CN101896598A - Fluid receiving cartridge and utilization of the same - Google Patents

Fluid receiving cartridge and utilization of the same Download PDF

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Publication number
CN101896598A
CN101896598A CN2008801208048A CN200880120804A CN101896598A CN 101896598 A CN101896598 A CN 101896598A CN 2008801208048 A CN2008801208048 A CN 2008801208048A CN 200880120804 A CN200880120804 A CN 200880120804A CN 101896598 A CN101896598 A CN 101896598A
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CN
China
Prior art keywords
mentioned
reactive tank
fluid
liquid
fluid receiving
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Pending
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CN2008801208048A
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Chinese (zh)
Inventor
村里真宽
山田和成
山本大
织部晃畅
丹羽孝介
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NGK Insulators Ltd
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NGK Insulators Ltd
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Publication of CN101896598A publication Critical patent/CN101896598A/en
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    • 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
    • B01L3/502715—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 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F33/00—Other mixers; Mixing plants; Combinations of mixers
    • B01F33/30—Micromixers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F33/00—Other mixers; Mixing plants; Combinations of mixers
    • B01F33/40—Mixers using gas or liquid agitation, e.g. with air supply tubes
    • B01F33/403—Mixers using gas or liquid agitation, e.g. with air supply tubes for mixing liquids
    • 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
    • B01L2200/04—Exchange or ejection of cartridges, containers or reservoirs
    • 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
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0627—Sensor or part of a sensor is integrated
    • B01L2300/0636—Integrated biosensor, microarrays
    • 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
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0681—Filter
    • 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
    • B01L2300/0803—Disc shape
    • 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
    • B01L2300/0864—Configuration of multiple channels and/or chambers in a single devices comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
    • 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
    • B01L2300/0867—Multiple inlets and one sample wells, e.g. mixing, dilution
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0409—Moving fluids with specific forces or mechanical means specific forces centrifugal forces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0475—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure
    • B01L2400/0487—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics
    • 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
    • B01L3/502753—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 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10T—TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION
    • Y10T137/00—Fluid handling
    • Y10T137/8593—Systems

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  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Health & Medical Sciences (AREA)
  • Dispersion Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Clinical Laboratory Science (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Physical Or Chemical Processes And Apparatus (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Liquid is received in liquid receiving sections (1-15, 17) formed in a first layer (50a) and a second layer (50b) of a cartridge and each having a specific volume determined according to a liquid to be received in the liquid receiving section. A reaction tank is connected to one of flow ports (1a-16a), and, with the reaction tank connected, pressure is applied to a liquid receiving section connected to the flow port to which the reaction tank is connected. This causes outside air to be supplied to that liquid receiving section from an outside-air flow section (21) communicating with the outside, and liquid received in the liquid receiving section is supplied to the reaction tank. Then, the cartridge is rotated to connect another flow port to the reaction tank, and as a result, liquids are supplied to the reaction tank. In this process, the outside air supplied from the outside-air flow section (21) is supplied to the reaction tank via the flow port connected to the reaction tank and via one of the liquid receiving sections, and as a result, the liquids received in the reaction tank are stirred by the inflow of the outside air.

Description

Fluid receiving cartridge and utilization thereof
Technical field
The present invention relates to a kind of fluid receiving cartridge (cartridge), fluid reaction unit, used this fluid receiving cartridge the gene analytic method, used the reaction unit of this fluid receiving cartridge and used the reaction method of this reaction unit.
Background technology
In the past, as fluid receiving cartridge, known have by constitute with the substrate of stiffener and container that elastomerics forms and in this container, be formed with by stream and be connected the structure (for example, with reference to patent documentation 1) that maybe can connect a plurality of chambers that dispose.Structure at this patent documentation 1 described fluid receiving cartridge is, by applying external force from the export-oriented elastomerics of container, flow-like material in stream and the chamber is moved and carries out chemical reaction.
Patent documentation 1:JP spy opens the 2005-313065 communique
Summary of the invention
Yet, in the fluid receiving cartridge of patent documentation 1, for example, make it under the situation of the easy structure that a direction moves etc. on limit pushing cylinder limit, when applying external force, sometimes having a plurality of fluids that carry out chemical reaction can not thorough mixing, and the situation that can not carry out chemical reaction fully takes place.
The present invention carries out in view of above-mentioned problem, main purpose be to provide a kind of make the fluid of accommodating more fully in fluid receiving cartridge, the fluid reaction unit of reactive tank internal reaction, used this fluid receiving cartridge the gene analytic method, used the reaction unit of this fluid receiving cartridge and used the reaction method of this reaction unit.
The present invention adopts following method for reaching above-mentioned purpose.
Fluid receiving cartridge of the present invention is the fluid receiving cartridge that fluid is accommodated removedly, and it has:
Be formed at respectively in the housing (housing) and can accommodate a plurality of resettlement sections of this fluidic with the specified volume that fluid was determined of accommodating;
Be communicated with above-mentioned resettlement section and outside, and can make extraneous air be passed to the extraneous air throughput of this resettlement section;
A plurality of connection throughput, these a plurality of connection throughput are provided in respectively by relatively moving and switching and are arranged at outside and can accommodate the link position of the regulation that above-mentioned fluidic reactive tank is connected with any one connection of above-mentioned resettlement section with above-mentioned fluid receiving cartridge and with this reactive tank, and the differential pressure of the fluid effect of accommodating by this resettlement section of subtend, the fluid that is contained in this resettlement section can be supplied to a plurality of connection throughput of above-mentioned reactive tank.
In this fluid receiving cartridge, with fluid receiving in being formed at housing and in a plurality of resettlement sections by the determined specified volume of fluid of accommodating, when reactive tank being connected under the state that connects throughput to resettlement section effect differential pressure, from with the extraneous air throughput of external communications when extraneous air is supplied with in the resettlement section, the fluid of accommodating is supplied to reactive tank.And mobile fluid receiving cartridge is connected other connection throughput and a plurality of fluids is supplied to this reactive tank with reactive tank.At this moment, because the extraneous air of supplying with from the extraneous air throughput is fed into reactive tank by resettlement section, connection throughput, therefore be contained in a plurality of fluids of reactive tank owing to the inflow of this extraneous air is foamed.Therefore, can make the fluid of accommodating more fully at the reactive tank internal reaction.
In fluid receiving cartridge of the present invention, above-mentioned a plurality of connection throughput also can be formed on the plane of above-mentioned housing, are set the round shape into the rotating shaft coaxle that moves with any one rotation of above-mentioned reactive tank and above-mentioned tube respectively.Like this, when switch connecting a plurality of connection throughput with reactive tank, rotation mobile response groove or tube get final product, are easier to thus switch connection, are easy to fluid receiving in reactive tank, so are easy to make it fully to react.At this moment, at least one of above-mentioned a plurality of resettlement sections is formed in the above-mentioned housing of circular plate shape, and forming the tubulose of the circuitous shape that increases from the circuitous amplitude of interior all side direction outer circumferential sides of the housing of above-mentioned circular plate shape, above-mentioned a plurality of connection throughput also can be arranged at interior all sides of the housing of above-mentioned circular plate shape.Like this, increase the space that can effectively utilize the housing of circular plate shape towards the circuitous amplitude of outer circumferential side, and, by forming tubulose, the fluid that is contained in the resettlement section can be supplied to reactive tank fully.Perhaps, above-mentioned a plurality of resettlement sections also can be formed in the above-mentioned housing of circular plate shape, and the more outer circumferential side in the many more housings that are formed at above-mentioned circular plate shape more of the fluidic amount of accommodating.Like this, be easy to form the more resettlement section of fluidic amount of accommodating.
In fluid receiving cartridge of the present invention, at least one of above-mentioned a plurality of resettlement sections also can form the tubulose that approaching more above-mentioned connection throughput has the tendency that attenuates more.Like this, be easy to from the resettlement section more whole fluid transfers to reactive tank, and then be easy to make it fully to react at reactive tank.
In fluid receiving cartridge of the present invention, at least one of above-mentioned a plurality of resettlement sections also can form the tubulose of dual tapered.Like this, can prevent that the fluid of accommodating from flowing out when circulation or during carrying, thereby make the fluid of accommodating fully at the reactive tank internal reaction.
In fluid receiving cartridge of the present invention, above-mentioned a plurality of resettlement sections also can form the many more long more tubuloses of the fluidic amount of accommodating.Like this, can effectively accommodate fluid.
In fluid receiving cartridge of the present invention, above-mentioned housing also can be made of a plurality of dividing layers, and above-mentioned resettlement section is formed at any one dividing layer in above-mentioned a plurality of dividing layer or crosses over plural dividing layer and form.Like this, by a plurality of dividing layers, can form more resettlement section.At this moment, above-mentioned a plurality of resettlement sections also can be formed at more from the formed plane of above-mentioned connection throughput away from dividing layer, it is more to form the fluidic amount of accommodating more.Like this, since few more away from the quantity that connects the throughput resettlement section more, therefore can further reduce the quantity that connects resettlement section and the path that is connected throughput, and then be easy to make tube.
In fluid receiving cartridge of the present invention, also can have: but be formed in the above-mentioned housing and the reservoir of store fluid in the mode that is communicated with extraneous air; Above-mentioned reactive tank and above-mentioned fluid receiving cartridge are relatively moved, and when switching in the mode of the position relation that forms regulation, be arranged at the regulation that above-mentioned reactive tank is communicated with above-mentioned reservoir binding site in conjunction with throughput.Like this, can carry out need be in reactive tank the reaction of store fluid.At this moment, also can have: be arranged at above-mentioned reservoir and above-mentioned, allow, and stop from the outflow restrictions of this reservoir to the fluidic circulation of this reactive tank from of the fluidic circulation of above-mentioned reactive tank to above-mentioned reservoir in conjunction with between the throughput; Be arranged at above-mentionedly, can be adsorbed on the deceleration column of the resultant that the reaction of above-mentioned reactive tank forms in conjunction with between throughput and the above-mentioned outflow restrictions; Be arranged between any one resettlement section and above-mentioned deceleration column of above-mentioned a plurality of resettlement sections, allow that the fluidic to this deceleration column circulates from this resettlement section, and stop from the inflow restrictions of this deceleration column to the fluidic circulation of this resettlement section.Like this, need not to switch reactive tank with in conjunction with being connected of throughput, can be adsorbed in deceleration column at the resultant that the reactive tank reaction generates after, make the fluid flow the resettlement section in arrive this deceleration column, and make this fluid turn back to reactive tank.At this moment, above-mentioned deceleration column also can contain ceramic column casing.And it is also harmless that the structure of above-mentioned resettlement section is used in reservoir.
In having the fluid receiving cartridge of the present invention of reservoir, above-mentioned reservoir also can have the absorbing material of absorption fluids in the inside harvesting.Like this, the fluid that temporarily is contained in reservoir can be remained in the reservoir more reliably.
In fluid receiving cartridge of the present invention, as above-mentioned fluid, above-mentioned resettlement section also can be accommodated and be carried out the liquid that amplification, fragmentation, hybridization, mark are used to being used for genomic dna that SNP, variation resolve, or carries out the liquid that reverse transcription, amplification, fragmentation, hybridization, the mark of cDNA are used from RNA.Carry out the liquid that amplification, fragmentation, hybridization, mark are used to being used for genomic dna that SNP, variation resolve, or carry out the liquid that reverse transcription, amplification, fragmentation, hybridization, the mark of cDNA use from RNA and need a variety of, and, it is fully reacted, and it is bigger therefore to be suitable for meaning of the present invention.
In fluid receiving cartridge of the present invention, as above-mentioned fluid, the genomic dna that above-mentioned resettlement section also can be accommodated being used for the chromosome abnormalty parsing carries out the liquid that amplification, fragmentation or mark are used.Being used for genomic dna that chromosome abnormalty resolves, to carry out liquid needs that amplification, fragmentation or mark use a variety of, and, it is fully reacted, therefore suitable meaning of the present invention is bigger.
In fluid receiving cartridge of the present invention, said external airiness portion also can have: have the extraneous air hole and be communicated with above-mentioned resettlement section and outside extraneous air circulation flow path; Be equipped on this extraneous air circulation flow path, porous material not by above-mentioned fluid but by the said external air.Like this, the fluid that is contained in the resettlement section is not flowed out to the outside from the resettlement section by the extraneous air throughput.At this moment, said external airiness portion also can be connected with a plurality of above-mentioned resettlement sections.Like this, can make the fluid that is contained in the resettlement section not pass through the extraneous air throughput, further the quantity in the extraneous air hole of minimizing and external communications to the effusive while of outside from the resettlement section.
And above-mentioned connection throughput also can be connected with above-mentioned reactive tank, by the differential pressure to the fluid effect that is contained in this reactive tank, the fluid that is contained in this reactive tank can be supplied with to above-mentioned resettlement section.Like this, the fluid receiving by will being contained in reactive tank can carry out new reaction etc. at reactive tank thus to the resettlement section.
Fluid reaction of the present invention unit has: above-mentioned any one fluid receiving cartridge; Can connect with a plurality of throughput that are connected that are equipped on above-mentioned fluid receiving cartridge, and can accommodate the fluidic reactive tank of supplying with from the connection throughput of this connection.
In this fluid reaction unit, owing to have above-mentioned any one fluid receiving cartridge, therefore played the effect that above-mentioned any one fluid receiving cartridge played, for example, can make effect that the fluid of accommodating reacts at reactive tank more fully and a plurality of connection throughput and reactive tank are switched when being connected, rotation mobile response groove or tube get final product, thereby are easier to switch connection, be easy to fluid receiving to be easy to make its effect of fully reacting etc. thus in reactive tank.At this moment, above-mentioned reactive tank also can form the thin more tubulose of approaching more above-mentioned connection throughput.Like this, both be easy to make whole fluids between fluid receiving portion and reactive tank, to circulate, and also be easy to make the fluid in the reactive tank to foam because of the extraneous air that connects the throughput circulation by connecting throughput.
Reaction unit of the present invention is the reaction unit that a plurality of fluids is mixed and make its reaction, and it has: the installation unit that above-mentioned any one fluid receiving cartridge can be installed; Can connect with a plurality of throughput that are connected of the fluid receiving cartridge that is equipped on above-mentioned installation, and can accommodate the fluidic reactive tank of supplying with from the connection throughput of this connection; At least one of the fluid receiving cartridge of above-mentioned reactive tank and above-mentioned installation moved to the mobile unit of above-mentioned reactive tank and the link position of the regulation of above-mentioned a plurality of any one connection that is connected throughput; To the resettlement section of above-mentioned fluid receiving cartridge effect differential pressure, and can pay the unit with the pressure that the fluid that is contained in this resettlement section is supplied with to above-mentioned reactive tank.
In this reaction unit, any one above-mentioned fluid receiving cartridge is installed, and at least one of the fluid receiving cartridge of reactive tank and installation moved the link position of any one regulation that is connected with reactive tank of a plurality of connection throughput that are connected to the fluid receiving cartridge that is equipped on installation, the resettlement section effect differential pressure of co-current flow body receiving cartridge, the fluid that will be contained in this resettlement section is thus supplied with to reactive tank.So, under reactive tank being connected in the state that connects throughput during to resettlement section effect differential pressure, from the extraneous air throughput of external communications when extraneous air is supplied with in the resettlement section, the fluid of accommodating is fed into reactive tank.And mobile fluid receiving cartridge connects other and connects throughput and reactive tank, and a plurality of fluids are supplied to this reactive tank.At this moment, because the extraneous air that is supplied to from the extraneous air throughput is supplied to reactive tank by resettlement section, connection throughput, therefore be contained in a plurality of fluids of reactive tank owing to the inflow of this extraneous air is foamed.Therefore, the fluid of accommodating is reacted more fully at reactive tank.
Reaction unit of the present invention is the reaction unit that a plurality of fluids is mixed and make its reaction, also can have: can connect the unitary connector element of above-mentioned any one fluid reaction; At least one of the fluid receiving cartridge of above-mentioned reactive tank and above-mentioned installation moved to the mobile unit of reactive tank in the fluid reaction unit of above-mentioned connection and the regulation link position of above-mentioned a plurality of any one connection that is connected throughput; To the resettlement section of above-mentioned fluid receiving cartridge effect differential pressure, and can pay the unit with the pressure that the fluid that is contained in this resettlement section is supplied with to above-mentioned reactive tank.Like this, the fluid of accommodating is reacted more fully at reactive tank.
In reaction unit of the present invention, above-mentioned reactive tank also can form the thin more tubulose of approaching more above-mentioned connection throughput.Like this, both be easy to make whole fluids between fluid receiving portion and reactive tank, to circulate, and also be easy to make the fluid in the reactive tank to foam because of the extraneous air that connects the throughput circulation by connecting throughput.
Reaction unit of the present invention also can have control unit, this control unit is contained in the series reaction step that the fluid of above-mentioned resettlement section of the fluid receiving cartridge of above-mentioned installation carries out based on use, so that any one of above-mentioned a plurality of resettlement sections and above-mentioned reactive tank ways of connecting when controlling above-mentioned mobile unit, are paid the unit to make the mode of fluid transfer control above-mentioned pressure to above-mentioned resettlement section effect differential pressure.Like this, based on a series of reactions steps, can transfer fluid and make its reaction.
In reaction unit of the present invention, above-mentioned pressure is paid the unit and also can be connected with above-mentioned reactive tank, by reduce air pressure in this reactive tank with above-mentioned fluid from above-mentioned resettlement section to above-mentioned reactive tank sucking-off, by the air pressure in this reactive tank that raises above-mentioned fluid is released to above-mentioned resettlement section from this reactive tank.Like this, the pressure that only changes in the reactive tank can be released fluid from resettlement section sucking-off fluid or to the resettlement section, therefore can transfer fluid with better simply structure.
In reaction unit of the present invention, above-mentioned installation unit also can be installed the fluid receiving cartridge of the housing with the circular plate shape that is formed with above-mentioned resettlement section, above-mentioned mobile unit is by rotating any one that moves above-mentioned reactive tank and above-mentioned fluid receiving cartridge, at least one of the fluid receiving cartridge of mobile thus above-mentioned reactive tank and above-mentioned installation around turning axle.Like this, when switching a plurality of connection throughput of connection and reactive tank, rotation mobile response groove or tube get final product, and are easier to thus switch connect.At this moment, above-mentioned pressure is paid the unit also can be by above-mentioned reactive tank to the above-mentioned resettlement section of above-mentioned fluid receiving cartridge applied pressure, and above-mentioned mobile unit is rotated around turning axle and moved above-mentioned fluid receiving cartridge.Like this, on fluid receiving cartridge, do not connect pressure and pay the unit, therefore be easy to make its rotation.Perhaps, above-mentioned installation unit is installed this fluid receiving cartridge by abutting part and fixed part, wherein, this abutting part is on the plane of the housing of the above-mentioned circular plate shape that is formed with above-mentioned connection throughput, with this plane butt that sets to the fluid receiving cartridge of the round shape of the rotating shaft coaxle that moves with the rotation of above-mentioned housing, and be provided with and the above-mentioned communicating pores that is connected the throughput connection, fixed part is rotatable this fluid receiving cartridge of fixing under the state of the above-mentioned plane butt that makes above-mentioned abutting part and above-mentioned fluid receiving cartridge, and above-mentioned reactive tank also can be connected with the communicating pores that is arranged at above-mentioned abutting part.Like this, can under the state of the more or less freely switching of carrying out resettlement section and reactive tank, fluid receiving cartridge be installed.
Reaction unit of the present invention also can have: the tube temperature adjustment unit that can regulate the temperature of the fluid storage tube that is installed on above-mentioned installation unit; Can regulate the reactive tank temperature adjustment unit of the temperature of above-mentioned reactive tank.Like this, by respectively the fluid storage tube of installing being adjusted to the nonreactive temperature of the liquid of accommodating, or reactive tank is adjusted to the temperature of suitable reaction, the Temperature Influence of the fluid storage tube of not installed thus can make fluid fully reaction in reactive tank of accommodating.
Reaction method of the present invention has been to use the reaction method that a plurality of fluids is mixed and make the reaction unit of its reaction, comprise: (a) can connect with a plurality of throughput that are connected on being provided in technical scheme 1～16 arbitrary described fluid receiving cartridge that is installed on above-mentioned reaction unit, and at least one of fluid receiving cartridge that can accommodate the fluidic reactive tank supplied with from the connection throughput of this connection and above-mentioned installation move to the stage of this reactive tank and the link position of the regulation of above-mentioned a plurality of any one connection that is connected throughput; (b), and will be contained in the stage of the fluid receiving of this resettlement section to above-mentioned reactive tank to the resettlement section of above-mentioned fluid receiving cartridge applied pressure.
In this reaction method, at least one of the fluid receiving cartridge of reactive tank and installation moved to the link position of reactive tank and the regulation of a plurality of any one connection that is connected throughput, the resettlement section applied pressure of co-current flow body receiving cartridge, to be contained in the fluid receiving of this resettlement section to reactive tank, wherein, this reactive tank can connect with a plurality of throughput that are connected on being provided in above-mentioned any one fluid receiving cartridge that is installed on reaction unit, and can accommodate the fluid of supplying with from the connection throughput of this connection.So, when reactive tank being connected under the state that connects throughput to resettlement section effect differential pressure, from the extraneous air throughput of external communications when extraneous air is supplied with in the resettlement section, the fluid of accommodating is supplied to reactive tank.And mobile fluid receiving cartridge connects throughput with other and is connected with reactive tank, and a plurality of fluids are supplied to this reactive tank.At this moment, the extraneous air of supplying with from the extraneous air throughput is fed into reactive tank by resettlement section, connection throughput, therefore is contained in a plurality of fluids of reactive tank owing to the inflow of this extraneous air is foamed.Therefore, can make the fluid of accommodating more fully at the reactive tank internal reaction.
Gene analytic method of the present invention, the fluid receiving cartridge that uses will carry out the liquid that amplification, fragmentation, hybridization, mark are used to being used for genomic dna that SNP, variation resolve, or carry out liquid that reverse transcription, amplification, fragmentation, hybridization, the mark of cDNA use as fluid from RNA, be contained in the resettlement section, detect the variation of SNP, DNA or the discovery amount of RNA (mRNA or miRNA etc.).
Gene analytic method of the present invention, the fluid receiving cartridge that uses will carry out liquid that amplification, fragmentation or mark use as fluid to being used for genomic dna that chromosome abnormalty resolves, be contained in the resettlement section, measure with the human chromosome variation of the chromosome copies number that is object.
Description of drawings
Fig. 1 is the formation schematic pie graph of expression reaction unit 90.
Fig. 2 is the explanatory view of the section after explanation tube installing mechanism 80 and tube are installed.
Fig. 3 is the explanatory view that is arranged at the degassing vessel of reactive tank 30.
Fig. 4 is the chamber of tube 50 inside and the explanatory view of stream.
Fig. 5 is the explanatory view of miniature check valve 160.
Fig. 6 is the explanatory view of position of the porous material of tube 50 inside.
Fig. 7 is the explanatory view of the thermal insulation structure of reaction unit 90.
Fig. 8 utilizes reaction unit 90 to obtain the explanatory view of the step of cDNA from mRNA.
Fig. 9 is the explanatory view of the stream when deceleration column 16 flows into mixing solutions.
Figure 10 is the explanatory view of the stream during by deceleration column 16 sucking-off elution damping fluids.
Figure 11 is the chamber of tube 150 inside and the explanatory view of stream.
Figure 12 is the explanatory view of generation step of the genomic dna of SNP being labeled that detect usefulness.
Figure 13 is the explanatory view of the annexation of column casing 106 and reactive tank 30, exhausted bath box 28.
Figure 14 is the explanatory view of the filling step of column casing.
Figure 15 is the sectional view of the formation of other reactive tank fixed parts 136 of explanation.
Figure 16 is the explanatory view of other bottom surfaces of tube 50.
Figure 17 is the chamber of tube 250 inside and the explanatory view of stream.
Figure 18 is the explanatory view of position of the porous material of tube 250 inside.
Figure 19 is the explanatory view of step of amplification that utilizes the DNA of reaction unit 90.
Figure 20 is the explanatory view of step of fragmentation that utilizes the DNA of reaction unit 90.
Figure 21 is the explanatory view of step of mark that utilizes the DNA of reaction unit 90.
Figure 22 is the explanatory view of the stream when deceleration column 206 flows into dna solution.
Figure 23 is the explanatory view of the stream during by deceleration column 206 sucking-off elution damping fluids.
Figure 24 is the outside drawing of tube 350.
Figure 25 is the explanatory view of the first layer 351a of tube 350.
Figure 26 is the explanatory view of the second layer 351b of tube 350.
Figure 27 is the explanatory view of the 3rd layer of 351c of tube 350.
Figure 28 is the explanatory view of the 4th layer of 351d of tube 350.
Figure 29 is vertical view and the front view of microarray 350b.
Figure 30 is the B-B ' phantom view of tube 350.
Figure 31 is the explanatory view that the genomic dna to rice carries out the step of amplification when adjusting.
Figure 32 is the explanatory view that makes the step of adjusted DNA and the reaction of DNA point.
Figure 33 is the outside drawing of tube 450.
Figure 34 is the explanatory view of the first layer 451a of a body 450a.
Figure 35 is the explanatory view of the second layer 451b of a body 450a.
Figure 36 is the explanatory view of the 3rd layer of 451c of a body 450a.
Figure 37 is the explanatory view of step of the aftertreatment of CGH array.
Embodiment
First embodiment
Secondly, use accompanying drawing to describe to being used to implement best mode of the present invention.Fig. 1 is that the embodiment that the present invention is specialized in expression is the schematic pie graph of the formation of reaction unit 90, Fig. 2 is the explanatory view of an installing mechanism 80, Fig. 2 (a) is an exploded perspective view, Fig. 2 (b) is a stereographic map, Fig. 2 (c) is the A-A ' sectional view of Fig. 2 (b), Fig. 3 is the sectional view of reactive tank 30, Fig. 4 is arranged at the first layer 50a of tube 50 and the fluid storage portion of accommodating liquid of second layer 50b, the explanatory view of the configuration of the stream of liquid and gas etc., Fig. 5 is the explanatory view that contains the section of miniature check valve 160, Fig. 6 is the explanatory view of receiving and keeping in the position of the porous material of tube 50 inside, and Fig. 7 is the explanatory view of the thermal insulation structure of reaction unit 90.And, though tube 50 is described in detail later, have and reactive tank 30 difference attachable opening 1a～16a, 18a and accepting hole 17a, for convenience of description, it is referred to as " hole ", and fluid storage portion 1～15,17,18 and the exhausted bath box 28 that can accommodate liquid are referred to as " chamber ".And in the present embodiment, reaction unit 90 describes as the device that obtains mark cDNA for the discovery amount that detects mRNA.
As shown in Figure 1, the reaction unit 90 of present embodiment has: can install can take out plurality of reagents (liquid) be contained in respectively each fluid storage portion and above have guide part 52 the tube 50 the tube installing mechanism 80; Can be connected with a plurality of holes that are equipped on the tube 50 that is mounted, and can accommodate the reactive tank 30 of the liquid of supplying with from the hole of this connection; On the position of the reactive tank 30 in any one hole of connecting cylinder 50, the rotating mechanism 32 that tube 50 rotation around its central shaft is moved; To the fluid storage portion of tube 50 effect differential pressure, the liquid that is contained in this fluid storage portion is supplied with to reactive tank 30, and the pump 34 that the liquid that is contained in reactive tank 30 is moved to tube 50; Reactive tank 30 is fixed in the reactive tank fixed part 36 of support component 92; Can regulate tube amber ear card (Peltier) the element 38a of the temperature of the tube of installing by tube installing mechanism 80 50; Can regulate the reactive tank Peltier's element 36a of the temperature of reactive tank 30; The not shown start button that the processing of user's Indicator Reaction device 90 begins; The controller 40 of control reaction unit 90 integral body.And this reaction unit 90 has: be disposed at the rectangular base 90a of its foot and be equipped on the support component 92 of side L word shape of the front side of base portion 90a.In this support component 92, be formed with stage casing face 92a and the upright upward 92b of wall portion that is located at the rear side of this stage casing face 92a.And, dispose pump 34 and controller 40 etc. in the rear side of support component 92.
As shown in Figure 2, tube installing mechanism 80 has: be inserted with reactive tank 30 and be embedded in the rotating disk 82 of tube 50 guide part 52; With this rotating disk 82 from the top press section 84 of the application of force downwards; The universal stage 38 (with reference to Fig. 1) of mounting tube 50.Consider watertightness and oil tightness, on rotating disk 82, used fluorine class material (for example Teflon (registered trademark, below also with)).And, consider that thermotolerance and heat insulating ability, rotating disk are easy to slip into etc., have used fluorine class material (for example Teflon) on press section 84.Shown in the A-A ' sectional view of Fig. 2 (c), the bearing surface 52a of interior all sides of guide part 52 that rotating disk 82 and circle shape are equipped with the tube 50 in a plurality of holes connects by the O shape circle 54a that is disposed at each hole, and has any one hole that is communicated with in each hole and a circulation flow path 82a of reactive tank 30.Here, O shape circle 54a sets with each hole is corresponding.This rotating disk 82 can be embedded in the guide part 52 of tube 50 with being rotated, and by the circulation flow path 82a that sets any one hole (being any one chamber) in each hole is communicated with reactive tank 30.And this rotating disk 82 has disconnected and other holes that are not communicated with circulation flow path 82a and the circulation of extraneous air under the state that is embedded in guide part 52, and can only transfer the liquid from the hole that is connected with circulation flow path 82a.Press section 84 is to make rotating disk 82 and mounting under the state that the bearing surface 52a of the tube 50 on the universal stage 38 connects, can make tube 50 rotatably to rotating disk 82 from the top parts of the application of force downwards.This press section 84 by with the mode of two top step portion 82b of clamping rotating disk 82 from the top application of force downwards, thereby the above-below direction of restriction tube 50 move and rotation is moved, even tube 50 rotations are moved also and the location dimension of the circulation flow path 82a of rotating disk 82 can be held in same position thus.Thus, by rotating cylinder 50, can form the state that any one hole of having only in each hole is communicated with reactive tank 30.And, by O shape circle 54a is clipped in the middle, in the bearing surface 52a of rotating disk 82 and tube 50, form the space thus, thereby can suck extraneous air to the inside of tube 50, or tube 50 gas inside are discharged by this space and between the first ventilating pit 21a described later.
As shown in Figure 1, rotating mechanism 32 has: the universal stage 38 of mounting tube 50; The motor 37 of this universal stage 38 is moved in interim rotation for the link position that is fixed in regulation.Universal stage 38 is discoideus, is supported in the stage casing face 92a of support component 92 by the rotatable earth's axis.The material of this universal stage 38 is the materials that copper carried out chemical nickel plating, and is formed with not shown a plurality of protuberances on the off side in the above.Bottom surface at tube 50, with the corresponding position of the configuration of each protuberance on, be formed with a plurality of recesses (not shown) that this protuberance inserts, with under this protuberance and the state that recess embeds mutually with tube 50 mountings on universal stage 38 time, can be on the initial stage link position of stipulating stationary magazine creel 50.On this universal stage 38, its inside is equipped with the tube Peltier's element 38a of the temperature that can regulate tube 50.By this temperature of regulating this universal stage 38, the tube 50 of mounting can be adjusted to certain temperature thus with Peltier's element 38a.And, also can use the material that aluminium has been carried out the pellumina processing as the material of universal stage 38.
The material of reactive tank fixed part 36 is copper have been carried out the material of chemical nickel plating, and is fixed in the central authorities of the 92b of wall portion of support component 92, and at mounting fixation reaction groove 30 removably above the tube 50 of universal stage 38.The reactive tank Peltier's element 36a of the temperature that can regulate reactive tank 30 is contained in the inside of this reactive tank fixed part 36.Temperature by this reactive tank is regulated this reactive tank fixed part 36 with Peltier's element 36a can be adjusted to certain temperature with reactive tank 30 thus.And, also can use the material that aluminium has been carried out the pellumina processing as the material of reactive tank fixed part 36.
Reactive tank 30 is parts that material is formed by polypropylene, as shown in Figure 1, 2, is more near lower side, the piped parts that promptly attenuate near Kong Zeyue more.This reactive tank 30 is equipped with rotating disk 82 (with reference to Fig. 2 (c)) in the lower end across O shape circle 54b, is connected with as shown in Figure 1 in the upper end and send vapor pipe 34a.In this reactive tank 30, the pressure that is produced by pump 34 running is by sending vapor pipe 34a effect, and its pressure acts on tins 50 any one chamber that connects by rotating disk 82.And this reactive tank 30 is accommodated from the liquid of fluid storage portion 1～15,17,18 sucking-offs, or makes the liquid foam of accommodating, or utilizes the liquid of accommodating to carry out various reactions.And as shown in Figure 3, in the inside of this reactive tank 30, being provided with longitudinally, groove is air discharge duct 30a.
Pump 34 is to carry air and to the so-called surge pump of the front end applied pressure that connects by membranous volume-variation.As shown in Figure 1, this pump 34 is connected with sending vapor pipe 34a, and send vapor pipe 34a and 30 pairs of reactive tanks to be contained in liquid effects pressure in tube 50 the chamber by this.This pump 34 constitute can in the suction opening of this pump 34 and the air taking port any with send vapor pipe 34a to be connected to switch.The suction opening of this pump 34 when sending vapor pipe 34a to be connected, has been reduced the air pressure in the reactive tank 30, thus can be from fluid storage portion 1～15,17,18 to reactive tank 30 sucking-off liquid.And when sending vapor pipe 34a to be connected, the air pressure in the reactive tank 30 that raise can extrude liquid to exhausted bath box 28 from reactive tank 30 thus with the air taking port of this pump 34.
It is the microprocessor at center that controller 40 constitutes with CPU42, has the flash rom 43 of the various handling procedures of storage and temporarily stores the RAM44 that data are preserved data.From this controller 40 to pump 34 output comprise with the control signal of the change signal of the connection state of sending vapor pipe 34a, to motor 37 output control signals, to reactive tank with Peltier's element 36a and tube with Peltier's element 38a output service voltage etc.
Tube 50 is parts that material is formed by cyclenes copolymer, as shown in Figure 2, have: form the first layer 50a of circular plate shape, equally form circular plate shape second layer 50b, be formed at the top of the first layer 50a and be used to make this 50 guide part 52 with central shaft rotation.This first layer 50a and second layer 50b have a plurality of chambeies respectively.As shown in Figure 4, this tube 50 has: a plurality of fluid storage portion 1～15,17,18 that can accommodate liquid with the specified volume of the liquid of accommodating decision; By making tube 50 rotations move and switch, thereby be equipped on opening 1a～16a, the 18a of any one the regulation link position that is connected with reactive tank 30 that makes in these chambeies respectively; Be communicated with fluid storage portion 1～15,17,18 and extraneous air, and in the chamber, introduce extraneous air or discharge the extraneous air throughput 21 of gas in the chamber; Be used for accommodating the accepting hole 17a of liquid to fluid storage portion 17; Can accommodate the exhausted bath box 28 of the waste liquid of reactive tank 30 supplies; Be arranged between exhausted bath box 28 and the opening 16a, homologation reaction groove 30 stops from the outflow limiting valve 16e of this exhausted bath box 28 to the waste liquid circulation of reactive tank 30 to the waste liquid circulation of exhausted bath box 28; Can be adsorbed on the deceleration column 16 of the production thing of reactive tank 30 reactions; Be arranged between fluid storage portion 17 and the deceleration column 16, allow the liquid communication to deceleration column 16, stop from the inflow limiting valve 16d of deceleration column 16 to the liquid communication of fluid storage portion 17 from fluid storage portion 17.
Form the parts of fine of accommodating than liquid by the throughput branch with liquid, fluid storage portion the 1～15,17, the 18th thus, forms the piped space of dual tapered.For example, shown in the fluid storage portion 4 relative with fluid storage portion 1, the amount of the liquid that this fluid storage portion 1～15,17,18 accommodates forms the longer pipe shape more at most more.And fluid storage portion 12～15,17 forms the tubulose of the circuitous shape that increases from the circuitous amplitude of tube interior all side direction outer circumferential sides of 50 of circular plate shape, circuitous tubular become thin more near each hole more.And, from the isolating second layer 50b of bearing surface 52a of the first layer 50a that is formed with each hole, for the amount that makes the liquid of accommodating more than other chambeies that are formed at the first layer 50a, and be formed with big volumetrical fluid storage portion 14,15.And, though each fluid storage portion 1～13,17,18 is communicated with by extraneous air circulation flow path 21d, but as described later, because harvesting has hydrophobic porous material, the liquid that can not that therefore is contained in this fluid storage portion 1～13,17,18 mixes by extraneous air circulation flow path 21d.
Extraneous air throughput 21 has: the extraneous air circulation flow path 21d that is connected with an end of the outer circumferential side of the fluid storage portion 1～13,17,18 of the first layer 50a; The extraneous air circulation flow path 21e that is connected with an end of the outer circumferential side of the fluid storage portion 14,15 of second layer 50b; The first ventilating pit 21a that these extraneous air circulation flow paths 21d, 21e are communicated with extraneous air; Receive and keep the hydrophobic porous material in extraneous air circulation flow path 21d, 21e.And, in Fig. 6, use grid representation extraneous air throughput 21.This hydrophobic porous material can not be by liquid but can be passed through extraneous air,, for example uses special teflon (テ Off ロ Application) porous material (day eastern electrician company make TEMISH) here.And, the end of extraneous air circulation flow path 21e is connected with fluid storage portion 14,15, the other end is connected with the first ventilating pit 21a to vertical stream that the stream position of the first layer 50a 22b forms by the stream position 22c from second layer 50b, and is communicated with extraneous air by this first ventilating pit 21a.
Opening 1a～16a, 18a are communicated with fluid storage portion 1～15,17,18 respectively, the hole of using during from fluid storage portion 1～15,17,18 feeding liquids, and be set at top interior all sides (bearing surface 52a) of the first layer 50a.Opening 1a～16a, 18a are provided in respectively and the turning axles of tube 50 by rotating mechanism 32 rotation promptly on the same plane of the round shape of the spigot shaft coaxle of tube 50.And the differential pressure by to the liquid effects that is contained in the fluid storage portion 1～15,17,18 that is connected with these openings can make the liquid that is contained in these fluid storage portions 1～15,17,18 supply to reactive tank 30.Here, opening 14a, 15a be by being formed at the stream longitudinally that connects the first layer 50a and the second layer, and the end with the fluid storage portion 14,15 of second layer 50b is that link position 14c, 15c link to each other respectively.
Accepting hole 17a is communicated with fluid storage portion 17, the hole of using when accommodating from liquid that the outside is transferred, and be set at top interior all sides (bearing surface 52a) of the first layer 50a.This accepting hole 17a is equipped on the circle that is equipped with opening 1a～16a, 18a.Here, fluid storage portion 17 is connected with inflow limiting valve 16d to vertical stream of the position of second layer 50b 24c by the stream position 24b from an end of this fluid storage portion 17.And the other end of this fluid storage portion 17 is connected with extraneous air circulation flow path 21d, though be described in detail later, harvesting has the hydrophobic porous material that does not pass through liquid in this extraneous air circulation flow path 21d.Thus, all can not in fluid storage portion 17, accommodate liquid, can be provided with this accepting hole 17a in the mode of in fluid storage portion 17, accommodating liquid thus from any end.And, in other fluid storage portion 1～15, can accommodate from opening 1a～15a.
Deceleration column 16 is set at opening 16a and flows out between the limiting valve 16e, contains column casing.Here, use ceramic column casing (for example, silica gel etc.) as column casing.And, move to reactive tank 30 by opening 16a owing to be contained in the liquid of fluid storage portion 17 respectively, therefore the opening 16a that is communicated with deceleration column 16 is communicated with and is set at top interior all sides (bearing surface 52a) of the first layer 50a with this fluid storage portion 17.
Flowing out limiting valve 16e and flowing into limiting valve 16d is so-called vacuum breaker.For example, can use miniature check valve shown in Figure 5 160.Miniature check valve 160 for example is made of with the cover body 162 of fixing this valve body 161 circular valve 161 of silicon rubber system.This valve body 161 is provided with circular-arc slit, and it is the structure that axle shakes that the baffle plate 163 of this slit inboard forms with axis of rocking 164.Cover body 162 has the valve body clamping part 162a of clamping valve body 161.This valve body clamping part 162a forms diameter less than baffle plate 163 with the diameter in the hole in 161 left sides of valve body among the figure, and the diameter in the hole on valve body 161 right sides is formed greater than the diameter of baffle plate 163 and less than the diameter of valve body 161.Thus, when fluid A direction in figure flows, be that axle bumper plate 163 tilts to occur the slit to the A direction with the axis of rocking 164 of valve body 161, fluid from this slit by mobile.On the other hand, when fluid was desired in figure the B direction and flowed, though be that axle bumper plate 163 is desired to tilt to the B direction with the axis of rocking 164 of valve body 161, baffle plate 163 did not have the slit with the end face butt of cover body 162, and fluid is mobile.This miniature check valve 160 is that the B direction is this reciprocal mode from the direction of deceleration column 16 towards stream position 23c as flowing out limiting valve 16e with the A direction among Fig. 5, is arranged at tube 50.And, as flowing into limiting valve 16d, be that the B direction is this reciprocal mode from the direction of position 24c towards deceleration column 16 with the A direction among Fig. 5, be arranged at tube 50.And miniature check valve 160 uses the parts in the gloomy research department exploitation of the tinkling of pieces of jade of engineering section of system of engineering portion of Okayama Univ..
As shown in Figure 4, exhausted bath box 28 is the spaces that are provided with in the mode around outermost week Monday of tube 50, forms a space crossing over the first layer 50a and second layer 50b.In the inside of this exhausted bath box 28, for the waste liquid that will temporarily flow into remains in the exhausted bath box 28 reliably, harvesting has the water-absorbent porous material of for example sponge that absorbs waste liquid etc.This water-absorbent porous material is along the outer circumferential side one week configuration (with reference to the oblique line position of Fig. 6) of tube 50.And, the stream that this exhausted bath box 28 forms by the position 23b from the stream of position 23d all sides in being arranged at of the stream end that is connected with this exhausted bath box 28 and from this position 23b to vertical stream that the position of the stream of second layer 50b 23c forms, be connected in deceleration column 16.That is, the fluid that passes through from deceleration column 16 and outflow limiting valve 16e is accommodated in this exhausted bath box 28.And, on the first layer 50a of the position of exhausted bath box 28, be provided with the second ventilating pit 28a that is communicated with extraneous air.
In the reaction unit 90 that so constitutes, the liquid that will contain the reaction institute with medicament etc. of regulation is contained in the fluid storage portion of tube 50 respectively with desirable amount, and rotate successively by motor 37 and to move tube 50 when changing the link position in hole, by pump 34 from the fluid storage portion 1～15,17,18 that connects to of the reaction of reactive tank 30 feeding liquids to stipulate at reactive tank 30, or transfer these reacted liquid to exhausted bath box 28 from reactive tank 30, form the formation of practicable various processing thus.Special when carrying out the purification of resultant of reaction, by making column casing absorption reaction resultant and the liquid of not wanting accommodated to exhausted bath box 28, from fluid storage portion 17 make liquid from this column casing by and supply to reactive tank 30 and carry out.And, this reaction unit 90 as shown in Figure 7, because reactive tank 30 is arranged at tube 50 outside, so the temperature variation of reactive tank 30 is difficult to transmit to tube 50, reactive tank 30 and tube 50 can be remained on different separately temperature (for example temperature of reaction and preservation with temperature etc.) thus.
Here, to reaction unit 90, especially to describing from the operation that obtains mark cDNA as the mRNA of sample.Fig. 8 is the explanatory view that obtains the step of mark cDNA from mRNA.In the figure, the expression of the pattern opening, the reactive tank 30 that tube 50 fluid storage portion 1～15,17 and exhausted bath box 28 are arranged, connect.Among the figure, in fluid storage portion 1～15,17 and exhausted bath box 28, record kind and the amount of accommodating and described each symbol that constitutes of Fig. 4 of liquid.Inner state of not accommodating liquid is represented in the chamber on empty hurdle.And, about reactive tank 30, contain the situation ellipse representation of liquid in the reactive tank 30, the liquid of accommodating to be carried out situation about handling represent with rectangle, the situations that what is not all accommodated in the reactive tank 30 are with the ellipse representation on empty hurdle.And, arrow express liquid among the figure or gas flow direction.And, for convenience of description, in the part additional phase numbering of reactive tank 30.
At first, the user will put into reactive tank 30 backs as the mRNA of sample and be connected with rotating disk 82, and it is embedded tube 50, thus as reaction member (with reference to Fig. 2 (b)).And, open the lateral not shown door that is arranged at reactive tank fixed part 36, the top of reactive tank 30 is communicated with sending vapor pipe 34a, rotating disk 82 with the portion of being pressed 84 downwards the mode of the application of force slide from the side, thus with this reaction member mounting in universal stage 38.At this moment, press section 84 is because material is special teflon, and on a plurality of protuberances of deflection on be arranged at universal stage 38, the mode that is formed at a plurality of recesses of 50 bottom surface with embedding is carried out mounting, and with press section 84 downwards the state of the application of force install.Secondly, press not shown start button.So the CPU42 of controller 40 reads and carries out the cDNA complex sign handling procedure that is stored in flash rom 43.When carrying out this program, CPU42 at first remains on specified temperature (for example 20 ℃) with Peltier's element 38a with tube 50 by tube.Secondly, make tube 50 rotations that opening 1a is communicated with reactive tank 30 by drive-motor 37, make the air pressure in the pump 34 running reduction reactive tanks 30, the liquid that will be contained in fluid storage portion 1 thus is sucked in the reactive tank 30 (stage S100).
Secondly, opening 2a is communicated with reactive tank 30, sucking-off is contained in the liquid (stage S110) of fluid storage portion 2.Then, when with Peltier's element 36a the temperature in the reactive tank 30 being remained on 70 ℃ by reactive tank, make the air pressure in pump 34 runnings the continuing reduction reactive tank 30, continue to introduce extraneous air in the reactive tank 30 by extraneous air throughput 21, fluid storage portion 2, opening 2a, circulation flow path 82a thus, made the liquid foam that is contained in the reactive tank 30 thus 10 minutes, thereby make the mixing solutionss in the reactive tank 30 react (stage S120).Continue to introduce extraneous air by extraneous air throughput 21, any one fluid storage portion 1～15,17, the opening that is arranged at an end of this fluid storage portion 1～15,17, circulation flow path 82a, can make the liquid foam in the reactive tank 30 thus.And, below, the situation of so-called " foaming " is all carried out this operation.And, as shown in Figure 3, in reactive tank 30, be provided with air discharge duct 30a, it is fashionable from the orifice flow that connects that air pressure in reactive tank 30 reduces extraneous air, by mobile gas in this air discharge duct 30a, the liquid level that prevents the liquid of accommodating rises and to adhering to of wall etc., thereby the liquid of accommodating is reacted more fully.Get back to the explanatory view of Fig. 8, then, foaming made its reaction (stage S130) in 3 minutes when the temperature in the reactive tank 30 was remained on 42 ℃.Reactive tank 30 is communicated with successively with opening 3a～8a, and sucking-off is contained in liquid (the stage S140～S190) of fluid storage portion 3～8 successively.Foaming made its reaction (stage S200) in 40 minutes when then, the temperature in the reactive tank 30 being remained on 42 ℃.Opening 9a is communicated with reactive tank 30, and sucking-off is contained in the liquid (stage S210) of fluid storage portion 9.Foaming made its reaction (stage S220) in 40 minutes when then, the temperature in the reactive tank 30 being remained on 42 ℃.Reactive tank 30 is communicated with successively with opening 10a～12a, and sucking-off is contained in liquid (the stage S230～S250) of fluid storage portion 10～12 successively.Foaming made its reaction (stage S260) in 60 minutes when then, the temperature in the reactive tank 30 being remained on 65 ℃.Reactive tank 30 is communicated with successively with opening 13a, 14a, and sucking-off is contained in the liquid (adsorption-buffering liquid) (stage S270, S280) of fluid storage portion 13～14 successively.
Then, opening 16a is communicated with reactive tank 30, the connection state of pump 34 is switched, make the air pressure in pump 34 runnings and the rising reactive tank 30, make the mixing solutions in the reactive tank 30 be passed to deceleration column 16 (stage S290).Use Fig. 9 that the appearance of this moment is described.Fig. 9 is the explanatory view of the stream of mixing solutions circulation.The opening 16a of mixing solutions by tube 50 the first layer 50a shown in Fig. 9 (a) flows into and when circulating in deceleration column 16, only underlined cDNA is by the absorption of the column casings in the deceleration column 16 in the reaction mixture.And the waste liquid that passes through from column casing passes through from the outflow limiting valve 16e of the second layer 50b shown in Fig. 9 (b), and passes through to vertical stream of the first layer 50a from the position 23c of stream, finally flows into exhausted bath box 28.At this moment, though be connected with fluid storage portion 17 in the stream of waste liquid circulation, owing to be arranged at the inflow limiting valve 16d in front of it, this waste liquid can influent resettlement section 17 yet.
Then, opening 15a is communicated with reactive tank 30, the connection state of pump 34 is switched, make the air pressure in pump 34 runnings and the reduction reactive tank 30, sucking-off is contained in the liquid (lavation buffer solution) (stage S300) of fluid storage portion 15.Then, opening 16a is communicated with reactive tank 30, the connection state of pump 34 is switched, make the air pressure in pump 34 runnings and the rising reactive tank 30, make reactive tank 30 interior flow of liquor lead to deceleration column 16 and thus column casing is washed (stage S310).In addition, identical with the stage 290, the lavation buffer solution after the washing flows into exhausted bath box 28.Then, make under opening 16a and the state that reactive tank 30 is communicated with, connection state to pump 34 is switched, make the air pressure in pump 34 runnings and the reduction reactive tank 30, the liquid (elution damping fluid) that is contained in fluid storage portion 17 is circulated and to reactive tank 30 sucking-offs (stage S320) from deceleration column 16, be contained in thus in the reactive tank 30 (stage S330).Use Figure 10 that the appearance of this moment is described.Figure 10 is the explanatory view of stream that is contained in the liquid communication of fluid storage portion 17.When reducing the air pressure in the reactive tank 30, the air pressure of the stream of deceleration column 16 and waste liquid flow side thereof also reduces.At this moment, though all can be by from exhausted bath box 28 sucking-offs by flowing out limiting valve 16e waste liquid and gas, but from fluid storage portion 17, the liquid that is contained in this fluid storage portion 17 passes through to the vertical stream of the position of second layer 50b 24c from the stream position 24b of an end of fluid storage portion 17, and from deceleration column 16, pass through, finally be sucked in the reactive tank 30.At this moment, because therefore the adsorbed mark cDNA stripping in the elution damping fluid of column casing forms the state that has entered the solution that contains this cDNA in the reactive tank 30.And, though on the stream of interior all sides of fluid storage portion 17, be provided with accepting hole 17a, be rotated dish 82 and block, can not take place that extraneous air flows into or liquid flows out by this accepting hole 17a.So, when making the resultant stripping of column casing absorption and sucking-off, from by flowing into the liquid stripping that fluid storage portion 17 that limiting valve 16d is connected with this column casing accommodated and being sucked in the reactive tank 30.Then, when continuing the operation of pump 34 temperature in the reactive tank 30 are remained on 80 ℃ 30 minutes, make the mixing solutionss that are sucked in the reactive tank 30 concentrate (stage S340), obtain mark cDNA thus.
So, in a plurality of fluid storage portion 1～15,17 any one is connected with reactive tank 30, the liquid effects differential pressure that operation by pump 34 is accommodated the fluid storage portion that connects, and with this fluid storage in reactive tank 30, continue the operation of pump 34 by original state, to be incorporated in the reactive tank 30 from the extraneous air that extraneous air throughput 21 flows into, be contained in foaming, the reaction of the liquid of reactive tank 30 thus.And, use the mark cDNA that obtains, can carry out analysis to the mRNA that puts into reactive tank 30 at first.For example, the mark cDNA that obtains is applied to common DNA chip.Through hybridization, washing procedure, with scanner be taken into the fluorescence that from the point of DNA chip obtains, and strength of signal carried out digitized processing thereafter.To be made as contrast from the signal that normal cell mRNA obtains, by with from checking the ratio of the signal that the object cell mRNA obtains, measure the variation of discovery amount.
Here, get the corresponding relation of the integrant and the integrant of the present invention of present embodiment clear.The tube 50 of present embodiment is equivalent to fluid receiving cartridge of the present invention, and opening 1a～16a, 18a are equivalent to connect throughput, and fluid storage portion 1～15,17 is equivalent to the resettlement section, and the combiner of the first layer 50a and second layer 50b is equivalent to housing.And, the first layer 50a and second layer 50b are equivalent to dividing layer respectively, exhausted bath box 28 is equivalent to reservoir, opening 16 is equivalent in conjunction with throughput, flow out limiting valve 16e and be equivalent to flow out restrictions, flow into limiting valve 16d and be equivalent to flow into restrictions, the water-absorbent porous material is equivalent to absorbing material, the first ventilating pit 21a is equivalent to the extraneous air hole, hydrophobic porous material is equivalent to porous material, and tube installing mechanism 80 is equivalent to installation unit, and rotating mechanism 32 is equivalent to mobile unit, pump 34 is equivalent to pressure and pays the unit, rotating disk 82 is equivalent to abutting part, and press section 84 is equivalent to fixed part, and circulation flow path 82a is equivalent to communicating pores, tube is equivalent to a temperature adjustment unit with Peltier's element 38a, and reactive tank is equivalent to reactive tank temperature adjustment unit with Peltier's element 36a.
Reaction unit 90 according to the present embodiment of above detailed description, at the first layer 50a that is formed at tube 50, second layer 50b, and in a plurality of fluid storage portion 1～15 of the specified volume of the decision of the liquid by accommodating, accommodate liquid in 17, under any one opening in a plurality of holes and the state that reactive tank 30 is connected, in the fluid storage portion 1～15 that is communicated with to the opening that is connected with reactive tank 30, during 17 applied pressures, extraneous air is supplied to this fluid storage portion 1～15 from the extraneous air throughput 21 with external communications, in the time of 17, the liquid of accommodating is fed in the reactive tank 30.And, tube 50 rotations, other openings are connected with reactive tank 30, and a plurality of liquid are fed in the reactive tank 30.At this moment, extraneous air from 21 supplies of extraneous air throughput, be fed in the reactive tank 30 by opening and any one the fluid storage portion 1～15,17 that is connected with reactive tank 30, therefore be contained in a plurality of liquid of reactive tank 30 owing to the inflow of this extraneous air is foamed.Therefore, the liquid of accommodating is reacted more fully in reactive tank 30.
And, owing to be formed on the bearing surface 52a of the first layer 50a, the hole set respectively into the round shape of the spigot shaft coaxle of tube 50, each hole and reactive tank 30 switched when being connected, tube 50 rotations are got final product, therefore be easier to switch connect, be easy to fluid storage in reactive tank 30, be easy to thus it is fully reacted.And, about forming the tubulose of the circuitous shape that increases from the circuitous amplitude of tube interior all side direction outer circumferential sides of 50 of circular plate shape in the fluid storage portion 1～15,17, because each hole is set at interior all sides of the tube 50 of circular plate shape, can increase circuitous amplitude towards outer circumferential side thus, effectively utilize the space of the tube 50 of circular plate shape, and,, the liquid that is contained in fluid storage portion 1～15,17 is supplied in the reactive tank 30 fully by forming tubulose.And, about forming the structure that becomes thin more near each hole more in the fluid storage portion 1～15,17, then be easier to whole liquid transportings, and then be easy to make its fully reaction in reactive tank 30 to reactive tank 30.And, about in the chamber owing to the throughput branch of liquid forms the piped parts that parts of fine forms dual tapered of accommodating than liquid, can prevent that the liquid of accommodating from flowing out when tube 50 circulations or during foaming, thereby be easy to make the liquid of accommodating reactive tank 30 in, to react fully.And,, then can accommodate liquid effectively about forming the many more long more piped parts of the amount of liquid of accommodating in the chamber.And tube 50 formations comprise the first layer 50a and second layer 50b, because the chamber is formed at any one deck among the first layer 50a and the second layer 50b or crosses over the first layer 50a and second layer 50b formation, can form more chamber thus.And, from the bearing surface 52a that is formed with each hole away from second layer 50b on, for the more big volumetrical chamber of the amount that forms the liquid of accommodating than the first layer 50a, because the quantity away from bearing surface 52a chamber is few more more, can reduce the quantity of vertical stream in connection chamber and hole thus, and then be easy to make tube 50.And, flow out limiting valve 16e, deceleration column 16 and flow into limiting valve 16d owing to be provided with, therefore need not to switch being connected of reactive tank 30 and opening 16a, can be adsorbed on the column casing of deceleration column 16 at the resultant that reactive tank 30 internal reactions generate, thereby after in this deceleration column 16, making the liquid communication in the fluid storage portion 17, this liquid is turned back in the reactive tank 30.And,, will remain on reliably in the exhausted bath box 28 from the waste liquid that reactive tank 30 temporarily is contained in exhausted bath box 28 thus because harvesting has the water-absorbent porous material that absorbs waste liquid in the inside of exhausted bath box 28.And because extraneous air throughput 21 has hydrophobic porous material, therefore the parts about being connected with extraneous air throughput 21 in the chamber can not make the liquid of accommodating in the chamber flow out to the outside by extraneous air throughput 21.And, about the parts that are connected with extraneous air throughput 21 in the chamber, can not make the liquid of accommodating in the chamber pass through extraneous air throughput 21 to the effusive while of outside, can further reduce the quantity of the first ventilating pit 21a.And, because reactive tank 30 is more near the thin more piped parts in hole, therefore be easy to make whole liquid between chamber and reactive tank 30, circulate by the hole, be easy to make the interior liquid foam of reactive tank 30 by the extraneous air that leads to by orifice flow.
And CPU42 carries out predefined cDNA complex sign handling procedure, and controls revolution mechanism 32 and pump 34 can be transferred liquid and make its reaction thus based on the step of cDNA complex sign handling procedure.Thus, with the user liquid is supplied to that the temperature with regulation are compared its reaction specified time in the reactive tank 30, can carry out each treatment process reliably, therefore inhibited reaction result's error more reliably with predetermined conditions.And, by pump 34 reduce air pressure in the reactive tanks 30 from fluid storage portion 1～15,17 to reactive tank 30 sucking-off liquid, air pressure in the rising reactive tank 30 is released waste liquid from reactive tank 30 to exhausted bath box 28, can transfer liquid by this fairly simple formation of pressure that changes in the reactive tank 30 thus.And, make tube 50 rotate and switch being connected of each hole and reactive tank 30 around turning axle, compare with the rotating disk 82 that send vapor pipe 34a that has been rotatably connected thus, be easier to make its rotation.And in the time of the rotating disk 82 of rotatable and bearing surface 52a butt and tube 50, by this rotating disk 82 is installed tube 50 to the press section 84 of the bearing surface 52a application of force, reactive tank 30 is connected with rotating disk 82, thus, tube 50 can be installed under the state of the switching that can be relatively easy to carry out chamber and reactive tank 30.And, owing to have tube Peltier's element 38a and reactive tank Peltier's element 36a, therefore can respectively the tube of installing 50 be adjusted to the nonreactive temperature of the liquid of accommodating, or reactive tank 30 is adjusted to the temperature of suitable reaction, the Temperature Influence of the tube of not installed thus 50 can make liquid fully reaction in reactive tank 30 of accommodating.
And the present invention is not subjected to any qualification of above-mentioned first embodiment, as long as belong to technical scope of the present invention, certainly implements in every way.
For example, in the above-described first embodiment,, also can be used as the device that other chemical reactions are used though tube 50 and reaction unit 90 are as the device that obtains mark cDNA for the discovery amount that detects mRNA from mRNA.That is, tube 50 is accommodated the liquid that other chemical reactions are used, and reaction unit 90 utilizes this tube 50 to carry out other reactions and gets final product.Under this kind situation, liquid reaction fully in reactive tank 30 that fluid storage portion 1～15,17 is accommodated.At this moment, the fluid storage portion 1～15,17 that forms in tube 50 can form the volume of the liquid that can accommodate other chemical reaction consumptions, and reaction unit 90 can be stored handling procedure that other chemical reactions use and carry out other chemical reactions according to this program.And, this point in second and third embodiment described later too.For example, tube accommodate be used to generate SNP detect with and the genomic dna of being labeled, reaction unit is that SNP variation handling procedure is carried out chemical reaction according to the program that the genomic dna of this being labeled generates usefulness.Figure 11 has accommodated the explanatory view of tube 150 that this SNP detects the liquid of usefulness, and Figure 12 is until the explanatory view with the step of genomic dna amplification and being labeled.As shown in figure 11, tube 150 is except tube 50 fluid storage portion 14,15,17 and deceleration column 16, connected stream, hole, the outflow limiting valve of not stated, the point that flows into limiting valve, exhausted bath box 28, and the volume of corresponding liquid of accommodating has designed outside these points of shape of fluid storage portion 1～13, the same with tube 50, thus identical integrant is added prosign and omitted detailed explanation.And reaction unit uses the device of the formation the same with the reaction unit stated 90 except storage SNP variation handling procedure in flash rom 43, and in the following description, use and reaction unit 90 the same symbols describe.
When using this tube 150 to detect SNP, the user at first will put into reactive tank 30 as the genomic dna of sample, and with the same mounting of above-mentioned first embodiment in universal stage 38.Secondly, press not shown start button.So the CPU42 of controller 40 reads and carries out the SNP variation handling procedure that is stored in flash rom 43.When carrying out this program, CPU42 at first remains on specified temperature (for example 20 ℃) with Peltier's element 38a with tube 50 by tube.Secondly, reactive tank 30 is communicated with opening 1a～3a, and sucking-off is contained in liquid (the stage S400～S420) of fluid storage portion 1～3 successively.Then, with the temperature in the reactive tank 30 remain on 95 ℃ and foam made its reaction in 15 minutes after, repeatedly 40 circulations make its carry out with the temperature in the reactive tank 30 remain on 95 ℃ and 30 seconds of foaming, 80 ℃ of temperature down 1 second of foaming, in 68 ℃ of circulations (stage S430) of 6 minutes of foaming down of temperature.Then, with the temperature in the reactive tank 30 remain on 10 ℃ 1 minute, make reaction stop (stage S440).Then, when making pump 34 running the temperature in the reactive tank 30 are remained on 80 ℃ 30 minutes, make the mixing solutionss that are sucked in the reactive tank 30 concentrate (stage S450), obtain amplification DNA dry substance thus.Then, reactive tank 30 is communicated with opening 4a～7a, and sucking-off is contained in liquid (the stage S460～S490) of fluid storage portion 4～7 successively.Then, make reactive tank 30 interior temperature remain on 95 ℃ of foaming 5 minutes, foam 1 minute down for 50 ℃, foam down for 58 ℃ in temperature and reacted (stage S500) in 60 minutes in temperature.Then, with the temperature in the reactive tank 30 remain on 10 ℃ 1 minute, make reaction stop (stage S510).Then, when making pump 34 running the temperature in the reactive tank 30 are remained on 80 ℃ 30 minutes, make the mixing solutionss that are sucked in the reactive tank 30 concentrate (stage S520), obtain to connect (ligation) DNA dry substance thus.Then, reactive tank 30 is communicated with opening 8a～13a, and sucking-off is contained in liquid (the stage S530～S580) of fluid storage portion 8～13 successively.Then, with the temperature in the reactive tank 30 remain on 95 ℃ and foam made its reaction in 1 minute after, repeatedly 25 circulations make its carry out with the temperature in the reactive tank 30 remain on 95 ℃ and the 30 seconds kinds of foaming, in the foaming 6 minutes down of 55 ℃ of temperature, in the circulations (stage S590) in 30 seconds of foaming down of 72 ℃ of temperature.Then, with the temperature in the reactive tank 30 remain on 10 ℃ 1 minute, make reaction stop (stage S600).Then, when making pump 34 running the temperature in the reactive tank 30 are remained on 80 ℃ 15 minutes, make the mixing solutionss that are sucked in the reactive tank 30 concentrate (stage S610), obtain the joint DNA of being labeled thus.Use the joint DNA of being labeled that so obtains, for example, the marker DNA that obtains is applied to common DNA chip.Through hybridization, washing procedure, with scanner be taken into the fluorescence that from the point of DNA chip obtains, and strength of signal carried out digitized processing thereafter.By double-colored signal ratio, distinguish SNP.
In the first above-mentioned embodiment, for example, even the less chamber of amount of liquid that fluid storage portion 1,2,8,9 etc. accommodates, also be formed on the outer circumferential side of the first layer 50a of circular plate shape, but also can be formed at interior all sides in the chamber that the amount of liquid of accommodating is less, that is, the many more chambeies of amount of liquid of accommodating are formed at more outer circumferential side more.Like this, be easy to form the more chamber of amount of the liquid of accommodating.And, this point in second and third embodiment described later too.
In the first above-mentioned embodiment,, also can form non-circular plate shape such as rectangular shape though tube 50 forms circular plate shape.And, this point in second and third embodiment described later too.
In the first above-mentioned embodiment,,, then be not limited to rotation and move as long as reactive tank 30 and tube 50 are relatively moved though make tube 50 rotations that reactive tank 30 and tube 50 are relatively moved.For example, reactive tank 30 and tube 50 straight lines are moved.At this moment, on tube 50, can have and switch a plurality of holes that set with the linearity that is connected of reactive tank 30.And, this point in second and third embodiment described later too.
In the first above-mentioned embodiment,, also can not have these Peltier's elements 38a and 36a though reaction unit 90 has tube with Peltier's element 38a and reactive tank Peltier's element 36a.Even under this kind situation, also can make the liquid reaction fully in reactive tank 30 that is contained in fluid storage portion 1～15,17.And, this point in second and third embodiment described later too.
In the first above-mentioned embodiment,, also can not have deceleration column 16 though tube 50 has deceleration column 16.And,, also can not have these valves though have the limiting valve 16d of inflow and flow out limiting valve 16e.And, this point in second and third embodiment described later too.
In the first above-mentioned embodiment,, thereby also can make mobile reactive tank 30 and tins 50 is relatively moved the hole that switching is connected with reactive tank 30 of reactive tank 30 rotations though thereby reaction unit 90 switches the hole is connected with reactive tank 30 by making tube 50 rotations that reactive tank 30 is relatively moved with tube 50.And, this point in second and third embodiment described later too.
In the first above-mentioned embodiment, though each hole set into the round shape of the spigot shaft coaxle of tube 50, the axle that each hole is adapted to the circle of round shape is not limited to the central shaft of tube 50.For example, the axle that will be different from its central shaft at tube 50 is as 30 rotations of turning axle relative response groove, and carry out under the situation of switching in the hole that is connected with reactive tank 30, the axle of the circle that this turning axle can be set as each hole, under the situation that reactive tank 30 moves with respect to tube 50 rotation, the axle of the circle that the turning axle that these reactive tank 30 rotations can be moved sets as each hole.And, this point in second and third embodiment described later too.
In the first above-mentioned embodiment, though externally among air flow path 21d, the 21e harvesting hydrophobic porous material is arranged, can not receive and keep yet.And,, can not receive and keep though harvesting has the water-absorbent porous material in exhausted bath box 28 yet.And, this point in second embodiment described later too.
In the first above-mentioned embodiment,, also can there be liquid to flow in the chamber beyond exhausted bath box 28 though only exhausted bath box 28 has liquid to flow into.For example, in the cDNA complex sign handling procedure of having stated, in not accommodating the fluid storage portion 18 of liquid, can there be liquid to flow into.At this moment, can chamber after liquid is flowed into this liquid transporting in, can have the water-absorbent porous materials of absorption liquid style such as sponge etc.And, this point in second embodiment described later too.
In the first above-mentioned embodiment, though the outer circumferential side of fluid storage portion 1～13,17,18 connects by extraneous air circulation flow path 21d, the outer circumferential side of fluid storage portion 1～13,17,18 can not connect by extraneous air circulation flow path 21d yet.And, this point in second and third embodiment described later too.
In the first above-mentioned embodiment, though pump 34 is a surge pump, thereby also can be by smooth out with the fingers with roller pipe to the so-called hose pump of the place ahead applied pressure of this pipe coupling.And, and, this point in second embodiment described later too.This kind situation, when adopting stepper-motor as the operation of smoothing out with the fingers pipe, the air capacity to carrying that can be more careful is controlled.Particularly adopted the hose pump and the intercepting basin of this stepper-motor, need not to use vacuum breaker can make the elution damping fluid to column casing circulation and be housed in (with reference to Figure 13) in the reactive tank 30 by use.Figure 13 is the explanatory view of the annexation of column casing 106 when using the hose pump that has adopted stepper-motor as pump 34 and reactive tank 30, exhausted bath box 28.As shown in the figure, column casing 106 is connected with opening 102 by stream 104, and is connected with exhausted bath box 28 by stream 108, intercepting basin 110 and stream 112.And the elution damping fluid is accommodated in the fluid storage portion 116 that is connected with other openings 114.Make the liquid that is contained in reactive tank 30 when column casing 106 circulations, after making opening 102 and reactive tank 30 is communicated with by rotating disk 82, make the hose pump running, make this liquid to column casing 106 circulations thus, and accommodate waste liquid to exhausted bath box 28.Make the elution damping fluid when column casing 106 circulations, temporarily opening 114 is communicated with reactive tank 30, make the hose pump running, thus the elution damping fluid in the fluid storage portion 116 is contained in the reactive tank 30 by rotating disk 82.And, opening 102 is communicated with reactive tank 30, the running hose pump makes the elution damping fluid to column casing 106 circulation and be accumulated in the intercepting basin 110.That is,, thus the elution damping fluid is not put into exhausted bath box 28 and is accumulated in the intercepting basin 110 by the air capacity that careful control is carried by hose pump.And, when hose pump running is attracted, accumulate in the elution damping fluid of intercepting basin 110 once more to column casing 106 circulations and be contained in reactive tank 30.So, need not to use vacuum breaker, can be after waste liquid be contained in exhausted bath box 28, make the elution damping fluid to column casing 106 circulations and be contained in reactive tank 30.And owing to be filled with water-absorbent porous material such as sponge in exhausted bath box 28, so waste liquid can not flow backwards and pollutes the elution damping fluid.
In the first above-mentioned embodiment, though be tube 50 and reaction unit 90, but both can be the reaction member that constitutes by tube 50 and rotating disk 82 and reactive tank 30, it also can be the corresponding type reaction unit in unit that utilizes this reaction member to react, as the corresponding type reaction unit in unit, for example can be the reaction unit that removes the formation of rotating disk 82 and reactive tank 30 from above-mentioned reaction unit 90.Even also can obtain and the same effect of above-mentioned first embodiment in this case.And, this point in second and third embodiment described later too.
In the first above-mentioned embodiment, though reaction unit 90 is carried out the cDNA complex sign handling procedure of having stated and liquid is reacted by controller 40, but also can have switch that makes pump 34 runnings and the switch that makes the rotating disk rotation, manually carry out a series of reaction by the user.Under this kind situation, can make the liquid reaction fully in reactive tank 30 that is contained in fluid storage portion 1～15,17.And, this point in second and third embodiment described later too.
In the first above-mentioned embodiment,, also can on the first layer 50a and second layer 50b, form the chamber according to the degree of temperature regulation necessity though on the first layer 50a and second layer 50b, formed the chamber according to the amount of accommodating liquid.That is, second layer 50b compares with the first layer 50a and keeps the universal stage 38 of tube with Peltier's element 38a in more approaching when tube 50 is installed on reaction unit 90, therefore compares with the first layer 50a and carries out more reliable temperature regulation.Thus, also can on the more approaching second layer 50b that carries out thermoregulator universal stage 38, form and accommodate the chamber that more needs thermoregulator liquid.At this moment, also can be approaching more the center of tube 50 form more and accommodate the chamber that more needs thermoregulator liquid.Thereby this is owing to be easy to carry out temperature regulation near the center from extraneous air is far away more more.At this moment, also can form empty chamber at the outer circumferential side of tube 50.Suppress the influence of extraneous air by the air in the chamber of sky, thereby can be easier to carry out temperature regulation.Here, empty chamber both can be to form adiabatic special-purpose chamber, also can be to accommodate the preceding exhausted bath box of waste liquid.
In the first above-mentioned embodiment,, also can be tube with a plurality of exhausted bath box though have the tube 50 of an exhausted bath box 28.For example, also can be tube, thereby can will use in the waste liquid that produces in the series reaction step of this tube that the back wants that the waste liquid that uses is contained in the exhausted bath box that is different from other waste liquids with a plurality of exhausted bath box.At this moment, also can want that exhausted bath box and other exhausted bath box of the waste liquid that uses are connected to other openings with accommodating the back.Perhaps, accommodate the back and want that the exhausted bath box of the waste liquid that uses and the exhausted bath box of accommodating other waste liquids also can use the common opening, use the switching valve that switches stream to switch the exhausted bath box of accommodating.
In the first above-mentioned embodiment, the column casing of deceleration column 16 can be filled by step shown below.Figure 14 is the explanatory view of the filling step of column casing.In this Figure 14, use the longitudinal section of column casing 16 to describe.At first, the side seal material 62 (for example, special teflon pipe, heat-shrinkable tube) with tubular embeds column casing (with reference to Figure 14 (a)).By doing like this, the liquid that flows into column casing is flowed out from the side of column casing.Then, will be pressed into the column casing that side seal material 62 forms as one in the hole of the deceleration column 16 that is formed at the first layer 50a and second layer 50b (with reference to Figure 14 (b)).Then, be that the discoid lid 64 in the rubber outside diameter aperture that is slightly larger than second layer 50b is keeping from the below embedding (Figure 14 (c)) with the gap of column casing bottom surface with material, finished the deceleration column 16 (with reference to Figure 14 (d)) of having filled column casing thus.So, owing to column casing is embedded, therefore can fill column casing with better simply operation by pressure.
In the first above-mentioned embodiment,, also can constitute by two fixing parts as shown in figure 15 though reactive tank fixed part 36 is made of parts with door of opening to the front.Figure 15 is the sectional view of the formation of other reactive tank fixed parts 136 of explanation.As shown in the figure, reactive tank fixed part 136 is made of upside fixing part 136a and downside fixing part 136b.This upside fixing part 136a is connected by axis of rocking A with downside fixing part 136b, and constitutes and can shake along C direction among the figure and D direction.On upside fixing part 136a, be connected with and send vapor pipe 34a.On downside fixing part 136b, be provided with the hole of inlaying reactive tank 30, and the built-in groove that responds Peltier's element 36a.The tube 50 of this kind situation and the installation steps of reactive tank 30 are as follows: the tube 50 of the rotating disk 82 that has been slidingly installed at first from the side, and be disposed on the universal stage 38.And, open upside fixing part 136a along the C direction, reactive tank 30 is embedded in after downside fixing part 136b goes up, close upside fixing part 136a along the D direction.After closing upside fixing part 136a along the D direction, with the state of pushing upside fixing part 136a is fixed in downside fixing part 136b by not shown hook.So the upside of reactive tank 30 and downside are all by O shape circle 136c, 54b sealing.So, by closing this operation of upside fixing part 136a, can be by the upside and the downside of O shape circle 136c, 54b sealed reaction groove 30.
In the first above-mentioned embodiment, also can be as shown in figure 16, will be used for application cylinder 50 position of rotation the bottom surface be recessed to form groove for the T word shape that constitutes by diametric groove 251 and perpendicular radial direction groove 252.In this case, rotating disk 38 top also corresponding to this bottom shape forms the projection of the T word shape that is made of diametric projection and perpendicular radial direction projection.
In the first above-mentioned embodiment, by motor 37 make universal stage 38 along opposite spin and with reactive tank 30 when among opening 1a～16a, the 18a any one is connected, also can with universal stage 38 opposite spins behind the position of the opening that surpasses this connection, universal stage 38 positive dirctions be rotated to the position of the opening of this connection.For example, before the processing of implementation stage S300, temporarily with universal stage 38 opposite spins until the opening 15a that will the connect opening 14a side of passing by.And, universal stage 38 positive dirctions are rotated to opening 15a and reactive tank 30 opposed positions.By doing like this, can suppress deterioration by the control accuracy of the position of rotation of the loosening universal stage 38 that causes of the gear mechanism of motor 37.
In the first above-mentioned embodiment, as shown in figure 15, also can be communicated with this breather valve that send vapor pipe 34a and extraneous air 134 sending on the vapor pipe 34a to be provided with, when operating pumps 34, with after opening breather valve 134 and making pump 34 running block gear mechanism loosening of motor of driving pump 34, that closes that thereby breather valve 134 carries out aim parameter send the deflated mode, carries out the operation of pump 34.Like this, can not produce the loosening error that causes of gear mechanism of the motor of driving pump 34, thereby send exhaust.
In the first above-mentioned embodiment, as shown in figure 15, also can be communicated with this breather valve that send vapor pipe 34a and extraneous air 134 sending on the vapor pipe 34a to be provided with, will be from after the fluid storage of sucking-offs in the tube 50 be reactive tank 30 in, with the 134 opening specified times of breather valve.Like this, can will be sucked in the reactive tank 30 and be raised to the bottom that the liquid of reactive tank 30 tops is fallen reactive tank 30.
In the first above-mentioned embodiment, make liquid when reactive tank 30 internal reactions, by extraneous air is continued to be incorporated in the reactive tank 30, and it is almost being reacted under the non-pressurized pressure, but also can make its add depress or reduce pressure under react.For example, also the opening that Kong Wei opens can be set on tube 50, depress at plus-minus and make its when reaction, with behind reactive tank 30 and the opposed mode rotating cylinder 50 of this opening, by making pump 34 runnings, pressurized state and the decompression state in the groove 30 of making a response thus also makes its reaction.Situation under the decompression further uses Peltier's element 36a to reactive tank 30 internal heating by reactive tank, can make the liquid concentration in the reactive tank 30 thus more rapidly.Add situation about depressing, can carry out compressive reaction.
In the first above-mentioned embodiment, as shown in figure 15, also can be provided with and to make magnet 70 motor rotating 72, and in reactive tank 30, put into the rotor 74 that contains magnet, make magnet 70 rotations make rotor 74 rotations thus by motor 72 on the next door of reactive tank fixed part 36.By doing like this, the liquid in can stirred tank 30 also makes its reaction.Here, magnet and magnet 70 as rotor 74 is contained can use neodium magnet.And, according to configuration shown in Figure 15, because rotor 74 is disposed at the top than magnet 70, so rotor 74 oblique rotations during magnet 70 rotation.And the magnet 70 that is disposed at the below than rotor 74 pulls rotor 74 downwards, can prevent the come-up of the rotor 74 that the surface tension of the liquid of accommodating causes thus.And, even rotor 74 come-ups also can stir the liquid of accommodating.
Second embodiment
Figure 17 is the explanatory view of configuration of stream etc. that is arranged at fluid storage portion, liquid and the gas of accommodating liquid of tube 250 the first layer 250a of present embodiment and second layer 250b, and Figure 18 is the explanatory view of receiving and keeping in the position of the porous material of tube 250 inside.And, tube 50 uses tube 250, will use this 250 DNA amplification handling procedure that carries out and dna fragmentation program the reaction unit of present embodiment except that replacing, the dna marker handling procedure is stored in the flash rom 43, is the formation the same with the reaction unit 90 of first embodiment.Thus, describe as reaction unit 90, and to the additional same-sign detailed of same integrant.And, though be described in detail later, but tube 250 has opening 201a～206a, 208a～211a, 213a～217a, 219a, the 220a that can be connected with reactive tank 30 respectively, with accepting hole 207a, 212a, 218a, but for convenience of description, it is referred to as " hole ", and fluid storage portion 201～205,207～210,212～216,218～220 and the exhausted bath box 228 that can accommodate liquid are referred to as " chamber ".And in the present embodiment, reaction unit 90 describes as the device to DNA amplification.
Tube 250 is parts that material is formed by cyclenes copolymer, as shown in figure 17, have: form circular plate shape the first layer 250a, second layer 250b, be formed at the top of the first layer and be used to make this 250 with the same not shown guide part of the guide part of having stated 52 of central shaft rotation.This first layer 250a and second layer 250b have a plurality of chambeies respectively.As shown in figure 17, this tube 250 has: a plurality of fluid storage portion 201～205,207～210,212～216,218～220 that can accommodate liquid with the specified volume of the liquid of accommodating decision; By making tube 250 rotations move and switch, thereby be equipped on opening 201a～206a, 208a～211a, 213a～217a, 219a, the 220a of any one the regulation link position that is connected with reactive tank 30 that makes in this fluid storage portion 201～205,207～210,212～216,218～220 respectively; Be communicated with fluid storage portion 201～205,207～210,212～216,218～220 and extraneous air, and to fluid storage portion 201～205,207～210,212～216,218～220 in, introduce extraneous air or the extraneous air throughput 221 of discharge gas in the fluid storage portion 201～205,207～210,212～216,218～220; Be used for accommodating accepting hole 207a, 212a, the 218a of liquid to fluid storage portion 207,212,218; Can accommodate the exhausted bath box 228 of the waste liquid of reactive tank 30 supplies; Be arranged between exhausted bath box 228 and each opening 206a, 211a, the 217a, allow from reactive tank 30 to the circulation of the waste liquid of exhausted bath box 228 and stop from outflow limiting valve 206e, 211e, the 217e of this exhausted bath box 228 to the waste liquid circulation of reactive tank 30; Can be adsorbed on the deceleration column 206,211,217 of the production thing of reactive tank 30 reactions; Be arranged between fluid storage portion 207,212,218 and the deceleration column 206,211,217, allow the liquid communication to deceleration column 206,211,217, and stop from inflow limiting valve 206d, 211d, the 217d of deceleration column 206,211,217 to the liquid communication of fluid storage portion 207,212,218 from fluid storage portion 207,212,218.
Form the parts of fine of accommodating than liquid by the throughput branch with liquid, fluid storage portion 201～205,207～210,212～216,218～220th thus, form the piped space of dual tapered.For example, shown in the fluid storage portion 204 relative with fluid storage portion 201, the many more longer pipe shapes that form more of the amount of the liquid that this fluid storage portion 201～205,207～210,212～216,218～220 accommodates.And fluid storage portion 202～205,207～210,212,213,215,218,219 forms the tubulose of the circuitous shape that increases from the circuitous amplitude of tube interior all side direction outer circumferential sides of 250 of circular plate shape, circuitous tubular become thin more near each hole more.And, from the not shown bearing surface isolating second layer 250b the same with the bearing surface 52a that has stated of interior all sides of the guide part of the first layer 250a that is formed with each hole, for the amount that makes the liquid of accommodating more than other chambeies that are formed at the first layer 250a, and be formed with big volumetrical fluid storage portion 204,205,216.And, though each fluid storage portion 201～203,207～210,212～215,218～220 is communicated with by extraneous air circulation flow path 221d, but as described later, because harvesting has hydrophobic porous material, the liquid that can not that therefore is contained in this fluid storage portion 201～203,207～210,212～215,218～220 mixes by extraneous air circulation flow path 221d.
Extraneous air throughput 221 has: the extraneous air circulation flow path 221d that is connected with an end of the outer circumferential side of the fluid storage portion 201～203,207～210,212～215,218～220 of the first layer 250a; The extraneous air circulation flow path 221e that is connected with an end of the outer circumferential side of the fluid storage portion 204,205,216 of second layer 250b; The first ventilating pit 221a that these extraneous air circulation flow paths 221d, 221e are communicated with extraneous air; Receive and keep the hydrophobic porous material in extraneous air circulation flow path 221d, 221e.And, in Figure 18, use grid representation extraneous air throughput 221.This hydrophobic porous material can not be by liquid but can be passed through extraneous air,, for example uses special teflon porous material (day eastern electrician company make TEMISH) here.And, the end of extraneous air circulation flow path 221e is connected with fluid storage portion 204,205,216, the other end is connected with the first ventilating pit 221a to vertical stream that the stream position of the first layer 250a 222b, 223b form by the stream position 222c from second layer 250b, and is communicated with extraneous air by this first ventilating pit 221a.
Opening 201a～206a, 208a～211a, 213a～217a, 219a, 220a are communicated with fluid storage portion 201～205,207～210,212～216,218～220 respectively, be used for from the hole of fluid storage portion 201～205,207～210,212～216,218～220 feeding liquids, and be set at top interior all sides (bearing surface) of the first layer 250a.This opening 201a～206a, 208a～211a, 213a～217a, 219a, 220a are provided in respectively and the turning axles of tube 250 by rotating mechanism 32 rotations promptly on the same plane of the round shape of the spigot shaft coaxle of tube 250.And, differential pressure by to the liquid effects that is contained in the fluid storage portion 201～205,207～210,212～216,218～220 that is connected with these openings can supply to reactive tank 30 with the liquid that is contained in these fluid storage portions 201～205,207～210,212～216,218～220.Here, opening 204a, 205a, 216a be by being formed at the stream longitudinally that connects the first layer 250a and second layer 250b, and the end with the fluid storage portion 204,205,216 of second layer 250b is that link position 204c, 205c, 216c link to each other respectively.
Accepting hole 207a, 212a, 218a are communicated with fluid storage portion 207,212,218, the hole of using when accommodating from liquid that the outside is transferred, and be set at top interior all sides (bearing surface) of the first layer 250a.This accepting hole 207a, 212a, 218a are equipped on the circle that is equipped with opening 201a～206a, 208a～211a, 213a～217a, 219a, 220a.Here, fluid storage portion 207,212,218 by from the stream position 224b～226b of an end of this fluid storage portion 207,212,218 to each stream longitudinally of the position of second layer 250b 224c～226c with flow into limiting valve 206d, 211d, 217d are connected respectively.And the other end of this fluid storage portion 207,212,218 is connected with extraneous air circulation flow path 221d, though be described in detail later, harvesting has the hydrophobic porous material that does not pass through liquid in this extraneous air circulation flow path 221d.Thus, all can not in fluid storage portion 207,212,218, accommodate liquid, be provided with this accepting hole 207a, 212a, 218a in the mode that can in fluid storage portion 207,212,218, accommodate liquid thus from any end.And, in other fluid storage portion 201～205,208～210,213～216,219,220, can accommodate from opening 201a～205a, 208a～210a, 213a～216a, 219a, 220a.
Deceleration column 206,211,217 is set between opening 206a, 211a, 217a and outflow limiting valve 206e, 211e, the 217e, contains column casing.Here, use ceramic column casing (for example, silica gel etc.) as column casing.And, move to reactive tank 30 by opening 206a, 211a, 217a owing to be contained in the liquid of fluid storage portion 207,212,218 respectively, therefore opening 206a, the 211a that is communicated with deceleration column 206,211,217,217a is communicated with and is set at the first layer 250a with this fluid storage portion 207,212,218 top interior all sides (bearing surface).
Outflow limiting valve 206e, 211e, 217e and inflow limiting valve 206d, 211d, 217d are so-called vacuum breaker.For example, can use miniature check valve shown in Figure 5 160.Miniature check valve 160 for example is made of with the cover body 162 of fixing this valve body 161 circular valve 161 of silicon rubber system.This valve body 161 is provided with circular-arc slit, and it is the structure that axle shakes that the baffle plate 163 of this slit inboard forms with axis of rocking 164.Cover body 162 has the valve body clamping part 162a of clamping valve body 161.This valve body clamping part 162a forms diameter less than baffle plate 163 with the diameter in the hole in 161 left sides of valve body among the figure, and the diameter in the hole on valve body 161 right sides is formed greater than the diameter of baffle plate 163 and less than the diameter of valve body 161.Thus, when fluid A direction in figure flows, be that axle bumper plate 163 tilts to occur the slit to the A direction with the axis of rocking 164 of valve body 161, fluid from this slit by mobile.On the other hand, when fluid was desired in figure the B direction and flowed, though be that axle bumper plate 163 is desired to tilt to the B direction with the axis of rocking 164 of valve body 161, baffle plate 163 did not have the slit fluid mobile with the end face butt of cover body 162.This miniature check valve 160 is that the B direction is this reciprocal mode from the direction of deceleration column 206 towards stream position 227c as flowing out limiting valve 206e with the A direction among Fig. 5, is arranged at tube 250.And, as flowing into limiting valve 206d, be that the B direction is this reciprocal mode from the direction of position 224c towards deceleration column 206 with the A direction among Fig. 5, be arranged at tube 250.About flowing into limiting valve 211d, 217d, outflow limiting valve 211e, 217e respectively with respect to the same configuration of deceleration column 211,217 quilts.And miniature check valve 160 uses the parts in the gloomy research department exploitation of the tinkling of pieces of jade of engineering section of system of engineering portion of Okayama Univ..
As shown in figure 17, exhausted bath box 228 is the spaces that are provided with in the mode around outermost week Monday of tube 250, and forms a space crossing over the first layer 250a and second layer 250b.In the inside of this exhausted bath box 228, for the waste liquid that will temporarily flow into remains in the exhausted bath box 228 reliably, harvesting has the water-absorbent porous material of for example sponge that absorbs waste liquid etc.This water-absorbent porous material is along the outer circumferential side one week configuration (with reference to the oblique line position of Figure 18) of tube 250.And, the stream that this exhausted bath box 228 forms by the position 227c from the stream of position 227d all sides in being arranged at of the stream end that is connected with this exhausted bath box 228 and from this position 227b to vertical stream that the position of the stream of second layer 250b 227c forms, be connected in deceleration column 206,211,217.That is, from deceleration column 206,211,217 with flow out the fluid that passes through limiting valve 206e, 211e, the 217e, accommodated in this exhausted bath box 228.And, on the first layer 250a of the position of exhausted bath box 228, be provided with the second ventilating pit 228a that is communicated with extraneous air.
In the reaction unit 90 that so constitutes, the liquid that will contain the reaction institute with medicament etc. of regulation is contained in the fluid storage portion of tube 250 respectively with desirable amount, and rotate successively by motor 37 and to move tube 250 when changing the link position in hole, by pump 34 from the fluid storage portion 201～205,207～210,212～216,218～220 that connects to of the reaction of reactive tank 30 feeding liquids to stipulate at reactive tank 30, or transfer these reacted liquid to exhausted bath box 228 from reactive tank 30, form the formation of practicable various processing thus.Special when carrying out the purification of resultant of reaction, by making column casing absorption reaction resultant and the liquid of not wanting accommodated to exhausted bath box 228, from fluid storage portion 207,212,218 make liquid from this column casing by and supply to reactive tank 30 and carry out.And, this reaction unit 90 and first embodiment of having stated shown in Figure 7 the same, because reactive tank 30 is arranged at the outside of tube 250, therefore the temperature variation of reactive tank 30 is difficult to transmit to tube 250, reactive tank 30 and tube 250 can be remained on different separately temperature (for example temperature of reaction and preservation with temperature etc.) thus.
Here, to the operation of the reaction unit 90 of present embodiment, especially to from describing as the genomic dna amplification of sample and the operation of mark.The explanatory view of the step when Figure 19 is amplification DNA, Figure 20 are to be the explanatory view that the DNA of fragmentation is carried out the step of mark with the amplification dna fragmentationization and explanatory view, the Figure 21 of step that will extract the dna fragmentationization of target part.In these figure, the fluid storage portion of the expression tube 250 of pattern and exhausted bath box 228, the opening, the reactive tank 30 that connect.Among these figure, in fluid storage portion 201～205,207～210,212～216,218 and exhausted bath box 228, record kind and the amount of accommodating and described each symbol that constitutes of Figure 17 of liquid.Inner state of not accommodating liquid is represented in the chamber on empty hurdle.And, about reactive tank 30, contain the situation ellipse representation of liquid in the reactive tank 30, the liquid of accommodating to be carried out situation about handling represent with rectangle, the situations that what is not all accommodated in the reactive tank 30 are with the ellipse representation on empty hurdle.And, arrow express liquid among the figure or gas flow direction.And, for convenience of description, in the part additional phase numbering of reactive tank 30.
At first, using Figure 19 that the amplification of DNA is handled describes.The user will put into reactive tank 30 as the DNA of the sample of thinking amplification and be connected with rotating disk 82, and with the same shown in Fig. 2 (b) of first embodiment of having stated, it be embedded tube 250, thus as reaction member.And, open the lateral not shown door that is arranged at reactive tank fixed part 36, the top of reactive tank 30 is communicated with sending vapor pipe 34a, rotating disk 82 with the portion of being pressed 84 downwards the mode of the application of force slide from the side, thus with this reaction member mounting in universal stage 38.At this moment, because press section 84 materials are special teflon, by deflection, on a plurality of protuberances on be arranged at universal stage 38, the mode that embeds a plurality of recesses that are formed at 250 bottom surface is carried out mounting, and with press section 84 downwards the state of the application of force install.Secondly, press not shown start button.So the CPU42 of controller 40 reads and carries out the DNA amplification handling procedure that is stored in flash rom 43.When carrying out this program, CPU42 at first remains on specified temperature (for example 20 ℃) with Peltier's element 38a with tube 250 by tube.Secondly, make tube 250 rotations that opening 201a is communicated with reactive tank 30 by drive-motor 37, make the air pressure in the pump 34 running reduction reactive tanks 30, the liquid that will be contained in fluid storage portion 201 thus is sucked in the reactive tank 30 (stage S700).
Secondly, when with Peltier's element 36a the temperature in the reactive tank 30 being remained on 4 ℃ by reactive tank, make the air pressure in pump 34 runnings the continuing reduction reactive tank 30, continue to introduce extraneous air in the reactive tank 30 by extraneous air throughput 221, fluid storage portion 201, opening 201a, circulation flow path 82a thus, made the liquid foam that is contained in the reactive tank 30 thus 10 minutes, thereby make the dna solutions reactions (stage S710) in the reactive tank 30.So, continue to introduce extraneous air by extraneous air throughput 221, any one fluid storage portion 201～205,207～210,212～216,218, the opening that is arranged at an end of this fluid storage portion 201～205,207～210,212～216,218, circulation flow path 82a, can make the liquid foam in the reactive tank 30 thus.And, below, the situation of so-called " foaming " is all carried out this operation.And, as shown in Figure 3, in reactive tank 30, be provided with air discharge duct 30a, it is fashionable from the orifice flow that connects that air pressure in reactive tank 30 reduces extraneous air, by mobile gas in this air discharge duct 30a, the liquid level that prevents the liquid of accommodating rises and to adhering to of wall etc., thereby the liquid of accommodating is reacted more fully.Get back to the explanatory view of Figure 19, then, opening 202a is communicated with reactive tank 30, and sucking-off is contained in the liquid (stage S720) of fluid storage portion 202, foaming was 30 minutes when the temperature in the reactive tank 30 was remained on 30 ℃, made the DNA reaction (stage S730) in the reactive tank 30.Opening 203a is communicated with reactive tank 30, and sucking-off is contained in the liquid (purification damping fluid) (stage S740) of fluid storage portion 203.Foaming made its reaction (stage S750) in 5 minutes when then, the temperature in the reactive tank 30 being remained on 25 ℃.Opening 204a is communicated with reactive tank 30, liquid (adsorption-buffering liquid) sucking-off that is contained in fluid storage portion 204 is mixed (stage S760).
Then, opening 206a is communicated with reactive tank 30, the connection state of pump 34 is switched, make the air pressure in pump 34 runnings and the rising reactive tank 30, make the amplification dna solution in the reactive tank 30 be passed to deceleration column 206 (stage S770).Use Figure 22 that the appearance of this moment is described.Figure 22 is the explanatory view of the stream of amplification dna solution circulation.The opening 206a of amplification dna solution by tube 250 the first layer 250a shown in Figure 22 (a) flows into and when circulating, have only DNA in the reaction mixture by the absorption of the column casings in the deceleration column 206 in deceleration column 206.And the waste liquid that passes through from column casing passes through from the outflow limiting valve 206e of the second layer 250b shown in Figure 22 (b), and passes through to vertical stream of the first layer 250a from the position 227c of stream, finally flows into exhausted bath box 228.At this moment, though be connected with fluid storage portion 207 in the stream of waste liquid circulation, owing to be arranged at the inflow limiting valve 206d in front of it, this waste liquid can influent resettlement section 207 yet.So, when the resultant that reactive tank 30 internal reactions are generated is adsorbed in any one column casing, except that the deceleration column that makes liquid communication and flow out limiting valve, opening, longitudinally the difference of stream position, with here the explanation the same, resultant is adsorbed by any one column casing, and the waste liquid that passes through from column casing flows into exhausted bath box 228.
Then, opening 205a is communicated with reactive tank 30, the connection state of pump 34 is switched, make the air pressure in pump 34 runnings and the reduction reactive tank 30, sucking-off is contained in the liquid (lavation buffer solution) (stage S780) of fluid storage portion 205.Then, opening 206a is communicated with reactive tank 30, the connection state of pump 34 is switched, make the air pressure in pump 34 runnings and the rising reactive tank 30, make reactive tank 30 interior flow of liquor lead to deceleration column 206 and thus column casing is washed (stage S790).And, the same exhausted bath box 228 that flows into of the washings after the washing with stage S730.Then, make under opening 206a and the state that reactive tank 30 is communicated with, connection state to pump 34 is switched, make the air pressure in pump 34 runnings and the reduction reactive tank 30, the liquid (elution damping fluid) that is contained in fluid storage portion 207 is circulated and to reactive tank 30 sucking-offs (stage S800) from deceleration column 206, be contained in thus in the reactive tank 30 (stage S810).Use Figure 12 that the appearance of this moment is described.Figure 12 is the explanatory view of stream that is contained in the liquid communication of fluid storage portion 207.When reducing the air pressure in the reactive tank 30, the air pressure of the stream of deceleration column 206 and waste liquid flow side thereof also reduces.At this moment, though all can be by from exhausted bath box 228 sucking-offs by flowing out limiting valve 206e waste liquid and gas, but from fluid storage portion 207, the liquid that is contained in this fluid storage portion 207 passes through to the vertical stream of the position of second layer 250b 224c from the stream position 224b of an end of fluid storage portion 207, and from deceleration column 206, pass through, finally be sucked in the reactive tank 30.At this moment, because therefore the adsorbed amplification DNA stripping in the elution damping fluid of column casing has formed the state that enters the solution that contains this amplification DNA in the reactive tank 30.And, though on the stream of interior all sides of fluid storage portion 207, be provided with accepting hole 207a, be rotated dish 82 and block, can not take place that extraneous air flows into or liquid flows out by this accepting hole 207a.So, when making the resultant stripping of any one column casing absorption and sucking-off, except that the deceleration column that makes liquid communication and flow into limiting valve, opening, longitudinally the difference of stream position, the same with explanation here is from the liquid stripping of being accommodated by the fluid storage portion that is connected with this column casing of inflow limiting valve and be sucked in the reactive tank 30.Then, when continuing the operation of pump 34 temperature in the reactive tank 30 are remained on 80 ℃ 30 minutes, make the amplification dna solutions that are sucked in the reactive tank 30 concentrate (stage S820), obtain amplification DNA thus.
Secondly, use Figure 20 that the fragmentation of DNA is described.The CPU42 of controller 40 reads and carries out the dna fragmentation handling procedure that is stored in flash rom 43.This program continues to be implemented after the DNA amplification handling procedure of having stated finishes.When carrying out this program, CPU42 at first makes opening 208a be communicated with reactive tank 30, and sucking-off is contained in the liquid (stage S900) of fluid storage portion 208.1 minute (stage S910) of foaming when secondly, the temperature in the reactive tank 30 being remained on 37 ℃.Then, opening 209a is communicated with reactive tank 30, liquid (adsorption-buffering liquid) sucking-off that fluid storage portion 209 is accommodated also mixes (stage S920).Then, opening 211a is communicated with reactive tank 30, makes the fragmentation dna solution in the reactive tank 30 be passed to deceleration column 211 (stage S930).At this moment, have only the fragmentation DNA in the reaction mixture to be adsorbed by the column casing of deceleration column 211, the waste liquid that passes through from column casing flows into exhausted bath box 228.Then, opening 210a is communicated with reactive tank 30, liquid (lavation buffer solution) sucking-off (stage S940) that fluid storage portion 210 is accommodated.Then, opening 211a is communicated with reactive tank 30, makes liquid communication in the reactive tank 30, column casing is washed (stage S950) to deceleration column 211.Then,, the liquid (elution damping fluid) that is contained in fluid receiving portion 212 is circulated in deceleration column 211 making under opening 211a and the state that reactive tank 30 is communicated with, sucking-off in reactive tank 30 (stage S960), and be contained in the reactive tank 30 (stage S970).At this moment, because therefore the adsorbed fragmentation DNA stripping in the elution damping fluid of column casing has formed the state that enters the solution that contains this fragmentation DNA in the reactive tank 30.Then, when continuing the operation of pump 34 temperature in the reactive tank 30 are remained on 80 ℃ 30 minutes, make the fragmentation dna solutions that are sucked in the reactive tank 30 concentrate (stage S980), obtain fragmentation DNA thus.
Secondly, use Figure 21 that the mark of DNA is described.The CPU42 of controller 40 reads and carries out the dna marker handling procedure that is stored in flash rom 43.This program continues to be implemented after the dna fragmentation handling procedure of having stated finishes.When carrying out this program, CPU42 at first makes opening 213a be communicated with reactive tank 30, and sucking-off is contained in the liquid (stage S1000) of fluid storage portion 213.5 minutes (stage S1010) of foaming when secondly, the temperature in the reactive tank 30 being remained on 95 ℃.3 minutes (stage S1020) of foaming when then, the temperature in the reactive tank 30 being remained on 0 ℃.Then, opening 214a is communicated with reactive tank 30, the liquid sucking-off (stage S1030) that fluid storage portion 214 is accommodated.40 minutes (stage S1040) of foaming when then, the temperature in the reactive tank 30 being remained on 37 ℃.Then, opening 215a is communicated with reactive tank 30, liquid (adsorption-buffering liquid) sucking-off that fluid storage portion 215 is accommodated also mixes (stage S1050).Then, opening 217a is communicated with reactive tank 30, makes the fragmentation dna solution in the reactive tank 30 be passed to deceleration column 217 (stage S1060).At this moment, have only the marker DNA in the reaction mixture to be adsorbed by the column casing of deceleration column 217, the waste liquid that passes through from column casing flows into exhausted bath box 228.Then, opening 216a is communicated with reactive tank 30, liquid (lavation buffer solution) sucking-off (stage S1070) that fluid storage portion 216 is accommodated.Then, opening 217a is communicated with reactive tank 30, makes liquid communication in the reactive tank 30, column casing is washed (stage S1080) to deceleration column 217.Then,, the liquid (elution damping fluid) that is contained in fluid receiving portion 218 is circulated in deceleration column 217 making under opening 217a and the state that reactive tank 30 is communicated with, sucking-off in reactive tank 30 (stage S1090), and be contained in the reactive tank 30.At this moment, because therefore the adsorbed marker DNA stripping in the elution damping fluid of column casing has formed the state that enters the solution that contains this marker DNA in the reactive tank 30.Obtain marker DNA thus.
So, in a plurality of fluid storage portion 201～205,207～210,212～216,218 any one is connected with reactive tank 30, the liquid effects differential pressure that operation by pump 34 is accommodated the fluid storage portion that connects, and with this fluid storage in reactive tank 30, continue the operation of pump 34 by original state, to be incorporated in the reactive tank 30 from the extraneous air that extraneous air throughput 221 flows into, be contained in foaming, the reaction of the liquid of reactive tank 30 thus.And, use the marker DNA that obtains, can carry out analysis to the DNA that puts into reactive tank 30 at first.For example, the marker DNA that obtains is applied to common DNA chip.Through hybridization, washing procedure, with scanner be taken into the fluorescence that from the point of DNA chip obtains, and strength of signal carried out digitized processing thereafter.To be made as contrast from the signal that normal cell DNA obtains, by with from checking the ratio of the signal that the object cell DNA obtains, measure the variation of chromosome copies number.
Here, get the corresponding relation of the integrant and the integrant of the present invention of present embodiment clear.The tube 250 of present embodiment is equivalent to fluid receiving cartridge of the present invention, fluid storage portion 201～205,207～210,212～216,218～220 is equivalent to the resettlement section, the combiner of the first layer 250a and second layer 250b is equivalent to housing, and opening 201a～206a, 208a～211a, 213a～217a, 219a, 220a are equivalent to connect throughput.And, the first layer 250a and second layer 250b are equivalent to dividing layer respectively, exhausted bath box 228 is equivalent to reservoir, opening 206a, 211a, 217a is equivalent in conjunction with throughput, flow out limiting valve 206e, 211e, 217e is equivalent to flow out restrictions, flow into limiting valve 206d, 211d, 217d is equivalent to flow into restrictions, the water-absorbent porous material is equivalent to absorbing material, the first ventilating pit 221a is equivalent to the extraneous air hole, hydrophobic porous material is equivalent to porous material, and tube installing mechanism 80 is equivalent to installation unit, and rotating mechanism 32 is equivalent to mobile unit, pump 34 is equivalent to pressure and pays the unit, rotating disk 82 is equivalent to abutting part, and press section 84 is equivalent to fixed part, and circulation flow path 82a is equivalent to communicating pores, tube is equivalent to a temperature adjustment unit with Peltier's element 38a, and reactive tank is equivalent to reactive tank temperature adjustment unit with Peltier's element 36a.
Reaction unit 90 according to the present embodiment of above detailed description, at the first layer 250a that is formed at tube 250, second layer 250b, and in a plurality of fluid storage portion 201～205 of the specified volume of the decision of the liquid by accommodating, 207～210,212～216, accommodate liquid in 218, under any one opening in a plurality of holes and the state that reactive tank 30 is connected, in the fluid storage portion 201～205 that is communicated with to the opening that is connected with reactive tank 30,207～210,212～216, during 218 applied pressures, extraneous air is supplied to this fluid storage portion 201～205 from the extraneous air throughput 221 with external communications, 207～210,212～216, in the time of 218, the liquid of accommodating is fed in the reactive tank 30.And, tube 250 rotations, other openings are connected with reactive tank 30, and a plurality of liquid are fed in the reactive tank 30.At this moment, extraneous air from 221 supplies of extraneous air throughput, be fed in the reactive tank 30 by opening and any one the fluid storage portion 201～205,207～210,212～216,218 that is connected with reactive tank 30, therefore be contained in a plurality of liquid of reactive tank 30 owing to the inflow of this extraneous air is foamed.Therefore, the liquid of accommodating is reacted more fully in reactive tank 30.
And, because on the bearing surface that is formed at the first layer 250a, the hole set respectively into the round shape of the spigot shaft coaxle of tube 250, each hole and reactive tank 30 switched when being connected, tube 250 rotations are got final product, therefore be easier to switch connect, be easy to fluid storage in reactive tank 30, be easy to thus it is fully reacted.And, about forming the tubulose of the circuitous shape that increases from the circuitous amplitude of tube interior all side direction outer circumferential sides of 250 of circular plate shape in the fluid storage portion 201～205,207～210,212～216,218, because each hole is set at interior all sides of the tube 250 of circular plate shape, can increase circuitous amplitude towards outer circumferential side thus, effectively utilize the space of the tube 250 of circular plate shape, and, by forming tubulose, the liquid that is contained in fluid storage portion 201～205,207～210,212～216,218 is supplied in the reactive tank 30 fully.And, about forming the structure that becomes thin more near each hole more in the fluid storage portion 201～205,207～210,212～216,218, then be easier to whole liquid transportings, and then be easy to make its fully reaction in reactive tank 30 to reactive tank 30.And, about in the chamber owing to the throughput branch of liquid forms the piped parts that parts of fine forms dual tapered of accommodating than liquid, can prevent that the liquid of accommodating from flowing out when tube 250 circulations or during foaming, thereby be easy to make the liquid of accommodating reactive tank 30 in, to react fully.And,, then can effectively accommodate liquid about forming the many more long more piped parts of the amount of liquid of accommodating in the chamber.And tube 250 formations comprise the first layer 250a and second layer 250b, because the chamber is formed at any one deck among the first layer 250a and the second layer 250b or crosses over the first layer 250a and second layer 250b formation, can form more chamber thus.And, from the bearing surface that is formed with each hole away from second layer 250b on, for the amount of the liquid of accommodating than the first layer 250a more manys and is formed with big volumetrical chamber, because the quantity away from the bearing surface chamber is few more more, can reduce the quantity of vertical stream in connection chamber and hole thus, and then be easy to make tube 250.And, flow out limiting valve 206e, 211e, 217e owing to be provided with, deceleration column 206,211,217 and inflow limiting valve 206d, 211d, 217d, therefore need not to switch being connected of reactive tank 30 and opening 206a, 211a, 217a, can be adsorbed in respectively on the column casing of deceleration column 206,211,217 at the resultant that reactive tank 30 internal reactions generate, thereby after in this deceleration column 206,211,217, making the liquid communication in the fluid storage portion 207,212,218 respectively, this liquid is turned back in the reactive tank 30.And,, will remain on reliably in the exhausted bath box 228 from the waste liquid that reactive tank 30 temporarily is contained in exhausted bath box 228 thus because harvesting has the water-absorbent porous material that absorbs waste liquid in the inside of exhausted bath box 228.And because extraneous air throughput 221 has hydrophobic porous material, therefore the parts about being connected with extraneous air throughput 221 in the chamber can not make the liquid of accommodating in the chamber flow out to the outside by extraneous air throughput 221.And, about the parts that are connected with extraneous air throughput 221 in the chamber, can not make the liquid of accommodating in the chamber pass through extraneous air throughput 221 to the effusive while of outside, can further reduce the quantity of the first ventilating pit 221a.And, because reactive tank 30 is more near the thin more piped parts in hole, therefore be easy to make whole liquid between chamber and reactive tank 30, circulate by the hole, be easy to make the interior liquid foam of reactive tank 30 by the extraneous air that leads to by orifice flow.
And, CPU42 carries out predefined DNA amplification handling procedure and dna fragmentation handling procedure, dna marker handling procedure, controls revolution mechanism 32 and pump 34 can be transferred liquid and make its reaction thus based on the step of DNA amplification handling procedure and dna fragmentation handling procedure, dna marker handling procedure.Thus, with the user liquid is supplied to that the temperature with regulation are compared its reaction specified time in the reactive tank 30, can carry out each treatment process reliably, therefore inhibited reaction result's error more reliably with predetermined conditions.And, by pump 34 reduce air pressure in the reactive tanks 30 from fluid storage portion 201～205,207～210,212～216,218 to reactive tank 30 sucking-off liquid, air pressure in the rising reactive tank 30 is released waste liquid from reactive tank 30 to exhausted bath box 228, can transfer liquid by this fairly simple formation of pressure that changes in the reactive tank 30 thus.And, make tube 250 rotate and switch being connected of each hole and reactive tank 30 around turning axle, compare with the rotating disk 82 that send vapor pipe 34a that has been rotatably connected thus, be easier to make its rotation.And in the time of the rotating disk 82 of rotatable and bearing surface butt and tube 250, by this rotating disk 82 is installed tube 250 to the press section 84 of the bearing surface application of force, reactive tank 30 is connected with rotating disk 82, thus, tube 250 can be installed under the state of the switching that can be relatively easy to carry out chamber and reactive tank 30.And, owing to have tube Peltier's element 38a and reactive tank Peltier's element 36a, therefore can respectively the tube of installing 250 be adjusted to the nonreactive temperature of the liquid of accommodating, or reactive tank 30 is adjusted to the temperature of suitable reaction, the Temperature Influence of the tube of not installed thus 250 can make liquid fully reaction in reactive tank 30 of accommodating.
The 3rd embodiment
Figure 24 is the outside drawing of the tube 350 of this enforcement line mode, Figure 25～Figure 28 is the vertical view of the first layer 351a～4th layer 351d of a body 350, Figure 29 is vertical view and the front view of microarray 350b, and Figure 30 is the B-B ' phantom view of tube 350 shown in Figure 24.Here, in the figure of Figure 25～Figure 28, what dotted line was represented is following structure.And, the reaction unit of present embodiment, remove in the reaction unit 90 of first embodiment of Fig. 1, replace tube 50 and use tube 350, and the kind that will use these 350 rice that carry out is specific adjusts handling procedure and reaction treatment procedure stores in the point of flash rom 43 with DNA, use the point of the hose pump of having stated as pump 34, be connected with this pump 34 send among the vapor pipe 34a installation and measuring this send the not shown manometric point of the pressure in the vapor pipe 34a, when the next door of reactive tank fixed part 36 is provided with the motor of having stated 72 (with reference to Figure 15), in reactive tank 30, put into the rotor 74 (with reference to Figure 15) stated and make rotor 74 rotations by motor 72 and beyond the point that stirs, be the same formation with the reaction unit 90 of first embodiment.Thus, in the present embodiment, to the additional prosign of the integrant the same and omit detailed explanation with the reaction unit 90 of first embodiment.And in tube 350, fluid storage portion 302～304,308,309,311,315～321,323,325 and the exhausted bath box 327,328 that can accommodate liquid are referred to as " chamber ".And in the present embodiment, reaction unit 90 describes as the device of discerning the kind of rice from DNA.
Because the pump 34 of present embodiment is a hose pump,, raises thus or be reduced in the pressure that the place ahead with pipe coupling acts on therefore by setting sense of rotation, step number (rotation number) and the speed of the stepper-motor that is connected with this pump 34.In the following description of present embodiment, to accommodating the operation of liquid and when the operation of tube 350 side direction reactive tanks 30 feeding liquids is switched to tube 350 sides from reactive tank 30, after setting sense of rotation, step number (rotation number) and the speed of the stepper-motor that is connected with this pump 34, pump 34 is turned round.And, in the time of need regulating the pressure that acts in the place ahead, represent the mode of goal pressure to be arranged at the not shown manometric output valve of sending vapor pipe 34a with pipe coupling, carry out the setting of sense of rotation, step number (rotation number) and the speed of stepper-motor.
As shown in figure 24, tube 350 has: accommodate the tube body 350a of liquid of the kind that is used to discern rice and the microarray 350b that is installed on this body that can freely dismantle.
Tube body 350a is the parts that material is formed by cyclenes copolymer, is made of four layers of the first layer 351a that forms circular plate shape～4th layer 351d, as shown in figure 25, has the guide part 352 of embedding rotating disk 82 (with reference to Fig. 2) on the first layer 351a.And, this body 350a has the filling orifice 341 that three grooves 342 (playing the effect identical with the groove 252 of first embodiment) and column casing are filled usefulness along radial direction below the 4th layer of 351d, have satellite hole 340c, the 340d that O shape circle is provided with usefulness on the 3rd layer of 351c and the 4th layer of 351b.The second layer 351b of this body 350a and the 3rd layer of 351c have a plurality of chambeies respectively.As Figure 25～shown in Figure 28, this body 350a has: a plurality of fluid storage portion 302～304,308,309,311,315～321,323,325 that can accommodate liquid with the specified volume of the liquid decision of accommodating; By making tube 350 rotations move and switch, thereby be equipped on opening 302a～304a, 308a, 309a, 311a, 315a～321a, 323a, the 325a of any one the regulation link position that is connected with reactive tank 30 that makes in this fluid storage portion respectively; Be communicated with fluid storage portion 302～304,308,309,311,315～321,323,325 and extraneous air, and to fluid storage portion 302～304,308,309,311,315～321,323,325 in, introduce extraneous air or the extraneous air throughput 326 of discharge gas in the fluid storage portion 302～304,308,309,311,315～321,323,325; Can accommodate from the exhausted bath box 327,328 of the waste liquid of reactive tank 30 supplies; Can be adsorbed on the deceleration column 306 of the production thing of reactive tank 30 reactions; By making tube 350 rotations move and switch, thus be equipped on respectively any one regulation link position that is connected with reactive tank 30 of making in the exhausted bath box 327,328 in conjunction with opening 306a, 313a; Obturator hernia 301a, 305a, 307a, 312a, 322a, 324a that Kong Wei opens; Be not communicated with the locking stream 310 that to accommodate liquid with extraneous air; Be used for injecting the filling orifice 310a that liquid that liquid maybe will be contained in locking stream 310 supplies to reactive tank 30 to locking stream 310.
Fluid storage portion the 302～304,308,309,311,315～321,323, the 325th forms the space of the shape of dual tapered.In these fluid storage portions, second layer 351b crosses in the more fluid storage portion 304,308,309,315,316,318,319,321,323 of the amount of liquid of accommodating and the 3rd layer of 351c forms a space, and the less fluid storage portion 302,303,311,317,320,325 of the amount of liquid of accommodating only is formed at the side of second layer 351b or the 3rd layer of 351c.And, fluid storage portion 302～304,308,309,311,315,316,318,319,321,323, one end of the central side of 325 approaching tubes 350, by the stream 302b～304b that is formed at the following of the 3rd layer of 351c and is connected with near the bottom surface of each fluid storage portion side, 308b, 309b, 311b, 315b, 316b, 318b, 319b, 321b, 323b, vertical stream of 325b and the 3rd layer of 351c and second layer 351b is respectively with opening 302a～304a, 308a, 309a, 311a, 315a, 316a, 318a, 319a, 321a, 323a, 325a connects.Fluid storage portion 317,320 is near an end of the central side of tube 350, by being formed at the following of second layer 351b and the stream 317b, the 320b that are connected with near the bottom surface of each fluid storage portion side and vertical stream of second layer 351b, be connected with opening 317a, 320a respectively.End than the distally is connected with extraneous air throughput 326 respectively from the center of tube 350 in fluid storage portion 302～304,308,309,311,315～321,323,325.And, about being described in detail later of extraneous air throughput 326.
Opening 302a～304a, 308a, 309a, 311a, 315a～321a, 323a, 325a are communicated with fluid storage portion 302～304,308,309,311,315～321,323,325 respectively, the hole of using during from fluid storage portion 302～304,308,309,311,315～321,323,325 feeding liquids, and be set at interior all sides (bearing surface) of the first layer 350a.This opening 302a～304a, 308a, 309a, 311a, 315a～321a, 323a, 325a are provided in respectively and the turning axles of tube 350 by rotating mechanism 32 rotations promptly on the same plane of the round shape of the spigot shaft coaxle of tube 350.And, differential pressure by to the liquid effects that is contained in the fluid storage portion 302～304,308,309,311,315～321,323,325 that is connected with these openings can supply to reactive tank 30 with the liquid that is contained in these fluid storage portions 302～304,308,309,311,315～321,323,325.
Extraneous air throughput 326 is to be formed at the following of the 3rd layer of 351c and from fluid storage portion 302,303,309,311,325 from the tube center than the end in the distally extraneous air circulation flow path 302c that direction is extended outside radius, 303c, 309c, 311c, 325c, with be formed at the following of second layer 351b and from fluid storage portion 317,320 from the tube center than the end in the distally extraneous air circulation flow path 317c that direction is extended outside radius, 320c and vertically be formed at ventilating pit 302d～304d of the first layer 351a, 308d, 309d, 311d, 315d～321d, 323d, the general name of 325d.In this ventilating pit 302d～304d, 308d, 309d, 311d, 315d～321d, 323d, 325d, ventilating pit 302d, 303d, 309d, 311d, 325d be respectively by extraneous air circulation flow path 302c, 303c, 309c, 311c, 325c be formed at second layer 351b and the stream longitudinally of the 3rd layer of 351c, and fluid storage portion 302,303,309,311,325 is communicated with extraneous air.And ventilating pit 317d, 320d be respectively by extraneous air circulation flow path 317c, 320c be formed at the stream longitudinally of second layer 351b, and fluid storage portion 317,320 is communicated with extraneous air.And ventilating pit 304d, 308d, 315d, 316d, 318d, 319d, 321d, 323d be by stream, and fluid storage portion 304,308,315,316,318,319,321,323 is communicated with extraneous air.And, in this extraneous air throughput 326, also can set not by liquid but the hydrophobic porous material that extraneous air is passed through.Like this, being contained in the liquid of fluid storage portion 302～304,308,309,311,315～321,323,325 can be by extraneous air throughput 326 and not flowing out to the outside.And, can use for example teflon porous material (day eastern electrician manufacturing TEMISH of company) as hydrophobic porous material.
Shown in Figure 26,27, exhausted bath box the 327, the 328th is arranged at the space of the most peripheral of tube 350, and forms a space crossing over second layer 351b and the 3rd layer of 351c.This exhausted bath box 327 is connected with deceleration column 306 by the waste liquid stream 327e that extends along radial direction that is connected with this exhausted bath box 327 and is formed at second layer 351b and stream that vertically connects among second layer 351b near an end of tube 350 central sides from this waste liquid stream 327e and the diffusion stream 327f along the radial direction extension that is connected with this stream.That is, be released to this exhausted bath box 327 from the fluid that deceleration column 306, passes through in conjunction with opening 306a.Here, as shown in figure 27, the width of diffusion stream 327f and the width of deceleration column 306 are same width roughly.On the other hand, the waste liquid stream 328e of exhausted bath box 328 by being connected with this exhausted bath box 328 is connected with the 328f of stream longitudinally that is arranged at second layer 351b.And the first layer 350a is provided with ventilating pit 327d, the 328d that is communicated with exhausted bath box 327,328 and extraneous air.And,,, also can receive and keep the water-absorbent porous material that absorbs waste liquid for the waste liquid that will temporarily flow into remains in the exhausted bath box 327,328 reliably in the inside of this exhausted bath box 327,328.Can use for example sponge etc. as the water-absorbent porous material.
Deceleration column 306 is set in conjunction with between opening 306a and the diffusion stream 327f, contains column casing.Here, use ceramic column casing (for example, silica gel etc.) as column casing.Make pump 34 running will be contained in liquid communication in the reactive tank 30 to deceleration column 306 and accumulate after spreading stream 327f to pressurizations in the reactive tank 30, the words of further pressurization, accumulate in the liquid of this diffusion stream 327f and accommodated in the exhausted bath box 327, decompression, once more from this deceleration column 306 by and be accommodated in the inside of reactive tank 30.To the filling of the column casing of this deceleration column 306, after side is filled column casing below the 4th layer of 351d by filling orifice 341, carry out in the lower cover upper cover of the 4th layer of 351d.
In conjunction with opening 306a, 313a is to be communicated with exhausted bath box 327,328 respectively, the hole of when this exhausted bath box 327,328 is accommodated liquid, using, and be set at top interior all sides (bearing surface) of the first layer 351a.This is provided in respectively and the turning axles of tube 350 by rotating mechanism 32 (with reference to Fig. 1) rotation promptly on the same plane of the round shape of the spigot shaft coaxle of tube 350 in conjunction with opening 306a, 313a.
Obturator hernia 301a, 305a, 307a, 312a, 322a, 324a are the parts that the Kong Wei of the first layer 351a opens, and stipulate its position by the sealing member 354 that is integrally formed as the shape that a plurality of O shapes circle connects.These obturator hernias since when being positioned in reactive tank 30 opposed states with the following locking of reactive tank 30, therefore pressurized operation and the decompression operation by pump 34 can pressurize or reduce pressure the liquid that are sucked in the reactive tank 30.
Locking stream 310 forms a groove on the 3rd layer of 351c, and by being formed at the stream 310b that extends along radial direction of the 3rd layer of 351c and the stream longitudinally that is formed at the 3rd layer of 351c and second layer 351b, 310a is connected with filling orifice.This locking stream 310 is different with the aforesaid liquid resettlement section away from an end of the central side of tube 350, is not connected with extraneous air throughput 326.Thus, when this locking stream 310 was not communicated with reactive tank 30, filling orifice 310a was blocked in the following meeting of rotating disk 82, and this locking stream 310 forms airtight space.
Filling orifice 310a is communicated with locking stream 310, accommodates the hole of using when liquid that liquid maybe will be contained in locking stream 310 supplies to reactive tank 30 to this locking stream 310, and is set at interior all sides (bearing surface) of the first layer 351a.This filling orifice 310a is with other holes, is provided in and the turning axles of tube 350 by rotating mechanism 32 rotations promptly on the same plane of the round shape of the spigot shaft coaxle of tube 350.
As shown in figure 24, microarray 350b forms and can extract insertion in the slot 330 that the 3rd layer of 351c of tube body 350a and the 4th layer of 351d form.And,, therefore be easy to microarray 350b be inserted in slot 330 or from slot 330, extract owing to be provided with handle 367 in the base segment portion of microarray 350b.
As Figure 29 and shown in Figure 30, this microarray 350b has: arrange first and second connecting hole 361,362 that is arranged at front, the reaction stream 365 of the U word shape that first and second connecting hole 361,362 of two ends and this is connected is arranged at the some zone 366 of two place's straight line portioies in this reaction stream 365.And, be provided with rake 368 in the front of this microarray 350b.
Microarray 350b is being inserted under the state of slot 330, as shown in figure 30, the sealing member 355 of the shape that first connecting hole 361 connects by two O shapes circles and is formed at second layer 351b and is connected with the 328g of stream longitudinally that is communicated with in conjunction with opening 313a.And second connecting hole 362 is connected with the 328f of stream longitudinally (with reference to Figure 26) that is formed at second layer 351b by sealing member 355.And Figure 30 (a) represents the sectional view with the slot 330 before the microarray 350b insertion slot 330.Here, because sealing member 355 is run into the rake 368 that is arranged at microarray 350b at first when microarray 350b inserts slot 330, therefore can not be stuck on this microarray 350b.And satellite hole 340c (with reference to Figure 27), the 340d (with reference to Figure 28) of sealing member 355 by being arranged at the 3rd layer of 351c and the 4th layer of 351d respectively installed by side below the 4th layer of 351d.
As shown in figure 29, in the some zone 366 of microarray 350b, be formed with at a plurality of DNA points that when reactive tank 30 flows into dna solution, make its hybridization by first connecting hole 361.Particularly, in a zone 366, the DNA point is arranged a plurality of the time at certain intervals along the width of reaction stream 365, also arranges a plurality of along its length at certain intervals.And, as Figure 29 and shown in Figure 30, because this microarray 350b forms in the mode that the time reaction stream 365 that is installed on slot 330 is positioned at the below, therefore is easy to Peltier's element 38a reaction stream 365 interior liquid be carried out temperature regulation by the tube of universal stage 38 inside.
In the reaction unit 90 of the present embodiment that so constitutes, use the tube 350 that in container body 350a, has inserted microarray 350b in advance.This 350 liquid that will contain the reaction institute with medicament etc. of regulation is contained in the fluid storage portion of tube 350 respectively with desirable amount, for from fluid storage portion 302～304,308,309,311,315～321,323,325 to reactive tank 30 feeding liquids and the reaction of stipulating at reactive tank 30, or transfer this reacted liquid to exhausted bath box 327,328 from reactive tank 30, rotate the position of moving tins 350 the hole that tube 350 is connected with reactive tank 30 with the change that suits successively by motor 37.Special when carrying out the purification of resultant of reaction, by making column casing absorption reaction resultant and the liquid of not wanting being released to exhausted bath box 327, or make the resultant of reaction stripping of column casing absorption and temporarily accumulate behind diffusion stream 327f by the liquid that any one fluid storage portion is accommodated, be fed to reactive tank 30, react thus.And, this reaction unit 90 is shown in Figure 7 identical with first embodiment of having stated, because reactive tank 30 is arranged at the outside of tube 350, therefore the temperature variation of reactive tank 30 is difficult to transmit to tube 350, reactive tank 30 and tube 350 can be remained on different separately temperature (for example temperature of reaction and preservation with temperature etc.) thus.
Here, to the operation of the reaction unit 90 of present embodiment, especially the genomic dna as the rice of sample is carried out the amplification adjustment, the operation before reacting of the DNA point in the some zone 366 that is formed at microarray 350b is described.Figure 31 is the explanatory view of the step of amplification when adjusting the genomic dna of rice, and Figure 32 is the explanatory view of genomic dna that makes adjustment and the DNA point that is formed at the some zone 366 of the microarray 350b step of reacting.In these figure, the expression of pattern has the fluid storage portion and the exhausted bath box 327,328 of tube 350, the opening of connection and filling orifice and reactive tank 30.Among these figure, in fluid storage portion 302～304,308,309,311,315～321,323,325 and exhausted bath box 327,328, record the kind and the amount of accommodating thereof of liquid, and each symbol that constitutes of being put down in writing of Figure 25～Figure 28.Inner state of not accommodating liquid is represented in the chamber on empty hurdle.And, about reactive tank 30, contain the situation ellipse representation of liquid in the reactive tank 30, the liquid of accommodating to be carried out situation about handling represent with rectangle, the situations that what is not all accommodated in the reactive tank 30 are with the ellipse representation on empty hurdle.And, arrow express liquid among the figure or gas flow direction.And, for convenience of description, in the part additional phase numbering of reactive tank 30.
At first, use Fig. 1, Fig. 2 and Figure 31 that amplification, the adjustment of genomic dna are handled and described.The user will put into reactive tank 30 and will be connected with rotating disk 82 as the genomic dna of the rice of wanting to discern kind, and with the same shown in Fig. 2 (b) of first embodiment of having stated, it be embedded tube 350, thus as reaction member.And, open the lateral not shown door that is arranged at reactive tank fixed part 36, the top of reactive tank 30 is communicated with sending vapor pipe 34a, rotating disk 82 with the portion of being pressed 84 downwards the mode of the application of force slide from the side, thus with this reaction member mounting in universal stage 38.At this moment, because press section 84 materials are special teflon, by deflection, on a plurality of protuberances on be arranged at universal stage 38, the mode that embeds a plurality of recesses that are formed at 250 bottom surface is carried out mounting, and with press section 84 downwards the state of the application of force install.Secondly, press not shown start button.So the CPU42 of controller 40 reads and carries out the DNA that is stored in flash rom 43 and adjusts handling procedure.When carrying out this program, CPU42 at first makes tube 350 rotations that opening 302a is communicated with reactive tank 30 by drive-motor 37, make the air pressure in the pump 34 running reduction reactive tanks 30, the liquid that will be contained in fluid storage portion 302 thus is sucked in the reactive tank 30 (stage S1100).
Secondly, opening 303a is communicated with reactive tank 30, makes pump 34 running sucking-offs be contained in the liquid (stage S1110) of fluid storage portion 303.Then, so that obturator hernia 305a and reactive tank 30 ways of connecting rotating cylinders 350, with the temperature in the reactive tank 30 remain on 95 ℃ stir made its reaction in 15 minutes after, repeatedly 40 circulations make its carry out with the temperature in the reactive tank 30 remain on 95 ℃ stir 1 minute, stir 1 fen half down 66 ℃ of temperature, in 72 ℃ of circulations of stirring for 30 seconds down of temperature, stir down for 72 ℃ in temperature at last and made its reaction (stage S1120) in 10 minutes.Here, what is called " stirring " is meant by motor 72 and makes rotor 47 rotations of putting into reactive tank 30, the solution in the hybrid reaction groove 30 thus.Then, opening 304a is communicated with reactive tank 30, makes pump 34 runnings and sucking-off be contained in the liquid (adsorption-buffering liquid (3.8mol/L ammonium sulfate)) (stage S1130) of fluid storage portion 304.Then, make in conjunction with opening 306a to be communicated with, make pump 34 runnings and make the mixing solutions in the reactive tank 30 be passed to deceleration column 306 (stage S1140) with reactive tank 30.The first layer 351a of mixing solutions by tube 350 shown in Figure 25 flows into and when circulating, have only DNA in the reaction mixture by the absorption of the column casings in the deceleration column 306 in deceleration column 306 in conjunction with opening 306a.And shown in stating, the waste liquid that passes through from column casing finally is released to exhausted bath box 327 by diffusion stream 327f shown in Figure 27.
Then, opening 323a is communicated with reactive tank 30, make pump 34 runnings and sucking-off be contained in the liquid (first lavation buffer solution (1.9mol/L ammonium sulfate)) of fluid storage portion 323, and stirred 1 minute when the temperature in the reactive tank 30 are remained on 25 ℃, (stage S1150) washed in the inside of reactive tank 30.Here, the inside of washing reaction groove 30 is to salt out in order to prevent.Then, make pump 34 running and with the fluid storage after the washing in the reactive tank 30 in fluid storage portion 323 (stage S1160).Then, opening 308a is communicated with reactive tank 30, makes pump 34 runnings and sucking-off be contained in the liquid (second lavation buffer solution (pH6.010mmol/L phosphoric acid alcohol mixeding liquid)) (stage S1170) of fluid storage portion 308.Then, make in conjunction with opening 306a to be communicated with, make pump 34 runnings and make second lavation buffer solution in the reactive tank 30 be passed to deceleration column 306, column casing is washed (stage S1180) with reactive tank 30.Then, opening 309a is communicated with reactive tank 30, makes pump 34 runnings and sucking-off be contained in the liquid (elution damping fluid (pH8.020mmol/L Tutofusin tris hydrochloric acid)) (stage S1190) of fluid storage portion 309.Then, make in conjunction with opening 306a to be communicated with, after making pump 34 running and making elution damping fluids in the reactive tank 30 be passed to deceleration column 306, do not make eluant flow out to exhausted bath box 327 and accumulate in diffusion stream 327f (stage S1200) with reactive tank 30.Particularly, make the elution damping fluid be passed to deceleration column 306 after, stop pump 34 (hose pump) smoothing out with the fingers to pipe.At this moment, because the amplification DNA stripping that is adsorbed in column casing, therefore becomes the state of having accumulated the solution that contains this amplification DNA in diffusion stream 327f in the elution damping fluid.
Here, shown in stating, the width of diffusion stream 327f and the width of deceleration column 306 about equally, the stripping after circulating in column casing is in the degree of the diffusion of the DNA of elution damping fluid, the situation of stream that is narrower than the width of this diffusion stream 327f with width is compared greatly.Thus, the distribution bias of DNA is less in the elution damping fluid.So, the width of the width of diffusion stream 327f and deceleration column 306 is made as about equally the reasons are as follows.That is, when making the elution damping fluid be passed to deceleration column 306, the adsorbed more stripping of DNA of elution damping fluid of initial circulation, the adsorbed DNA of Liu Tong elution damping fluid has had a lot of part strippings at last, thereby stripping hardly.Its result, the width of supposing to spread stream 327f is made as when narrower than deceleration column 306, and is lower near the local DNA concentration of deceleration column 306 after the stripping of elution damping fluid finishes, higher away from the local DNA concentration of the deceleration column 306 of diffusion stream 327f.In the time of elution damping fluid and DNA will being drawn back in the reactive tank 30 with this state, the elution damping fluid is opposite direction circulation with stripping DNA the time in deceleration column 306, and the concentration of the DNA of Liu Tong elution damping fluid uprises at last.So, the elution damping fluid is being returned the moment that finishes in reactive tank 30, be easy to residual DNA in the deceleration column 306.Relative therewith, when being made as with deceleration column 306 width that spreads stream 327f about equally, even the concentration of the DNA of the elution damping fluid of initial circulation is higher from deceleration column 306, the concentration of the DNA of the elution damping fluid that circulates later is lower, and the deviation of the concentration distribution of diffusion back DNA is difficult to exist.Thus, can improve the rate of recovery of DNA.
And, behind stage S1200, make pump 34 runnings and accumulate to be drawn back into (stage S1210) in the reactive tank 30 in the elution damping fluid of the DNA stripping of spreading stream 327f.At this moment, the width of diffusion stream 327f and the width of deceleration column 306 so are compared with the narrower situation of width about equally, and the diffusion of the DNA of stripping is bigger, with the DNA of stripping in reactive tank 30 during together with the sucking-off of elution damping fluid, the DNA that is difficult to residual stripping in the deceleration column 306.Then, filling orifice 310a is communicated with reactive tank 30, makes pump 34 runnings and the elution damping fluid in the reactive tank 30 is injected into locking stream 310 interior (stage S1220).At this moment, be filled in the liquid compression that the air of locking stream 310 is injected into, thereby form the higher state of pressure.Here,, can regulate the amount that flows into the mixing solutions in the locking stream 310 in the pressure of reactive tank 30 by pump 34 regulating effects from the volume and the applied pressure of locking stream 310.For example, pressure is made as 202kpa (two normal atmosphere), can injects the mixing solutions of half amount of volume of locking stream 310.Then, opening 309a is communicated with reactive tank 30, the mixing solutions that remains in the reactive tank 30 is released to fluid storage portion 309 (stage S1230).Here, because the pressure when stage S1220 is injected into locking stream 310 with mixing solutions residues in reactive tank 30 inside, therefore when opening 309a was communicated with reactive tank 30, by this pressure, the mixing solutions in the reactive tank 30 was released to fluid storage portion 309.Then, filling orifice 310a is communicated with reactive tank 30, the mixing solutions that is injected into reaction stream 310 is supplied to reactive tank 30 (stage S1240), obtain adjusted DNA thus.At this moment, owing to by the pressure reactive tank 30 in mixing solutions is released to fluid storage portion 309 at stage S1240, so the pressure in the reactive tank 30 are when descending, the pressure the when air in the locking stream 310 remain on stage S1220 injection mixing solutions.Thus, the mixing solutions that is injected into locking stream 310 is fed in the reactive tank 30 by this pressure difference.And,, also can make pump 34 runnings and mixing solutions is supplied in the reactive tank 30 in order reliably the mixing solutionss in the fluid storage portion 309 to be supplied to reactive tank 30.
Secondly, use Figure 32 to describe to making adjusted DNA and the step that the DNA point that is formed at the some zone 366 of microarray 350b reacts.The CPU42 of controller 40 reads and carries out the reaction treatment program that is stored in flash rom 43.This program is adjusted at the DNA that has stated and is continued to be implemented after handling procedure finishes.When carrying out this program, CPU42 at first makes opening 311a be communicated with the reactive tank 30 with adjusted DNA, and makes pump 34 sucking-off thus of turning round be contained in the liquid (stage S1300) of fluid storage portion 311.Secondly, so that obturator hernia 312a and reactive tank 30 ways of connecting rotating cylinders 350 stir 5 minutes (stage S1310) when the temperature in the reactive tank 30 is remained on 90 ℃.Stir 5 minutes (stage S1320) when then, the temperature in the reactive tank 30 being remained on 10 ℃.Then, make in conjunction with opening 313a and be communicated with reactive tank 30, the mixing solutions that will be contained in reactive tank 30 by the running of adjusting pump 34 is temporarily accumulated in the reaction stream 365 of microarray 350b, and by tube with Peltier's element 38a should react remain in the stream 365 42 ℃ 60 minutes, and after making dna probe that the DNA that is arranged at a zone 366 orders and the DNA in the mixing solutions carry out hybridization, make the air pressure in pump 34 runnings and the raising reactive tank 30 once more, the liquid of temporarily accumulating in reaction stream 365 is released to exhausted bath box 328 (stage S1330).At this moment, the mixing solutions that passes through in microarray 350b is accommodated in exhausted bath box 328 after passing through from the path of having stated.
Then, opening 315a is communicated with reactive tank 30, and makes pump 34 sucking-off thus of turning round be contained in the liquid (stage S1340) of fluid storage portion 315.Then, make in conjunction with opening 313a and be communicated with reactive tank 30, the washings that will be contained in reactive tank 30 by the running of adjusting pump 34 is temporarily accumulated in the reaction stream 365 of microarray 350b, and by tube with Peltier's element 38a should react remain in the stream 365 25 ℃ 5 minutes, and after a zone 366 washed, make the air pressure in pump 34 runnings and the raising reactive tank 30 once more, the washings of temporarily accumulating in reaction stream 365 is released to exhausted bath box 328 (stage S1350).Then, use the liquid that is contained in fluid storage portion 316 to carry out and stage S1340 and the same processing of stage S1350, thus (stage S1360～stage S1370) washed in the some zone 366 of microarray 350b.Then, opening 317a is communicated with reactive tank 30, and makes pump 34 sucking-off thus of turning round be contained in the liquid (stage S1380) of fluid storage portion 317.Then, make in conjunction with opening 313a and be communicated with reactive tank 30, the liquid that will be contained in reactive tank 30 by the running of adjusting pump 34 is temporarily accumulated in the reaction stream 365 of microarray 350b, and should react remain in the stream 365 25 ℃ 30 minutes, after making the DNA in a zone 366 carry out chemiluminescence reaction, make the air pressure in pump 34 runnings and the raising reactive tank 30 once more, the liquid of temporarily accumulating in reaction stream 365 is released to exhausted bath box 328 (stage S1390).Then, use the liquid that is contained in fluid storage portion 318,319 to carry out respectively and stage S1340 and the same processing of stage S1350, thus (stage S1400 and stage S1430) washed in the some zone 366 of microarray 350b.Then, opening 320a is communicated with reactive tank 30, and makes pump 34 sucking-off thus of turning round be contained in the liquid (stage S1440) of fluid storage portion 320.Then, make in conjunction with opening 313a and be communicated with reactive tank 30, the liquid that will be contained in reactive tank 30 by the running of adjusting pump 34 is temporarily accumulated in the reaction stream 365 of microarray 350b, and should react remain in the stream 365 25 ℃ 30 minutes, after making the DNA in a zone 366 carry out the pigementation reaction, make the air pressure in pump 34 runnings and the raising reactive tank 30 once more, the liquid of temporarily accumulating in reaction stream 365 is released to exhausted bath box 328 (stage S1450).Then, opening 321a is communicated with reactive tank 30, and makes pump 34 sucking-off thus of turning round be contained in the liquid (stage S1460) of fluid storage portion 321.Then, make in conjunction with opening 313a to be communicated with, the liquid that is contained in reactive tank 30 is circulated in the reaction stream 365 of microarray 350b, and the pigementation that the DNA in arrests zone 366 carries out reacts (stage S1470) with reactive tank 30.So, in microarray 350b, obtain pigementation DNA point (stage S1480).Use OA this microarray 350b to be carried out image scanning, and carry out the judgement of rice variety from the pigementation pattern of scan image performance with scanner (GF-8700F, Seiko Epson Corporation makes).And, also can carry out the judgement of pigementation pattern by vision.
Here, get the corresponding relation of the integrant and the integrant of the present invention of present embodiment clear.The tube 350 of present embodiment is equivalent to fluid receiving cartridge of the present invention, opening 302a～304a, 308a, 309a, 311a, 315a～321a, 323a, 325a are equivalent to connect throughput, fluid storage portion 302～304,308,309,311,315～321,323,325 is equivalent to the resettlement section, and the combiner of the first layer 351a～4th layer 351d is equivalent to housing.And, the first layer 351a～4th a layer 351d is equivalent to dividing layer respectively, exhausted bath box 327,328 are equivalent to reservoir, in conjunction with opening 306a, 313a is equivalent in conjunction with throughput, ventilating pit 302d～304d, 308d, 309d, 311d, 315d～321d, 323d, 325d is equivalent to the extraneous air hole, tube installing mechanism 80 is equivalent to installation unit, rotating mechanism 32 is equivalent to mobile unit, pump 34 is equivalent to pressure and pays the unit, rotating disk 82 is equivalent to abutting part, and press section 84 is equivalent to fixed part, and circulation flow path 82a is equivalent to communicating pores, tube is equivalent to a temperature adjustment unit with Peltier's element 38a, and reactive tank is equivalent to reactive tank temperature adjustment unit with Peltier's element 36a.
Reaction unit 90 according to the present embodiment of above detailed description, each hole that will set with the round shape of the spigot shaft coaxle of tube 350 and reactive tank 30 switch when being connected, and rotating cylinder 350 get final product, therefore is easier to the switching connection, be easy to fluid storage in reactive tank 30, be easy to thus it is fully reacted.And, about forming the structure that becomes thin more near each hole more in the fluid storage portion 302～304,308,309,311,315～321,323,325, then be easier to whole liquid transportings to reactive tank 30, and then be easy to make its fully reaction in reactive tank 30.And,, then can effectively accommodate liquid about forming the many more long more piped parts of the amount of liquid of accommodating in the chamber.And tube 350 constitutes and to comprise the first layer 351a～4th layer 351d, because the chamber is formed at any one deck among second layer 351b and the 3rd layer of 351c or crosses over second layer 351b and the 3rd layer of 351c forms, can form more chamber thus.
And CPU42 carries out predefined DNA and adjusts handling procedure and reaction treatment program, and controls revolution mechanism 32 and pump 34 can be adjusted the step handover liquid of handling procedure and reaction treatment program and make its reaction thus based on DNA.Thus, with the user liquid is supplied to that the temperature with regulation are compared its reaction specified time in the reactive tank 30, can carry out each treatment process reliably, therefore inhibited reaction result's error more reliably with predetermined conditions.And, by pump 34 reduce air pressure in the reactive tanks 30 from fluid storage portion 302～304,308,309,311,315～321,323,325 to reactive tank 30 sucking-off liquid, air pressure in the rising reactive tank 30 is released waste liquid from reactive tank 30 to exhausted bath box 327,328, can transfer liquid by this fairly simple formation of pressure that changes in the reactive tank 30 thus.And, make tube 350 rotate and switch being connected of each hole and reactive tank 30 around turning axle, compare with the rotating disk 82 that send vapor pipe 34a that has been rotatably connected thus, be easier to make its rotation.And in the time of the rotating disk 82 of rotatable and bearing surface butt and tube 350, by this rotating disk 82 is installed tube 350 to the press section 84 of the bearing surface application of force, reactive tank 30 is connected with rotating disk 82, thus, tube 350 can be installed under the state of the switching that can be relatively easy to carry out chamber and reactive tank 30.And, owing to have tube Peltier's element 38a and reactive tank Peltier's element 36a, therefore can respectively the tube of installing 350 be adjusted to the nonreactive temperature of the liquid of accommodating, or reactive tank 30 is adjusted to the temperature of suitable reaction, the Temperature Influence of the tube of not installed thus 350 can make liquid fully reaction in reactive tank 30 of accommodating.
And the present invention is not subjected to any qualification of above-mentioned the 3rd embodiment, as long as belong to technical scope of the present invention, certainly implements in every way.
For example, in the 3rd above-mentioned embodiment, tube 350 and reaction unit 90 are used to discern the kind of rice, use but also can be used as other chemical reactions.For example, also can accommodate the liquid of the aftertreatment that is used for the CGH array by tube, the program that reaction unit is used according to the aftertreatment of this CGH array is that the CGH finishing sequence is carried out chemical reaction.Figure 33 is tube 450 the outside drawing that the aftertreatment of CGH array is used, Figure 34～Figure 36 is the vertical view of the first layer 451a～3rd layer 451c of a body 450a, and Figure 37 is the explanatory view that makes the step that the DNA point in marker DNA and the some zone that is formed at the microarray 450b that the CGH aftertreatment uses reacts.Here, among the figure of Figure 34～Figure 36, what dotted line was represented is following structure.
As shown in figure 33, tube 450 has: accommodate the tube body 450a of the liquid that the aftertreatment of CGH array uses, the microarray 450b that is installed on this body that can freely dismantle.
Tube body 450a is the parts that material is formed by cyclenes copolymer, is made of three layers of the first layer 451a that forms circular plate shape～3rd layer 351d, as shown in figure 34, has the guide part 452 of embedding rotating disk 82 (with reference to Fig. 2) on the first layer 451a.And this body 450a has three grooves 442 (playing the effect identical with the groove 252 of first embodiment) along radial direction below the 4th layer of 351d, have the satellite hole 440c that O shape circle is provided with usefulness on the 3rd layer of 451c.As Figure 34～shown in Figure 36, this body 450a has: fluid storage portion 412,415,417,419,421,423; Opening 412a, 415a, 417a, 419a, 421a, 423a; Ventilating pit 412d, 415d, 417d, 419d, 421d, 423d, 428d; Exhausted bath box 428; In conjunction with opening 413a; Obturator hernia 401a～411a, 416a, 418a, 420a, 422a, 424a, 425a.Fluid storage portion 412,415,417,419,421,423 is communicated with opening 412a, 415a, 417a, 419a, 421a, 423a respectively near an end of the central side of a body 450a, is communicated with ventilating pit 412d, 415d, 417d, 419d, 421d, 423d respectively away from an end of the central side of tube body 450a.One end of exhausted bath box 428 by waste liquid stream 428a and the microarray 450b that is installed on slot 430 be communicated with in conjunction with opening 413a, the other end directly is communicated with ventilating pit 428d.Obturator hernia 401a～411a, 416a, 418a, 420a, 422a, 424a, 425a are the parts that the Kong Wei of the first layer 451a opens, and stipulate its position by the sealing member 454 that is integrally formed as the shape that a plurality of O shapes circle connects.
Use Fig. 1, Fig. 2 and Figure 37 that the aftertreatment of the CGH array that used tube body 450a is described.The user will put into reactive tank 30 as the marker DNA of sample, and the same with the first above-mentioned embodiment, and mounting is in universal stage 38.Secondly, press not shown start button.So the CPU42 of controller 40 reads and carries out the CGH finishing sequence that is stored in flash rom 43.When carrying out this program, CPU42 at first makes opening 412a be communicated with reactive tank 30, makes pump 34 runnings, will be contained in the liquid sucking-off (stage S1500) of fluid storage portion 412 thus.Then, so that obturator hernia 411a and reactive tank 30 ways of connecting rotating cylinders 450 stir 5 minutes (stage S1510) when the temperature in the reactive tank 30 is remained on 90 ℃.Stir 5 minutes (stage S1520) when then, the temperature in the reactive tank 30 being remained on 10 ℃.Then, make in conjunction with opening 413a and be communicated with reactive tank 30, the mixing solutions that will be contained in reactive tank 30 by the running of adjusting pump 34 is temporarily accumulated in the reaction stream of microarray 450b, and should react remain in the stream 42 ℃ 240 minutes, after making dna probe that the DNA in the some zone that is arranged at microarray 450b orders and the DNA in the mixing solutions carry out hybridization, make the air pressure in pump 34 runnings and the raising reactive tank 30 once more, will temporarily accumulate in the fluid storage that reacts stream to exhausted bath box 428 (stage S1530).At this moment, the mixing solutions that passes through in microarray 450b is accommodated in exhausted bath box 428 after passing through from the path of having stated.
Then, opening 415a is communicated with reactive tank 30, and makes pump 34 sucking-off thus of turning round be contained in the liquid (stage S1540) of fluid storage portion 415.Then, make in conjunction with opening 413a and be communicated with reactive tank 30, the washings that will be contained in reactive tank 30 by the running of adjusting pump 34 is temporarily accumulated in the reaction stream 365 of microarray 450b, and by tube with Peltier's element 38a should react remain in the stream 42 ℃ 1 minute, and after the some zone of microarray 450b washed, make the air pressure in pump 34 runnings and the raising reactive tank 30 once more, the washings of temporarily accumulating in the reaction stream is released to exhausted bath box 428 (stage S1550).Then, use the liquid that is contained in fluid storage portion 417 to carry out and stage S1540 and the same processing of stage S1550, thus (stage S1560～stage S1570) washed in the some zone of microarray 450b.Then, except that by tube with remaining on 42 ℃ in the reaction stream of Peltier's element 38a with microarray 450b 2 minutes, the liquid that use is contained in fluid storage portion 419 carries out and stage S1540 and the same processing of stage S1550, thus (stage S1580～stage S1590) is washed in the some zone of microarray 450b.Then, use the liquid that is contained in fluid storage portion 421,423 to carry out respectively and stage S1540 and the same processing of stage S1550, thus (stage S1600～stage S1630) washed in the some zone of microarray 450b.So, the DNA point (stage S1640) in microarray 450b behind the acquisition hybridization.Have the microarray 450b that the DNA behind the hybridization of acquisition like this is ordered, for example be configured in the adapter corresponding, use scanner to read, strength of signal is carried out digitized processing and is utilized from the fluorescence of DNA point acquisition with the dedicated scan instrument.
In the 3rd above-mentioned embodiment, though after the processing of the stage of Figure 31 S1210, the mixing solutions that uses locking stream 310 to regulate amount is supplied in the reactive tank 30, obtain adjusted DNA (stage S1220～stage S1240) thus, but also can be after the processing of stage S1210, so that obturator hernia 312a and reactive tank 30 ways of connecting rotation column casing 350, set the sense of rotation of the stepper-motor that is connected with pump 34, step number (rotation number) and speed, make pump 34 runnings that the air pressure in the reactive tank 30 is made as 70kpa, with the temperature in the reactive tank 30 remain on 80 ℃ 50 minutes, and the mixing solutions in the reactive tank 30 is concentrated, obtain adjusted DNA thus.
This part is with 2007-323914 number basis as the opinion right of priority of Japan's patent application of Japan's patent application application on December 14th, 2008-172659 number 1 of Japan's patent application application on July 1st, 2008-061051 number 1 of Japan's patent application application on March 11st, 2007-323915 number 1 of application on December 14th, 2007, and its full content is contained within this specification by reference.
The present invention can be used for various chemical reactions except that the parsing of gene.

Claims (28)

1. fluid receiving cartridge, it is to accommodate the fluid receiving cartridge that fluid and fluid can therefrom take out, and it is characterized in that having:
Be formed at respectively in the housing and can accommodate a plurality of resettlement sections of this fluidic with the specified volume that fluid was determined of accommodating;
Be communicated with above-mentioned resettlement section and outside, and can make extraneous air be passed to the extraneous air throughput of this resettlement section;
A plurality of connection throughput, described a plurality of connection throughput is provided in respectively by relatively moving and switching to be arranged at outside and can to accommodate above-mentioned fluidic reactive tank and above-mentioned fluid receiving cartridge and makes the link position of the regulation that this reactive tank is connected with any one connection of above-mentioned resettlement section, and, the fluid that is contained in this resettlement section can be supplied with to above-mentioned reactive tank by differential pressure to the fluid effect of accommodating this resettlement section.
2. fluid receiving cartridge according to claim 1 is characterized in that,
Above-mentioned a plurality of connection throughput is being formed on the plane of above-mentioned housing, is set the round shape into the rotating shaft coaxle that moves with any one rotation of above-mentioned reactive tank and above-mentioned housing respectively.
3. fluid receiving cartridge according to claim 2 is characterized in that,
At least one of above-mentioned a plurality of resettlement sections is formed in the above-mentioned housing of circular plate shape, and forms the tubulose of the circuitous shape that increases from the circuitous amplitude of interior all side direction outer circumferential sides of the housing of above-mentioned circular plate shape,
Above-mentioned a plurality of connection throughput is arranged at interior all sides of the housing of above-mentioned circular plate shape.
4. according to claim 2 or 3 described fluid receiving cartridges, it is characterized in that,
Above-mentioned a plurality of resettlement section is formed in the above-mentioned housing of circular plate shape, and the fluidic amount of accommodating is formed at the more outer circumferential side in the housing of above-mentioned circular plate shape more at most more.
5. according to arbitrary described fluid receiving cartridge in the claim 1～4, it is characterized in that,
At least one of above-mentioned a plurality of resettlement sections forms the tubulose that approaching more above-mentioned connection throughput has the tendency that attenuates more.
6. according to arbitrary described fluid receiving cartridge in the claim 1～5, it is characterized in that,
At least one of above-mentioned a plurality of resettlement sections forms the tubulose of dual tapered.
7. according to arbitrary described fluid receiving cartridge in the claim 1～6, it is characterized in that,
Above-mentioned a plurality of resettlement section forms the long more more at most tubulose of fluidic amount of accommodating.
8. according to arbitrary described fluid receiving cartridge in the claim 1～7, it is characterized in that,
Above-mentioned housing is made of a plurality of dividing layers,
Above-mentioned resettlement section is formed at any one dividing layer in above-mentioned a plurality of dividing layer or crosses over plural dividing layer and form.
9. fluid receiving cartridge according to claim 8 is characterized in that,
Above-mentioned a plurality of resettlement section be formed at more from the formed plane of above-mentioned connection throughput away from dividing layer, it is more to form the fluidic amount of accommodating more.
10. according to arbitrary described fluid receiving cartridge in the claim 1～9, it is characterized in that having:
But be formed in the above-mentioned housing and the reservoir of store fluid in the mode that is communicated with extraneous air;
Above-mentioned reactive tank and above-mentioned fluid receiving cartridge are relatively moved, and when switching in the mode of the position relation that forms regulation, be arranged at the regulation that above-mentioned reactive tank is communicated with above-mentioned reservoir binding site in conjunction with throughput.
11. fluid receiving cartridge according to claim 10 is characterized in that, has:
Be arranged at above-mentioned reservoir and above-mentioned, allow, and stop from the outflow restrictions of this reservoir to the fluidic circulation of this reactive tank from of the fluidic circulation of above-mentioned reactive tank to above-mentioned reservoir in conjunction with between the throughput;
Be arranged at above-mentionedly, can be adsorbed on the deceleration column of the resultant that the reaction of above-mentioned reactive tank forms in conjunction with between throughput and the above-mentioned outflow restrictions;
Be arranged between any one resettlement section and above-mentioned deceleration column of above-mentioned a plurality of resettlement sections, allow that the fluidic to this deceleration column circulates from this resettlement section, and stop from the inflow restrictions of this deceleration column to the fluidic circulation of this resettlement section.
12. fluid receiving cartridge according to claim 11 is characterized in that,
Above-mentioned deceleration column has ceramic column casing.
13. according to claim 11 or 12 described fluid receiving cartridges, it is characterized in that,
Above-mentioned reservoir has the absorbing material of absorption fluids in the inside harvesting.
14. according to arbitrary described fluid receiving cartridge in the claim 1～13, it is characterized in that,
As above-mentioned fluid, above-mentioned resettlement section is accommodated and is carried out the liquid that amplification, fragmentation, hybridization, mark are used to being used for genomic dna that SNP, variation resolve, or carries out the liquid that reverse transcription, amplification, fragmentation, hybridization, the mark of cDNA are used from RNA.
15. according to arbitrary described fluid receiving cartridge in the claim 1～13, it is characterized in that,
As above-mentioned fluid, the genomic dna that accommodate being used for the chromosome abnormalty parsing above-mentioned resettlement section carries out the liquid that amplification, fragmentation or mark are used.
16., it is characterized in that described extraneous air throughput has according to arbitrary described fluid receiving cartridge in the claim 1～15:
Have the extraneous air hole and be communicated with above-mentioned resettlement section and outside extraneous air circulation flow path;
Be equipped in this extraneous air circulation flow path, make above-mentioned fluid can not by but porous material that the said external air can be passed through.
17. fluid receiving cartridge according to claim 16 is characterized in that,
Said external airiness portion is connected with a plurality of above-mentioned resettlement sections.
18. a fluid reaction unit is characterized in that having:
Arbitrary described fluid receiving cartridge in the claim 1～17;
Can connect with a plurality of throughput that are connected that are equipped on above-mentioned fluid receiving cartridge, and can accommodate the fluidic reactive tank of supplying with from the connection throughput of this connection.
19. fluid reaction according to claim 18 unit is characterized in that,
Above-mentioned reactive tank forms the then thin more tubulose of approaching more above-mentioned connection throughput.
20. a reaction unit, it mixes a plurality of fluids and makes fluid reaction, and this reaction unit is characterised in that to have:
The installation unit of arbitrary described fluid receiving cartridge in the claim 1～17 can be installed;
Can connect with a plurality of throughput that are connected of the fluid receiving cartridge that is equipped on above-mentioned installation, and can accommodate the fluidic reactive tank of supplying with from the connection throughput of this connection;
At least one of the fluid receiving cartridge of above-mentioned reactive tank and above-mentioned installation moved to the mobile unit of above-mentioned reactive tank and the link position of the regulation of above-mentioned a plurality of any one connection that is connected throughput;
To the resettlement section of above-mentioned fluid receiving cartridge effect differential pressure, and can pay the unit with the pressure that the fluid that is contained in this resettlement section is supplied with to above-mentioned reactive tank.
21. reaction unit according to claim 20 is characterized in that, has:
Be contained in the series reaction step that the fluid of above-mentioned resettlement section of the fluid receiving cartridge of above-mentioned installation carries out based on use, so that any one of above-mentioned a plurality of resettlement sections and above-mentioned reactive tank ways of connecting when controlling above-mentioned mobile unit, are paid unitary control unit to make the mode of fluid transfer control above-mentioned pressure to above-mentioned resettlement section effect differential pressure.
22. according to claim 20 or 21 described reaction units, it is characterized in that,
Above-mentioned pressure is paid the unit and is connected with above-mentioned reactive tank, by reduce air pressure in this reactive tank with above-mentioned fluid from above-mentioned resettlement section to above-mentioned reactive tank sucking-off, by the air pressure in this reactive tank that raises above-mentioned fluid is released to above-mentioned resettlement section from this reactive tank.
23. according to arbitrary described reaction unit in the claim 20～22, it is characterized in that,
Above-mentioned installation unit is installed fluid receiving cartridge, and above-mentioned fluid receiving cartridge has the housing of the circular plate shape that is formed with above-mentioned resettlement section,
Above-mentioned mobile unit is rotated around turning axle by any one that makes above-mentioned reactive tank and above-mentioned fluid receiving cartridge and is moved, and moves at least one of fluid receiving cartridge of above-mentioned reactive tank and above-mentioned installation thus.
24. reaction unit according to claim 23 is characterized in that,
Above-mentioned installation unit is installed this fluid receiving cartridge by abutting part and fixed part, wherein,
This abutting part sets this plane butt into the fluid receiving cartridge of the round shape of the rotating shaft coaxle that moves with the rotation of above-mentioned housing with the above-mentioned throughput that is connected on the plane of the housing that is formed at above-mentioned circular plate shape, and be provided with and the above-mentioned communicating pores that is connected the throughput connection
This fixed part is rotatably fixed this fluid receiving cartridge under the state of the above-mentioned plane butt that makes above-mentioned abutting part and above-mentioned fluid receiving cartridge;
Above-mentioned reactive tank is connected with the communicating pores that is arranged at above-mentioned abutting part.
25., it is characterized in that having according to arbitrary described reaction unit in the claim 20～24:
Can regulate the tube temperature adjustment unit of the temperature of the fluid storage tube that is installed on above-mentioned installation unit;
Can regulate the reactive tank temperature adjustment unit of the temperature of above-mentioned reactive tank.
26. a reaction method, it is to have used the reaction method that a plurality of fluids is mixed and make the reaction unit of fluid reaction, and this reaction method is characterised in that, comprising:
(a) can be provided in the claim 1～17 that is installed on above-mentioned reaction unit in a plurality of on arbitrary described fluid receiving cartridge be connected that throughput connects and at least one of fluid receiving cartridge that can accommodate the fluidic reactive tank supplied with from the connection throughput of this connection and above-mentioned installation moves to stage of this reactive tank and the link position of the regulation of above-mentioned a plurality of any one connection that is connected throughput;
(b), and will be contained in the stage of the fluid receiving of this resettlement section to above-mentioned reactive tank to the resettlement section of above-mentioned fluid receiving cartridge applied pressure.
27. a gene analytic method is characterized in that,
Use the described fluid receiving cartridge of claim 14, detect the variation of SNP, DNA or the discovery amount of RNA.
28. a gene analytic method is characterized in that,
Use the described fluid receiving cartridge of claim 15, measuring with the human chromosome is the variation of the chromosome copies number of object.
CN2008801208048A 2007-12-14 2008-12-12 Fluid receiving cartridge and utilization of the same Pending CN101896598A (en)

Applications Claiming Priority (9)

Application Number Priority Date Filing Date Title
JP2007-323915 2007-12-14
JP2007323915 2007-12-14
JP2007-323914 2007-12-14
JP2007323914 2007-12-14
JP2008-061051 2008-03-11
JP2008061051 2008-03-11
JP2008-172659 2008-07-01
JP2008172659A JP2009240296A (en) 2007-12-14 2008-07-01 Fluid receiving cartridge and utilization of the same
PCT/JP2008/072638 WO2009078352A1 (en) 2007-12-14 2008-12-12 Fluid receiving cartridge and utilization of the same

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EP2241613A4 (en) 2011-09-21
EP2241613A1 (en) 2010-10-20
US20100276309A1 (en) 2010-11-04
WO2009078352A1 (en) 2009-06-25
JP2009240296A (en) 2009-10-22

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