WO2022024477A1 - 微小粒子分取装置及び微小粒子分取方法 - Google Patents

微小粒子分取装置及び微小粒子分取方法 Download PDF

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Publication number
WO2022024477A1
WO2022024477A1 PCT/JP2021/015969 JP2021015969W WO2022024477A1 WO 2022024477 A1 WO2022024477 A1 WO 2022024477A1 JP 2021015969 W JP2021015969 W JP 2021015969W WO 2022024477 A1 WO2022024477 A1 WO 2022024477A1
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Prior art keywords
sheath liquid
fine particle
flow path
sorting device
particle sorting
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PCT/JP2021/015969
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English (en)
French (fr)
Japanese (ja)
Inventor
正英 古川
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Sony Group Corp
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Sony Group Corp
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Application filed by Sony Group Corp filed Critical Sony Group Corp
Priority to JP2022540018A priority Critical patent/JP7694568B2/ja
Priority to DE112021003987.3T priority patent/DE112021003987T5/de
Priority to CN202180059309.6A priority patent/CN116134304B/zh
Priority to US18/016,510 priority patent/US20230296488A1/en
Publication of WO2022024477A1 publication Critical patent/WO2022024477A1/ja
Anticipated expiration legal-status Critical
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1404—Handling flow, e.g. hydrodynamic focusing
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads or physically stretching molecules
    • 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
    • B01L3/502769—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 multiphase flow arrangements
    • B01L3/502776—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 multiphase flow arrangements specially adapted for focusing or laminating flows
    • 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
    • B01L3/502769—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 multiphase flow arrangements
    • B01L3/502784—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 multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0647—Handling flowable solids, e.g. microscopic beads, cells, particles
    • B01L2200/0652—Sorting or classification of particles or molecules
    • 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
    • B01L2200/14—Process control and prevention of errors
    • B01L2200/143—Quality control, feedback systems
    • 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
    • B01L2300/088—Channel loops
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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    • 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
    • B01L2400/0433—Moving fluids with specific forces or mechanical means specific forces vibrational 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1456—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals
    • G01N15/1459—Optical investigation techniques, e.g. flow cytometry without spatial resolution of the texture or inner structure of the particle, e.g. processing of pulse signals the analysis being performed on a sample stream
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1484—Optical investigation techniques, e.g. flow cytometry microstructural devices
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/149—Optical investigation techniques, e.g. flow cytometry specially adapted for sorting particles, e.g. by their size or optical properties
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N2015/1006—Investigating individual particles for cytology
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1404—Handling flow, e.g. hydrodynamic focusing
    • G01N2015/1406—Control of droplet point
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
    • G01N15/14—Optical investigation techniques, e.g. flow cytometry
    • G01N15/1404—Handling flow, e.g. hydrodynamic focusing
    • G01N15/1409—Handling samples, e.g. injecting samples
    • G01N2015/1411—Features of sheath fluids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
    • G01N15/10—Investigating individual particles
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    • G01N15/1404—Handling flow, e.g. hydrodynamic focusing
    • G01N2015/142—Acoustic or ultrasonic focussing

Definitions

  • This technology relates to a fine particle sorting device and a fine particle sorting method.
  • cell sorter In a cell sorter, generally, a vibrating element or the like vibrates a flow cell or a microchip to atomize the fluid discharged from the flow path. The droplets separated from the fluid are charged with a plus (+) or minus (-) charge, and then their traveling direction is changed by a polarizing plate or the like, and the droplets are collected in a predetermined container or the like.
  • Patent Document 1 discloses that a fraction is formed by using a microchip and a vibration is applied to an orifice of the microchip by a vibrating element.
  • the control technology for stably forming droplets is one of the important factors for improving the accuracy of sorting.
  • the formation of droplets is unstable, such as the fluid discharged from the discharge port of the flow path becoming droplets or the break-off point (BOP) being unstable, it is said that the formation of the droplets is unstable. It is known that the time for charging a droplet to be charged becomes unstable, and as a result, the distribution of fine particles becomes unstable.
  • the microchip has many high-order eigenvalues and corresponding mode shapes. There can be multiple affected frequencies depending on their eigenvalues. Therefore, when vibration is applied to the orifice of the microchip, the vibration intensity of the microchip changes in a complicated manner depending on the frequency, which may lead to destabilization of the break-off point.
  • the main purpose of this technology is to provide a technology that enables the formation of stable droplets.
  • a main flow path through which a liquid containing fine particles flows a sheath liquid flow path that communicates with the main flow path and allows the sheath liquid to flow, and a sheath liquid introduction portion that introduces the sheath liquid.
  • a microparticle sorting device having a microchip provided with a microchip and vibrating the sheath liquid passing through the sheath liquid introduction portion.
  • a connecting member that can be attached to the microchip and has a sheath liquid introduction connecting portion that is connected to the sheath liquid introduction portion may be further provided.
  • a vibrating element may be attached to the connecting member.
  • the drive frequency of the vibrating element may be different from the resonance frequency of the flow path in the microchip. In the present technology, the drive frequency of the vibrating element may be within ⁇ 10% from the resonance frequency of the flow path in the microchip.
  • the sheath liquid introduction connecting portion may have a sheath liquid converging portion whose width gradually or partially narrows from the vibrating element side toward the sheath liquid introduction portion side. In the present technology, the height of the sheath liquid converging portion may gradually or partially decrease from the vibrating element side toward the sheath liquid introduction portion side.
  • a connecting portion having a tubular portion communicating with the tip of the sheath liquid converging portion may be provided between the sheath liquid converging portion and the sheath liquid introducing portion.
  • a tubular member may be inserted inside the tubular portion.
  • the tubular portion and / or at least a portion of the tubular member may consist of any one or more selected from the group consisting of elastomers, resins, and metals.
  • the sheath liquid converging portion may be a substantially conical shape, a substantially polygonal pyramid shape, or an exponential function or a parabolic rotating body.
  • the sheath liquid converging portion may be made of a resin, a metal, or a transparent member.
  • an electrode may be inserted in the sheath liquid converging portion.
  • a swirling flow that swirls the sheath liquid may be generated at the sheath liquid converging portion.
  • the sheath liquid introduction port for introducing the sheath liquid into the sheath liquid converging portion may be located at a position away from the center of the sheath liquid converging portion.
  • a main flow path through which a liquid containing fine particles flows a sheath liquid flow path that communicates with the main flow path and supplies a sheath liquid, and a sheath liquid introduction portion that introduces the sheath liquid are provided. It has a microchip, a light irradiation unit that irradiates the fine particles with light, a light detection unit that detects light from the fine particles, and a processing unit that processes a signal obtained from the light detection unit.
  • the present invention provides a fine particle sorting device that vibrates the sheath liquid that flows through the sheath liquid introduction portion.
  • At least a main flow path through which a liquid containing fine particles flows, a sheath liquid flow path that communicates with the main flow path and supplies a sheath liquid, and a sheath liquid introduction portion that introduces the sheath liquid are at least. Also provided is a method for separating fine particles in which the sheath liquid flowing through the sheath liquid introduction portion is vibrated in the provided microchip.
  • FIGS. A to C are diagrams showing a configuration example of the orifice of the microchip. It is a figure which shows the image of the standing wave generated in the flow path. It is a figure which shows the relationship between the fluctuation pressure in a nozzle and a drive frequency. It is a figure which shows the structural example of the applicable piezoelectric element part. It is a figure which shows the structural example when the bending type piezoelectric element is used. It is a figure which shows the electrode arranged near the sheath liquid converging part.
  • a and B are diagrams showing a configuration example of the fine particle sorting device according to the second embodiment. It is a figure which shows the structural example of the fine particle sorting apparatus which concerns on 3rd Embodiment. It is a graph which shows the result of having measured the frequency characteristic of the break-off point when it was vibrated by using the prior art, and the break-off point when it was vibrated by using this technique. It is a figure which shows the form example 1 of the connection part schematically. It is a graph which shows the relationship between the drive voltage and the BOP length of 27 to 31 kHz. It is a figure which shows the form example 2 of the connection part schematically. It is a figure which shows the form example 3 of the connection part schematically. It is a figure which shows the form example 4 of the connection part schematically.
  • Fourth Embodiment (fine particle sorting apparatus 100) (1) Light irradiation unit 103 (2) Photodetector 104 (3) Processing unit 105 (4) Sorting unit 106 (including charged unit 106c) (5) Storage unit 107 (6) Display unit 108 (7) Input unit 109 (8) Control unit 110 5.
  • Fifth Embodiment small particle sorting method
  • FIG. 1 is a diagram showing a configuration example of the fine particle sorting device 100 according to the first embodiment.
  • the fine particle sorting device 100 according to the present embodiment has a main flow path M2 through which a liquid containing fine particles flows, a sheath liquid flow path M41 communicating with the main flow path and supplying a sheath liquid, and the sheath liquid. It has a microchip M including a sheath liquid introduction unit M4 to be introduced, and shakes the sheath liquid flowing through the sheath liquid introduction unit M4.
  • a sheath liquid introduction unit M4 to be introduced, and shakes the sheath liquid flowing through the sheath liquid introduction unit M4.
  • FIG. 2 is a diagram showing a configuration example of the microchip M
  • FIG. 3 is a diagram showing a configuration example of the orifice M1 of the microchip M.
  • a in FIG. 2 is a schematic top view
  • B in FIG. 2 is a schematic cross-sectional view corresponding to the PP cross-section in A.
  • a in FIG. 3 is a top view
  • B in FIG. 3 is a sectional view
  • C in FIG. 3 is a front view.
  • the microchip M includes a sheath liquid flow path M41 that communicates with the main flow path M2 and allows the sheath liquid to flow through, a sheath liquid introduction unit M4 that introduces the sheath liquid, and the sheath liquid introduction unit M4.
  • the sample liquid flow path M31 that communicates with the main flow path M2 and allows the sample liquid containing fine particles to flow through, the sample liquid introduction unit M3 that introduces the sample liquid, and the confluence part where the sample flow is introduced and merges with the sheath liquid. It is formed.
  • the sheath liquid introduced from the sheath liquid introduction unit M4 is divided into two directions and then fed, and then the sample liquid is sandwiched from two directions at the confluence with the sample liquid introduced from the sample liquid introduction unit M3. And join the sample solution.
  • a three-dimensional laminar flow in which the sample laminar flow is located in the center of the sheath laminar flow is formed at the confluence portion.
  • the M51 shown in FIG. 2A is a suction for clearing the clogging or air bubbles by applying a negative pressure to the main flow path M2 to temporarily reverse the flow when the main flow path M2 is clogged or bubbles are generated. Shows the flow path.
  • a suction opening M5 connected to a negative pressure source such as a vacuum pump is formed at one end of the suction flow path M51. Further, the other end of the suction flow path M51 is connected to the main flow path M2 at the communication port M52.
  • the narrowing portions M61 (see A in FIG. 2) and M62 (see FIG. 3) are formed so that the area of the cross section perpendicular to the liquid feeding direction gradually or gradually decreases from the upstream to the downstream in the liquid feeding direction.
  • the laminar flow width is narrowed down in (see A and B). After that, the three-dimensional laminar flow is discharged as a fluid stream from the orifice M1 provided at one end of the flow path.
  • the fluid stream ejected from the orifice M1 is atomized by the vibrating element applying vibration to the orifice M1, but in the present technology, the fluid stream ejected from the orifice M1 is formed into droplets. Is made into droplets by applying vibration to the sheath liquid passing through the sheath liquid introduction portion M4, as will be described later.
  • the orifice M1 is open in the direction of the end faces of the substrate layers Ma and Mb, and a notch M11 is provided between the opening position and the end face of the substrate layer.
  • the notch M11 is formed by cutting out the substrate layers Ma and Mb between the opening position of the orifice M1 and the end face of the substrate so that the diameter L1 of the notch M11 is larger than the opening diameter L2 of the orifice M1. (See C in FIG. 3).
  • the diameter L1 of the notch M11 is preferably formed to be at least twice as large as the opening diameter L2 of the orifice M1 so as not to hinder the movement of the droplets ejected from the orifice M1.
  • micro means that at least a part of the flow path included in the microchip M has a dimension of ⁇ m order, particularly a cross-sectional dimension of ⁇ m order. That is, in the present technology, the “microchip” refers to a chip including a flow path on the order of ⁇ m, particularly a chip including a flow path having a cross-sectional dimension of the order of ⁇ m. For example, a chip including a particle sorting portion composed of a flow path having a cross-sectional dimension of ⁇ m order can be called a microchip according to the present technology.
  • the microchip M can be manufactured by a method known in the art.
  • the microchip M is formed by laminating the substrate layers Ma and Mb on which the main flow path M2 is formed.
  • the formation of the main flow path M2 on the substrate layers Ma and Mb can be performed, for example, by injection molding of a thermoplastic resin using a mold.
  • the flow path may be formed on all of two or more substrates, or may be formed only on a part of two or more substrates.
  • the microchip M may be formed of three or more substrates by further bonding the substrates from the upward, downward, or both directions with respect to the plane of the substrate on which each flow path is formed.
  • the material for forming the microchip M a material known in the art can be used.
  • examples thereof include, but are not limited to, polycarbonate (PC), cycloolefin polymer, polypropylene, PDMS (polydimethylsiloxane), polymethylmethacrylate (PMMA), polyethylene, polystyrene, glass, silicon and the like.
  • PC polycarbonate
  • PDMS polydimethylsiloxane
  • PMMA polymethylmethacrylate
  • polyethylene polystyrene
  • glass silicon and the like.
  • silicon silicon and the like.
  • polymer materials such as polycarbonate, cycloolefin polymer, and polypropylene are particularly preferable because they are excellent in processability and can be inexpensively manufactured using a molding apparatus.
  • the microchip M is preferably transparent.
  • at least a portion through which light (laser light and scattered light) passes may be transparent, and the entire microchip M may be transparent.
  • the "sample” contained in the sample liquid is particularly fine particles, and the fine particles may be particles having dimensions capable of flowing in the flow path in the microchip M.
  • fine particles may be appropriately selected by those skilled in the art.
  • examples of the fine particles include biological fine particles such as cells, cell clumps, microorganisms, and ribosomes, and synthetic fine particles such as gel particles, beads, latex particles, polymer particles, and industrial particles.
  • biological microparticles also referred to as "biological particles” can include chromosomes, ribosomes, mitochondria, organelles (organelles), etc. that make up various cells.
  • the cells may include animal cells (eg, blood cell lineage cells, etc.), plant cells.
  • the cell can be, in particular, a blood-based cell or a tissue-based cell.
  • the blood line cell may be, for example, a floating line cell such as a T cell or a B cell.
  • the tissue-based cells may be, for example, adherent cultured cells or adherent cells separated from the tissue.
  • the cell mass may include, for example, spheroids, organoids and the like.
  • Microorganisms may include bacteria such as Escherichia coli, viruses such as tobacco mosaic virus, and fungi such as yeast.
  • the biological microparticles may also include biological macromolecules such as nucleic acids, proteins, and complexes thereof.
  • Synthetic fine particles can be, for example, fine particles made of an organic or inorganic polymer material, a metal, or the like.
  • Organic polymer materials may include polystyrene, styrene / divinylbenzene, polymethylmethacrylate and the like.
  • Inorganic polymer materials may include glass, silica, magnetic materials and the like.
  • the metal may include colloidal gold, aluminum and the like.
  • the synthetic microparticles may be, for example, gel particles, beads, etc., in particular gel particles or beads to which one or more combinations selected from oligonucleotides, peptides, proteins, and enzymes are bound. It's okay.
  • the shape of the fine particles may be spherical or substantially spherical, or may be non-spherical.
  • the size and mass of the fine particles can be appropriately selected by those skilled in the art depending on the size of the flow path of the microchip M.
  • the size of the flow path of the microchip M can also be appropriately selected depending on the size and mass of the fine particles.
  • the microparticles may be optionally attached with chemical or biological labels such as fluorescent dyes, fluorescent proteins and the like.
  • the label may facilitate the detection of the microparticles.
  • the fine particles are preferably biological particles, and may be cells in particular.
  • the fine particle sorting device 100 further includes a connecting member that can be attached to the microchip M and has a sheath liquid introduction connecting portion C2 that is connected to the sheath liquid introducing portion M4.
  • the connecting member C shown in FIG. 1 has at least a sample liquid introduction connecting portion C1 connected to the sample liquid introducing portion M3 and a sheath liquid introducing connecting portion C2 connected to the sheath liquid introducing portion M4.
  • the connecting member C that can be attached to and detached from the microchip M, when a large number of different fine particles are continuously separated by using one device, a part of the constituent articles of the device can be removed. Become. Therefore, even if the fine particles contained in the previously separated fluid flow remain in the component, the component can be removed together with the component, and the risk of contamination can be reduced. Further, by disposing of the microchip M and the connecting member C for each sample, the labor of the cleaning operation performed when changing the sample can be saved, and the burden on the operator can be reduced.
  • the sheath liquid introduction connecting portion C2 may have a liquid feeding tube capable of feeding liquid from the sheath liquid feeding portion 101. Further, the liquid feeding tube may have a tube-to-tube connecting portion that is directly connected to the sheathed liquid feeding portion 101. In this case, it is preferable that the tube-to-tube connecting portion is configured so that the liquid in the liquid feeding tube does not come into contact with the outside air. As a result, the cleanliness of the sheath liquid can be ensured.
  • the sample liquid introduction connecting portion C1 may have a tube fixing portion for fixing a liquid feeding tube capable of feeding liquid from the sample liquid feeding unit 102.
  • the liquid feeding tube can be formed integrally with the connecting member C, but can also be formed separately.
  • the liquid feeding tube and the tube fixing portion capable of feeding liquid from the sample liquid feeding portion 102 are formed so as to be removable from the connecting member C, and are arranged at a place different from the sheath liquid feeding portion 101. It is possible to facilitate the connection with the sample liquid feeding unit 102.
  • the vibration element C3 is attached to the connection member C. As a result, it is possible to propagate the vibration to the sheath liquid passing through the sheath liquid introduction portion M4 of the microchip M, and induce the formation of droplets after being ejected from the nozzle.
  • the drive frequency of the vibrating element C3 is different from the resonance frequency of the flow path in the microchip M. The reason for this will be described in detail below.
  • a standing wave is generated between the vibrating element C3 and the nozzle, but the fluctuating pressure in the vicinity of the nozzle changes depending on the shape and dimensions of the flow path, the material, the drive frequency, and the like.
  • the state in which the fluctuating pressure is maximized is the so-called resonance (see FIG. 5).
  • the resonance frequency can be lowered by increasing the length of the flow path, and by combining a plurality of such properties.
  • the resonance frequency can be adjusted.
  • the detailed frequency characteristics of the fluctuating pressure can be estimated by using commercially available acoustic analysis software or the like.
  • the drive frequency is set as a frequency band having a certain width for adjustment.
  • the amount of shift is not particularly limited as long as the drive frequency band does not straddle the resonance frequency even if there is a disturbance such as a temperature change, and it may be set in consideration of the balance between the efficiency improvement effect to be obtained and the stability in the frequency band. ..
  • the drive frequency of the vibrating element can be set to be within ⁇ 10% from the resonance frequency of the flow path in the microchip M, for example.
  • the sheath liquid introduction connecting portion C2 has a sheath liquid converging portion C21 whose width gradually or partially narrows from the side to which the vibration element C3 is attached toward the sheath liquid introduction portion M4 side. Is preferable.
  • the thickness of the flow path in the sheath liquid introduction connecting portion C2 is gradually narrowed from the thickness of the vibration element C3 to the thickness of the sheath liquid introduction portion M4, and the scale and flow are as large as the vibration element C3. It is possible to connect the scale of the size of a path, concentrate the vibration energy of the vibration element C3 in the vicinity of the sheath liquid introduction portion M4, and efficiently send the vibration energy to the flow path in the microchip M with a small drive voltage. ..
  • how the vibration is propagated to the sheath liquid in the vicinity of the sheath liquid introduction portion M4 of the microchip M will be described in detail.
  • the sheath liquid is supplied from the sheath liquid feeding unit 101 to the sheath liquid converging unit C21, and the sheath liquid is vibrated by the vibrating element C3 arranged upstream of the converging unit C21.
  • the vibrating element C3 is composed of, for example, a piezoelectric element portion and a piston portion, each of which is firmly bonded by an adhesive or the like.
  • the structure of the piezoelectric element portion does not matter as long as the vibration finally taken out is in the X direction (see B in FIG. 1) and can be vibrated with the required amplitude at the target vibration frequency.
  • a laminated type, a square plate type, a disk type, a tube type, or the like can be considered (see A to C in FIG. 6).
  • a magnetic force such as a permanent magnet and a solenoid may be used.
  • a structure in which the bent type piezoelectric element is attached to the top surface of the convergent portion C21 as shown in FIG. 7 may be used.
  • the sheath liquid is sent into the chip from the sheath liquid introduction portion M4 of the microchip M, and the vibration of the vibrating element C3 propagates through the sheath liquid and induces droplet formation after being ejected from the nozzle.
  • the vibrating element C3 for example, a piezoelectric element such as a piezo element can be used, but as described above, a vibrating element C3 that converts electric energy into vibration via a magnetic force such as a permanent magnet and a solenoid can be used. .. Further, the frequency is not limited to the ultrasonic region of 20 kHz or more, and can be appropriately set according to the size of the droplet to be formed.
  • the shape of the sheath liquid converging portion C21 gradually or partially decreases in height from the vibrating element C3 side toward the sheath liquid introduction portion M4 side.
  • it can be, for example, a substantially conical shape, a substantially polygonal pyramid shape, or an exponential function or a parabolic rotating body.
  • the shape is not limited to these shapes, but in general, a form in which the flow path is gradually narrowed is more advantageous (for example). Since the amplitude of the vibrating element C3 may be small, droplets can be formed with a low drive voltage of the piezoelectric element, and the degree of freedom in selection and design of the piezoelectric element is increased).
  • a substantially polygonal pyramid shape is selected as the shape of the sheath liquid converging portion C21, a flat surface portion can be provided. Therefore, in particular, when the sheath liquid converging portion C21 is formed of a transparent member, there is an advantage that the inside of the converging portion C21 can be easily seen and bubbles and the like can be easily confirmed. It should be noted that a substantially polygonal weight having a small internal angle such as a substantially triangular weight is not desirable because bubbles are easily caught in each corner and it is difficult to remove the bubbles. .. Further, in order to enhance the effect without obstructing the swirling flow described later, a polygonal weight having a large internal angle is desirable.
  • R includes, for example, adding R of about 1/10 or more of the radius of the inscribed circle of the polygon.
  • the material for forming the sheath liquid converging portion C21 a material known in the art can be used, but in the present technology, it is preferable to form the sheath liquid converging portion C21 with a resin, a metal, or a transparent member.
  • the resin for example, polyetheretherketone (PEEK) or the like can be used.
  • the transparent member for example, polymethyl methacrylate (PMMA), polycarbonate (PC) or the like can be used.
  • PMMA polymethyl methacrylate
  • PC polycarbonate
  • the sheath liquid converging portion C21 By forming the sheath liquid converging portion C21 with the transparent member, the inside of the sheath liquid converging portion C21 can be observed.
  • the metal for example, stainless steel, aluminum alloy, titanium alloy and the like can be used. By forming the sheath liquid converging portion C21 from metal, it is possible to omit the electrode for droplet charging.
  • FIG. 8 is a diagram showing an electrode C4 arranged near the sheath liquid converging portion C21.
  • the sheath liquid converging portion C21 is formed of an insulator such as a resin when the formed droplets are charged
  • the electrode C4 is inserted into the sheath liquid converging portion C21 as shown in FIG. Then, the droplet can be charged through the sheath liquid.
  • the purpose of this is to make the distance between the droplet splitting point and the electrode C4 as close as possible, and to perform charging at a timing closer to the ideal.
  • the sheath liquid converging portion C21 hits a portion of the entire flow path system in which the flow path becomes thick, and air bubbles tend to stay inside the flow path.
  • the sheath liquid introduction port C20 for introducing the sheath liquid into the sheath liquid converging portion C21 is located at a position away from the center of the sheath liquid converging portion C21.
  • the position of the sheath liquid introduction port C20 may be offset upward from the center of the sheath liquid converging portion C21 and further arranged upstream.
  • the offset may be in the lower direction instead of the upper direction of the sheath liquid converging portion C21.
  • the electrode C4 is inserted in this portion in order to charge the droplet (see FIG. 8)
  • fine bubbles may be caught in the boundary between the hole formed in the sheath liquid converging portion C21 and the electrode C4. Therefore, the effect of removing air bubbles adhering to the periphery of the electrode C4 by the swirling flow can be expected.
  • the microchip M and the connecting member C can be appropriately removed as needed, and may be disposable. Further, the vibrating element C3 attached to the connecting member C may also be distributed while being attached to the connecting member C in advance. In this case, the vibrating element C3 may be disposable.
  • FIG. 9A and 9B are diagrams showing a configuration example of the fine particle sorting device according to the second embodiment.
  • a of FIG. 9 is a schematic top view
  • B of FIG. 9 is a schematic cross-sectional view.
  • the sample flow is introduced into the microchip M to form a confluence portion that merges with the sheath liquid, and the sample liquid is introduced into the sheath liquid in the microchip M.
  • the sample liquid is introduced into the sheath liquid converging portion C21, and the core stream containing the sample liquid is converged by hydrodynamic focusing.
  • the present embodiment is the same as the above-mentioned first embodiment except that the method of introducing the sample liquid into the sheath liquid is different.
  • the flow flows by the straightening vane formed inside the sheath liquid converging portion C21 upstream from the position of the sample liquid introduction port for introducing the sample liquid into the sheath liquid converging portion C21.
  • the sample liquid is introduced. This can prevent the core stream from being disturbed by the swirling flow.
  • the straightening vane can be formed of, for example, a thin plate.
  • it is preferable that the straightening vane is not arranged in the thickest portion of the flow path so as not to obstruct the swirling flow.
  • connection portion C22 is a morphological example of the connection portion C22 in the fine particle sorting device 100 according to the third embodiment.
  • the vibration element C3 is omitted.
  • the sheath liquid introduction connecting portion C2 has a sheath liquid converging portion C21 whose width and height gradually or partially narrow from the side to which the vibration element C3 is attached toward the sheath liquid introduction portion M4 side.
  • a connecting portion C22 having a tubular portion C221 communicating with the tip C211 of the sheath liquid converging portion C21 is provided between the sheath liquid converging portion C21 and the sheath liquid introducing portion M4. It should be noted that this embodiment is the same as the first embodiment described above except that the connection portion C22 is provided.
  • connection portion C22 examples include an elastomer, a resin, a metal, or a combination of two or more of these.
  • thermosetting elastomers examples include thermosetting elastomers and thermoplastic elastomers.
  • thermosetting elastomer examples include vulcanized rubber such as natural rubber and synthetic rubber; and resin-based elastomer such as silicone rubber and fluororubber.
  • thermoplastic elastomer examples include polystyrene-based, olefin / alken-based, polyvinyl chloride-based, polyurethane-based, polyester-based, and polyamide-based thermoplastic elastomers.
  • thermoplastic resins examples include thermoplastic resins, thermosetting resins and the like.
  • thermoplastic resin examples include polyolefins such as polystyrene, polyethylene, and polypropylene; polyvinyl chloride; acrylic resin; ABS resin; AS resin; polyamide, polycarbonate, polyacetal, and engineering plastics such as polyethylene terephthalate; polyethersulfone and poly.
  • superengineering plastics such as ether etherketone (PEEK) and thermoplastic polyimides.
  • thermosetting resin examples include phenol resin, melamine resin, polyurethane, unsaturated polyester resin, epoxy resin and the like.
  • Examples of the metal include aluminum alloys, titanium alloys, stainless steel and the like.
  • connection portion C22 a specific example of the form of the connection portion C22 will be described in detail.
  • FIG. 12 is a diagram schematically showing a form example 1 of the connection portion C22.
  • the tubular member C222 is inserted inside the tubular portion C221 made of a thermosetting elastomer.
  • the vibration when it comes into contact with the sheath liquid, it can deviate from the acoustic impedance of the sheath liquid (when the acoustic impedance is about the same, the vibration is diffused and the vibration energy is dissipated), and as a result, it is generated from the vibration element C3.
  • the generated vibration can be prevented from being diffused to the connection portion C22, and the vibration energy can be efficiently transmitted to the flow path in the microchip M.
  • a shorter BOP length can be obtained or the same as shown in FIG.
  • the BOP length can be obtained with a lower drive voltage.
  • Examples of the material forming the tubular member C222 include the above-mentioned elastomer, resin, metal, or a combination of two or more of these.
  • polystyrene, acrylic resin, aluminum alloy, titanium alloy, or stainless steel is preferable, aluminum alloy, titanium alloy, or stainless steel is more preferable, and stainless steel is further preferable.
  • the tubular member C222 is made of the above-mentioned material, but it is more preferable that all of the tubular member C222 is made of the above-mentioned material.
  • the tubular portion C221 may be formed of a thermosetting elastomer to function as a sealing.
  • the length in the longitudinal direction of the tubular member C222 may be the same as the length in the longitudinal direction of the tubular portion C221, but it may be slightly shorter in consideration of crushing for sealing. ..
  • the size of the inner and outer diameters of the tubular member C222 is not particularly limited, but the outer diameter of the tubular member C222 can be, for example, the same as the flow path diameter of the tubular portion C221 or slightly larger than the flow path diameter.
  • FIG. 14 is a diagram schematically showing a form example 2 of the connection portion C22.
  • the connecting portion C22 including the tubular portion 221 is formed by two-color molding with the resin C22a and the thermosetting elastomer C22b.
  • the thermosetting elastomer C22b for sealing is fused to the resin C22a by two-color molding.
  • the tubular portion C221 of the connecting portion C22 with the resin C22a, when the tubular portion 221 is in contact with the sheath liquid as compared with the case where the entire tubular portion 221 is formed of the thermosetting elastomer.
  • examples of the resin C22a include those described above, but in the second embodiment, polystyrene or acrylic resin is particularly preferable.
  • FIG. 15 is a diagram schematically showing a form example 3 of the connection portion C22.
  • the entire connecting portion C22 including the tubular portion 221 is formed of metal, and the ⁇ ring C22c for sealing is arranged at both ends of the connecting portion C22.
  • the metal include those described above, but in the third embodiment, aluminum alloys, titanium alloys, or stainless steels are particularly preferable, and stainless steels are more preferable.
  • the material for forming the ⁇ ring C22c a material known in the art can be used. For example, the above-mentioned elastomer, resin, or a combination of two or more of these may be mentioned.
  • FIG. 16 is a diagram schematically showing a form example 4 of the connection portion C22.
  • a part of the connecting portion C22 including the sheath liquid converging portion C21 and the tubular portion 221 is formed of metal, and then the ⁇ ring C22c and the thermosetting elastomer C22b are arranged at both ends thereof for sealing. ing.
  • the sheath liquid converging portion C21 up to the sheath liquid converging portion C21 with metal it is possible to further suppress the diffusion of the vibration generated from the vibrating element C3.
  • examples of the metal forming a part of the sheath liquid converging portion C21 and the connecting portion C22 include those described above, but in the fourth embodiment, aluminum alloy, titanium alloy, or stainless steel is particularly preferable. , Stainless steel is more preferred. Further, examples of the material forming the ⁇ ring C22c include those described above.
  • FIG. 10 is a diagram showing a configuration example of the fine particle sorting device according to the fourth embodiment.
  • the fine particle sorting device 100 according to the present embodiment has a main flow path M2 through which a liquid containing fine particles flows, a sheath liquid flow path M41 that communicates with the main flow path M2 and supplies a sheath liquid, and the sheath liquid.
  • a microchip including a sheath liquid introduction unit M4 for introducing the above, a light irradiation unit 103 for irradiating the fine particles with light, a light detection unit 104 for detecting light from the fine particles, and the light detection unit 104.
  • It has a processing unit 105 for processing the signal obtained from the above, and vibrates the sheath liquid flowing through the sheath liquid introduction unit. Further, if necessary, a sorting unit 106, a storage unit 107, a display unit 108, an input unit 109, a control unit 110, and the like may be provided.
  • microchip M Since the microchip M is the same as the one described above, the explanation is omitted here. Further, since the method of vibrating the sheath liquid passing through the sheath liquid introduction portion M4 of the microchip M is the same as that described above, the description thereof is omitted here.
  • the light irradiation unit 103 irradiates the fine particles to be sorted with light (for example, excitation light).
  • the light irradiation unit 103 may include a light source that emits light and an objective lens that collects excitation light for fine particles flowing in the detection region.
  • the light source may be appropriately selected by those skilled in the art depending on the purpose of sorting, and may be, for example, a laser diode, a SHG laser, a solid-state laser, a gas laser, or a high-intensity LED, or two or more of them. It may be a combination of.
  • the light irradiation unit 103 may include other optical elements, if necessary, in addition to the light source and the objective lens.
  • the photodetection unit 104 detects light (scattered light and / or fluorescence) generated from the fine particles by irradiation by the light irradiation unit 103.
  • the photodetector 104 may include a condenser lens that collects fluorescence and / or scattered light generated from fine particles, and a photodetector.
  • a PMT, a photodiode, a CCD, a CMOS, or the like can be used, but the present technology is not limited to these.
  • the light detection unit 104 may include other optical elements, if necessary, in addition to the condenser lens and the photodetector.
  • the photodetector 104 may further include, for example, a spectroscopic unit.
  • a spectroscopic unit examples include a grating, a prism, an optical filter, and the like.
  • the spectroscopic unit can, for example, detect light having a wavelength to be detected separately from light having another wavelength.
  • the fluorescence detected by the light detection unit 104 may be fluorescence generated from the fine particles themselves or a substance labeled with the fine particles, for example, a fluorescent substance, but the present technology is not limited to these.
  • the scattered light detected by the light detection unit 104 may be forward scattered light, side scattered light, Rayleigh scattering, or Mie scattering, or may be a combination thereof.
  • the processing unit 105 is connected to the photodetection unit 104 and processes the signal obtained from the photodetection unit 104. For example, the detected value of the light received from the photodetector 104 can be corrected, and the feature amount of each fine particle can be calculated. More specifically, the feature amount indicating the size, morphology, internal structure, etc. of the fine particles is calculated from the detected values of the received fluorescence, the forward scattered light, and the backscattered light. It is also possible to generate a preparative control signal by making a preparative determination based on the calculated feature amount and the preparative conditions received from the input unit in advance.
  • the processing unit 105 can also analyze the state of fine particles using an external analysis device or the like based on the detection value of the light detected by the photodetection unit 104.
  • the processing unit 105 may be implemented by a personal computer or a CPU, and is further stored as a program in a hardware resource including a recording medium (nonvolatile memory (USB memory, etc.), HDD, CD, etc.). , It is also possible to make it function by a personal computer or a CPU. Further, the processing unit 105 may be connected to each unit of the fine particle sorting device 100 via a network.
  • Sorting unit 106 (including charged unit 106c)
  • the preparative unit 106 has at least a polarizing plate 106a that changes the charged droplets in a desired direction, and a collection container 106b that collects the droplets.
  • the charging unit 106c is separately defined in FIG. 4, it is a part of the preparative unit 106, and is charged based on the preparative control signal generated by the processing unit 105.
  • the vibrating element C3 attached to the connecting member C forms droplets by propagating vibration to the sheath liquid as described above.
  • the charged portion 106c is connected to the electrode C4 inserted into the sheath liquid converging portion C21 described above, and the droplets ejected from the orifice M1 of the microchip M are positively or negatively added based on the preparative control signal generated by the processing unit 105. Charged to. Then, the path of the charged droplet is changed in a desired direction by the deflection plate (opposite electrode) 106a to which the voltage is applied, and the droplet is separated.
  • the storage unit 107 stores all matters related to measurement such as the value detected by the optical detection unit 103, the feature amount calculated by the processing unit 105, the preparative control signal, and the preparative conditions input by the input unit. do.
  • the storage unit 107 is not essential, and an external storage device may be connected.
  • the storage unit 107 for example, a hard disk or the like can be used.
  • the recording unit 107 may be connected to each unit of the fine particle sorting device 100 via a network.
  • the display unit 108 can display all items related to measurement such as the value detected by the light detection unit 103 and the feature amount calculated by the processing unit 105.
  • the display unit 108 preferably displays the feature amount for each fine particle calculated by the processing unit 105 as a scattergram.
  • the display unit 108 is not essential, and an external display device may be connected.
  • the display unit 110 for example, a display, a printer, or the like can be used.
  • the display unit 108 may be connected to each unit of the fine particle sorting device 100 via a network.
  • the input unit 109 is a part for a user such as an operator to operate.
  • the user can access the control unit 110 described later through the input unit 109 and control each unit of the fine particle sorting device 100.
  • the input unit 109 preferably sets a region of interest for the scattergram displayed on the display unit 108, and determines the preparative conditions.
  • the input unit 109 is not essential, and an external operating device may be connected.
  • an external operating device for example, a mouse, a keyboard, or the like can be used. Further, the input unit 109 may be connected to each unit of the fine particle sorting device 100 via a network.
  • the control unit 110 is configured to be able to control each of the light irradiation unit 103, the light detection unit 104, the analysis unit 105, the preparative unit 106, the charge unit 106c, the recording unit 107, the display unit 108, and the input unit 109.
  • the control unit 110 may be separately arranged for each unit of the fine particle sorting device 100, or may be provided outside the fine particle sorting device 100. For example, it may be carried out by a personal computer or a CPU, and further, it may be stored as a program in a hardware resource including a recording medium (nonvolatile memory (USB memory, etc.), HDD, CD, etc.), and the personal computer or CPU may be used. It is also possible to make it work by. Further, the control unit 110 may be connected to each unit of the fine particle sorting device 100 via a network.
  • the method for separating fine particles according to the present technology includes a main flow path through which a liquid containing fine particles flows, a sheath liquid flow path that communicates with the main flow path and supplies a sheath liquid, and a sheath liquid that introduces the sheath liquid.
  • the sheath liquid flowing through the sheath liquid introduction portion is vibrated.
  • fine particle sorting method according to the present technology is the same as the method performed by the fine particle sorting device according to the present technology described above, the description thereof is omitted here.
  • FIG. 11 is a graph showing the results of measuring the frequency characteristics of the break-off point when vibrated using this conventional technique and the break-off point when vibrated using this technique.
  • the following configurations can also be adopted.
  • It has a microchip provided with a main flow path through which a liquid containing fine particles flows, a sheath liquid flow path that communicates with the main flow path and allows the sheath liquid to flow, and a sheath liquid introduction portion that introduces the sheath liquid. death, A fine particle sorting device that vibrates the sheath liquid that flows through the sheath liquid introduction portion.
  • the fine particle sorting device according to [1] further comprising a connecting member that can be attached to the microchip and has a sheath liquid introduction connecting portion that is connected to the sheath liquid introduction portion.
  • the fine particle sorting apparatus according to [9], wherein at least a part of the tubular portion and / or the tubular member is made of any one or more selected from the group consisting of an elastomer, a resin, and a metal.
  • the fine particle sorting device according to any one of [6] to [10], wherein the sheath liquid converging portion is a substantially conical shape, a substantially polygonal pyramid shape, or a rotating body having an exponential function or a parabola.
  • the fine particle sorting device according to any one of [6] to [11], wherein the sheath liquid converging portion is made of a transparent member or a metal.
  • a microchip including a main flow path through which a liquid containing fine particles flows, a sheath liquid flow path that communicates with the main flow path and supplies a sheath liquid, and a sheath liquid introduction portion that introduces the sheath liquid.
  • a light irradiation unit that irradiates the fine particles with light
  • a photodetector that detects light from the fine particles
  • a processing unit that processes the signal obtained from the light detection unit, and Have
  • a fine particle sorting device that vibrates the sheath liquid that flows through the sheath liquid introduction portion.
  • a microchip having at least a main flow path through which a liquid containing fine particles flows, a sheath liquid flow path that communicates with the main flow path and supplies a sheath liquid, and a sheath liquid introduction portion that introduces the sheath liquid.
  • Fine particle sorting device 101 Sheath liquid feeding unit 102
  • Sample liquid feeding unit 103 Light irradiation unit 104: Light detection unit 105: Processing unit 106: Sorting unit 107: Storage unit 108: Display unit 109: Input Unit 110: Control unit M: Microchip Ma, Mb: Substrate layer M1: Orifice M11: Notch M2: Main flow path M3: Sample liquid introduction part M31: Sample liquid flow path M4: Sheath liquid introduction part M41: Sheath liquid flow path M5: Suction opening M51: Suction flow path M52: Communication port M61, 62: Narrowing part M7: Straight part L1: Notch M11 diameter L2: Orifice M1 opening diameter C: Connecting member C1: Sample liquid introduction connecting part C2: Sheath liquid introduction connecting part C20: Sheath liquid introduction port C21: Sheath liquid converging part C211: Tip of sheath liquid converging part C22: Connection part C22a: Resin C22b: The

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