main surface 100 c.
- the magnet storage chamber 120 has a space in which the magnet 121 can be stored. By inserting the magnet 121 into the magnet storage chamber 120 through the opening 120a, the magnet 121 can be loaded on the substrate 100 '.
- the opening 120a of the magnet storage chamber 120 may be provided on the main surface 100d, or may be provided on a side surface located between the two main surfaces 100c and 100d.
- the turntable 201a of the sample analyzer 200 may include a magnet unit including the magnet 121.
- the magnet 121 when the user arranges the sample analysis substrate 100 at a predetermined position of the turntable 201 a (magnet unit), the magnet 121 is arranged at a position where the magnetic particles can be captured on the wall surface of the main chamber 107.
- the sample analyzer 200 may include a magnet 121 and a drive mechanism that moves the magnet 121.
- the sample analysis substrate 100 includes a storage chamber for holding the magnet 121, and the drive mechanism inserts the magnet 121 into the storage chamber of the sample analysis substrate 100 according to B / F separation, and the magnet 121 in the storage chamber. May be taken out.
- the complex 310 in the reaction solution and the unreacted magnetic particle-immobilized antibody 305 (hereinafter, when referring to both of these, simply Magnetic particles 311) are collected and captured on the outermost peripheral side surface 107a side by the magnetic force of the magnet 121 arranged in the vicinity of the outermost peripheral side surface 107a.
- the space of the main chamber 107 may include a second region 107e adjacent to and connected to the first region 107f and the first region 107f.
- the first region 107f is a non-capillary space in which a liquid can move by gravity
- the second region 107e is a capillary space in which a capillary force works.
- the thickness of the first region 107f is larger than the thickness of the second region 107e
- the first region 107f has a larger space than the second region 107e.
- the thicknesses of the first region 107f and the second region 107e are values within the above-described range described as the thickness of the flow path.
- the second region 107e is in contact with the outermost peripheral side surface 107a, and at least a part of the first region 107f is closer to the rotating shaft 110 than the second region 107e. Further, the opening 117h of the second flow path 112 is provided on one of the side surfaces in contact with the first region 107f.
- the liquid in the main chamber 107 is transferred to the recovery chamber 108 via the third flow path 113.
- the opening 113g of the third flow path 113 is provided on a side surface in contact with the second region 102e so as to be connected to the space of the second region 102e.
- the first region 107f is a space in which the liquid can move by gravity, it is possible to secure a space having a size as required.
- the second region 107e is a capillary space, the second region 107e is always filled with a part of the liquid held in the main chamber 107. For this reason, when the 3rd flow path 113 contacts the 2nd area
- a cleaning solution and a substrate solution are introduced into the main chamber 107, so that the main chamber 107 has a sufficient space for holding these liquids, and the held liquid is collected as necessary.
- the reliable transfer to 108 is an important feature.
- the third flow path 113 has an opening 113g and an opening 113h, the opening 113g is connected to the main chamber 107, and the opening 113h is connected to the recovery chamber 108.
- the opening 113g of the third channel 113 is the outermost peripheral side surface 107a located on the side farthest from the rotation shaft 110 among the side surfaces of the main chamber 107, or an adjacent side surface adjacent to the outermost peripheral side surface 107a, and the outermost peripheral surface. It is preferable to be provided at a position including a connection portion with the side surface 107a.
- FIG. 3D shows an example in which the opening 113g is provided on the adjacent side surface adjacent to the outermost peripheral side surface 107a. As described above, the opening 113g is connected to the second region 107e of the main chamber 107.
- the opening 113h of the third flow path 113 is located on the side farther from the rotation shaft 110 than the opening 113g.
- the opening 113h is the innermost peripheral side surface 108b located on the side closest to the rotating shaft 110 or the adjacent side surface adjacent to the innermost peripheral side surface 108b, and is the innermost peripheral side surface. It is preferably provided at a position close to 108b.
- FIG. 3D shows an example in which the opening 113h is provided in a part of the innermost peripheral side surface 108b.
- the third channel 113 can also suck the liquid held in the main chamber 107 by capillary action.
- the thickness of the third flow path 113 is smaller than the thickness of the second region 107 e of the main chamber 107.
- a capillary force stronger than the second region 107e of the main chamber 107 can be applied to the third channel 113, and a part of the liquid in the second region 107e of the main chamber 107 is transferred to the third channel 113. Sucked.
- the third channel 113 can further control the movement of the liquid by the siphon principle. Therefore, as a siphon structure, the third flow path 113 has a first bent portion 113n and a second bent portion 113m.
- the first bent portion 113n has a convex shape on the side opposite to the rotating shaft 110
- the second bent portion 113m has a convex shape on the rotating shaft 110 side.
- 113 n of 1st bending parts are located between the main chamber 107 located in the side near the rotating shaft 110 among the main chamber 107 and the collection
- the principle of siphon refers to liquid feeding control based on the balance between the centrifugal force applied to the liquid by the rotation of the sample analysis substrate 100 and the capillary force of the flow path.
- the third channel 113 is a capillary channel without a siphon structure
- it is transferred from the reaction chamber 106 to the main chamber 107 via the second channel 112 by the centrifugal force generated by the rotation of the sample analysis substrate 100.
- the liquid transferred to the main chamber 107 is filled in the third channel 113 by the capillary force of the third channel 113.
- the sample analysis substrate 100 continues to rotate, the liquid is not held in the main chamber 107 but is transferred to the recovery chamber 108 via the third flow path 113.
- the sample analysis substrate 100 is rotating at a rotation speed at which a centrifugal force stronger than the capillary force of the third channel 113 can be applied.
- the third flow path 113 has a siphon structure
- the liquid transferred from the reaction chamber 106 to the main chamber 107 is liquid in the third flow path 113 due to the capillary force of the third flow path 113.
- the centrifugal force is greater than the capillary force applied to the liquid. Since this is stronger, the entire third flow path 113 is not filled with the liquid. That is, the third flow path 113 is filled with the liquid only to the same height as the distance of the liquid surface of the liquid existing in the main chamber 107 with respect to the rotation shaft 110.
- the sample analysis substrate 100 is rotating at a rotational speed that applies a centrifugal force that is weaker than the capillary force of the third channel 113, the third channel 113 is filled with the liquid by the capillary force, and the capillary tube No further movement of the liquid by force.
- the sample analysis substrate 100 When it is desired to transfer the liquid in the main chamber 107 to the recovery chamber 108, the sample analysis substrate 100 is rotated at a rotational speed (including a rotation stop) at which a centrifugal force equal to or lower than the capillary force of the third flow path 113 can be applied. By rotating, all of the third flow path 113 is filled with the liquid by the capillary force. Thereafter, when the sample analysis substrate 100 is rotated at a rotation speed at which a centrifugal force stronger than the capillary force of the third channel 113 can be applied, the liquid in the main chamber 107 can be transferred to the recovery chamber 108. .
- the reaction solution, the cleaning solution, and the substrate solution can be once held in the main chamber 107.
- B / F separation, magnetic particle cleaning and The reaction with the substrate solution can be appropriately performed.
- the rotation shaft 110 and the innermost peripheral side surface 108b closest to the rotation shaft 110 of the recovery chamber 108 located far from the rotation shaft 110 are provided.
- R1> R2 condition 1
- the capillary for applying the sample analysis substrate 100 to the liquid in the third flow path 113 When rotating at a rotational speed at which a centrifugal force stronger than the force is applied, the reaction liquid or the cleaning liquid transferred to the main chamber 107 can be prevented from being transferred to the recovery chamber 108 as it is.
- the recovery chamber 108 stores a reaction liquid other than the magnetic particles 311 transferred from the main chamber 107 via the third flow path 113 and a used cleaning liquid.
- the collection chamber 108 has a space with a capacity larger than the total amount of the above-described reaction liquid and the total used cleaning liquid according to the number of times of cleaning.
- a main part for holding the liquid is located farther from the rotation shaft 110 than the main chamber 107.
- the first storage chamber 104 stores a cleaning liquid used for cleaning in the B / F separation.
- the complex 310 can be washed multiple times during the B / F separation. Therefore, the first storage chamber 104 has a space that can hold a total volume of cleaning liquid corresponding to the number of cleanings.
- the cleaning liquid in the first storage chamber 104 is transferred to the first holding chamber 101 via the fourth flow path 114.
- the fourth flow path 114 has an opening 114g and an opening 114h.
- the opening 114g of the fourth flow path 114 is the outermost peripheral side surface 104a located on the side farthest from the rotation shaft 110 among the side surfaces of the first storage chamber 104, or an adjacent side surface adjacent to the outermost peripheral side surface 104a, It is preferable to be provided at a position including a connection portion with the outermost peripheral side surface 104a.
- FIG. 3F shows an example in which the opening 114g is provided at the connection portion between the outermost peripheral side surface 104a and the adjacent side surface.
- the opening 114h of the fourth flow path 114 is located on the side farther from the rotating shaft 110 than the opening 114g.
- the opening 114h is connected to the side surface of the first holding chamber 101 as described below.
- the cleaning liquid in the first storage chamber 104 is subjected to the fourth flow by centrifugal force. It is transferred to the first holding chamber 101 via the path 114.
- the fourth channel 114 may be a capillary channel or a channel that can be transferred by gravity.
- the first holding chamber 101 holds the entire cleaning liquid stored in the first storage chamber 104. Thereafter, in order to clean the composite 310 in the main chamber 107, a part of the cleaning liquid is transferred to the reaction chamber 106 and the rest is retained. The amount of the cleaning liquid used for one cleaning is weighed by the first flow path 111 as described below. For this reason, the first holding chamber 101 has a volume equal to or larger than the first flow path 111 and has a volume equal to or larger than the total amount of the cleaning liquid corresponding to the number of times of cleaning (for example, the first flow path 111 for two cleanings). The volume of the first flow path 111 is 3 times or more if the cleaning is performed three times.
- the opening 114h of the fourth flow path 114 is provided on one inner peripheral side surface that opposes the outermost peripheral side surface 103a of the first holding chamber 101 with a space for holding the liquid therebetween.
- First flow path 111 As described above, the first flow path 111 connects the first holding chamber 101 and the reaction chamber 106. For this reason, when the cleaning liquid is introduced into the first storage chamber 104, the cleaning liquid is once transferred to the reaction chamber 106 and then transferred to the main chamber 107.
- the first flow path 111 includes a first portion 111q and a second portion 111r connected to the first portion 111q.
- the second portion 111r is a capillary channel.
- the first portion 111q includes an opening 111g and is connected to the first holding chamber 101.
- a part of the first holding chamber 101 and a part of the first flow path 111 are located in the radial direction about the rotation axis 110 with the opening 111g interposed therebetween.
- the second portion 111r has a second opening 111h and is connected to the first region 106qf of the first portion 106q of the reaction chamber 106.
- the second opening 111h is closer to the rotating shaft 110 than the opening 112g of the second flow path 112.
- the first portion 111q of the first flow path 111 includes a first region 111qe and a second region 111qf.
- the first portion 111q has a shape extending in an oblique direction with respect to the radial direction of the sample analysis substrate 100.
- the second region 111qf is located closer to the rotation shaft 110 than the first region 111qe in the first portion 111q.
- the first region 111qe is a capillary space
- the second region 111qf is not a capillary space that can be filled with liquid by a capillary phenomenon.
- the thickness of the second region 111qf is larger than the thickness of the first region 111qe. For this reason, the amount of the cleaning liquid to be weighed can be increased by filling the second region 11qf with the liquid.
- the air holes 122 are provided in the second region 111qf, and the second region 111qf also functions as an airway that ensures air movement. Specifically, by providing the second region 111qf, when bubbles are generated in the liquid held in the first region 111qe for any reason, the bubbles move to the second region 111qf, and the liquid Air bubbles inside are easily removed. Thereby, when the sample analysis substrate 100 is rotated, in particular, it is possible to suppress the bubbles from entering the second portion 111r and hindering the movement of the liquid.
- the second region 116qf is removed.
- One flow path 111 is filled with a cleaning liquid by capillary action.
- the sample analysis substrate 100 is rotated at a rotation speed at which a centrifugal force stronger than the capillary force applied to the cleaning liquid in the first channel 111 is applied.
- the cleaning liquid transferred to the first holding chamber 101 on the plane perpendicular to the rotating shaft 110 and the first flow with respect to the straight line db connecting the rotating shaft 110 and the position z, The cleaning liquid returns to the path 111.
- the reference position z is two side surfaces s1 and s2 that are located farther from the rotation axis 110 than the space of the first holding chamber 101 or the space of the first flow path 111. , Defined by a boundary position between a surface s1 inclined toward the first holding chamber 101 side and a surface s2 inclined toward the second holding chamber side with respect to the tangential direction dt of the arc ar around the rotation axis 110. . Thereby, the cleaning liquid for one time is weighed, and the cleaning liquid is transferred to the main chamber 107. As described above, the cleaning liquid transferred to the main chamber 107 is transferred to the recovery chamber 108 via the third flow path 113.
- the second storage chamber 105 stores the substrate solution at the start of analysis using the sample analysis system. There is no restriction
- the fifth flow path 115 connects the second storage chamber 105 and the second holding chamber 102.
- the fifth flow path 115 extends in the radial direction about the rotation shaft 110 and is configured by a capillary path.
- the fifth flow path 115 has an opening 115g and an opening 115h.
- the opening 115g is the outermost peripheral side surface 105a farthest from the rotation shaft 110 or the adjacent side surface adjacent to the outermost peripheral side surface 105a among the side surfaces of the second storage chamber 105, and at a position close to the outermost peripheral side surface 105a. It is preferable to be provided.
- an opening 115g is provided on the outermost peripheral side surface 105a.
- the opening 115 h is connected to the second holding chamber 102.
- the second holding chamber 102 holds the substrate solution transferred from the second storage chamber 105 during B / F separation including washing after the start of analysis using the sample analysis system.
- the second holding chamber 102 is located farther from the rotation shaft 110 than the second storage chamber 105.
- the second holding chamber 102 has a first outer peripheral side surface 102a1 and a second outer peripheral side surface 102a2, and a first inner peripheral side surface 102b1 and a second inner peripheral side surface 102b2, which sandwich a space for holding the substrate solution.
- the first outer peripheral side surface 102a1 and the second outer peripheral side surface 102a2 do not overlap in the radial direction, and the first outer peripheral side surface 102a1 is located farther from the rotating shaft 110 than the second outer peripheral side surface 102a2.
- the first inner peripheral side surface 102b1 and the second inner peripheral side surface 102b2 do not overlap in the radial direction, and the first inner peripheral side surface 102b1 is located farther from the rotating shaft 110 than the second inner peripheral side surface 102b2. .
- the second holding chamber 102 further includes an adjacent side surface 102c adjacent to the first outer peripheral side surface 102a1 and the first inner peripheral side surface 102b1, and an adjacent side surface 102d adjacent to the second outer peripheral side surface 102a2 and the second inner peripheral side surface 102b2.
- the space of the second holding chamber 102 is sandwiched between the adjacent side surface 102c and the adjacent side surface 102d and has a shape extending in the circumferential direction.
- the opening 116g of the sixth flow path 116 is provided in the first inner peripheral side surface 102b1. Further, the opening 116g of the sixth flow path 116, which will be described in detail below, is located adjacent to the connection position of the adjacent side surface 102d with the second inner peripheral side surface 102b2. That is, the sixth flow path 116 is provided on the side closer to the rotating shaft 110 of the adjacent side surface 102d. With this structure, most of the space of the second holding chamber 102 is located farther from the rotating shaft 110 than the opening 116 h of the sixth flow path 116. Therefore, even when the sample analysis substrate 100 is held at various rotation angles, the substrate solution held in the second holding chamber 102 is transferred to the third holding chamber 103 via the sixth channel 116. Can be suppressed.
- the first outer peripheral side surface 102a1 is located far from the rotating shaft 110, and the space of the second holding chamber 102 includes a convex portion 102t that protrudes toward the outer peripheral side in contact with the first outer peripheral side surface 102a1. Therefore, by holding the substrate solution in the convex portion 102t of the space of the second holding chamber 102, the liquid level of the substrate solution held in the second holding chamber 102 is separated from the opening 111g of the sixth channel 116. It is possible to suppress the transfer to the third holding chamber 103 via the sixth flow path 116 more reliably.
- the sixth flow path 116 connects the second holding chamber 102 and the third holding chamber 103.
- the sixth flow path 116 has an opening 116 g and an opening 116 h, and the opening 116 g is provided on the adjacent side surface 102 d of the second holding chamber 102.
- the opening 116 h is provided on one of the side surfaces of the third holding chamber 103.
- the sixth flow path 116 is a flow path capable of moving the liquid by gravity.
- the third holding chamber 103 holds the substrate solution transferred from the second holding chamber 102 via the sixth channel 116.
- the third holding chamber 103 includes a first portion 103q adjacent to the second holding chamber 102 in the circumferential direction and a second portion 103r adjacent to the seventh flow path 117 in the circumferential direction.
- the first portion 103q and the second portion 103r are arranged in the radial direction. Further, the third holding chamber 103 is close to the adjacent side surface 101 d of the second holding chamber 102.
- the third holding chamber 103 has an outermost peripheral side surface 103a farthest from the rotating shaft 110 and a second adjacent side surface 103c2 adjacent to the outermost peripheral side surface 103a. Further, it has an innermost peripheral side surface 103b closest to the rotating shaft 110 and a first adjacent side surface 103c1 adjacent to the innermost peripheral side surface 103b. With respect to the space of the third holding chamber 103, the first adjacent side surface 103 c 1 and the second adjacent side surface 103 c 2 are arranged on the same side, that is, the side facing the second holding chamber 102 and the seventh flow path 117. A recess 103s is formed between the first adjacent side surface 103c1 and the second adjacent side surface 103c2, and the first adjacent side surface 103c1 and the second adjacent side surface 103c2 are separated by the recess 103s.
- the first portion 103q of the third holding chamber 103 includes an innermost peripheral side surface 103b and a first adjacent side surface 103c1, and the second portion 103r includes an outermost peripheral side surface 103a and a second adjacent side surface 103c2.
- an opening 116h of the sixth channel 116 is provided at a position of the first adjacent side surface 103c1 close to the innermost peripheral side surface 103b.
- the opening 117g of the seventh flow path 117 is provided at a position of the second adjacent side surface 103c2 that is separated from the outermost peripheral side surface 103a, more specifically, at a position that is closest to the rotating shaft 110. It has been.
- the second portion 103r is a capillary space that connects the outermost peripheral side surface 103a and the portion where the opening 117g of the seventh channel 117 is located.
- 103re may be provided. In this case, it is preferable that the capillary space 103re is located along the second adjacent side surface 103c2.
- the seventh flow path 117 has a first portion 117q and a second portion 117r, and an opening 117g and an opening 117h. One end of the first portion 117q and one end of the second portion 117r are connected to each other.
- the opening 117g is located at the other end of the first portion 117q, and is connected to the second adjacent side surface 103c2 of the third holding chamber 103 as described above.
- An opening 117 h is located at the other end of the second portion 117 r and is connected to the main chamber 107.
- the second portion 117r is a capillary channel.
- the first portion 117q includes a first region 117qe and a second region 117qf.
- the first portion 117q has a shape extending in the circumferential direction.
- the second region 117qf is located closer to the rotation shaft 110 than the first region 117qe.
- the first region 117qe is a capillary space, and the second region 117qf is not a capillary space that can be filled with liquid by capillary action.
- the thickness of the second region 117qf is larger than the thickness of the first region 117qe, and when the first region 117qe is filled with the liquid by capillary action, the second region 117qf is not filled with the liquid.
- An air hole 122 is provided in the second region 117qf.
- the thickness of the first region 117qe is preferably smaller than the thickness of the capillary space 103re of the third holding chamber 103. Thereby, the seventh channel 117 can draw the substrate solution held in the capillary space 103re of the third holding chamber 103.
- the opening 117g is located closer to the rotating shaft 110 than the opening 117h.
- each part of the seventh flow path 117 is located at the same position as the opening 117 g from the rotation shaft 110 or opened from the rotation shaft 110. It is preferable to be located farther than 117 g. Accordingly, when a centrifugal force stronger than the capillary force applied to the substrate solution in the seventh flow path 117 is applied to the substrate solution in a state where the seventh flow path 117 is filled with the substrate solution, all of the seventh flow paths 117 are The substrate solution is transferred to the main chamber 107.
- the total volume of the first region 117qe and the second portion 117r of the first portion 117q corresponds to the amount of the substrate solution used for the analysis, and when these portions are filled with the substrate solution by capillary force, the substrate solution is weighed. Done.
- the second region 117qf of the first portion 117q functions as an airway. For some reason, when bubbles are generated in the substrate solution held in the first region 117qe of the first portion 117q, the bubbles move to the second region 117qf, and the bubbles in the substrate solution are easily excluded. Become. Thereby, when the sample analysis substrate 100 is rotated, in particular, it is possible to suppress the bubbles from entering the second portion 117r and hindering the movement of the substrate solution.
- the first region 117qe and the second portion 117r of the first portion 117q have been described as a capillary space and a capillary channel, but these spaces are a space and a flow channel in which liquid moves by gravity. It may be.
- FIG. 4 is a flowchart showing the operation of the sample analysis system 501.
- a program that defines the procedure for controlling each part of the sample analysis system 501 for operating the sample analysis system 501 is stored in, for example, the memory of the control circuit 205.
- the execution of the program by the arithmetic unit causes the following operations. Realize. Prior to the following steps, the sample analysis substrate 100 is loaded into the sample analysis apparatus 200, and the origin of the sample analysis substrate 100 is detected.
- a drying reagent 125 including a magnetic particle-immobilized antibody 305 and a labeled antibody 308 is held in advance.
- the sample analysis substrate is all rotated clockwise, for example.
- the rotation direction of the sample analysis substrate is not limited to clockwise, and may be counterclockwise.
- the cleaning solution and the substrate solution are introduced into the first storage chamber 104 and the second storage chamber 105, respectively.
- the substrate solution contains a substrate that generates a change in light emission, fluorescence, or absorption wavelength due to reaction with the labeling substance 307 or catalysis by the labeling substance 307.
- the sample solution is introduced into the first portion 106 q of the reaction chamber 106.
- the sample may be injected into the first portion 106q of the reaction chamber 106 by a syringe or the like.
- the sample solution When the sample solution is introduced into the first portion 106q of the reaction chamber 106, the sample solution fills the first region 106qf that is the non-capillary space of the first portion 106q and the second region 106qe that is the capillary space. Since the second region 106qe is connected to the second region 106re, which is the capillary space of the second portion 106r, as shown in FIG. 6, the sample solution is sucked into these capillary spaces by the capillary force. As a result, the sample solution located in the first region 106qf of the first portion 106q moves to the second region 106re of the second portion 106r.
- the drying reagent 125 and the sample solution come into contact with each other, the magnetic particle-immobilized antibody 30 is released into the sample solution, and the labeled antibody 308 is eluted into the sample solution.
- the magnetic particle-immobilized antibody 305, the antigen 306 in the sample, and the labeled antibody 308 are bound by an antigen-antibody reaction, and a complex 310 is generated. Since the magnetic particle-immobilized antibody 30 in the sample solution promotes the release and elution of the labeled antibody 308, the sample analysis substrate 100 may be swung.
- the fifth flow path 115, the fourth flow path 114, and the second flow path 112 are filled with the reaction solution including the substrate solution, the cleaning solution, and the complex 310, respectively, by capillary action.
- Step S12 After the complex 310 is generated, the sample analysis substrate 100 is rotated, and the reaction solution containing the complex 310 is moved from the second region 106re of the second portion 106r of the reaction chamber 106 to the main chamber 107. As described above, the second flow path 112 is filled with the reaction solution by capillary action. For this reason, when a centrifugal force stronger than the capillary force applied to the reaction solution in the second flow path 112 due to the rotation of the sample analysis substrate 100 acts on the reaction solution including the complex 310 of the reaction chamber 106, the reaction solution becomes the main solution. It is transferred to the chamber 107.
- the reaction solution transferred to the main chamber 107 is not subsequently transferred to the recovery chamber 108 in a state where the sample analysis substrate 100 is rotating. This is because, as described above, since the third flow path 113 forms a siphon, the liquid does not move in the direction toward the rotating shaft 110 against the centrifugal force against the centrifugal force. Of the reaction liquid containing the composite 310 transferred to the main chamber 107, most of the magnetic particles 311 are captured by the outermost peripheral side surface 107 a by the magnetic force of the magnet 121.
- the rotation speed of the sample analysis substrate 100 is set such that a centrifugal force is generated by the rotation, so that a liquid such as a reaction solution does not move by gravity, and a centrifugal force stronger than the capillary force of each capillary channel can be applied. Is done.
- this rotation speed is set for rotation using centrifugal force.
- the rotation direction of the sample analysis substrate 100 is clockwise.
- the cleaning liquid is transferred from the first storage chamber 104 to the first holding chamber 101 through the fourth flow path 114. Further, the substrate solution is transferred from the second storage chamber 105 to the second holding chamber 102 through the fifth flow path 115.
- the sample analysis substrate 100 is stopped at a predetermined first angle.
- the predetermined first angle means that in the sample analysis substrate 100, the cleaning liquid transferred to the first holding chamber 101 exceeds the opening 111 g of the first flow path 111, and the first portion 111q, the substrate solution in the second holding chamber 102 does not contact the opening 116g of the sixth channel 116, and the reaction solution in the main chamber 107 does not contact the opening 113g of the third channel 113.
- the angle at which contact can be made.
- This angle depends on the shape of the second holding chamber 102, the main chamber 107, and the first holding chamber 101, the position in the substrate 100 ′, the amount of the cleaning solution, the substrate solution, and the reaction solution, the inclination angle ⁇ of the sample analysis substrate 100, and the like.
- the gravity direction (indicated by an arrow) in the sample analysis system 501 projected onto a plane parallel to the sample analysis substrate 100 may be within the angle range indicated by ⁇ 1 of the sample analysis substrate 100. .
- the reaction liquid in the main chamber 107 fills the third channel 113 by capillary force by contacting the opening 113g of the third channel 113.
- Step S13 The sample analysis substrate 100 is rotated. A centrifugal force is generated with the rotation, and acts on the reaction solution and the magnetic particles 311 (complex 310 and unreacted magnetic particles) in the main chamber 107. This centrifugal force works so that the liquid and the composite 310 move to the outermost peripheral side surface 107 a side of the main chamber 107. For this reason, the magnetic particles 311 are pressed against the outermost peripheral side surface 107a.
- the reaction solution subjected to the centrifugal force is discharged from the third flow path 113 and transferred to the recovery chamber 108. Due to the sum of the centrifugal force and the attractive force of the magnet 121, the magnetic particles 311 are strongly pressed against the outermost peripheral side surface 107 a and captured.
- the cleaning liquid in the first holding chamber 101 receives centrifugal force due to rotation, but remains in the first holding chamber 101 because it is pressed against the outermost peripheral side surface 101 a of the first holding chamber 101.
- the substrate solution in the second holding chamber 102 also receives centrifugal force due to rotation, but remains in the second holding chamber 102 because it is pressed against the first outer peripheral side surface 102a1.
- the reaction solution and the magnetic particles 311 are separated. Specifically, the reaction liquid moves to the recovery chamber 108 and the magnetic particles 311 remain in the main chamber 107. Even if the rotation of the sample analysis substrate 100 stops, the magnetic particles 311 can remain in the state of being collected on the outermost peripheral side surface 107a by the attractive force received from the magnet 121.
- the stop angle at this time may be the first angle, the second angle of the next step, or another angle.
- Step S14 As shown in FIG. 9, when the sample analysis substrate 100 is not stopped at the second angle in the previous step, the sample analysis substrate 100 is slightly rotated and stopped at the predetermined second angle.
- the second angle is an angle at which the cleaning liquid transferred to the first holding chamber 101 comes into contact with the opening 111 g of the first flow path 111.
- the gravity direction is an angle within the angle range indicated by ⁇ ⁇ b> 2 of the sample analysis substrate 100.
- the cleaning liquid comes into contact with the first portion 111q of the first channel 111 through the opening 111g, the cleaning liquid is sucked into the entire first region 111qe of the first portion 111q by the capillary force, and the first portion 111q of the first channel 111 is drawn.
- the second portion 111r is filled with the cleaning liquid.
- the second region 111qf is also filled with the cleaning liquid moved by gravity. Thereby, the washing
- the first flow path 111 In order to ensure that the first flow path 111 is filled with the cleaning liquid, it may be rotated about several degrees alternately around the second angle in the clockwise direction and counterclockwise, that is, may be swung. Since capillary force acts on the first flow path 111, the cleaning liquid does not move from the second portion 111 r of the first flow path 111 to the main chamber 107 at this time.
- Step S15 Subsequently, the sample analysis substrate 100 is rotated. The centrifugal force due to the rotation acts on the cleaning liquid in the first flow path 111 and the first holding chamber 101. As described with reference to FIG. 3G, the cleaning liquid located on the first flow path 111 side with respect to the straight line db moves to the first portion 106q of the reaction chamber 106 via the first flow path 111. Further, the cleaning liquid located on the first holding chamber 101 side with respect to the straight line db is returned to the first holding chamber 101 by centrifugal force. Accordingly, as shown in FIG. 10, only the cleaning liquid weighed by the first flow path 111 is transferred to the first portion 106 q of the reaction chamber 106.
- the substrate solution is pressed against the first outer peripheral side surface 102a1 in the second holding chamber 102 by centrifugal force. For this reason, the substrate solution remains in the second holding chamber 102.
- the cleaning liquid in the first holding chamber 101 is pressed against the outermost peripheral side surface 101a in the first holding chamber 101 by centrifugal force. For this reason, the cleaning liquid remains in the first holding chamber 101.
- the sample analysis substrate 100 is stopped at a predetermined third angle as shown in FIG.
- the third angle is an angle at which the cleaning liquid in the first holding chamber 101 does not contact the opening 111g.
- the gravity direction in the sample analysis system 501 projected onto a plane parallel to the sample analysis substrate 100 may be within the angle range indicated by ⁇ 3 on the sample analysis substrate 100.
- the cleaning liquid fills the first region 106qf that is the non-capillary space of the first portion 106q and the second region 106qe that is the capillary space. Since the second region 106qe is connected to the second region 106re, which is a capillary space of the second portion 106r, the cleaning liquid is sucked into these capillary spaces by the capillary force. As a result, the cleaning liquid located in the first region 106qf of the first portion 106q moves to the second region 106re of the second portion 106r. As a result, the reaction solution remaining in the first portion 106q and the second portion 106r of the reaction chamber 106 is mixed with the cleaning liquid. That is, the reaction chamber 106 is cleaned with the cleaning liquid.
- the cleaning liquid in the reaction chamber 106 contacts the opening 112g of the second flow path 112, thereby filling the second flow path 112 by capillary action.
- Step S16 The sample analysis substrate 100 is rotated, and the cleaning liquid is moved from the second region 106re of the second portion 106r of the reaction chamber 106 to the main chamber 107. Since the second flow path 112 is filled with the cleaning liquid, if the centrifugal force stronger than the capillary force acts on the cleaning liquid in the second flow path 112 by the rotation of the sample analysis substrate 100, the cleaning liquid is stored in the main chamber 107. It is transferred to. Accordingly, the reaction solution remaining in the first portion 106q and the second portion 106r of the reaction chamber 106 is transferred to the main chamber 107 together with the cleaning liquid. Further, the magnetic particles 311 in the main chamber 107 come into contact with the cleaning liquid, and the first cleaning is performed.
- the cleaning liquid transferred to the main chamber 107 is not subsequently transferred to the recovery chamber 108 while the sample analysis substrate 100 is rotating. This is because the third flow path 113 constitutes a siphon, so that the cleaning liquid does not move in the direction toward the rotation shaft 110 against the centrifugal force.
- the cleaning liquid in the first holding chamber 101 and the substrate solution in the second holding chamber 102 remain in the respective chambers.
- the sample analysis substrate 100 is stopped at a predetermined fourth angle.
- the fourth angle is an angle at which the cleaning liquid in the first holding chamber 101 does not contact the opening 111g and the cleaning liquid transferred to the main chamber 107 can contact the opening 113g of the third flow path 113.
- the gravity direction in the sample analysis system 501 projected onto a plane parallel to the sample analysis substrate 100 may be within the angle range indicated by ⁇ 4 on the sample analysis substrate 100.
- the cleaning liquid in the main chamber 107 is in contact with the opening 113g of the third channel 113 and fills the third channel 113 by capillary action.
- Step S17 The sample analysis substrate 100 is rotated. A centrifugal force is generated with the rotation and acts on the cleaning liquid and the magnetic particles 311 in the main chamber 107. This centrifugal force works so that the cleaning liquid and the magnetic particles 311 move toward the outermost peripheral side surface 107 a of the main chamber 107, and the magnetic particles 311 are captured on the outermost peripheral side surface 107 a by the centrifugal force and the attractive force by the magnet 121.
- the cleaning liquid subjected to the centrifugal force is discharged from the third flow path 113 and transferred to the recovery chamber 108. For this reason, only the cleaning liquid is discharged from the third flow path 113, and the magnetic particles 311 remain in the main chamber 107.
- the cleaning liquid in the first holding chamber 101 and the substrate solution in the second holding chamber 102 are pressed against the outermost peripheral side surface 101a and the first outer peripheral side surface 102a1, respectively, and remain in the first holding chamber 101 and the second holding chamber 102.
- the rotation of the sample analysis substrate 100 is stopped. As a result, the cleaning liquid and the magnetic particles 311 are separated. Specifically, the cleaning liquid moves to the recovery chamber 108 and the magnetic particles 311 remain in the main chamber 107. Even if the rotation of the sample analysis substrate 100 stops, the magnetic particles 311 can remain in the state of being collected on the outermost peripheral side surface 107a by the attractive force received from the magnet 121.
- the stop angle at this time may be the fourth angle or the fifth angle of the next step.
- Step S18 As shown in FIG. 14, when not stopping at the fifth angle in the previous step, the sample analysis substrate 100 is slightly rotated and stopped at the predetermined fifth angle.
- the fifth angle is an angle at which the cleaning liquid transferred to the first holding chamber 101 comes into contact with the opening 111 g of the first flow path 111.
- the gravity direction is an angle within the angle range indicated by ⁇ 5 of the sample analysis substrate 100. Since the amount of the cleaning liquid remaining in the first holding chamber 101 in step S4 is different, the angle range ⁇ 5 may be different from the angle range ⁇ 2.
- the cleaning liquid is sucked into the first flow path 111 from the first holding chamber 101 by the capillary force in the first portion 111q of the first flow path 111, and the first portion 111q and the second portion 111r of the first flow path 111 are the cleaning liquid. It is filled. Thereby, the cleaning liquid for one time is weighed again.
- the sample analysis substrate 100 may be swung around the fifth angle so that the first flow path 111 is surely filled with the cleaning liquid. Since capillary force acts on the first flow path 111, the cleaning liquid does not move from the first flow path 111 to the reaction chamber 106 at this time.
- Step S19 Subsequently, the sample analysis substrate 100 is rotated. The centrifugal force due to the rotation acts on the cleaning liquid in the first flow path 111 and the first holding chamber 101. Similar to the first cleaning, the cleaning liquid positioned on the first flow path 111 side moves to the first portion 106q of the reaction chamber 106 via the first flow path 111 with reference to the straight line db shown in FIG. 3G. Further, the cleaning liquid located on the first holding chamber 101 side with respect to the straight line db is returned to the first holding chamber 101 by centrifugal force. Therefore, as shown in FIG. 15, only the cleaning liquid weighed by the first flow path 111 is transferred to the first portion 106 q of the reaction chamber 106.
- the substrate solution is pressed against the first outer peripheral side surface 102a1 in the second holding chamber 102 by centrifugal force. For this reason, the substrate solution remains in the second holding chamber 102.
- the cleaning liquid in the first holding chamber 101 is pressed against the outermost peripheral side surface 101a in the first holding chamber 101 by centrifugal force. For this reason, the cleaning liquid remains in the first holding chamber 101.
- the sample analysis substrate 100 is stopped at a predetermined sixth angle.
- the sixth angle is an angle at which the cleaning liquid in the first holding chamber 101 does not contact the opening 111g.
- the gravity direction in the sample analysis system 501 projected onto a plane parallel to the sample analysis substrate 100 may be within the angle range indicated by ⁇ 6 on the sample analysis substrate 100.
- the cleaning liquid is the first region 106qf that is the non-capillary space of the first portion 106q and the second space that is the capillary space.
- the region 106qe is filled. Since the second region 106qe is connected to the second region 106re, which is a capillary space of the second portion 106r, the cleaning liquid is sucked into these capillary spaces by the capillary force. As a result, the cleaning liquid located in the first region 106qf of the first portion 106q moves to the second region 106re of the second portion 106r. As a result, the reaction solution that may remain in the first portion 106q and the second portion 106r of the reaction chamber 106 is mixed with the cleaning liquid. That is, the reaction chamber 106 is cleaned again with the cleaning liquid.
- the cleaning liquid in the reaction chamber 106 contacts the opening 112g of the second flow path 112, thereby filling the second flow path 112 by capillary action.
- Step S20 As in the first cleaning, the sample analysis substrate 100 is rotated, and the cleaning liquid is moved from the second region 106re of the second portion 106r of the reaction chamber 106 to the main chamber 107. Since the second flow path 112 is filled with the cleaning liquid, if the centrifugal force stronger than the capillary force acts on the cleaning liquid in the second flow path 112 by the rotation of the sample analysis substrate 100, the cleaning liquid is stored in the main chamber 107. It is transferred to. Thereby, the reaction solution that may remain in the first portion 106q and the second portion 106r of the reaction chamber 106 is transferred to the main chamber 107 together with the cleaning liquid. Further, the magnetic particles 311 in the main chamber 107 come into contact with the cleaning liquid, and the second cleaning is performed.
- the cleaning liquid transferred to the main chamber 107 is not subsequently transferred to the recovery chamber 108 when the sample analysis substrate 100 is rotating as described above.
- the cleaning liquid in the first holding chamber 101 and the substrate solution in the second holding chamber 102 remain in the respective chambers.
- the sample analysis substrate 100 is stopped at a predetermined seventh angle.
- the seventh angle is an angle at which the cleaning liquid in the first holding chamber 101 does not come into contact with the opening 111 g and the cleaning liquid transferred to the main chamber 107 can come into contact with the opening 113 g in the third flow path 113.
- the gravity direction in the sample analysis system 501 projected onto a plane parallel to the sample analysis substrate 100 may be within the angle range indicated by ⁇ 7 on the sample analysis substrate 100.
- the cleaning liquid in the main chamber 107 is in contact with the opening 113g of the third channel 113 and fills the third channel 113 by capillary action.
- Step S21 The sample analysis substrate 100 is rotated. A centrifugal force is generated with the rotation and acts on the cleaning liquid and the magnetic particles 311 in the main chamber 107. This centrifugal force works so that the cleaning liquid and the magnetic particles 311 move toward the outermost peripheral side surface 107 a of the main chamber 107, and the magnetic particles 311 are captured on the outermost peripheral side surface 107 a by the centrifugal force and the attractive force by the magnet 121.
- the cleaning liquid that has received the centrifugal force is discharged from the third flow path 113 and transferred to the collection chamber 108. For this reason, only the cleaning liquid is discharged from the third flow path 113, and the magnetic particles 311 remain in the main chamber 107.
- the cleaning liquid in the first holding chamber 101 is pressed against the outermost peripheral side surface 103 a and stays in the first holding chamber 101.
- the substrate solution is also pressed against the first outer peripheral side surface 102 a 1 and remains in the second holding chamber 102.
- the rotation of the sample analysis substrate 100 is stopped. Thereby, as shown in FIG. 18, the cleaning liquid and the magnetic particles 311 are separated. Specifically, the cleaning liquid moves to the recovery chamber 108 and the magnetic particles 311 remain in the main chamber 107. Even if the rotation of the sample analysis substrate 100 stops, the magnetic particles 311 can remain in the state of being collected on the outermost peripheral side surface 107a by the attractive force received from the magnet 121.
- the stop angle at this time may be the seventh angle or the eighth angle of the next step. B / F separation and cleaning are completed by the above steps.
- Step S22 First, the substrate solution is moved from the second holding chamber 102 to the third holding chamber 103.
- the sample analysis substrate 100 is slightly rotated and stopped at the predetermined eighth angle. At this time, the sample analysis substrate 100 is rotated clockwise.
- the eighth angle is an angle at which the substrate solution in the second holding chamber 102 comes into contact with the opening 116g of the sixth flow path 116 and the entire amount of the substrate solution can move to the third holding chamber 103 by gravity.
- the angle is approximately at which the sixth flow path 116 is disposed along the direction of gravity. Thereby, the substrate solution in the second holding chamber 102 is transferred to the third holding chamber 103.
- the sample analysis substrate 100 is rotated clockwise so that the substrate solution comes into contact with the capillary space 103re of the second portion 103r in the third holding chamber 103 at a predetermined ninth angle. Stop.
- the substrate solution is sucked into the capillary space 103re by the contact between the capillary space 103re and the substrate solution.
- the substrate solution that fills the capillary space 103re is sucked into the seventh flow channel 117 from the opening 117g by the capillary force, and the first region 117qe and the second region of the first portion 117q of the seventh flow channel.
- Portion 117r is filled with substrate solution. Thereby, the substrate solution is weighed.
- the seventh flow path 117 In order to ensure that the seventh flow path 117 is filled with the substrate solution, it may be rotated about the ninth angle alternately about several degrees clockwise or counterclockwise, that is, may be swung. Since capillary force acts on the seventh flow path 117, the cleaning liquid does not move from the second portion 112r of the seventh flow path 117 to the main chamber 107 at this time.
- Step S23 Subsequently, the sample analysis substrate 100 is rotated.
- the centrifugal force due to the rotation acts on the cleaning liquid in the seventh flow path 117 and the first holding chamber 101.
- the substrate solution in the seventh flow path 117 moves to the main chamber 107 by centrifugal force.
- the substrate solution located closer to the third holding chamber 103 than the opening 112 g is pressed against the outermost peripheral side surface 102 a of the third holding chamber 103 by the centrifugal force, and remains in the third holding chamber 103.
- the substrate solution moved to the main chamber 107 contains a substrate.
- This substrate reacts with the labeling substance 307 contained in the labeling antibody 308 in the magnetic particles 311 held in the main chamber 107, or causes a change in light emission, fluorescence or absorption wavelength by the catalytic reaction of the labeling substance 307. .
- the rotation of the sample analysis substrate 100 is stopped at the tenth angle as shown in FIG.
- the tenth angle indicates that the main chamber 107 is optically measured so that a change in the emission, fluorescence or absorption wavelength of the substrate in the main chamber 107 can be detected, for example, the light receiving element of the optical measurement unit 207 is close to the main chamber 107. This is an angle arranged with a predetermined positional relationship with respect to the unit 207.
- the optical measurement unit 207 performs optical measurement of the liquid held in the main chamber 107. Specifically, the optical measurement unit 207 detects a signal such as a dye, luminescence, or fluorescence of a substrate corresponding to the labeling substance 307 of the labeled antibody 308 bound to the complex 310 included in the magnetic particle 311. Thereby, detection of the antigen 306, determination of the concentration of the antigen 306, and the like can be performed.
- a signal such as a dye, luminescence, or fluorescence of a substrate corresponding to the labeling substance 307 of the labeled antibody 308 bound to the complex 310 included in the magnetic particle 311.
- Optical measurement by the optical measurement unit 207 may be performed with the sample analysis substrate 100 rotated.
- the substrate analysis substrate 100 may be rotated to detect signals such as the dye, luminescence, and fluorescence of the substrate.
- the first holding chamber 101 that holds the cleaning liquid is connected to the reaction chamber 106 by the first flow path 111, and the cleaning liquid weighed by the first flow path 111 is the reaction chamber.
- the composite body 310 to be cleaned is transferred to the main chamber 107 through 106. Therefore, even when the reaction solution remains in the reaction chamber 106, it can be washed with the cleaning solution, and the remaining reaction solution is transferred to the chamber holding the complex 310 and reacted with the substrate solution. It is possible to suppress the occurrence of errors in the measurement of the specimen. Therefore, it is possible to analyze the detection of the sample with high accuracy.
- the first holding chamber 101 in which the substrate solution is held during the B / F separation and washing process has a space extending in the circumferential direction and is adjacent to the outermost peripheral side surface. Out of the two adjacent side surfaces, the adjacent side surfaces that are adjacent to the reaction chamber 106, the first holding chamber 101 that holds the cleaning liquid, and the adjacent side surfaces that are close to the main chamber 107, are located farther from these, and An opening of the sixth flow path 116 is provided on the adjacent side. For this reason, even if the sample analysis substrate 100 is rotated at various angles, the substrate solution is unlikely to contact the opening 111g of the sixth flow path 116, and the substrate solution is prevented from being transferred to the third holding chamber 103. Can do.
- sample analysis substrate 100 (Another embodiment of sample analysis substrate 100) Various modifications can be made to the sample analysis substrate 100 of the above embodiment.
- reaction chamber 106 has the first portion 106q and the second portion 106r, but the first portion 106q may not be provided.
- the reaction chamber 106 ' has only the second region 106re' of the second portion.
- the second portion 111r of the first flow path 111 is opposed to the outermost peripheral side surface 106ra of the reaction chamber 106 'and is connected to the innermost peripheral side surface 106rb closest to the rotating shaft 110.
- the drying reagent 125 is held in the second region 106re '.
- the second region 106re ' may be a capillary space or a non-capillary space. Further, the drying reagent 125 may not be held in the second region 106re '.
- the magnetic particle-immobilized antibody 305 and the labeling substance 307 may be introduced into the reaction chamber 106 ′ together with the sample solution, or they may be transferred from another chamber.
- the second channel 112 may be a channel that can be transferred by capillary force, or may be a channel that can be transferred by gravity.
- the reaction chamber 106 ′′ has the first region 106rf ′ and the second region 106re ′ of the second portion, and does not have the first portion.
- the first region 106rf ' is a non-capillary space
- the second region 106re' is a capillary space.
- the drying reagent 125 is disposed in the second region 106re '. However, the drying reagent 125 may not be provided.
- the second channel 112 may be a channel that can be transferred by capillary force, or may be a channel that can be transferred by gravity.
- the first flow path 111 has a capillary space in which the cleaning liquid can be weighed, but the first flow path may not have a function of weighing.
- the sample analysis substrate 163 shown in FIG. 24A is different from the sample analysis substrate 100 in that the first flow path 211 is a flow path capable of transferring a liquid by gravity.
- the amount of the cleaning liquid flowing from the first holding chamber 101 to the first flow path 211 is controlled by controlling the rotation angle of the sample analysis substrate 163.
- the cleaning liquid can be transferred to the reaction chamber 106 in batches.
- the first holding chamber 101 has an outermost peripheral side surface 101a farthest from the rotary shaft 110 and an adjacent side surface 101c adjacent to the outermost peripheral side surface 101a, and has an opening on the rotary shaft 110 side by the outermost peripheral side surface 101a and the adjacent side surface 101c.
- a recess space is formed.
- the first flow path 211 is a flow path having an opening 211g and an opening 211h and capable of transferring a liquid by gravity, and the opening 211g is provided at a position near the rotating shaft 110 of the adjacent side surface 101c.
- the opening 211h of the first flow path 211 is connected to the first region 106qf of the first portion 106q of the reaction chamber.
- the cleaning liquid starts to flow into the first flow path 211 due to gravity, and the movement of the cleaning liquid causes The cleaning liquid is transferred until the surface of the cleaning liquid moves backward and coincides with the opening 211g. Therefore, the cleaning liquid can be transferred to the reaction chamber 106 a plurality of times by changing the angular position for holding the sample analysis substrate 163.
- a sample analysis substrate 164 shown in FIG. 24B and a sample analysis substrate 165 shown in FIG. 24C are further modified examples of the sample analysis substrate 163, and the reaction chamber 106 ′ shown in FIG. 23A and the reaction chamber shown in FIG. It differs from the sample analysis substrate 163 in that it includes 106 ′′. Even with these sample analysis substrates, the effect of cleaning the reaction chamber described above can be obtained.
- the first flow path may be a flow path capable of transferring liquid by capillary force.
- the sample analysis substrates 166, 167, and 168 shown in FIGS. 25A to 25C can transfer liquid by capillary force instead of the first flow path 211 of the sample analysis substrates 163, 164, and 165 of FIGS. 24A to 24C.
- 1st flow path 211 ' is provided.
- the first flow path 211 ' constitutes a siphon. Due to the siphon structure, when the reaction liquid held in the reaction chamber is transferred to the main chamber, the cleaning liquid transferred from the first storage chamber 104 to the first holding chamber 101 remains as it is in the first flow path. Transfer to the reaction chamber via 211 ′ can be prevented.
- the cleaning liquid is transferred from the first holding chamber 101 to the reaction chambers 106, 106 ′, 106 ′′ by the centrifugal force generated by the rotation of the sample analysis substrate. For this reason, it is difficult to transfer the cleaning liquid to the reaction chambers 106, 106 ', 106 "in a plurality of times.
- a sample analysis substrate 169 shown in FIG. 26 includes a first holding chamber 133 including a first portion 133q, a second portion 133r, and a connecting portion 133p that connects the second portion 133r and the first portion 133q.
- the second portion 133r and a part of the first portion 133q are generally arranged in the circumferential direction around the rotation shaft 110.
- a wall portion 100f constituted by the inner surface of the substrate 100 ' is located between the second portion 133r and the first portion 133q.
- the wall portion 100f separates the second portion 133r and the first portion 133q.
- the connecting portion 133p is located on the same radial direction as the wall portion 100f of the substrate 100 ', and is located closer to the rotating shaft 110 than the wall portion 100f.
- the connecting portion 133p is not filled with liquid by capillary action, and moves the liquid between the first portion 133q and the second portion 133r by gravity.
- the second portion 133r is located on the outer side of an arc ca having a radius that is a line segment connecting the rotation shaft 110 and the point 100e closest to the rotation axis of the wall portion 100f with the rotation shaft 110 as a center (the rotation shaft 110). Part 133re (located away from). With this portion 133re, a predetermined amount of cleaning liquid used for one cleaning can be weighed.
- the distance from the rotating shaft 110 to the opening 111g of the first flow path 111 in the second portion 133r is longer than the distance from the rotating shaft 110 to the point 100e closest to the rotating shaft of the wall portion 100f. For this reason, the cleaning liquid weighed by the portion 133re can be transferred from the first flow path 111 to the reaction chamber 106 by centrifugal force due to rotation.
- the first portion 133q of the first holding chamber 133 includes a side portion 133qt and a bottom portion 133qs.
- the side portion 133qt is located on the side of the first storage chamber 104 in the circumferential direction around the rotation shaft 110.
- the bottom 133qs is located farther from the rotation shaft 110 than the first storage chamber 104. Further, a part of the side portion 133qt and the entire bottom portion 133qs of the first portion 133q are located farther from the rotation shaft 110 than the second portion 133r.
- the side portion 133qt preferably includes a portion 133qt 'located on the rotating shaft 110 side with respect to the arc ca and a portion 133qt' 'located on the outer side. As described above, the portion 133qt 'is adjacent to the first portion 133q in the circumferential direction, and is connected to the connecting portion 133p.
- the portion located outside the arc ca (far from the rotating shaft 110), that is, the total volume of the portion 133qt '' and the bottom 133qs is the first storage chamber. It is preferable that it is larger than the total amount of the cleaning liquid retained in 104.
- the space of the first holding chamber 133 includes the bottom portion 133qs, when the sample analysis substrate 169 is stopped at a predetermined angle, a part of the cleaning liquid stored in the first storage chamber 104 is caused by capillary action.
- the fourth flow path 114 is filled. Then, by rotating the sample analysis substrate 169 in a state where the fourth flow path 114 is filled with the cleaning liquid, the centrifugal liquid causes the cleaning liquid in the first storage chamber 104 to the bottom 133qs via the fourth flow path 114. Be transported.
- the volume of the portion of the second portion 133r located outside the arc ca having a radius connecting the rotation axis 110 and the point 100e closest to the rotation axis 110 of the wall portion 100f is the first holding chamber 133. It is 1/2 or less of the volume.
- the first portion 133q is configured to include a part of the side portion 133qt and the bottom portion 133qs. However, the first portion 133q is centered on the rotation shaft 110, and the rotation shaft 110 of the rotation shaft 110 and the wall portion 100f. What is necessary is just to include the part located in the outer side of the circular arc which makes the line segment which connects the point nearest to to the radius.
- the cleaning liquid weighed in a certain amount fills the first flow path 111 by a capillary phenomenon, and then the sample analysis substrate 169 is more than the capillary force applied to the liquid in the first flow path 111.
- the centrifugal force is transferred to the main chamber 107 through the first flow path 111.
- the sample analysis substrate 169 shown in FIG. 26 includes the reaction chamber 106 shown in FIG. 3B or the like, but may include the reaction chamber 106 ′ or 106 ′′ shown in FIGS. 23A and 23B.
- the first holding chamber 101 holds the cleaning liquid for a plurality of times, but it may include a plurality of chambers for holding the cleaning liquid for one time.
- FIG. 27 includes a first storage chamber 104A, a third storage chamber 104B, a fourth flow path 114A, an eighth flow path 114B, a first holding chamber 101A, a fourth holding chamber 101B, and a first flow.
- a path 111A and a tenth flow path 111B are provided.
- the first storage chamber 104A, the fourth flow path 114A, the first holding chamber 101A and the first flow path 111A, and the third storage chamber 104B, the eighth flow path 114B, the fourth holding chamber 101B and the tenth flow path 111B In this way, the cleaning liquid for one independent time is held and transferred to the reaction chamber 106.
- the fourth flow path 114A and the eighth flow path 114B connect the first storage chamber 104A and the third storage chamber 104B to the first holding chamber 101A and the fourth holding chamber 101B, respectively, and the first flow path 111A and the tenth flow path 114B.
- the flow path 111B connects the first holding chamber 101A and the fourth holding chamber 101B and the reaction chamber 106.
- the fourth channel 114A and the eighth channel 114B are capillary channels and have a siphon structure.
- the first channel 111A and the tenth channel 111B have a structure capable of transferring a liquid by gravity.
- the first holding chamber 101A and the first flow path 111A are located farther from the rotating shaft 110 than the first storage chamber 104A and the third storage chamber 104B, respectively.
- the first channel 111A and the tenth channel 111B can transfer liquid by gravity.
- the first holding chamber 101A has an outermost peripheral side surface 101Aa and an adjacent side surface 101Ac adjacent to the outermost peripheral side surface.
- a tapered surface, a curved surface, or the like may be provided between the outermost peripheral side surface 101Aa and the adjacent side surface 101Ac for smooth corners (ridges).
- An opening 111Ag of the first flow path 111A is disposed at an end where the outermost peripheral side surface 101Aa is not located, at both ends of the adjacent side surface 101Ac.
- a concave portion having an opening is formed on the rotating shaft 110 side by the outermost peripheral side surface 101Aa and the adjacent side surface 101Ac, and a single cleaning liquid is held.
- the fourth holding chamber 101B has an outermost peripheral side surface 101Ba and an adjacent side surface 101Bc adjacent to the outermost peripheral side surface.
- a tapered surface, a curved surface, or the like may be provided between the outermost peripheral side surface 101Ba and the adjacent side surface 101Bc for smoothing the corners (ridges).
- the opening 111Bg of the tenth flow path 111B is arranged at the end where the outermost peripheral side surface 101Ba is not located.
- the outermost peripheral side surface 101Ba and the adjacent side surface 101Bc form a recess having an opening on the rotating shaft 110 side to hold a single cleaning liquid.
- the first holding chamber 101A and the fourth holding chamber 101B are located in the same one of the two regions divided by a straight line connecting the center of the reaction chamber 106 and the rotating shaft 110.
- the rotation axis 110 of the sample analysis substrate 170 is inclined at an angle greater than 0 ° and less than 90 ° with respect to the direction of gravity so that the recesses of the first holding chamber 101A and the recesses of the fourth holding chamber 101B can hold liquid. Support to do. Further, the sample analysis substrate 170 is held at a predetermined rotation angle so that the reaction chamber 1067 is positioned below the first holding chamber 101A and the fourth holding chamber 101B in the gravity direction. In this case, since the adjacent side surface 101Ac of the first holding chamber 101A and the adjacent side surface 101Bc of the fourth holding chamber 101B are non-parallel when viewed from the direction parallel to the rotation shaft 110, they are held from either one of the chambers.
- the other chamber can hold at least a part of the cleaning liquid. Therefore, by appropriately selecting the rotation angle of the sample analysis substrate 170, the cleaning liquid can be selectively transferred from the first holding chamber 101A and the fourth holding chamber 101B to the reaction chamber 106 at different timings.
- the angle ⁇ formed by the adjacent side surface 101Ac with respect to the straight line connecting the center of the reaction chamber 106 and the rotation shaft 110 when viewed from the direction parallel to the rotation shaft 110 is Bigger than. Therefore, the rotation angle of the sample analysis substrate 170 in which the first holding chamber 101A and the fourth holding chamber 101B are positioned below the reaction chamber 106 in the direction of gravity (P1 shown in FIG. 27 coincides with the 6 o'clock direction).
- the adjacent side surface 101Ac is first parallel (horizontal) to the direction orthogonal to the direction of gravity, so that the inside of the first holding chamber 101A is The entire amount of the cleaning liquid can be selectively transferred to the reaction chamber 106, and then the cleaning liquid held in the fourth holding chamber 101 ⁇ / b> B can be selectively transferred to the main chamber 107.
- each of the first portion 106q and the second portion 106r has a capillary space and a non-capillary space.
- the capillary space and non-capillary space in the reaction chamber are not limited to this, and various modifications are possible.
- the reaction chamber may include a first portion 106q having only the first region 106qf, and a second portion 106r having the first region 106rf and the second region 106re. That is, in the reaction chamber 106 illustrated in FIG. 3C and the like, the second region 106qe of the first portion 106q may be omitted.
- the first region 106rf is positioned closer to the rotation shaft 110 than the second region 106re, and the first region 106rf is connected to the first region 106qf of the first portion 106q. Yes.
- the first region 106qf and the first region 106rf are non-capillary spaces, and the second region 06re is a capillary space.
- the second portion 106r may not include the first region 106rf.
- the second portion 106r is configured only by the second region 106re that is a capillary space, and the second region 106re is connected to the second region 106qe of the first portion 106q.
- the second region 106qe in the first portion 106q may include 106s that is located closer to the rotating shaft 110 than the wall portion 126 and is connected to the second portion 106r.
- the first portion 106q may include only the first region 106qf that is a non-capillary space
- the second portion 106r may include only the second region 106re that is a capillary space.
- the boundary (connection portion) between the first portion 106q and the second portion 106r is located at a radial position connecting the rotation shaft 110 and the point 126p closest to the rotation shaft 110 of the wall portion 126.
- the reaction chamber may not have the first portion.
- the first portion 106q may include a first region 106qf and a second region 106qe, and the first region 106qf may be located closer to the rotation shaft 110 than the second region 106qe.
- the measurement system using magnetic particles has been described.
- the sample analysis substrate, the sample analysis device, the sample analysis system, and the sample analysis system program according to one aspect of the present application are provided with magnetic particles.
- the target to which the primary antibody is immobilized may be a wall surface in the chamber instead of the magnetic particles. That is, when the chamber is made of a material such as polystyrene or polycarbonate, the primary antibody can be immobilized on the wall surface in the chamber by physical adsorption, and sandwiched with antigen or labeled antibody in the chamber. The reaction can be triggered.
- a primary antibody has a functional group (for example, amino group or carboxyl group) that can bind to the primary antibody on the wall surface in the chamber, and the primary antibody can be immobilized by chemical bonding. And a sandwich-type binding reaction.
- a primary antibody can be couple
- the primary antibody When the primary antibody is immobilized on the wall surface of the chamber by physical adsorption or chemical bonding, it is mainly used in a system for detecting a dye, chemiluminescent or fluorescent signal. On the other hand, when the primary antibody is immobilized on a metal substrate, it is mainly used as a signal in a system for detecting an electrochemical signal (for example, current) or an electrochemiluminescence signal. In this case, the magnet 121 shown in FIG. 3B is unnecessary. Further, the reaction field for forming the complex 310 is not the reaction chamber 106 but the main chamber 107. Therefore, the primary antibody needs to be immobilized on the wall surface of the main chamber 107.
- sample analysis substrate, sample analysis apparatus, sample analysis system, and sample analysis system program of the present disclosure can be applied not only to non-competitive methods (sandwich immunoassay methods) but also to competitive methods and gene detection methods by hybridization. It is.
- the cleaning is performed twice, but may be performed three or more times as necessary.
- sample analysis substrate, sample analysis apparatus, sample analysis system, and sample analysis system program disclosed in the present application can be applied to analysis of specific components in a sample using various reactions.
- Sample analysis substrate 100 ′ Substrate 100a Base substrate 100b Cover substrate 100c, 100d Main surface 100f Wall portion 101, 101A First holding chamber 101B Fourth holding chamber 101a, 102a Outermost peripheral side surface 101c, 101d Adjacent side surface 102 Second holding chamber 103 third holding chamber 104 first storage chamber 105 second storage chamber 106 main chamber 106q first portion 106qa outermost peripheral side surface 106qe second region 106qf first region 106r second portion 106ra outermost peripheral side surface 106rb innermost peripheral side surface 106re second Area 106rf first area 107 main chamber 108 recovery chamber 110 rotating shaft 111 first flow path 112 second flow path 113 third flow path 114 first 4th channel 115 5th channel 116 6th channel 117 7th channel 120 Magnet storage chamber 121 Magnet 122 Air hole 123 Opening 125 Drying reagent 126 Wall parts 161-169 Sample analysis substrate 200 Sample analysis device 201 Motor 201a Turntable 203 Origin detector 203a Light source 203b Light receiving element
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Abstract
Description
回転運動によって、液体の移送を行う試料分析用基板であって、
回転軸を有する基板と、
前記基板内に位置し、第1液体を保持するための第1空間を有する第1保持チャンバーと、
前記基板内に位置し、検体を含む液体試料を保持するための空間を有する反応チャンバーと、
前記基板内に位置しており、第1開口および第2開口を有する第1流路であって、前記第1開口および前記第2開口がそれぞれ前記第1保持チャンバーおよび前記反応チャンバーに接続された第1流路と、
前記基板内に位置し、前記検体を含む液体試料および表面にリガンドが固定された磁性粒子を保持するための空間を有するメインチャンバーと、
前記基板内に位置しており、第3開口および第4開口を有する第2流路であって、前記第3開口および前記第4開口がそれぞれ前記反応チャンバーおよび前記メインチャンバーに接続された第2流路と、
前記基板内に位置し、磁石を収納することが可能な磁石収納室と、を備え、
前記第1開口は、前記第2開口よりも回転軸に近い側に位置し、
前記第2開口は、前記第3開口よりも回転軸に近い側に位置し、
前記磁石収納室は、前記磁石収納室に磁石が収納された場合、前記磁石によって前記メインチャンバー中の前記磁性粒子を前記メインチャンバー内に捕捉することができる位置に配置されている、
試料分析用基板。
[項目2]
前記反応チャンバーの空間内に配置された、ドライ化試薬をさらに備え、
前記ドライ化試薬は、前記磁性粒子を含む、項目1に記載の試料分析用基板。
[項目3]
前記反応チャンバーは、非毛細管空間を含む、項目1または2に記載の試料分析用基板。
[項目4]
前記反応チャンバーは、毛細管空間を含む、項目1または2に記載の試料分析用基板。
[項目5]
前記反応チャンバーは、非毛細管空間および毛細管空間を含み、
前記非毛細管空間に前記第1開口が接しており、
前記毛細管空間に第3開口が接している、
項目1または2に記載の試料分析用基板。
[項目6]
前記非毛細管空間は、前記毛細管空間よりも回転軸に近接して位置する部分を有する、項目5に記載の試料分析用基板。
[項目7]
前記前記反応チャンバーは第1部分および第2部分を含み、
前記基板は前記反応チャンバーの前記第1部分と前記第2部分との間に位置する壁部分を有し、
前記壁部分は、回転軸に向かう方向に凸部を形成し、
前記第1部分および前記第2部分において、前記壁部分の回転軸から最も近い点と、前記回転軸とを結ぶ線分を半径とする円弧よりも遠い側に前記毛細管空間の一部および前記非毛細管空間の一部がそれぞれ位置する、
項目5に記載の試料分析用基板。
[項目8]
前記第1部分側に位置する前記壁部分の一部又は全部には、前記第1部分と前記第2部分を接続する前記毛細管空間の一部が位置している、
項目7に記載の試料分析用基板。
[項目9]
前記基板内に位置しており、空間を有する回収チャンバーと、
前記基板内に位置しており、第5開口および第6開口を有する第3流路であって、前記第5開口および前記第6開口がそれぞれ前記メインチャンバーおよび前記回収チャンバーに接続された第3流路と、
をさらに備え、
前記第5開口は前記第6開口よりも前記回転軸に近接して位置する、
項目1から8のいずれかに記載の試料分析用基板。
[項目10]
前記第1流路は、非毛細管路である、項目1から9のいずれかに記載の試料分析用基板。
[項目11]
前記第1チャンバーは、前記回転軸から最も離れて位置する最外周側面と、前記最外周側面に隣接する隣接側面とを有し、
前記最外周側面と前記隣接側面とで凹部を形成し、
前記試料分析用基板を所定の角度位置で保持した場合に、前記凹部で前記第1液体を保持する、項目10に記載の試料分析用基板。
[項目12]
前記第1流路は、毛細管路である、項目1から11のいずれかに記載の試料分析用基板。
[項目13]
前記第1流路は、サイフォン構造を有する項目12に記載の試料分析用基板。
[項目14]
前記第1流路の一部は、前記第1開口を挟んで前記第1保持チャンバーの一部よりも前記回転軸に近接して位置している、項目12に記載の試料分析用基板。
[項目15]
前記第1保持チャンバーの空間は、第1部分および第2部分と、前記第1部分および前記第2部分の間に位置しており前記第1部分および前記第2部分を連結する連結部分とを有し、
前記基板は、前記第1保持チャンバーの前記空間の前記第1部分および前記第2部分を区切る壁部分を有し、
前記反応チャンバーは前記第1保持チャンバーの前記第2部分よりも前記回転軸から遠くに位置し、
前記第1保持チャンバーの前記空間の前記連結部分は、前記基板の前記壁部分よりも前記回転軸側に位置し、
前記第1流路は、前記第1保持チャンバーの前記空間の前記第2部分と接続されている、項目12に記載の試料分析用基板。
[項目16]
前記基板内に位置し、第2液体を保持するための空間を有する第4保持チャンバーと、
前記第4保持チャンバーと前記反応チャンバーとを接続し、前記第2液体を移送する第8流路と、
前記第1保持チャンバーは、前記回転軸から最も遠くに位置する最外周側面と、前記最外周側面に隣接する隣接側面と、前記最外周側面および前記隣接側面とで形成される凹部とを有し、
前記第4保持チャンバーは、前記回転軸から最も遠くに位置する最外周側面と、前記最外周側面に隣接する隣接側面と、前記最外周側面および前記隣接側面とで形成される凹部とを有し、前記第1保持チャンバーの前記隣接側面と前記第4保持チャンバーの隣接側面とは、前記回転軸に平行な方向からみて非平行である、
項目1から14のいずれかに記載の試料分析用基板。
[項目17]
項目1から16のいずれかに記載の試料分析用基板と、
前記試料分析用基板を前記回転軸周りに回転させるモータ、
前記モータの回転軸の回転角度を検出する回転角度検出回路、
前記回転角度検出回路の検出結果に基づき、前記モータの回転および停止時の回転角度を制御する駆動回路、および
演算器、メモリおよびメモリに記憶され、前記演算器に実行可能なように構成されたプログラムを含み、前記プログラムに基づき、前記モータ、前記回転角度検出回路および前記駆動回路の動作を制御する制御回路
を有する試料分析装置と、
を備えた試料分析システムであって、
前記プログラムは、
前記第1保持チャンバーおよび関反応チャンバーに前記第1液体および前記液体試料が導入された試料分析用基板が前記試料分析装置に装填された場合において、
(a)前記試料分析用基板を回転させることによって、前記反応チャンバーの前記液体試料を前記メインチャンバーへ移送させ、
(b)前記試料分析用基板を回転させることによって、前記工程(a)の後に、前記第1保持チャンバーの前記第1液体を前記反応チャンバーへ移送させ、
(c)前記試料分析用基板を回転させることにより、反応チャンバーの前記第1液体を、前記メインチャンバーへ移送させる、
試料分析システム。
[項目18]
項目1から16のいずれかに記載の試料分析用基板を回転軸回りに回転させるモータ、
前記モータの回転軸の回転角度を検出する回転角度検出回路、
前記回転角度検出回路の検出結果に基づき、前記モータの回転および停止時の回転角度を制御する駆動回路、および
演算器、メモリおよびメモリに記憶され、前記演算器に実行可能なように構成されたプログラムを含み、前記プログラムに基づき、前記モータ、前記回転角度検出回路および前記駆動回路の動作を制御する制御回路
を備え、
前記プログラムは、
前記第1保持チャンバーおよび関反応チャンバーに前記第1液体および前記液体試料が導入された試料分析用基板が前記試料分析装置に装填された場合において、
(a)前記試料分析用基板を回転させることによって、前記反応チャンバーの前記液体試料を前記メインチャンバーへ移送させ、
(b)前記試料分析用基板を回転させることによって、前記工程(a)の後に、前記第1保持チャンバーの前記第1液体を前記反応チャンバーへ移送させ、
(c)前記試料分析用基板を回転させることにより、反応チャンバーの前記第1液体を、前記メインチャンバーへ移送させる、
試料分析装置。
[項目19]
項目1から16のいずれかに記載の試料分析用基板と、
前記試料分析用基板を前記回転軸周りに回転させるモータ、
前記モータの回転軸の回転角度を検出する回転角度検出回路、
前記回転角度検出回路の検出結果に基づき、前記モータの回転および停止時の回転角度を制御する駆動回路、および
演算器、メモリおよびメモリに記憶され、前記演算器に実行可能なように構成されたプログラムを含み、前記プログラムに基づき、前記モータ、前記回転角度検出回路および前記駆動回路の動作を制御する制御回路
を有する試料分析装置と、
を備えた試料分析システム用のプログラムであって、
前記第1保持チャンバーおよび関反応チャンバーに前記第1液体および前記液体試料が導入された試料分析用基板が前記試料分析装置に装填された場合において、
(a)前記試料分析用基板を回転させることによって、前記反応チャンバーの前記液体試料を前記メインチャンバーへ移送させ、
(b)前記試料分析用基板を回転させることによって、前記工程(a)の後に、前記第1保持チャンバーの前記第1液体を前記反応チャンバーへ移送させ、
(c)前記試料分析用基板を回転させることにより、反応チャンバーの前記第1液体を、前記メインチャンバーへ移送させる、
試料分析システム用プログラム。
[項目20]
回転運動によって、液体の移送を行う試料分析用基板であって、
回転軸を有する基板と、
前記基板内に位置し、第1液体を保持するための第1空間を有する第1保持チャンバーと、
前記基板内に位置し、アナライトを含む液体試料を保持するための空間を有する反応チャンバーと、
前記基板内に位置しており、第1開口および第2開口を有する第1流路であって、前記第1開口および前記第2開口がそれぞれ前記第1保持チャンバーおよび前記反応チャンバーに接続された第1流路と、
前記基板内に位置し、前記アナライトを含む液体試料および表面にリガンドが固定された磁性粒子を保持するための空間を有するメインチャンバーと、
前記基板内に位置しており、第3開口および第4開口を有する第2流路であって、前記第3開口および前記第4開口がそれぞれ前記反応チャンバーおよび前記メインチャンバーに接続された第2流路と、
前記基板内に位置し、磁石を収納することが可能な磁石収納室と、
を備え、
前記第1開口は、前記第2開口よりも回転軸に近い側に位置し、
前記第2開口は、前記第3開口よりも回転軸に近い側に位置し、
前記磁石収納室は、前記磁石収納室に磁石が収納された場合、前記磁石によって前記メインチャンバー中の前記磁性粒子を前記メインチャンバー内に捕捉することができる位置に配置されている試料分析用基板を用いた送液方法であって、
(a)前記第1保持チャンバーおよび前記反応チャンバーに第1液体および液体試料をそれぞれ導入し、
(b)前記反応チャンバー中の前記液体試料を前記メインチャンバーへ移送し、
(c)前記工程(b)の後に、前記第1保持チャンバーの前記第1液体を、前記反応チャンバーへ移送し、
(d)前記反応チャンバー中の前記第1液体を、前記メインチャンバーへ移送する、
試料分析用基板の使用方法。
試料分析装置200は、モータ201と、原点検出器203と、回転角度検出回路204と、制御回路205と、駆動回路206と、光学測定ユニット207とを備える。
[1.全体の構成]
図3Aは、試料分析用基板100の分解斜視図である。試料分析用基板100は、回転軸110および回転軸110に平行な方向に所定の厚さを有する板形状の基板100’を備える。試料分析用基板100の基板100’は、ベース基板100aとカバー基板100bによって構成されている。本実施形態では、試料分析用基板100の基板100’は円形形状を有しているが、例えば、多角形形状、楕円形状、扇形形状等を有していてもよい。基板100’は、2つの主面100c、100dを有している。本実施形態では、主面100cおよび主面100dは互いに平行であり、主面100cおよび主面100dの間隔で規定される基板100’の厚さ(2つの主面の間の距離)は、基板100’のどの位置でも同じである。しかし、主面100c、100dは、平行でなくてもよい。例えば、2つの主面の一部分が非平行または平行であってもよいし、全体的に非平行であってもよい。また、基板100’の主面100cおよび100dの少なくも一方に凹部ないし凸部を有する構成を備えていてもよい。
図3Cに示すように、反応チャンバー106が試料分析用基板100に設けられている。反応チャンバー106は、図1を参照して説明したように、磁性粒子固定化抗体305と、抗原306を含む検体と、標識抗体308とを反応させて、複合体310を形成させる反応場である。
反応チャンバー106内の複合体310を含む溶液は、第2流路112を介して、メインチャンバー107へ移送される。第2流路112は、開口112gおよび開口112hを有する。第2流路112の開口112gは、反応チャンバー106の第2部分106rの第2領域106reを規定する側面のうち、回転軸110から最も遠い側に位置する最外周側面106ra、または、最外周側面106raに隣接する隣接側面であって、最外周側面106aとの接続部分を含む位置に設けられることが好ましい。反応チャンバー106中の液体をメインチャンバー107へ移送させるにあたり、反応チャンバー106中に液残りが生じることを抑制できるからである。図3Cは、開口112gが最外周側面106aの一部に設けられた例を示している。
図3Dに示すように、メインチャンバー107は、複合体310を含む溶液のB/F分離を行う場である。B/F分離のために、試料分析用基板100は、基板100’内に位置する磁石収納室120と磁石収納室120内に配置された磁石121を含む。
第3流路113は開口113gおよび開口113hを有し、開口113gがメインチャンバー107に接続され、開口113hが回収チャンバー108に接続されている。
回収チャンバー108は、第3流路113を介してメインチャンバー107から移送される磁性粒子311以外の反応液および使用済みの洗浄液を貯蔵する。回収チャンバー108は、前述の反応液と洗浄回数に応じた合計の使用済み洗浄液との合計量よりも大きな容量の空間を有する。回収チャンバー108は液体を保持する主要な部分が、メインチャンバー107よりも回転軸110に対して遠くに位置していることが好ましい。
図3Fを参照する。第1貯蔵チャンバー104は、B/F分離の際の洗浄に用いる洗浄液を貯蔵する。以下において詳細に説明するように、本実施形態の試料分析システムでは、B/F分離の際、複合体310を複数回洗浄することができる。このため、第1貯蔵チャンバー104は、洗浄回数に応じた合計容量の洗浄液を保持し得る空間を有している。
第1貯蔵チャンバー104の洗浄液は、第4流路114を介して、第1保持チャンバー101へ移送される。第4流路114は、開口114gおよび開口114hを有する。第4流路114の開口114gは、第1貯蔵チャンバー104の側面のうち、回転軸110から最も遠い側に位置する最外周側面104a、または、最外周側面104aに隣接する隣接側面であって、最外周側面104aとの接続部分を含む位置に設けられることが好ましい。図3Fは、開口114gが最外周側面104aと隣接側面との接続部分に設けられた例を示している。
第1保持チャンバー101は、第1貯蔵チャンバー104に貯蔵されていた全洗浄液を保持する。その後、メインチャンバー107で複合体310を洗浄するために、洗浄液の一部を反応チャンバー106へ移送させ、残りを保持する。一回の洗浄に用いる洗浄液の量は以下において説明するように、第1流路111によって秤量される。このため、第1保持チャンバー101は、第1流路111以上の容積を有しており、洗浄回数分の合計の洗浄液量以上の容積(例えば、2回の洗浄であれば第1流路111の2倍以上の容積、3回の洗浄であれば第1流路111の3倍以上の容積)を有している。
前述したように、第1流路111は、第1保持チャンバー101と反応チャンバー106とを接続している。このため、第1貯蔵チャンバー104に洗浄液が導入される場合、洗浄液は、いったん、反応チャンバー106へ移送され、その後、メインチャンバー107へ移送される。
図3Hを参照する。第2貯蔵チャンバー105は、試料分析システムを用いた分析の開始時に基質溶液を貯蔵する。第2貯蔵チャンバー105の形状に特に制限はなく、任意の形状を有していてもよい。
第5流路115は、第2貯蔵チャンバー105と、第2保持チャンバー102とを接続している。第5流路115は、例えば、回転軸110を中心とする半径方向に伸びており、毛細管路によって構成されている。第5流路115は、開口115gおよび開口115hを有する。開口115gは、第2貯蔵チャンバー105の側面のうち、回転軸110から最も離れた最外周側面105a、または、最外周側面105aに隣接する隣接側面であって、最外周側面105aに近接する位置に設けられることが好ましい。本実施形態では、最外周側面105aに開口115gが設けられている。開口115hは、第2保持チャンバー102に接続されている。
第2保持チャンバー102は、試料分析システムを用いた分析の開始後、洗浄を含むB/F分離の間、第2貯蔵チャンバー105から移送された基質溶液を保持する。第2保持チャンバー102は、第2貯蔵チャンバー105より回転軸110から遠くに位置している。第2保持チャンバー102は、基質溶液を保持する空間を挟む、第1外周側面102a1および第2外周側面102a2と、第1内周側面102b1および第2内周側面102b2とを有する。第1外周側面102a1と第2外周側面102a2とは、半径方向に重なっておらず、また、第1外周側面102a1は第2外周側面102a2よりも回転軸110から遠くに位置している。第1内周側面102b1と第2内周側面102b2とは半径方向に重なっておらず、また、第1内周側面102b1は第2内周側面102b2よりも回転軸110から遠くに位置している。
第6流路116は、第2保持チャンバー102と第3保持チャンバー103とを接続している。第6流路116は、開口116gおよび開口116hを有し、開口116gは第2保持チャンバー102の隣接側面102dに設けられている。また、開口116hは、第3保持チャンバー103の側面の1つに設けられている。第6流路116は重力によって液体の移動が可能な流路である。
第3保持チャンバー103は、第6流路116を介して第2保持チャンバー102から移送される基質溶液を保持する。第3保持チャンバー103は第2保持チャンバー102と周方向において隣接する第1部分103qおよび第7流路117と周方向に隣接する第2部分103rを含む。第1部分103qと第2部分103rとは半径方向に配置されている。また、第3保持チャンバー103は、第2保持チャンバー102の隣接側面101dに近接している。
第7流路117は、第1部分117qおよび第2部分117rと、開口117gおよび開口117hとを有する。第1部分117qの一端と第2部分117rの一端とは互いに接続されている。第1部分117qの他端に開口117gが位置しており、前述したように、第3保持チャンバー103の第2隣接側面103c2に接続されている。第2部分117rの他端に開口117hが位置しており、メインチャンバー107に接続されている。第2部分117rは毛細管路である。
試料分析システム501の動作を説明する。図4は、試料分析システム501の動作を示すフローチャートである。試料分析システム501を動作させるための、試料分析システム501の各部を制御する手順を規定したプログラムが、例えば制御回路205のメモリに記憶されており、演算器によるプログラムの実行により、以下の動作が実現する。以下の工程に先立ち、試料分析用基板100を試料分析装置200に装填し、試料分析用基板100の原点を検出する。また、反応チャンバー106の第2部分106rの第2領域106reには、磁性粒子固定化抗体305および標識抗体308を含むドライ化試薬125があらかじめ保持されている。以下の手順において、試料分析用基板は、例えば、すべて時計回りで回転させる。ただし、試料分析用基板の回転方向は、時計回りに限られず、半時計回りであってもよい。
まず、図5に示すように、洗浄液および基質溶液を第1貯蔵チャンバー104および第2貯蔵チャンバー105にそれぞれ導入する。基質溶液は、標識物質307との反応または標識物質307による触媒作用によって、発光、蛍光、あるいは、吸収波長の変化を生じる基質を含む。また、反応チャンバー106の第1部分106qに検体溶液を導入する。検体は、シリンジなどによって反応チャンバー106の第1部分106qに注入してもよい。
複合体310が生成した後、試料分析用基板100を回転させ、複合体310を含む反応液を反応チャンバー106の第2部分106rの第2領域106reからメインチャンバー107へ移動させる。前述したように第2流路112は、毛細管現象によって、反応液で満たされている。このため、反応チャンバー106の複合体310を含む反応液に、試料分析用基板100の回転により第2流路112内の反応液にかかる毛細管力よりも強い遠心力が働くと、反応液はメインチャンバー107へ移送される。メインチャンバー107へ移送された反応液は、試料分析用基板100が回転している状態では、続いて回収チャンバー108へ移送されることはない。前述したように第3流路113がサイフォンを構成しているため、遠心力に逆らって、液体が第3流路113を回転軸110に向かう方向へ移動しないからである。メインチャンバー107へ移送された複合体310を含む反応液のうち、磁性粒子311の多くは、磁石121の磁力により最外周側面107aに捕捉される。
試料分析用基板100を回転させる。回転にともない遠心力が発生し、メインチャンバー107内の反応液および磁性粒子311(複合体310および未反応の磁性粒子)に働く。この遠心力は、液体および複合体310がメインチャンバー107の最外周側面107a側へ移動するように働く。このため、磁性粒子311は、最外周側面107aに押し付けられる。
図9に示すように、前のステップで第2の角度で停止させない場合には、試料分析用基板100を少し回転させ、所定の第2の角度で停止させる。第2の角度は第1保持チャンバー101へ移送された洗浄液が、第1流路111の開口111gと接触する角度である。例えば図9に示す例では、試料分析用基板100のδ2で示す角度範囲内に重力方向が位置する角度である。
続いて、試料分析用基板100を回転させる。回転による遠心力が第1流路111および第1保持チャンバー101内の洗浄液に働く。図3Gを参照して説明したように、直線dbを基準として第1流路111側に位置する洗浄液は第1流路111を介して反応チャンバー106の第1部分106qへ移動する。また、直線dbを基準として第1保持チャンバー101側に位置する洗浄液は、遠心力によって、第1保持チャンバー101へ戻される。よって、図10に示すように、第1流路111によって秤量された洗浄液だけが反応チャンバー106の第1部分106qへ移送される。
試料分析用基板100を回転させ、洗浄液を反応チャンバー106の第2部分106rの第2領域106reからメインチャンバー107へ移動させる。第2流路112が、洗浄液で満たされているため、試料分析用基板100の回転により、第2流路112内の洗浄液に、毛細管力よりも強い遠心力が働くと、洗浄液はメインチャンバー107へ移送される。これにより、反応チャンバー106の第1部分106qおよび第2部分106rに残っていた反応溶液は、洗浄液と一緒にメインチャンバー107へ移送される。また、メインチャンバー107内の磁性粒子311が洗浄液と接触し、1回目の洗浄が行われる。
試料分析用基板100を回転させる。回転にともない遠心力が発生し、メインチャンバー107内の洗浄液および磁性粒子311に働く。この遠心力は、洗浄液及び磁性粒子311がメインチャンバー107の最外周側面107a側へ移動するように働き、磁性粒子311は遠心力および磁石121による吸引力によって最外周側面107aにおいて捕捉される。
図14に示すように、前のステップで第5の角度で停止させない場合には、試料分析用基板100を少し回転させ、所定の第5の角度で停止させる。第5の角度は第1保持チャンバー101へ移送された洗浄液が、第1流路111の開口111gと接触する角度である。例えば図14に示す例では、試料分析用基板100のδ5で示す角度範囲内に重力方向が位置する角度である。ステップS4における第1保持チャンバー101内に残っている洗浄液の量が異なるため、角度範囲δ5は角度範囲δ2と異なり得る。
続いて、試料分析用基板100を回転させる。回転による遠心力が第1流路111および第1保持チャンバー101内の洗浄液に働く。1回目の洗浄と同様、図3Gに示す直線dbを基準として第1流路111側に位置する洗浄液は第1流路111を介して反応チャンバー106の第1部分106qへ移動する。また、直線dbを基準として第1保持チャンバー101側に位置する洗浄液は、遠心力によって、第1保持チャンバー101へ戻される。よって、図15に示すように、第1流路111によって秤量された洗浄液だけが反応チャンバー106の第1部分106qへ移送される。
1回目の洗浄と同様、試料分析用基板100を回転させ、洗浄液を反応チャンバー106の第2部分106rの第2領域106reからメインチャンバー107へ移動させる。第2流路112が、洗浄液で満たされているため、試料分析用基板100の回転により、第2流路112内の洗浄液に、毛細管力よりも強い遠心力が働くと、洗浄液はメインチャンバー107へ移送される。これにより、反応チャンバー106の第1部分106qおよび第2部分106rに残っているかもしれない反応溶液は、洗浄液と一緒にメインチャンバー107へ移送される。また、メインチャンバー107内の磁性粒子311が洗浄液と接触し、2回目の洗浄が行われる。
試料分析用基板100を回転させる。回転にともない遠心力が発生し、メインチャンバー107内の洗浄液および磁性粒子311に働く。この遠心力は、洗浄液及び磁性粒子311がメインチャンバー107の最外周側面107a側へ移動するように働き、磁性粒子311は遠心力および磁石121による吸引力によって最外周側面107aにおいて捕捉される。
基質溶液をまず第2保持チャンバー102から第3保持チャンバー103へ移動させる。図19に示すように、前のステップで第8の角度で停止させない場合には、試料分析用基板100を少し回転させ、所定の第8の角度で停止させる。この時、試料分析用基板100は、時計回りに回転させる。第8の角度は第2保持チャンバー102内の基質溶液が第6流路116の開口116gと接触し、かつ、基質溶液の全量が重力によって第3保持チャンバー103へ移動し得る角度である。概ね第6流路116が重力方向に沿って配置される角度である。これにより、第2保持チャンバー102内の基質溶液が第3保持チャンバー103へ移送される。
続いて、試料分析用基板100を回転させる。回転による遠心力が第7流路117および第1保持チャンバー101内の洗浄液に働く。第7流路117内の基質溶液は、遠心力によって、メインチャンバー107へ移動する。開口112gよりも第3保持チャンバー103側に位置している基質溶液は、遠心力によって、第3保持チャンバー103の最外周側面102aへ押し付けられ、第3保持チャンバー103内に留まる。
光学測定ユニット207は、メインチャンバー107に保持された液体の光学的測定を行う。具体的には、光学測定ユニット207は、磁性粒子311に含まれる複合体310に結合した標識抗体308の標識物質307に応じた基質の色素、発光、蛍光等のシグナルを検出する。これにより、抗原306の検出、抗原306の濃度の定量等を行うことができる。
上記実施形態の試料分析用基板100には種々の改変が可能である。
上記実施形態において、反応チャンバー106は、第1部分106qおよび第2部分106rを有していたが、第1部分106qはなくてもよい。
上記実施形態において、第1流路111は、洗浄液を秤量することができる毛細管空間を有していたが、第1流路は秤量する機能を備えていなくてもよい。
上記実施形態では、第1流路111によって洗浄液を秤量していたが、第1保持チャンバーによって洗浄液を秤量してもよい。図26に示す試料分析用基板169は、第1部分133qと、第2部分133rと、第2部分133rおよび第1部分133qとを接続する連結部分133pとを含む第1保持チャンバー133を備える。
上記実施形態では、複数回分の洗浄液を第1保持チャンバー101が保持していたが、1回分の洗浄液を保持する複数のチャンバーを備えていてもよい。
図3C等を参照して説明した試料分析用基板100の反応チャンバー106は、第1部分106qおよび第2部分106rのそれぞれが、毛細管空間と非毛細管空間を有していた。反応チャンバーにおける毛細管空間と非毛細管空間はこれに限られず、種々の変形が可能である。
本実施形態では、磁性粒子を用いた測定系を想定した説明を行ったが、本願の一態様に係る試料分析用基板、試料分析装置、試料分析システムおよび試料分析システム用プログラムは、磁性粒子を用いた測定系に限定されるものではない。例えば、1次抗体が固定化される対象は、磁性粒子に替えて、チャンバー内の壁面であってもよい。すなわち、チャンバーがポリスチレンやポリカーボネートといった素材で構成されている場合には、チャンバー内の壁面に物理吸着により1次抗体を固定化させることができ、チャンバー内で抗原や標識抗体とのサンドイッチ型の結合反応をせしめることができる。また、チャンバー内の壁面に1次抗体と結合可能な官能基(例えば、アミノ基やカルボキシル基)を有し、化学結合により1次抗体を固定化させることができ、チャンバー内で抗原や標識抗体とのサンドイッチ型の結合反応をせしめることができる。また、チャンバー内の壁面に金属基板を備える構成であれば、例えば、SAMを用いて1次抗体を金属基板に結合して固定化させることができ、チャンバー内で抗原や標識抗体とのサンドイッチ型の結合反応をせしめることができる。一次抗体をチャンバー壁面に物理吸着または化学結合で固定化させる場合は、主に色素、化学発光または蛍光のシグナルを検出する系に使用される。一方、一次抗体を金属基板に固定化させる場合は、シグナルとして、主に電気化学的シグナル(例えば、電流)、電気化学発光のシグナルを検出する系に使用される。この場合、図3Bに示した磁石121は不要である。また、複合体310形成の反応場は反応チャンバー106ではなく、メインチャンバー107になる。したがって、一次抗体は、メインチャンバー107の壁面に固定化する必要がある。
100' 基板
100a ベース基板
100b カバー基板
100c、100d 主面
100f 壁部分
101、101A 第1保持チャンバー
101B 第4保持チャンバー
101a、102a 最外周側面
101c、101d 隣接側面
102 第2保持チャンバー
103 第3保持チャンバー
104 第1貯蔵チャンバー
105 第2貯蔵チャンバー
106 メインチャンバー
106q 第1部分
106qa 最外周側面
106qe 第2領域
106qf 第1領域
106r 第2部分
106ra 最外周側面
106rb 最内周側面
106re 第2領域
106rf 第1領域
107 メインチャンバー
108 回収チャンバー
110 回転軸
111 第1流路
112 第2流路
113 第3流路
114 第4流路
115 第5流路
116 第6流路
117 第7流路
120 磁石収納室
121 磁石
122 空気孔
123 開口
125 ドライ化試薬
126 壁部分
161~169 試料分析用基板
200 試料分析装置
201 モータ
201a ターンテーブル
203 原点検出器
203a 光源
203b 受光素子
203c 原点検出回路
204 回転角度検出回路
205 制御回路
206 駆動回路
207 光学測定ユニット
210 マーカ
210a エッジ
210b エッジ
302 磁性粒子
304 一次抗体
305 磁性粒子固定化抗体
306 抗原
307 標識物質
308 標識抗体
310 複合体
311 磁性粒子
501 試料分析システム
Claims (20)
- 回転運動によって、液体の移送を行う試料分析用基板であって、
回転軸を有する基板と、
前記基板内に位置し、第1液体を保持するための第1空間を有する第1保持チャンバーと、
前記基板内に位置し、検体を含む液体試料を保持するための空間を有する反応チャンバーと、
前記基板内に位置しており、第1開口および第2開口を有する第1流路であって、前記第1開口および前記第2開口がそれぞれ前記第1保持チャンバーおよび前記反応チャンバーに接続された第1流路と、
前記基板内に位置し、前記検体を含む液体試料および表面にリガンドが固定された磁性粒子を保持するための空間を有するメインチャンバーと、
前記基板内に位置しており、第3開口および第4開口を有する第2流路であって、前記第3開口および前記第4開口がそれぞれ前記反応チャンバーおよび前記メインチャンバーに接続された第2流路と、
前記基板内に位置し、磁石を収納することが可能な磁石収納室と、
を備え、
前記第1開口は、前記第2開口よりも前記回転軸に近い側に位置し、
前記第2開口は、前記第3開口よりも前記回転軸に近い側に位置し、
前記磁石収納室は、前記磁石収納室に磁石が収納された場合、前記磁石によって前記メインチャンバー中の前記磁性粒子を前記メインチャンバー内に捕捉することができる位置に配置されている、
試料分析用基板。 - 前記反応チャンバーの空間内に配置された、ドライ化試薬をさらに備え、
前記ドライ化試薬は、前記磁性粒子を含む、請求項1に記載の試料分析用基板。 - 前記反応チャンバーは、非毛細管空間を含む、請求項1または2に記載の試料分析用基板。
- 前記反応チャンバーは、毛細管空間を含む、請求項1または2に記載の試料分析用基板。
- 前記反応チャンバーは、非毛細管空間および毛細管空間を含み、
前記非毛細管空間に前記第1開口が接しており、
前記毛細管空間に前記第3開口が接している、
請求項1または2に記載の試料分析用基板。 - 前記非毛細管空間は、前記毛細管空間よりも回転軸に近接して位置する部分を有する、請求項5に記載の試料分析用基板。
- 前記前記反応チャンバーは第1部分および第2部分を含み、
前記基板は前記反応チャンバーの前記第1部分と前記第2部分との間に位置する壁部分を有し、
前記壁部分は、前記回転軸に向かう方向に凸部を形成し、
前記第1部分および前記第2部分において、前記壁部分の前記回転軸から最も近い点と、前記回転軸とを結ぶ線分を半径とする円弧よりも遠い側に前記毛細管空間の一部および前記非毛細管空間の一部がそれぞれ位置する、
請求項5に記載の試料分析用基板。 - 前記第1部分側に位置する前記壁部分の一部又は全部には、前記第1部分と前記第2部分を接続する前記毛細管空間の一部が位置している、
請求項7に記載の試料分析用基板。 - 前記基板内に位置しており、空間を有する回収チャンバーと、
前記基板内に位置しており、第5開口および第6開口を有する第3流路であって、前記第5開口および前記第6開口がそれぞれ前記メインチャンバーおよび前記回収チャンバーに接続された第3流路と、
をさらに備え、
前記第5開口は前記第6開口よりも前記回転軸に近接して位置する、
請求項1から8のいずれかに記載の試料分析用基板。 - 前記第1流路は、非毛細管路である、請求項1から9のいずれかに記載の試料分析用基板。
- 前記第1保持チャンバーは、前記回転軸から最も離れて位置する最外周側面と、前記最外周側面に隣接する隣接側面とを有し、
前記最外周側面と前記隣接側面とで凹部を形成し、
前記試料分析用基板を所定の角度位置で保持した場合に、前記凹部で前記第1液体を保持する、請求項10に記載の試料分析用基板。 - 前記第1流路は、毛細管路である、請求項1から11のいずれかに記載の試料分析用基板。
- 前記第1流路は、サイフォン構造を有する請求項12に記載の試料分析用基板。
- 前記第1流路の一部は、前記第1開口を挟んで前記第1保持チャンバーの一部よりも前記回転軸に近接して位置している、請求項12に記載の試料分析用基板。
- 前記第1保持チャンバーの空間は、第1部分および第2部分と、前記第1部分および前記第2部分の間に位置しており前記第1部分および前記第2部分を連結する連結部分とを有し、
前記基板は、前記第1保持チャンバーの前記空間の前記第1部分および前記第2部分を区切る壁部分を有し、
前記反応チャンバーは前記第1保持チャンバーの前記第2部分よりも前記回転軸から遠くに位置し、
前記第1保持チャンバーの前記空間の前記連結部分は、前記基板の前記壁部分よりも前記回転軸側に位置し、
前記第1流路は、前記第1保持チャンバーの前記空間の前記第2部分と接続されている、請求項12に記載の試料分析用基板。 - 前記基板内に位置し、第2液体を保持するための空間を有する第4保持チャンバーと、
前記4保持チャンバーと前記反応チャンバーとを接続し、前記第2液体を移送する第8流路と、
前記第1保持チャンバーは、前記回転軸から最も遠くに位置する最外周側面と、前記最外周側面に隣接する隣接側面と、前記最外周側面および前記隣接側面とで形成される凹部とを有し、
前記第4保持チャンバーは、前記回転軸から最も遠くに位置する最外周側面と、前記最外周側面に隣接する隣接側面と、前記最外周側面および前記隣接側面とで形成される凹部とを有し、前記第1保持チャンバーの前記隣接側面と前記第4保持チャンバーの隣接側面とは、前記回転軸に平行な方向からみて非平行である、
請求項1から14のいずれかに記載の試料分析用基板。 - 請求項1から16のいずれかに記載の試料分析用基板と、
前記試料分析用基板を前記回転軸周りに回転させるモータ、
前記モータの回転軸の回転角度を検出する回転角度検出回路、
前記回転角度検出回路の検出結果に基づき、前記モータの回転および停止時の回転角度を制御する駆動回路、および
演算器、メモリおよびメモリに記憶され、前記演算器に実行可能なように構成されたプログラムを含み、前記プログラムに基づき、前記モータ、前記回転角度検出回路および前記駆動回路の動作を制御する制御回路
を有する試料分析装置と、
を備えた試料分析システムであって、
前記プログラムは、
前記第1保持チャンバーおよび関反応チャンバーに前記第1液体および前記液体試料が導入された試料分析用基板が前記試料分析装置に装填された場合において、
(a)前記試料分析用基板を回転させることによって、前記反応チャンバーの前記液体試料を前記メインチャンバーへ移送させ、
(b)前記試料分析用基板を回転させることによって、前記工程(a)の後に、前記第1保持チャンバーの前記第1液体を前記反応チャンバーへ移送させ、
(c)前記試料分析用基板を回転させることにより、反応チャンバーの前記第1液体を、前記メインチャンバーへ移送させる、
試料分析システム。 - 請求項1から16のいずれかに記載の試料分析用基板を回転軸回りに回転させるモータ、
前記モータの回転軸の回転角度を検出する回転角度検出回路、
前記回転角度検出回路の検出結果に基づき、前記モータの回転および停止時の回転角度を制御する駆動回路、および
演算器、メモリおよびメモリに記憶され、前記演算器に実行可能なように構成されたプログラムを含み、前記プログラムに基づき、前記モータ、前記回転角度検出回路および前記駆動回路の動作を制御する制御回路
を備え、
前記プログラムは、
前記第1保持チャンバーおよび関反応チャンバーに前記第1液体および前記液体試料が導入された試料分析用基板が前記試料分析装置に装填された場合において、
(a)前記試料分析用基板を回転させることによって、前記反応チャンバーの前記液体試料を前記メインチャンバーへ移送させ、
(b)前記試料分析用基板を回転させることによって、前記工程(a)の後に、前記第1保持チャンバーの前記第1液体を前記反応チャンバーへ移送させ、
(c)前記試料分析用基板を回転させることにより、反応チャンバーの前記第1液体を、前記メインチャンバーへ移送させる、
試料分析装置。 - 請求項1から16のいずれかに記載の試料分析用基板と、
前記試料分析用基板を前記回転軸周りに回転させるモータ、
前記モータの回転軸の回転角度を検出する回転角度検出回路、
前記回転角度検出回路の検出結果に基づき、前記モータの回転および停止時の回転角度を制御する駆動回路、および
演算器、メモリおよびメモリに記憶され、前記演算器に実行可能なように構成されたプログラムを含み、前記プログラムに基づき、前記モータ、前記回転角度検出回路および前記駆動回路の動作を制御する制御回路
を有する試料分析装置と、
を備えた試料分析システム用のプログラムであって、
前記第1保持チャンバーおよび関反応チャンバーに前記第1液体および前記液体試料が導入された試料分析用基板が前記試料分析装置に装填された場合において、
(a)前記試料分析用基板を回転させることによって、前記反応チャンバーの前記液体試料を前記メインチャンバーへ移送させ、
(b)前記試料分析用基板を回転させることによって、前記工程(a)の後に、前記第1保持チャンバーの前記第1液体を前記反応チャンバーへ移送させ、
(c)前記試料分析用基板を回転させることにより、反応チャンバーの前記第1液体を、前記メインチャンバーへ移送させる、
試料分析システム用プログラム。 - 回転運動によって、液体の移送を行う試料分析用基板であって、
回転軸を有する基板と、
前記基板内に位置し、第1液体を保持するための第1空間を有する第1保持チャンバーと、
前記基板内に位置し、アナライトを含む液体試料を保持するための空間を有する反応チャンバーと、
前記基板内に位置しており、第1開口および第2開口を有する第1流路であって、前記第1開口および前記第2開口がそれぞれ前記第1保持チャンバーおよび前記反応チャンバーに接続された第1流路と、
前記基板内に位置し、前記アナライトを含む液体試料および表面にリガンドが固定された磁性粒子を保持するための空間を有するメインチャンバーと、
前記基板内に位置しており、第3開口および第4開口を有する第2流路であって、前記第3開口および前記第4開口がそれぞれ前記反応チャンバーおよび前記メインチャンバーに接続された第2流路と、
前記基板内に位置し、磁石を収納することが可能な磁石収納室と、
を備え、
前記第1開口は、前記第2開口よりも回転軸に近い側に位置し、
前記第2開口は、前記第3開口よりも回転軸に近い側に位置し、
前記磁石収納室は、前記磁石収納室に磁石が収納された場合、前記磁石によって前記メインチャンバー中の前記磁性粒子を前記メインチャンバー内に捕捉することができる位置に配置されている試料分析用基板を用いた送液方法であって、
(a)前記第1保持チャンバーおよび前記反応チャンバーに第1液体および液体試料をそれぞれ導入し、
(b)前記反応チャンバー中の前記液体試料を前記メインチャンバーへ移送し、
(c)前記工程(b)の後に、前記第1保持チャンバーの前記第1液体を、前記反応チャンバーへ移送し、
(d)前記反応チャンバー中の前記第1液体を、前記メインチャンバーへ移送する、
試料分析用基板の使用方法。
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JPWO2020009023A1 (ja) * | 2018-07-02 | 2021-06-24 | Phcホールディングス株式会社 | 試料分析用基板および試料分析方法 |
JP6994113B2 (ja) | 2018-07-02 | 2022-01-14 | Phcホールディングス株式会社 | 試料分析用基板および試料分析方法 |
WO2020162431A1 (ja) * | 2019-02-05 | 2020-08-13 | ユニバーサル・バイオ・リサーチ株式会社 | 検体処理装置 |
JPWO2020162431A1 (ja) * | 2019-02-05 | 2021-12-09 | ユニバーサル・バイオ・リサーチ株式会社 | 検体処理装置 |
JP7458573B2 (ja) | 2019-02-05 | 2024-04-01 | ユニバーサル・バイオ・リサーチ株式会社 | 検体処理装置 |
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JP6768002B2 (ja) | 2020-10-14 |
CN108431610B (zh) | 2022-04-05 |
US11262356B2 (en) | 2022-03-01 |
JPWO2017115733A1 (ja) | 2018-11-01 |
EP3399319A1 (en) | 2018-11-07 |
US20190018007A1 (en) | 2019-01-17 |
CN108431610A (zh) | 2018-08-21 |
EP3399319A4 (en) | 2019-01-02 |
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