WO2016031844A1 - マイクロチップ、マイクロチップ制御装置及びマイクロチップ制御システム - Google Patents
マイクロチップ、マイクロチップ制御装置及びマイクロチップ制御システム Download PDFInfo
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- WO2016031844A1 WO2016031844A1 PCT/JP2015/073960 JP2015073960W WO2016031844A1 WO 2016031844 A1 WO2016031844 A1 WO 2016031844A1 JP 2015073960 W JP2015073960 W JP 2015073960W WO 2016031844 A1 WO2016031844 A1 WO 2016031844A1
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- Prior art keywords
- microchip
- lysis
- cells
- lysate
- tank
- Prior art date
Links
- 238000006243 chemical reaction Methods 0.000 claims abstract description 75
- 230000002934 lysing effect Effects 0.000 claims abstract description 19
- 230000009089 cytolysis Effects 0.000 claims description 87
- 239000006166 lysate Substances 0.000 claims description 44
- 239000007788 liquid Substances 0.000 claims description 20
- 238000004090 dissolution Methods 0.000 claims description 18
- 230000007246 mechanism Effects 0.000 claims description 12
- 238000010438 heat treatment Methods 0.000 claims description 8
- 238000007789 sealing Methods 0.000 claims description 5
- 230000002265 prevention Effects 0.000 claims description 4
- 238000009434 installation Methods 0.000 claims description 2
- 230000003247 decreasing effect Effects 0.000 claims 1
- 239000012488 sample solution Substances 0.000 abstract description 30
- 238000004458 analytical method Methods 0.000 abstract description 3
- 239000000243 solution Substances 0.000 description 41
- 238000007400 DNA extraction Methods 0.000 description 27
- 238000003752 polymerase chain reaction Methods 0.000 description 25
- 238000002360 preparation method Methods 0.000 description 17
- 238000001962 electrophoresis Methods 0.000 description 13
- 108020004414 DNA Proteins 0.000 description 12
- 102000053602 DNA Human genes 0.000 description 12
- 239000000523 sample Substances 0.000 description 12
- 239000011324 bead Substances 0.000 description 9
- 239000011534 wash buffer Substances 0.000 description 9
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 8
- 229920000742 Cotton Polymers 0.000 description 7
- 238000005303 weighing Methods 0.000 description 7
- 239000012149 elution buffer Substances 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 239000011347 resin Substances 0.000 description 6
- 229920005989 resin Polymers 0.000 description 6
- 229910001172 neodymium magnet Inorganic materials 0.000 description 4
- 229920000642 polymer Polymers 0.000 description 4
- 239000000377 silicon dioxide Substances 0.000 description 4
- 239000002585 base Substances 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 108091093088 Amplicon Proteins 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- 238000005251 capillar electrophoresis Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- XIXADJRWDQXREU-UHFFFAOYSA-M lithium acetate Chemical compound [Li+].CC([O-])=O XIXADJRWDQXREU-UHFFFAOYSA-M 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 210000003296 saliva Anatomy 0.000 description 2
- 230000007723 transport mechanism Effects 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- 229910013594 LiOAc Inorganic materials 0.000 description 1
- 229940122426 Nuclease inhibitor Drugs 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000004590 computer program Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 239000005547 deoxyribonucleotide Substances 0.000 description 1
- 125000002637 deoxyribonucleotide group Chemical group 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229910052736 halogen Inorganic materials 0.000 description 1
- 150000002367 halogens Chemical class 0.000 description 1
- 238000010030 laminating Methods 0.000 description 1
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 1
- 229910052753 mercury Inorganic materials 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- NJPPVKZQTLUDBO-UHFFFAOYSA-N novaluron Chemical compound C1=C(Cl)C(OC(F)(F)C(OC(F)(F)F)F)=CC=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F NJPPVKZQTLUDBO-UHFFFAOYSA-N 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 235000011178 triphosphate Nutrition 0.000 description 1
- 239000001226 triphosphate Substances 0.000 description 1
- UNXRWKVEANCORM-UHFFFAOYSA-N triphosphoric acid Chemical compound OP(O)(=O)OP(O)(=O)OP(O)(O)=O UNXRWKVEANCORM-UHFFFAOYSA-N 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/06—Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M1/00—Apparatus for enzymology or microbiology
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
- G01N35/08—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N37/00—Details not covered by any other group of this subclass
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/02—Adapting objects or devices to another
- B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
- B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/06—Fluid handling related problems
- B01L2200/0689—Sealing
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/16—Reagents, handling or storing thereof
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
- B01L2300/087—Multiple sequential chambers
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5029—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures using swabs
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
Definitions
- the present invention is based on the priority claim of Japanese patent application: Japanese Patent Application No. 2014-176340 (filed on Aug. 29, 2014), the entire description of which is incorporated herein by reference. Shall.
- the present invention relates to a microchip, a microchip control device, and a microchip control system.
- Patent Document 1 discloses a microchip formed by laminating sheets (plates) including an elastic member.
- the microchip disclosed in Patent Document 1 does not have a configuration for preparing a sample solution. Therefore, in order to perform DNA (Deoxyribo Nucleic Acid) analysis using the microchip disclosed in Patent Document 1, it is necessary for an operator to prepare a sample solution manually in advance. The preparation of such a sample solution is a factor that increases the operator's work.
- An object of the present invention is to provide a microchip, a microchip control device, and a microchip control system that contribute to reducing the work of an operator.
- a cell receiving portion that receives cells collected from a subject, a lysis tank containing a lysis solution for lysing the cells, and a lysis reaction in which the lysis reaction of the cells is performed. And a microchip having a bath.
- a cell receiving portion for receiving a subject's cells, a lysis tank containing a lysis solution for lysing the cells, and a lysis reaction tank in which the lysis reaction of the cells is performed.
- a microchip placement portion on which the microchip is placed, a first lysis solution delivery portion for delivering the lysis solution from the lysis solution tank to the cell reception portion, and the cells from the cell reception portion to the lysis reaction vessel.
- a microchip control device comprising: a second lysate delivery unit that sends out the mixed lysate; and a lysis reaction unit that performs the lysis reaction by heating the lysate mixed with the cells in the lysis reaction tank.
- a cell receiving portion that receives a subject's cells, a lysis tank containing a lysis solution for lysing the cells, a lysis reaction tank in which a lysis reaction of the cells is performed,
- a microchip comprising: a first lysate delivery part for delivering the lysate from the lysate tank to the cell receiving part; and a lysate mixed with the cells from the cell acceptor to the lysis reaction tank.
- a microchip control system comprising: a second lysis solution delivery unit; and a lysis reaction unit that performs the lysis reaction by heating the lysis solution mixed with the cells in the lysis reaction tank.
- a microchip, a microchip control device, and a microchip control system that contribute to reducing the work of an operator are provided.
- FIG. 1A is a perspective view of the microchip 1000.
- 1B is an X-axis cross-sectional view of the microchip 1000
- FIGS. 1C and 1D are a Z1-axis cross-sectional view and a Z2-axis cross-sectional view of the microchip 1000, respectively.
- the microchip 1000 is a structure in which a plurality of films 1001 and a resin plate 1002 are laminated, a cell receiving portion 101 that receives cells collected from a subject, and a lysis solution tank 102 that contains a lysis solution for lysing cells. And a lysis reaction tank 103 in which a cell lysis reaction is performed.
- the dissolution reaction tank 103 is a non-adhered portion between the films 1001 and is in a collapsed state until the liquid enters, but expands to become a solution tank when the liquid flows in.
- the cell receiving portion 101 for example, a cotton swab to which the subject's cells are attached is inserted. At this time, the lysate is moved from the lysate tank 102 to the cell receiving portion 101 to detach the subject's cells from the cotton swab. Then, the lysis solution is moved from the cell receiving unit 101 to the lysis reaction tank 103 to heat the lysis solution.
- the liquid transport on the microchip 1000 will be described in detail later, the flow path in which the dissolved liquid is not moved during the liquid transport is in a closed state when compressed air or the like is injected into the pressure path. Therefore, for example, when the cell receiving unit 101 is pressurized and the lysate in the cell receiving unit 101 is moved to the lysis reaction tank 103, the lysate does not flow back into the lysis solution tank 102.
- the sample solution can be prepared on the microchip 1000.
- the sample solution is automatically prepared by a simple operation of inserting a cotton swab with the subject's cells attached to the cell receiving portion 101. it can. That is, the operator's work is reduced.
- a microchip, a microchip control device, and a microchip control system will be described with a specific example with reference to the drawings.
- a table 12 is disposed on a pedestal 11, and a sample solution preparation unit 13, a PCR unit 14, and an electrophoresis unit 15 are disposed on the table 12.
- the base 11 and the lid 16 are connected via a hinge, and the lid 16 can be opened and closed.
- the microchip 1000 is placed at a predetermined position on the table 12 by fitting a pin provided on the table 12 into a pin hole provided on the microchip 1000.
- the sample solution preparation unit 100 of the microchip 1000 comes into contact with the sample solution preparation unit 13.
- the DNA extraction / PCR unit 200 is in contact with the PCR unit 14, and the electrophoresis unit 300 is in contact with the electrophoresis unit 15.
- an electrode is inserted into the electrode tank of the microchip 1000 through the electrode hole provided in the microchip 1000.
- the lid 16 is provided with a plurality of pressure holes 17.
- the region of the lid 16 corresponding to these pressurizing holes 17 penetrates, and the pressurizing hole 17 is connected to the electromagnetic valve 19 via a tube 18. Further, the pressure hole 17 and various control holes on the microchip 1000 are connected by closing the lid 16.
- the pressure hole 17 and the control hole are preferably in close contact with each other with a sealing mechanism such as an O-ring 20 interposed.
- a pressure medium such as compressed air is sealed in the accumulator 21, and the controller 22 controls the electromagnetic valve 19 so that the pressure medium is taken in and out of the control hole on the microchip 1000 through the pressure hole 17. .
- the internal pressure of the pressure accumulator 21 is controlled to maintain a predetermined pressure by a pressure sensor and a pump (not shown).
- the lid 16 is provided with a DNA extraction unit 23 for extracting sample DNA or template DNA from the sample solution.
- the DNA extraction unit 23 includes, for example, a neodymium magnet that adsorbs magnetic beads when sample DNA is extracted using magnetic beads (silica).
- the DNA extraction unit 23 brings a magnet (for example, a neodymium magnet) close to the DNA extraction unit 244 or moves the magnet away from the DNA extraction unit 244 under the control of the controller 22.
- the sample solution preparation unit 13 and the PCR unit 14 include a temperature sensor, a heat transfer material, a Peltier element (thermoelectric element), a heat sink, and the like.
- the sample solution preparation unit 13 performs the lysis reaction by heating the lysate containing the cells of the subject, and the PCR unit 14 executes the PCR.
- the electrophoresis unit 15 is a mechanism that executes capillary electrophoresis and detection of a fluorescent label, and includes a fluorescent lamp detection mechanism, an excitation device such as a halogen lamp, a mercury lamp, and laser light, a filter, a camera, and the like.
- a DC voltage is applied to the electrode via the power supply unit 24 and capillary electrophoresis starts, the electrophoresis unit 15 monitors the fluorescent label flowing through the capillary and detects the change in fluorescence luminance over time in a graph. The result is output via the display unit 25.
- the controller 22 can also be realized by a computer program that causes a computer mounted on the microchip control apparatus 10 to execute the processing of the controller 22 described in detail later using its hardware.
- the microchip 1000 includes a sample solution preparation unit 100, a DNA extraction / PCR unit 200, and an electrophoresis unit 300, as shown in FIG. FIG. 3 shows a microchip 1000 having a triple structure for processing three samples simultaneously, but only a single structure will be described below for the sake of simplicity.
- the sample solution preparation unit 100 and the DNA extraction / PCR unit 200 are configured to be sandwiched between a plurality of elastic sheet resin plates.
- the elastic sheets are bonded to each other except for a part, and the non-bonded portion can be inflated by injecting a medium such as liquid or air.
- the sample solution preparation unit 100 has a trapezoidal protrusion 110 having a predetermined height on the resin plate.
- the elastic sheet has stretchability, heat resistance, acid resistance and alkali resistance.
- the resin plate is desirably hard enough to control the extension of the elastic sheet.
- the lower resin plate can also be provided on the base 11 of the microchip control device 10.
- the microchip 1000 is provided with various control holes such as medium entry / exit holes.
- FIGS. 4B, 4C, 4D, and 4E are an X-axis cross-sectional view, a Z1-axis cross-sectional view, and a Z2-axis, respectively. It is sectional drawing and Y-axis sectional drawing.
- the sample solution preparation unit 100 includes a cell receiving unit 101, lysis solution tanks 102 ⁇ / b> A and 102 ⁇ / b> B, and a lysis reaction tank 103 connected by flow paths 104 ⁇ / b> A, 104 ⁇ / b> B, 105, and 106.
- a pressurizing path 108 is disposed below each of the flow paths 104A, 104B, 105, 106.
- the flow paths 104 ⁇ / b> A, 104 ⁇ / b> B, 105, 106 can be opened and closed by injecting or releasing injection of the pressurized medium into the pressurizing path 108.
- the pressurizing path 108 is omitted for the sake of clarity.
- the cell receiving portion 101 is disposed on the protruding portion 110, has an opening on the upper side, passes through the protruding portion 110, and reaches the elastic sheet disposed on the bottom of the protruding portion 110.
- the cell receiving unit 101 is connected to the lysing solution tanks 102A and 102B via first flow paths 104A and 104B disposed between the elastic sheets, and also has a second flow path 105 disposed between the elastic sheets.
- To the dissolution reaction tank 103 For example, a cotton swab to which a subject's cells are attached is inserted into the cell receiving portion 101 from above the opening.
- the upper opening of the cell receiving portion 101 has a tapered shape in which the diameter increases toward the upper side, and in this way, for example, when the subject's saliva is attached to a cotton swab in a large amount, etc. The risk of liquid overflowing from the cell receiving portion 101 is reduced.
- the diameter of the cell receiving portion 101 becomes narrower as it goes downward. In this way, for example, even if the amount of saliva of the subject attached to the cotton swab is small, the subject has a high yield. Cells can be collected.
- the cell receiving portion 101 may be provided with a scattering prevention mechanism for preventing the lysing solution from scattering.
- a scattering prevention film 109 having slits is arranged so as to cover the cell receiving portion 101.
- the anti-scattering film 109 is provided, the projecting portion 110 is configured to be separable such as an upper projecting portion 110a and a lower projecting portion 110b, and the two projecting portions 110a and 110b are separated from the anti-scattering film 109. So that
- a sealing mechanism such as an O-ring 20 may be provided at the edge of the upper opening of the cell receiving portion 101. In this way, unevenness on the lid 16 side of the microchip control device 10 can be reduced, and cleaning becomes easy.
- a lysing solution eg, SDS / LiOAc solution (sodium dodecyl sulfate / lithium acetate solution)
- the lysing solution tank 102B contains proteinase K, etc.
- the nuclease inhibitor is similarly encapsulated.
- a pressure hole 107 is provided above the solution tanks 102A and 102B. When a pressurized medium such as compressed air is applied through the pressure hole 107, the solution tanks 102A and 102B contract, A lysate or the like moves to the cell receiving unit 101 via the first flow paths 104A and 104B.
- FIG. 6A to 6C are Z1-axis cross-sectional views of the microchip 1000
- FIG. 6D is an X-axis cross-sectional view
- FIG. 6E is a Z2-axis cross-sectional view.
- a solution is sealed in a solution tank 102A formed between elastic sheets.
- the pressurizing path 108 disposed below the first flow path 104A is inflated by injecting a pressurizing medium, and the first flow path 104A is closed.
- the dissolution reaction tank 103 is formed by expanding when liquid flows from the cell receiving portion 101.
- the first flow paths 104A and 104B and the third flow path 106 are closed, and the second flow path 105 is open.
- the upper part of the dissolution reaction tank 103 is opened, and the pressurized medium is taken in and out.
- the lower part of the dissolution reaction tank 103 is also opened, and comes into contact with a part of the sample solution preparation unit 13 (specifically, a heat transfer material that transfers heat from the Peltier element).
- the dissolution reaction tank 103 between the elastic sheets has a configuration in which a liquid flows in and expands, and therefore, when the dissolution reaction tank 103 expands, the dissolution reaction tank 103 comes into close contact with the sample solution preparation unit 13.
- the dissolution reaction tank 103 is connected to the DNA extraction unit 244 via the third channel 106. Therefore, when the second flow path 105 is closed and the third flow path 106 is opened and the lysis reaction tank 103 is pressurized after the heat treatment, the sample solution in the lysis reaction tank 103 is transferred via the third flow path 106 to the DNA. It flows into the extraction / PCR section 200.
- capacitance of the dissolution reaction tank 103 is smaller than the total amount of the solution with which the solution tank 102A, 102B is filled. According to such a configuration, the amount of the sample solution can be regulated to the capacity of the dissolution reaction tank 103, and air can be prevented from being mixed into the dissolution reaction tank 103.
- the solution may be reciprocated between the cell receiving unit 101 and the lysis reaction tank 103 (in other words, shuttling). By doing in this way, more cells can be collect
- FIG. 7 is a schematic plan view of the DNA extraction / PCR unit 200.
- the DNA extraction / PCR unit 200 includes a washing buffer tank 242, an elution buffer tank 243, a DNA extraction unit 244, a PCR unit 245, and a weighing unit 246.
- the washing buffer tank 242 and the elution buffer tank 243 have the same configuration as the dissolution liquid tanks 102A and 102B, and the washing buffer and the elution buffer are enclosed in advance.
- the washing buffer is, for example, a Tris (tris (hydroxymethyl) aminomethane) buffer
- the elution buffer also includes a polymerase that performs a primer extension reaction and a dNTP mix (deoxyribonucleotide triphosphate mix).
- the DNA extraction unit 244 is a mechanism provided for extracting DNA from the sample solution.
- magnetic beads silicon are enclosed in the DNA extraction unit 244 in advance, and sample DNA is extracted from the sample solution under the control of the controller 22 and the DNA extraction unit 23.
- the microchip control apparatus 10 has a neodymium magnet as the DNA extraction unit 23, and magnetic beads coated with silica are encapsulated in the DNA extraction unit 244 in advance.
- the microchip control apparatus 10 moves the sample solution from the dissolution reaction tank 103 to the DNA extraction unit 244, and adsorbs the DNA to the magnetic beads (silica) enclosed in the DNA extraction unit 244. Then, the magnetic beads are washed with the washing buffer in the washing buffer tank 242 to extract the DNA.
- the microchip control device 10 discharges the sample solution and the washing buffer from a drain port (not shown). At this time, the magnetic beads are adsorbed by the neodymium magnet, so that the magnetic beads are put together with the sample solution and the washing buffer. Is prevented from being discharged.
- the DNA extraction method can be modified, for example, by increasing the number of washings with reference to various protocols. Further, the DNA extraction method is not limited to the method using magnetic beads, and for example, a method using a column may be adopted.
- the PCR unit 245 executes PCR under temperature control by the PCR unit 14. Specifically, a primer set is enclosed in the PCR unit 245 in advance, and a desired base sequence in the sample DNA (template DNA) extracted by the DNA extraction unit 244 is amplified by the action of a polymerase contained in the elution buffer.
- the weighing unit 246 is a mechanism for weighing a solution containing an amplicon. Specifically, the weighing unit 246 is configured to be smaller than the PCR unit 245, and the liquid transportation is completed in a state where the solution in the PCR unit 245 has not completely moved to the weighing unit 246. In other words, the microchip control device 10 weighs the solution containing the amplicon by leaving a part of the solution in the PCR unit 245.
- the electrophoresis unit 300 includes a sample flow path 301, a capillary 302, and a polymer tank 303 as shown in FIG.
- the sample channel 301 is connected to the weighing unit 246 via the electrode tank 304 and is connected to the reservoir 305 at the opposite end.
- the reservoir 305 is a mechanism for preventing an overflow of the sample flowing into the sample channel 301.
- the capillary 302 is connected to the polymer tank 303 via the electrode tank 304.
- the sample channel 301 and the capillary 302 extend in parallel and are connected by a bridge 306 orthogonal to the sample channel 301 and the capillary 302. An electrode attached to the lid 16 is inserted into the electrode tank 304.
- the microchip control device 10 performs electrophoresis after filling the capillary 302 and the bridge 306 with a polymer and performing sample injection. At the time of the electrophoresis, the microchip control device 10 monitors the label flowing in the capillary by the electrophoresis unit 15, and displays the detection result showing the change in fluorescence luminance over time via the display unit 25. Output.
- [Appendix 1] A cell receptor for receiving cells collected from the subject; A lysis bath containing a lysis solution for lysing the cells; A lysis reaction tank in which the lysis reaction of the cells is performed; Having a microchip.
- the cell receiving part has openings in the vertical direction, a rod-like instrument to which the cells are attached is inserted from the upper opening, and the lower opening is connected to the lysis solution tank and the lysis reaction tank.
- [Appendix 3] The microchip according to appendix 1 or 2, wherein the upper opening has a tapered shape in which the diameter increases toward the upper side, and the lower opening has a tapered shape in which the diameter decreases toward the lower side.
- [Appendix 4] The microchip according to any one of appendices 1 to 3, wherein the cell receiving portion has a scattering prevention mechanism that prevents the dissolution solution from scattering.
- [Appendix 5] The microchip according to any one of appendices 1 to 4, wherein the lysis reaction tank is expanded by a liquid flowing from the cell receiving portion.
- [Appendix 6] The microchip according to appendix 5, wherein the volume of the lysis reaction tank is smaller than the volume of the cell receiving portion.
- [Appendix 7] The microchip according to any one of appendices 1 to 6, further comprising a sealing mechanism that provides a sealing state with a microchip control device that controls the microchip.
- a microchip installation unit in which a microchip having a cell receiving unit for receiving a subject's cells, a lysis solution tank containing a lysis solution for lysing the cells, and a lysis reaction vessel in which the lysis reaction of the cells is performed is installed
- a first lysate delivery part for delivering the lysate from the lysate tank to the cell receiving part
- a second lysate delivery part for delivering a lysate mixed with the cells from the cell receiving part to the lysis reaction tank
- a lysis reaction part for performing the lysis reaction by heating the lysate mixed with the cells in the lysis reaction tank
- a microchip control device A microchip control device.
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Abstract
Description
本発明は、日本国特許出願:特願2014-176340号(2014年8月29日出願)の優先権主張に基づくものであり、同出願の全記載内容は引用をもって本書に組み込み記載されているものとする。
本発明は、マイクロチップ、マイクロチップ制御装置及びマイクロチップ制御システムに関する。
以下では、図面を参照しつつ具体的な一例を挙げて、マイクロチップ、マイクロチップ制御装置及びマイクロチップ制御システムを説明する。図2に示すように、マイクロチップ制御装置10は、台座11にテーブル12が配置され、テーブル12には、サンプル溶液調製ユニット13、PCRユニット14、電気泳動ユニット15が配置される。さらに、台座11と蓋16は、ヒンジを介して接続されており、蓋16の開閉が可能である。
被験者から採取された細胞を受容する細胞受容部と、
前記細胞を溶解する溶解液を容れた溶解液槽と、
前記細胞の溶解反応が行われる溶解反応槽と、
を有するマイクロチップ。
[付記2]
前記細胞受容部は上下方向に各々開口部を有し、上部開口部から前記細胞が付着した棒状器具が挿入され、下部開口部は前記溶解液槽及び前記溶解反応槽と接続される、付記1に記載のマイクロチップ。
[付記3]
前記上部開口部は、上方に向かうにつれて口径が広がるテーパ形状を有し、前記下部開口部は下方に向かうにつれて口径が狭まるテーパ形状を有する、付記1又は2に記載のマイクロチップ。
[付記4]
前記細胞受容部は、前記溶解液の飛散を防ぐ飛散防止機構を有する、付記1~3のいずれか1つに記載のマイクロチップ。
[付記5]
前記溶解反応槽は、前記細胞受容部から流入した液体によって膨張する、付記1~4のいずれか1つに記載のマイクロチップ。
[付記6]
前記溶解反応槽の容積が、前記細胞受容部の容積よりも小さい、付記5に記載のマイクロチップ。
[付記7]
マイクロチップを制御するマイクロチップ制御装置との間の密封状態をもたらす密封機構を有する、付記1~6のいずれか1つに記載のマイクロチップ。
[付記8]
被験者の細胞を受容する細胞受容部と、前記細胞を溶解する溶解液を容れた溶解液槽と、前記細胞の溶解反応が行われる溶解反応槽とを有するマイクロチップが設置されるマイクロチップ設置部と、
前記溶解液槽から前記細胞受容部へ前記溶解液を送出する第1溶解液送出部と、
前記細胞受容部から前記溶解反応槽へ前記細胞が混入した溶解液を送出する第2溶解液送出部と、
前記溶解反応槽内の前記細胞が混入した溶解液を加熱して前記溶解反応を実行する溶解反応部と、
を有するマイクロチップ制御装置。
[付記9]
前記第2溶解液送出部は、前記溶解液槽と前記細胞受容部の間で前記溶解液を複数回往復させる、付記8に記載のマイクロチップ制御装置。
[付記10]
被験者の細胞を受容する細胞受容部と、
前記細胞を溶解する溶解液を容れた溶解液槽と、
前記細胞の溶解反応が行われる溶解反応槽と、
を有するマイクロチップと、
前記溶解液槽から前記細胞受容部へ前記溶解液を送出する第1溶解液送出部と、
前記細胞受容部から前記溶解反応槽へ前記細胞が混入した溶解液を送出する第2溶解液送出部と、
前記溶解反応槽内の前記細胞が混入した溶解液を加熱して前記溶解反応を実行する溶解反応部と、
を有するマイクロチップ制御装置と、
からなるマイクロチップ制御システム。
なお、付記8の形態及び付記10の形態は、付記1の形態と同様に、付記2の形態~付記7の形態に展開することが可能である。
11 台座
12 テーブル
13 サンプル溶液調製ユニット
14 PCRユニット
15 電気泳動ユニット
16 蓋
17 加圧穴
18 チューブ
19 電磁弁
20 O-リング
21 蓄圧器
22 コントローラ
23 DNA抽出ユニット
24 電源部
25 表示部
100 サンプル溶液調製部
101 細胞受容部
102、102A、102B 溶解液槽
103 溶解反応槽
104A、104B 第1流路
105 第2流路
106 第3流路
107 加圧孔
108 加圧路
109 飛散防止フィルム
110、110a、110b 突出部
200 DNA抽出・PCR部
242 洗浄バッファ槽
243 溶出バッファ槽
244 DNA抽出部
245 PCR部
246 秤量部
300 電気泳動部
301 サンプル流路
302 キャピラリ
303 ポリマ槽
304 電極槽
305 リザーバ
306 ブリッジ
1000 マイクロチップ
1001 フィルム
1002 樹脂プレート
Claims (10)
- 被験者から採取された細胞を受容する細胞受容部と、
前記細胞を溶解する溶解液を容れた溶解液槽と、
前記細胞の溶解反応が行われる溶解反応槽と、
を有するマイクロチップ。 - 前記細胞受容部は上下方向に各々開口部を有し、上部開口部から前記細胞が付着した棒状器具が挿入され、下部開口部は前記溶解液槽及び前記溶解反応槽と接続される、請求項1に記載のマイクロチップ。
- 前記上部開口部は、上方に向かうにつれて口径が広がるテーパ形状を有し、前記下部開口部は下方に向かうにつれて口径が狭まるテーパ形状を有する、請求項1又は2に記載のマイクロチップ。
- 前記細胞受容部は、前記溶解液の飛散を防ぐ飛散防止機構を有する、請求項1~3のいずれか1項に記載のマイクロチップ。
- 前記溶解反応槽は、前記細胞受容部から流入した液体によって膨張する、請求項1~4のいずれか1項に記載のマイクロチップ。
- 前記溶解反応槽の容積が、前記細胞受容部の容積よりも小さい、請求項5に記載のマイクロチップ。
- マイクロチップを制御するマイクロチップ制御装置との間の密封状態をもたらす密封機構を有する、請求項1~6のいずれか1項に記載のマイクロチップ。
- 被験者の細胞を受容する細胞受容部と、前記細胞を溶解する溶解液を容れた溶解液槽と、前記細胞の溶解反応が行われる溶解反応槽とを有するマイクロチップが設置されるマイクロチップ設置部と、
前記溶解液槽から前記細胞受容部へ前記溶解液を送出する第1溶解液送出部と、
前記細胞受容部から前記溶解反応槽へ前記細胞が混入した溶解液を送出する第2溶解液送出部と、
前記溶解反応槽内の前記細胞が混入した溶解液を加熱して前記溶解反応を実行する溶解反応部と、
を有するマイクロチップ制御装置。 - 前記第2溶解液送出部は、前記溶解液槽と前記細胞受容部の間で前記溶解液を複数回往復させる、請求項8に記載のマイクロチップ制御装置。
- 被験者の細胞を受容する細胞受容部と、
前記細胞を溶解する溶解液を容れた溶解液槽と、
前記細胞の溶解反応が行われる溶解反応槽と、
を有するマイクロチップと、
前記溶解液槽から前記細胞受容部へ前記溶解液を送出する第1溶解液送出部と、
前記細胞受容部から前記溶解反応槽へ前記細胞が混入した溶解液を送出する第2溶解液送出部と、
前記溶解反応槽内の前記細胞が混入した溶解液を加熱して前記溶解反応を実行する溶解反応部と、
を有するマイクロチップ制御装置と、
からなるマイクロチップ制御システム。
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| US15/506,015 US11066636B2 (en) | 2014-08-29 | 2015-08-26 | Microchip, microchip controlling apparatus and microchip controlling system technical field |
| EP15834976.1A EP3187577A4 (en) | 2014-08-29 | 2015-08-26 | Microchip, microchip control device and microchip control system |
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