WO2014141861A1 - マイクロ流路装置及びこれに関する方法 - Google Patents

マイクロ流路装置及びこれに関する方法 Download PDF

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Publication number
WO2014141861A1
WO2014141861A1 PCT/JP2014/054299 JP2014054299W WO2014141861A1 WO 2014141861 A1 WO2014141861 A1 WO 2014141861A1 JP 2014054299 W JP2014054299 W JP 2014054299W WO 2014141861 A1 WO2014141861 A1 WO 2014141861A1
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Prior art keywords
upstream
liquid
downstream
liquid plug
plug
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Ceased
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PCT/JP2014/054299
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English (en)
French (fr)
Japanese (ja)
Inventor
福田　淳二
博章 鈴木
安奈 山岸
詢子 榎本
雅俊 横川
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University of Tsukuba NUC
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University of Tsukuba NUC
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Priority to ES14763586.6T priority Critical patent/ES2664761T3/es
Priority to EP14763586.6A priority patent/EP2975133B1/en
Priority to US14/775,285 priority patent/US10071378B2/en
Publication of WO2014141861A1 publication Critical patent/WO2014141861A1/ja
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/02—Form or structure of the vessel
    • C12M23/16—Microfluidic devices; Capillary tubes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F23/00—Mixing according to the phases to be mixed, e.g. dispersing or emulsifying
    • B01F23/40—Mixing liquids with liquids; Emulsifying
    • B01F23/405—Methods of mixing liquids with liquids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
    • B01F25/00—Flow mixers; Mixers for falling materials, e.g. solid particles
    • B01F25/14—Mixing drops, droplets or bodies of liquid which flow together or contact each other
    • 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
    • B01F33/301—Micromixers using specific means for arranging the streams to be mixed, e.g. channel geometries or dispositions
    • 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
    • B01F33/302—Micromixers the materials to be mixed flowing in the form of droplets
    • 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
    • B01F33/302—Micromixers the materials to be mixed flowing in the form of droplets
    • B01F33/3021—Micromixers the materials to be mixed flowing in the form of droplets the components to be mixed being combined in a single independent droplet, e.g. these droplets being divided by a non-miscible fluid or consisting of independent droplets
    • 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
    • B01F33/302—Micromixers the materials to be mixed flowing in the form of droplets
    • B01F33/3022—Micromixers the materials to be mixed flowing in the form of droplets the components being formed by independent droplets which are alternated, the mixing of the components being achieved by diffusion between droplets
    • 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
    • B01L3/02—Burettes; Pipettes
    • B01L3/0289—Apparatus for withdrawing or distributing predetermined quantities of fluid
    • B01L3/0293—Apparatus for withdrawing or distributing predetermined quantities of fluid for liquids
    • 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
    • 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
    • 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
    • B01L3/502792—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 for moving individual droplets on a plate, e.g. by locally altering surface tension
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/38—Diluting, dispersing or mixing samples
    • 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
    • B01L2200/0605—Metering of fluids
    • 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
    • B01L2200/0621—Control of the sequence of chambers filled or emptied
    • 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
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/06—Valves, specific forms thereof
    • B01L2400/0688—Valves, specific forms thereof surface tension valves, capillary stop, capillary break

Definitions

  • the present invention relates to a microchannel device and a method related thereto, and more particularly to effective operation of a trace amount liquid in the microchannel device.
  • Patent Document 1 describes a cell culture apparatus in which at least two cell culture chambers having a channel structure in which cells are fixed and a culture medium is perfused are stacked.
  • Non-Patent Document 1 and Non-Patent Document 2 describe a technique for flowing a liquid plug through a flow path of a micro flow path device.
  • a stimulating substance for cell bodies (animal cells, microorganisms, etc.) is added to the chamber while maintaining a constant volume of the culture solution held in the cell culture chamber. It has been difficult to change the cell culture environment in the culture solution over time (for example, increasing the concentration of the stimulating substance in the culture solution stepwise).
  • a stimulating substance when brought into contact with a cell body cultured in a small amount of culture medium in a cell culture chamber, the response of the cell body to the stimulating substance is evaluated. Furthermore, it was difficult to efficiently sample a small amount of the culture solution multiple times at short time intervals.
  • the present invention has been made in view of the above problems, and an object of the present invention is to provide a microchannel device and a method related thereto that realize effective operation of a minute amount of liquid.
  • a method according to an embodiment of the present invention for solving the above-described problem includes the following (a) to (g): (a) an upstream flow path section, a downstream flow path section, and A liquid holding portion provided between a downstream end portion of the upstream flow passage portion and an upstream end portion of the downstream flow passage portion; and the upstream end of the downstream flow passage portion from the downstream end portion of the upstream flow passage portion.
  • a gas bypass channel part provided so as to bypass the liquid holding part up to a part; and (b) filling the gas in the gas bypass channel part with the gas, Holding a main liquid plug in the liquid holding part; (c) sequentially flowing a first upstream liquid plug and a second upstream liquid plug in the upstream flow path part toward the liquid holding part; ) By causing the first upstream liquid plug to flow into the liquid holding portion, the first Fusing an upstream liquid plug with the main liquid plug, and extruding a part of the fused main liquid plug to the downstream flow path; and (e) a gas following the first upstream fluid plug as the gas
  • the one of the main liquid plugs pushed out by the step (d) is made to flow into the bypass channel and push a part of the gas in the gas bypass channel to the upstream end of the downstream channel.
  • the method may further include the following (h): (h) collecting the first downstream liquid plug and the second downstream liquid plug.
  • the second downstream liquid plug is formed in (g) within a time of less than 1 second from the formation of the first downstream liquid plug in (e).
  • the first upstream liquid plug and the second upstream liquid plug may flow.
  • the second upstream liquid plug is connected to the main liquid plug in (f) within a time of less than 1 second after the first upstream liquid plug is fused with the main liquid plug in (d).
  • the first upstream liquid plug and the second upstream liquid plug may be allowed to flow in (c) so as to be fused.
  • each volume of said 1st upstream liquid plug, said 2nd upstream liquid plug, said 1st downstream liquid plug, and said 2nd downstream liquid plug is good also as being less than 1 microliter.
  • the main liquid plug held in the liquid holding part in (b) includes a first factor, and the first upstream liquid plug and the first flow in the upstream flow path part in (c).
  • One or both of the two upstream liquid plugs may include a second factor that acts on the first factor.
  • the first factor may be a cell body
  • the second factor may be a substance that acts on the cell body.
  • a microchannel device includes an upstream liquid plug and an upstream channel portion for flowing gas, and a downstream liquid plug and a downstream channel portion for flowing gas.
  • a liquid holding portion for holding a main liquid plug provided between the downstream end portion of the upstream flow passage portion and the upstream end portion of the downstream flow passage portion, and the downstream end portion of the upstream flow passage portion.
  • a gas bypass flow path section that is provided so as to bypass the liquid holding section to the upstream end of the downstream flow path section, and for flowing gas in a state where the liquid holding section holds the main liquid plug; It is characterized by having.
  • ADVANTAGE OF THE INVENTION According to this invention, the microchannel apparatus which implement
  • the upstream channel section, the liquid holding section, and the gas bypass channel section flow through the upstream channel section toward the liquid holding section that holds the main liquid plug.
  • the upstream liquid plug flows into the liquid holding part without flowing into the gas bypass flow path part, and the gas flowing through the upstream flow path part toward the liquid holding part holding the main liquid plug is retained in the liquid. It is good also as providing so that it may flow in into the said gas bypass channel part, without flowing in into a part.
  • the upstream channel portion, the liquid holding portion, and the gas bypass channel portion are filled with the gas, and the main liquid plug is filled in the liquid holding portion.
  • the upstream liquid plug is moved from the upstream flow path portion to the gas bypass flow path portion. It is good also as providing so that the free energy change in the case of flowing in into may become large.
  • FIG. 1 shows an example of a microchannel device 1 according to the present embodiment (hereinafter referred to as “the present device 1”).
  • 2A and 2B show operations included in an example of a method according to the present embodiment (hereinafter referred to as “the present method”).
  • the present apparatus 1 includes an upstream flow path portion 10 for flowing upstream liquid plugs PLu1, PLu2 and gases Gu1, Gu2, Gu3, downstream liquid plugs PLd1, PLd2 and gas.
  • a main liquid plug PLm provided between the downstream flow path part 20 for flowing Gd1, Gd2, Gd3, the downstream end part 11 of the upstream flow path part 10 and the upstream end part 21 of the downstream flow path part 20 is provided.
  • the method includes the following (a) to (g): (a) the upstream flow path section 10, the downstream flow path section 20, and the downstream of the upstream flow path section 10 A liquid holding portion 30 provided between the end portion 11 and the upstream end portion 21 of the downstream flow channel portion 20, and the upstream end of the downstream flow channel portion 20 from the downstream end portion 11 of the upstream flow channel portion 10.
  • this device 1 including a gas-bypass channel portion 40 provided so as to bypass the liquid holding unit 30 up to the end portion 21; (b) the gas-bypass Filling the flow path part 40 with the gas Gb and holding the main liquid plug PLm in the liquid holding part 30; (c) the first upstream liquid plug PLu1 and the second upstream liquid in the upstream flow path part 10; Flowing the plug PLu2 sequentially toward the liquid holding unit 30; ) By causing the first upstream liquid plug PLu1 to flow into the liquid holding unit 30, the first upstream liquid plug PLu1 is fused with the main liquid plug PLm, and one of the main liquid plugs PLm after merging.
  • the direction indicated by the arrow X indicates the direction in which the liquid and gas flow in the apparatus 1 (the same applies to other drawings). That is, the direction indicated by the arrow X is the downstream direction, and the direction opposite to the direction indicated by the arrow X is the upstream direction. Moreover, the arrow Y shown to FIG. 2A and 2B shows the gas flow in the gas bypass flow-path part 40 (it is the same also in another figure).
  • the upstream flow path portion 10 of the apparatus 1 is a flow path through which the upstream liquid plugs PLu1 and PLu2 that flow into the liquid holding section 30 and the gases Gu1, Gu2, and Gu3 that flow into the gas bypass flow path section 40 flow.
  • the upstream flow path section 10 is provided upstream of the liquid holding section 30 and is connected to the liquid holding section 30 at the downstream end 11 thereof.
  • An upstream end 41 of the gas bypass channel 40 is opened at the downstream end 11 of the upstream channel 10.
  • the downstream flow path unit 20 is a flow path through which the downstream liquid plugs PLd1, PLd2 formed from the main liquid plug PLm held in the liquid holding unit 30 and the gases Gd1, Gd2, Gd3 flowing out from the gas bypass flow path unit 40 flow. is there.
  • the downstream flow path unit 20 is provided downstream of the liquid holding unit 30 and is connected to the liquid holding unit 30 at the upstream end 21 thereof.
  • a downstream end 42 of the gas bypass channel 40 is opened at the upstream end 21 of the downstream channel 20.
  • the liquid holding unit 30 is provided between the upstream channel unit 10 and the downstream channel unit 20. That is, the liquid holding unit 30 is provided between the downstream end portion 11 of the upstream flow path portion 10 and the upstream end portion 21 of the downstream flow path portion 20.
  • the liquid holding unit 30 includes an upstream end 41 of the gas bypass channel 40 connected to the downstream end 11 of the upstream channel 10 (more specifically, the upstream channel 10.
  • the downstream end of the gas bypass channel 40 connected to the upstream end 21 of the downstream channel 20 downstream of the upstream end 41 of the gas bypass channel 40 at the downstream end 11. It is provided upstream of the portion 42 (more specifically, the opening portion of the downstream end portion 42 of the gas bypass flow passage portion 40 in the upstream end portion 21 of the downstream flow passage portion 20). It can be said that the liquid holding unit 30 is an intermediate channel that connects the upstream channel unit 10 and the downstream channel unit 20 in the main channel including the upstream channel unit 10 and the downstream channel unit 20.
  • the shape of the liquid holding part 30 is not particularly limited, for example, as shown in FIGS. It may be larger than the area and the cross-sectional area of the upstream end portion 21 of the downstream flow path portion 20.
  • the gas bypass flow path section 40 allows the gas flowing through the upstream flow path section 10 to flow into the downstream flow path section 30 without flowing into the liquid holding section 30 while the liquid holding section 30 holds the main liquid plug PLm. It is a flow path for.
  • the gas bypass flow path portion 40 bypasses the liquid holding portion 30 to allow the gas flowing in the upstream flow path portion 10 in a state where the liquid holding portion 30 holds the main liquid plug PLm, so that the downstream flow path portion 20 It is the flow path which leads to. Therefore, unlike the upstream flow path portion 10 and the downstream flow path portion 20, no liquid flows through the gas bypass flow path portion 40.
  • the gas bypass flow path section 40 has an upstream end 41 that opens to the downstream end section 11 of the upstream flow path section 10 and a downstream end section 42 that flows downstream from (c) to (g). It is good also as being sealed except opening to the upstream end part 21 of the path part 20. FIG. That is, in this case, during the period from (c) to (g) above, in the gas bypass flow path section 40, in addition to the flow of the gas Gb from the upstream end 41 to the downstream end 42, the flow of the gas Gb There is no.
  • the gas bypass channel 40 may be provided so as not to have an opening other than the upstream end 41 and the downstream end 42 as in the examples shown in FIGS. 1, 2A and 2B. Not limited to this, for example, in addition to the upstream end portion 41 and the downstream end portion 42, other openings are provided, but the other openings are closed during the period from (c) to (g). It is good.
  • the gas bypass flow path portion 40 has an upstream end 41 which is an enlarged flow path. That is, the cross-sectional area of the upstream end portion 41 of the gas bypass channel portion 40 increases toward the downstream side of the gas bypass channel portion 40. Further, the cross-sectional area of the opening at the downstream end portion 11 of the upstream flow passage portion 10 of the upstream end portion 41 of the gas bypass flow passage portion 40 is smaller than the cross-sectional area of the downstream end portion 11 of the upstream flow passage portion 10. Yes.
  • the upstream end portion 41 of the gas bypass flow passage portion 40 flows from the downstream end portion 11 of the upstream flow passage portion 10 in the upstream flow passage portion 10. It extends in a direction substantially perpendicular to the direction.
  • the method of manufacturing this apparatus 1 is a method of manufacturing the microchannel apparatus provided with the above upstream flow path parts 10, the downstream flow path parts 20, the liquid holding
  • the present apparatus 1 includes, for example, a substrate and the upstream flow path portion 10, the downstream flow path portion 20, the liquid holding portion 30, and the gas bypass flow path portion 40 formed as grooves on the surface of the substrate. It is good also as providing.
  • the material constituting the substrate is not particularly limited, but may be made of, for example, resin (for example, PDMS) or glass.
  • maintenance part 30, and the gas bypass flow path part 40 are good also as being formed in the surface of a board
  • the apparatus 1 is particularly preferably used for operating a small amount of liquid and gas. That is, the volume of the liquid holding unit 30 (that is, the volume of the main liquid plug PLm held in the liquid holding unit 30) may be, for example, 0.1 ⁇ L to 1000 ⁇ L, or 0.1 ⁇ L to 500 ⁇ L. It may be 0.1 ⁇ L to 100 ⁇ L. Further, the cross-sectional areas of the upstream flow path section 10, the downstream flow path section 20, and the gas bypass flow path section 40 may be, for example, 0.01 mm 2 to 1.0 mm 2 , and 0.01 mm 2 to 0. It may be 1 mm 2 .
  • the upstream flow path part 10, the liquid holding part 30, and the gas bypass flow path part 40 are upstream liquid plugs that flow through the upstream flow path part 10 toward the liquid holding part 30 that holds the main liquid plug PLm.
  • PLu1 and PLu2 flow into the liquid holding section 30 without flowing into the gas bypass flow path section 40, and flow through the upstream flow path section 10 toward the liquid holding section 30 holding the main liquid plug PLm.
  • Gu1, Gu2, and Gu3 may be provided so as to flow into the gas bypass channel 40 without flowing into the liquid holding unit 30.
  • the upstream liquid plugs PLu ⁇ b> 1 and PLu ⁇ b> 2 that flow through the upstream flow path part 10 toward the liquid holding part 30 that holds the main liquid plug PLm are downstream end parts 42 of the gas bypass flow path part 40. Is opened, flows into the liquid holding section 30 without flowing into the gas bypass flow path section 40, and toward the liquid holding section 30 holding the main liquid plug PLm.
  • the gas Gu1, Gu2, and Gu3 flowing in the gas flow into the gas bypass flow path section 40 without flowing into the liquid holding section 30 with the downstream end portion 32 of the liquid holding section 30 being opened.
  • a method for realizing such selective inflow of the upstream liquid plugs PLu1, PLu2, and PLu3 into the liquid holding unit 30 and selective inflow of the gases Gu1, Gu2, and Gu3 into the gas bypass channel unit 40 is particularly limited.
  • the gas bypass flow path section 40 is filled with the gas Gb, and the main liquid plug PLm is filled in the liquid holding section 30.
  • the upstream liquid plugs PLu1 and PLu2 are more likely to change in the upstream flow path portion 10 than the free energy change when the upstream liquid plugs PLu1 and PLu2 flow into the liquid holding portion 30 from the upstream flow path portion 10. It is good also as providing so that the free energy change at the time of flowing in into the said gas bypass flow-path part 40 may become large.
  • the upstream end 41 of the gas bypass channel 40 is an enlarged channel (that is, the cross-sectional area of the upstream end 41 is downstream).
  • the gases Gu1, Gu2, Gu3 flowing through the upstream flow path 10 and the gas Gb in the gas bypass flow path 40 are included in the gas.
  • the contact angle of the surface 41a with respect to the liquid constituting the upstream liquid plugs PLu1 and PLu2 is 90 ° or more, and the upstream liquid plugs PLu1 and PLu2 flow into the liquid holding part 30 from the downstream end part 11 of the upstream flow path part 10.
  • the increase rate of the area of the gas-liquid interface is that the upstream liquid plugs PLu1 and PLu2 are at the downstream end.
  • the ratio of the area of the gas-liquid interface when the upstream liquid plugs PLu1 and PLu2 are held by the upstream end 31 of the holding unit 30 to the area of the gas-liquid interface when held by the part 11 is there.
  • the rate of increase in the area of the gas-liquid interface is determined by the upstream liquid plugs PLu1 and PLu2 being downstream.
  • the upstream liquid plugs PLu1 and PLu2 with respect to the area of the gas-liquid interface in the state of being held at the end portion 11 are the gas-liquid interface in the state of being held at the upstream end portion 41 of the gas bypass flow path portion 40. It is the area ratio.
  • the upstream end 41 of the gas bypass passage 40 is not an enlarged passage (for example, when the cross-sectional area of the upstream end 41 is constant toward the downstream), the gas (specifically, the upstream flow 41 The contact angle of the inner surface 41a of the upstream end 41 with respect to the liquid constituting the upstream liquid plugs PLu1, PLu2 in the gases Gu1, Gu2, Gu3 flowing through the passage 10 and the gas Gb in the gas bypass channel 40)
  • the wet side length when the upstream liquid plugs PLu1 and PLu2 flow into the liquid holding unit 30 from the downstream end portion 11 of the upstream flow path portion 10 is 90 ° or more, and the upstream liquid plugs PLu1 and PLu2 are It is good also as providing so that it may become larger than the wet side length in the case of flowing in into the gas bypass flow path part 40 from the said downstream end part 11 of the upstream flow path part 10.
  • the wet side length when the upstream liquid plugs PLu1 and PLu2 flow into the liquid holding part 30 from the downstream end part 11 of the upstream flow path part 10 is, for example, the most upstream of the upstream end part 31 of the liquid holding part 30.
  • the cross-sectional shape of the side portion is a rectangle having a width W1 ( ⁇ m) and a height H1 ( ⁇ m), it is 2 ⁇ W1 ⁇ H1 ( ⁇ m 2 ).
  • the wet side length when the upstream liquid plugs PLu1 and PLu2 flow into the gas bypass channel 40 from the downstream end 11 of the upstream channel 10 is, for example, the upstream end 41 of the gas bypass channel 40.
  • the cross-sectional shape of the most upstream portion is a rectangle having a width W2 ( ⁇ m) and a height H2 ( ⁇ m), it is 2 ⁇ W2 ⁇ H2 ( ⁇ m 2 ).
  • the contact angle of the inner surface 41a of the upstream end portion 41a of the gas bypass flow path portion 40 with respect to the liquid constituting the upstream liquid plugs PLu1 and PLu2 is not particularly limited as long as it is 90 ° or more and less than 180 °. For example, it may be 100 ° or more.
  • the method for adjusting the contact angle of the inner surface 41a of the upstream end 41 of the gas bypass channel 40 with respect to the liquid is not particularly limited.
  • the liquid constituting the upstream liquid plugs PLu1 and PLu2, the upstream channel It is good also as adjusting with the combination of the material which comprises gas Gu1, Gu2, Gu3 which flows through the part 10, and gas Gb in the gas bypass flow path part 40, and the said inner surface 41a.
  • the gas bypass channel 40 is provided by forming a groove on the surface of the substrate, selection of a material constituting the substrate and / or upstream of the gas bypass channel 40 It is good also as adjusting the contact angle with respect to the liquid of the said inner surface 41a to the range mentioned above by the hydrophobization process with respect to the inner surface 41a of the edge part 41.
  • the gas bypass channel 40 is filled with the gas Gb, and the main liquid plug PLm is held in the liquid holding unit 30.
  • the main liquid plug PLm is a liquid plug sandwiched between the gas held at the downstream end portion 11 of the upstream flow path portion 10 and the gas held at the upstream end portion 21 of the downstream flow path portion 20.
  • the method of filling the gas bypass flow path portion 40 with the gas Gb and holding the main liquid plug PLm in the liquid holding portion 30 is not particularly limited.
  • the operations shown in FIGS. 3A and 3B are performed. It is good also as using the method of including.
  • the first upstream liquid plug PLu1 and the second upstream liquid plug PLu2 are sequentially flowed toward the liquid holding unit 30 through the upstream flow path unit 10. That is, as shown in FIG. 2A (ii), the first upstream liquid plug PLu1 sandwiched between the downstream gas Gu1 and the upstream gas Gu2 and the downstream of the first upstream liquid plug PLu1 A downstream gas Gu2 and a second upstream liquid plug PLu2 sandwiched between the upstream gas Gu3 are sequentially flowed.
  • the volume of the first upstream liquid plug PLu1 and the second upstream liquid plug PLu2 is such that a part of the main liquid plug PLm pushed out to the downstream flow path portion 20 in (d) and (f) is the downstream flow.
  • it is not particularly limited as long as the upstream end portion 21 of the path portion 20 is satisfied (that is, the portion closes the opening of the downstream end portion 42 of the gas bypass flow path portion 40 in the upstream end portion 21).
  • it may be less than or equal to the volume of the main liquid plug PLm held in the liquid holding unit 30, or may be smaller than the volume of the main liquid plug PLm.
  • the volume of the first upstream liquid plug PLu1 and the second upstream liquid plug PLu2 may be, for example, less than 1 ⁇ L, may be 500 nL or less, and may be 200 nL or less.
  • the volumes of the first upstream liquid plug PLu1 and the second upstream liquid plug PLu2 may be, for example, 1 nL or more.
  • the second downstream liquid plug PLd2 is formed in (g) within a time of less than 1 second after the formation of the first downstream liquid plug PLd1 in (e).
  • the first upstream liquid plug PLu1 and the second upstream liquid plug PLu2 may be allowed to flow.
  • time interval may be, for example, 500 milliseconds or less.
  • said time interval is good also as being 1 millisecond or more, for example.
  • the time intervals for forming the plurality of downstream liquid plugs PLd1 and PLd2 are, for example, the volume of the gas Gu2 between the first upstream liquid plug PLu1 and the second upstream liquid plug PLu2, the second upstream liquid plug PLu2 It is good also as adjusting to the above-mentioned range by one or more selected from the group which consists of the volume of this, and the flow rate of the said 2nd upstream liquid plug PLu2.
  • the second upstream liquid plug in (f) is within a period of less than 1 second after the first upstream liquid plug PLu1 is fused with the main liquid plug PLm in (d).
  • the first upstream liquid plug PLu1 and the second upstream liquid plug PLu2 may be flowed so that PLu2 is fused with the main liquid plug PLm.
  • fusion of the plurality of upstream liquid plugs PLu1, PLu2 and the main liquid plug PLm at a time interval of less than 1 second is realized.
  • the time interval may be, for example, 500 milliseconds or less.
  • the said time interval is good also as being 1 millisecond or more, for example.
  • the time interval for fusing the plurality of upstream liquid plugs PLu1, PLu2 and the main liquid plug PLm is, for example, the volume of the first upstream liquid plug PLu1, the first upstream liquid plug PLu1 and the second upstream liquid plug PLu2. It is good also as adjusting to the above-mentioned range by one or more selected from the group which consists of the volume of gas Gu2 between these, and the flow velocity of the said upstream liquid plug PLu1, PLu2.
  • the method of flowing the first upstream liquid plug PLu1 and the second upstream liquid plug PLu2 in the upstream flow path section 10 is not particularly limited.
  • the first upstream liquid plug PLu1 and the second upstream liquid plug PLu2 are flowed. It is good also as flowing by applying a pressure from upstream, and good also as flowing by reducing pressure from downstream.
  • the number of liquid plugs flowing through the upstream flow path section 10 is not particularly limited as long as it is two or more, and three or more liquid plugs may be flowed. That is, in the above (c), a plurality of upstream liquid plugs that are spaced apart from each other via a gas may be flowed.
  • the first upstream liquid plug PLu1 is caused to flow into the liquid holding unit 30, thereby fusing the first upstream liquid plug PLu1 with the main liquid plug PLm, and A part of the main liquid plug PLm is pushed out to the downstream flow path part 20.
  • the first upstream liquid plug PLu1 is caused to flow into the liquid holding part 30 without flowing into the gas bypass flow path part 40, thereby the first upstream liquid plug.
  • PLu1 is fused with the main liquid plug PLm.
  • the gas Gu2 following the first upstream liquid plug PLu1 is caused to flow to the downstream end portion 11 of the upstream flow path portion 10, thereby causing a part of the main liquid plug PLm to flow downstream. It pushes out from the upstream end 21 of the flow path part 20 to the downstream.
  • the volume of a part of the main liquid plug PLm pushed out from the liquid holding unit 30 is the same as the volume of the first upstream liquid plug PLu1. Further, the volume of the main liquid plug PLm after the fusion is the sum of the volume of the main liquid plug PLm before the fusion and the volume of the first upstream liquid plug PLu1.
  • the fusion of the first upstream liquid plug PLu1 and the main liquid plug PLm mixes the liquid constituting the first upstream liquid plug PLu1 and the liquid constituting the main liquid plug PLm.
  • the gas Gu2 following the first upstream fluid plug PLu1 is caused to flow into the gas bypass channel portion 40, and a part of the gas Gb in the gas bypass channel portion 40 is transferred to the downstream channel.
  • a part of the main liquid plug PLm extruded in (d) is cut to form the first downstream liquid plug PLd1.
  • the gas Gu2 following the first upstream fluid plug PLu1 is caused to flow into the gas bypass channel 40 without flowing into the liquid holding unit 30, thereby the gas bypass channel.
  • a part of the gas Gb held in the portion 40 is pushed out to the upstream end portion 21 of the downstream flow path portion 20.
  • the volume of the first downstream liquid plug PLd1 is the same as the volume of the first upstream liquid plug PLu1. That is, for example, as described above, when the volume of the first upstream liquid plug PLu1 is less than 1 ⁇ L, the volume of the first downstream liquid plug PLd1 is also less than 1 ⁇ L, and the first upstream liquid plug PLu1 Is the same volume.
  • the second upstream liquid plug PLu2 is caused to flow into the liquid holding unit 30 so as to fuse the second upstream liquid plug PLu2 with the main liquid plug PLm and A part of the main liquid plug PLm is pushed out to the downstream flow path part 20.
  • the liquid holding portion is allowed to flow without causing the second upstream liquid plug PLu2 to flow into the gas bypass passage portion 40.
  • the second upstream liquid plug PLu2 is fused with the main liquid plug PLm by flowing into the main liquid plug 30.
  • the volume of a part of the main liquid plug PLm pushed out from the liquid holding unit 30 is the same as the volume of the second upstream liquid plug PLu2. Further, the volume of the main liquid plug PLm after the fusion is the sum of the volume of the main liquid plug PLm before the fusion and the volume of the second upstream liquid plug PLu2.
  • the fusion of the second upstream liquid plug PLu2 and the main liquid plug PLm mixes the liquid constituting the second upstream liquid plug PLu2 and the liquid constituting the main liquid plug PLm.
  • the liquid constituting the main liquid plug PLm before merging with the first upstream liquid plug PLu1 and the first upstream liquid plug PLu1 are The liquid which comprises and the liquid which comprises the said 2nd upstream liquid plug PLu2 are mixed.
  • the fusion of the second upstream liquid plug PLu2 and the main liquid plug PLm is less than one second after the first upstream liquid plug PLu1 is fused with the main liquid plug PLm in the above (d). It may be performed within.
  • the gas Gu3 following the second upstream liquid plug PLu2 is caused to flow into the gas bypass channel portion 40, and a part of the gas Gb in the gas bypass channel portion 40 is transferred to the downstream channel.
  • a part of the main liquid plug PLm extruded in (f) is cut to form a second downstream liquid plug PLd2.
  • the volume of the second downstream liquid plug PLd2 is the same as the volume of the second upstream liquid plug PLu2. That is, for example, as described above, when the volume of the second upstream liquid plug PLu2 is less than 1 ⁇ L, the volume of the second downstream liquid plug PLd2 is also less than 1 ⁇ L, and the second upstream liquid plug PLu2 Is the same volume.
  • the second downstream liquid plug PLd2 may be formed within a time of less than 1 second after the first downstream liquid plug PLd1 is formed in (e). In this case, as described above, the formation of the plurality of downstream liquid plugs PLd1 and PLd2 at a time interval of less than 1 second is realized.
  • the apparatus 1 and the method described above effective operation of a trace amount liquid in the microchannel apparatus is realized. That is, in particular, using the present apparatus 1 including the gas bypass flow path section 40, the plurality of upstream liquid plugs PLu1, PLu2 are intermittently and sequentially fused with the main liquid plug PLm, and the plurality of upstream liquid plugs
  • the gas Gu2 and Gu3 following each of PLu1 and PLu2 are diverted from the upstream flow path part 10 to the downstream flow path part 20 without flowing into the liquid holding part 30, thereby corresponding to the plurality of upstream liquid plugs PLu1 and PLu2.
  • the plurality of downstream liquid plugs PLd1 and PLd2 can be intermittently formed sequentially to maintain the volume of the main liquid plug PLm constant.
  • the first upstream liquid plug PLu1 is fused with the main liquid plug PLm in (d) by adjusting the conditions for flowing the plurality of upstream liquid plugs PLu1 and PLu2 in the upstream flow path section 10.
  • the second upstream liquid plug PLu2 can be fused with the main liquid plug PLm in (f) above.
  • the upstream liquid plugs PLu1, PLu2 and the main liquid plug PLm are fused a plurality of times with extremely high time resolution (that is, the liquid constituting the upstream liquid plugs PLu1, PLu2 and the liquid constituting the main liquid plug PLm). Can be mixed).
  • the second downstream liquid plug PLd2 may be formed within a time of less than 1 second after the formation of the first downstream liquid plug PLd1 in (e). . That is, by adjusting the conditions for flowing the plurality of upstream liquid plugs PLu1, PLu2 in the upstream flow path section 10, within the time less than 1 second from the formation of the first downstream liquid plug PLd1 in (e) above, In the above (g), the second downstream liquid plug PLd2 can be formed. In this case, a plurality of samplings are realized at a time interval of less than 1 second. That is, sampling can be performed a plurality of times with extremely high time resolution.
  • the volumes of the first upstream liquid plug PLu1, the second upstream liquid plug PLu2, the first downstream liquid plug PLd1, and the second downstream liquid plug PLd2 are less than 1 ⁇ L. It is possible to realize an effective operation of a very small amount of liquid.
  • the method may further include the following (h): (h) collecting the first downstream liquid plug PLd1 and the second downstream liquid plug PLd2.
  • the first downstream liquid plug PLd1 and the second downstream liquid plug PLd2 are recovered. That is, as shown in FIGS. 2B (ix) and (x), the first downstream liquid plug PLd1 and the second downstream liquid plug PLd2 formed as described above are recovered from the downstream flow path portion 20.
  • the recovered first downstream liquid plug PLd1 and second downstream liquid plug PLd2 may be analyzed.
  • the main liquid plug PLm held in the liquid holding unit 30 in (b) includes the first factor, and the first upstream liquid that flows to the upstream flow path in (d) above.
  • One or both of the plug PLu1 and the second upstream liquid plug PLu2 may include a second factor that acts on the first factor.
  • the second factor can act on the first factor by fusing the upstream liquid plugs PLu1 and PLu2 with the main liquid plug PLm in (d) and / or (f) above.
  • the first factor may be fixed to the liquid holding unit 30.
  • the second factor may act directly on the first factor in the main liquid plug PLm, or may act indirectly on the first factor.
  • the indirect action of the second factor on the first factor is, for example, in the main liquid plug PLm, where the third factor is first generated from the second factor, and then the third factor is the first factor. It is good also as being performed by acting on this factor.
  • the first factor may be a cell body
  • the second factor may be a substance that acts on the cell body (for example, a stimulating substance).
  • the cell body may be fixed to the liquid holding unit 30.
  • the cell body is not particularly limited, but may be, for example, an animal cell.
  • the animal cell may be, for example, a mammalian cell. That is, the animal cell may be, for example, a cell of a human or non-human animal (eg, monkey, pig, dog, rat or mouse).
  • the animal cell may be a primary cell collected from a human or non-human animal, or may be an established cell line.
  • the animal cell may be a differentiated cell or an undifferentiated cell.
  • the animal cell may be an embryonic stem cell or a cell derived therefrom, or may be an iPS (induced Pluripotent Stem) cell or a cell derived therefrom.
  • the cell body may be an animal cell other than a mammal.
  • the cell body may be, for example, a microorganism.
  • the cell body may be, for example, a plant cell.
  • a plurality of upstream liquid plugs PLu1 and PLu2 each including a very small amount of liquid containing a rare stimulating substance are intermittently formed. Sequentially fused to a small amount of main liquid plug PLm containing a small amount of rare cell bodies, and a plurality of downstream liquid plugs PLd1, PLd2 corresponding to the plurality of upstream liquid plugs PLu1, PLu2 are intermittently formed sequentially.
  • the concentration of the stimulating substance contained in the main liquid plug PLm is increased stepwise to accurately change the culture environment of the cell body. Can be controlled.
  • the present apparatus 1 and the present method perform effective multiple sampling by intermittently sequentially forming the plurality of downstream liquid plugs PLd1 and PLd2 corresponding to the plurality of upstream liquid plugs PLu1 and PLu2.
  • the response of a small amount of rare cell bodies contained in the main liquid plug PLm to a stimulating substance can be effectively evaluated.
  • the main liquid plug PLm held in the liquid holding unit 30 in (b) includes a cell body, and the first upstream liquid plug PLu1 that flows into the upstream flow path unit 10 in (c) and
  • One or both of the second upstream liquid plugs PLu2 is a stimulus that changes the secretion amount of the cell body (amount of the specific substance secreted by the cell body) and / or the secretion rate (speed at which the cell body secretes the specific substance). It is good also as containing a substance.
  • the cell body included in the main liquid plug PLm held in the liquid holding unit 30 in (b) is a Langerhans cell, and the first flowing through the upstream flow path unit 10 in (c) above.
  • the upstream liquid plug PLu1 and the second upstream liquid plug PLu2 may include glucose.
  • the specific substance secreted by the Langerhans cells is insulin
  • glucose is a stimulating substance that changes the secreted amount and / or rate of insulin of the Langerhans cells.
  • the main liquid plug PLm held in the liquid holding unit 30 in (b) includes a cell body, and the first upstream liquid plug PLu1 and the second upstream liquid flowing into the upstream flow path unit 10 in (c).
  • the plugs PLu2 contain a stimulating substance that changes the secretion amount and / or the secretion rate of the cell body
  • the upstream liquid plugs PLu1 and PLu2 are connected to the main liquid plug in (d) and / or (f) above.
  • the stimulating substance is allowed to act on the cell body in the liquid holding unit 30, and the amount and / or speed changed by the action of the stimulating substance in (e) and / or (g) above.
  • the first downstream liquid plug PLd1 and / or the second downstream liquid plug PLd2 containing the specific substance secreted from the cell body can be formed.
  • the response of the cell body to the stimulating substance can be obtained with high time resolution as described above. Can be evaluated.
  • the response of the cell body to the stimulating substance may be evaluated without sampling. That is, for example, in the present apparatus 1 arranged on the sample stage of a microscope, a plurality of upstream liquid plugs PLu1, PLu2 containing a stimulating substance are fused with a main liquid plug PLm containing a cell body, and a plurality of downstream liquid plugs PLd1, By forming PLd2, the response of the cell body to the stimulating substance can be observed and evaluated under the microscope.
  • the first factor is not limited to the cell body. That is, for example, the first factor may be a first substance, and the second factor may be a second substance that generates a third substance by interacting with the first substance. .
  • the first factor may be an enzyme fixed to the liquid holding unit 30, and the second factor may be a substrate of the enzyme.
  • the upstream liquid plugs PLu1 and PLu2 containing the substrate are fused with the main liquid plug PLm containing the enzyme, so that the substrate in the liquid holding unit 30
  • An enzyme reaction is performed by contacting with an enzyme, and in (e) and / or (g), the first downstream liquid plug PLd1 and / or the second downstream liquid plug PLd2 containing a product produced by the enzyme reaction is formed. can do.
  • the amount of product produced by the enzyme reaction and the enzyme reaction rate are as described above. It can be evaluated with a high temporal resolution.
  • FIG. 4 shows another example of the device 1.
  • the upstream flow path section 10 of the present apparatus 1 cuts the liquid by supplying a liquid supply section 12 for supplying the liquid constituting the upstream liquid plugs PLu1, PLu2 (see FIG. 2A).
  • the upstream flow path section 10 of the apparatus 1 is provided upstream of the liquid supply section 12, the gas supply section 13, the liquid supply section 12, and the gas supply section 13.
  • a composition adjusting unit 15 for adjusting the composition of the upstream liquid plugs PLu1 and PLu2 by adding factors to the liquid plugs PLu1 and PLu2, and the upstream liquid plugs PLu1 and PLu2 whose compositions are adjusted are directed to the liquid holding unit 30.
  • a merging flow path portion 14 for flowing Since the upstream flow path unit 10 further includes the composition adjustment unit 15, the plurality of upstream liquid plugs PLu1 and PLu2 whose compositions are adjusted in the upstream flow path unit 10 can be efficiently formed.
  • the main liquid plug PLm held in the liquid holding unit 30 in (b) includes a cell body, and the first upstream liquid that flows to the upstream flow path unit 10 in (c) above.
  • the upstream flow path section 10 is the first upstream liquid plug PLu1.
  • / or the first upstream liquid plug PLu1 and the second upstream liquid plug having different concentrations of the stimulating substance by having the composition adjusting unit 15 for adding the stimulating substance to the second upstream liquid plug PLu2.
  • PLu2 can be formed efficiently.
  • FIG. 5 shows still another example of the device 1.
  • the gas bypass channel 40 has an upstream end 41 that is not an enlarged channel.
  • the cross-sectional area of the upstream end portion 41 of the gas bypass channel portion 40 is constant toward the downstream side of the gas bypass channel portion 40.
  • the cross-sectional area of the opening at the downstream end portion 11 of the upstream flow passage portion 10 of the upstream end portion 41 of the gas bypass flow passage portion 40 is smaller than the cross-sectional area of the downstream end portion 11 of the upstream flow passage portion 10. Yes.
  • the gas bypass flow path section 40 has an upstream end 41 including a short expanded flow path as compared with the above-described examples shown in FIGS. That is, the upstream end portion 41 opens to the downstream end portion 11 of the upstream flow passage portion 10 and extends substantially in a direction perpendicular to the flow direction from the downstream end portion 11. And a portion having a constant cross-sectional area extending downstream of the enlarged flow path portion in the direction.
  • the present apparatus 1 has two gas bypass flow path portions 40. That is, in this example, the device 1 bypasses the liquid holding unit 30 from the downstream end 11 of the upstream flow path 10 to the upstream end 21 of the downstream flow path 20 on one side of the upstream flow path 10. On the other side of the first gas bypass flow path portion 40 and the upstream flow path portion 10 provided to do so, the upstream end portion 21 of the downstream flow path portion 20 extends from the downstream end portion 11 of the upstream flow path portion 10. And a second gas bypass channel 50 provided so as to bypass the liquid holding unit 30.
  • the apparatus 1 as shown in FIG. 6 was manufactured by microfabrication technology. That is, as shown in FIG. 6I, the liquid supply unit 12 and the gas supply unit 13 are formed on the surface of a substrate 100 made of PDMS (polydimethylsiloxane) having a thickness of about 4 mm by replica molding using a photoresist.
  • channel which comprises the upstream flow path part 10 containing, the downstream flow path part 20, the liquid holding
  • the device 1 was obtained by laminating and bonding a PDMS (polydimethylsiloxane) substrate (not shown) having a thickness of about 2 mm to the PDMS substrate 100.
  • FIG. 6 (ii) shows the dimensions of each part of the manufactured apparatus 1.
  • the width of the merging channel portion 14 of the upstream channel portion 10 is 600 ⁇ m
  • the width of the tip portion having a small cross-sectional area) is 120 ⁇ m
  • the width of the downstream end portion 42 of the gas bypass channel portion 40 is 600 ⁇ m
  • the upstream end portion 41 and the downstream end of the gas bypass channel portion 40 The width of the portion between the portions 42 was 500 ⁇ m
  • the width of the downstream flow path portion 20 was 600 ⁇ m.
  • the liquid holding unit 30 was a circle having a diameter of 2 mm. Moreover, all the heights of the upstream flow path part 10, the downstream flow path part 20, the liquid holding
  • Examplementation of a method using a microchannel device The method was carried out using the apparatus 1 manufactured as described above.
  • the liquid constituting the liquid plug water to which a dye for enhancing visibility was added was used. Air was used as the gas.
  • the flow of the liquid and gas in the flow path was performed by applying pressure from upstream.
  • FIG. 7 shows a result of observing the flow of the liquid plug in the apparatus 1 in the present method under a phase contrast microscope.
  • the upstream flow path section 10 is filled with the gas bypass flow path section 40 filled with the gas Gb and the liquid holding section 30 holding the main liquid plug PLm.
  • the three upstream liquid plugs PLu 1, PLu 2, PLu 3 were sequentially flowed toward the liquid holding part 30.
  • the volume of the main liquid plug PLm was about 1.5 ⁇ L.
  • the volume of each of the three upstream liquid plugs PLu1, PLu2, and PLu3 was about 100 nL.
  • the first upstream liquid plug PLu1 is caused to flow into the liquid holding unit 30, thereby fusing the first upstream liquid plug PLu1 with the main liquid plug PLm. A part of the later main liquid plug PLm was pushed out to the downstream flow path portion 20.
  • the gas Gu2 following the first upstream fluid plug PLu1 is caused to flow into the gas bypass channel 40, and a part of the gas Gb in the gas bypass channel 40 is downstream.
  • a part of the main liquid plug PLm extruded to the downstream flow path portion 20 as described above was cut to form the first downstream liquid plug PLd1.
  • the second upstream liquid plug PLu2 is caused to flow into the liquid holding unit 30 so as to fuse the second upstream liquid plug PLu2 with the main liquid plug PLm. A part of the later main liquid plug PLm was pushed out to the downstream flow path portion 20.
  • the gas Gu3 following the second upstream fluid plug PLu2 is caused to flow into the gas bypass channel 40, and a part of the gas Gb in the gas bypass channel 40 is downstream.
  • a part of the main liquid plug PLm extruded to the downstream flow path portion 20 as described above was cut to form a second downstream liquid plug PLd2.
  • the third upstream liquid plug PLu3 is caused to flow into the liquid holding unit 30 so as to fuse the third upstream liquid plug PLu3 with the main liquid plug PLm, and after the fusion. A part of the main liquid plug PLm was pushed out to the downstream flow path portion 20.
  • the gas Gu4 following the third upstream fluid plug PLu3 is caused to flow into the gas bypass channel 40, and a part of the gas Gb in the gas bypass channel 40 is downstream.
  • a part of the main liquid plug PLm pushed to the downstream flow passage portion 20 as described above was cut to form a third downstream liquid plug PLd3.
  • the three downstream liquid plugs PLd1, PLd2, and PLd3 were recovered from the downstream flow path portion 20.

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US20160024451A1 (en) 2016-01-28
US10071378B2 (en) 2018-09-11
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EP2975133A1 (en) 2016-01-20
ES2664761T3 (es) 2018-04-23

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