WO2020063864A1 - 微流控芯片系统及液滴的制备方法 - Google Patents

微流控芯片系统及液滴的制备方法 Download PDF

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Publication number
WO2020063864A1
WO2020063864A1 PCT/CN2019/108536 CN2019108536W WO2020063864A1 WO 2020063864 A1 WO2020063864 A1 WO 2020063864A1 CN 2019108536 W CN2019108536 W CN 2019108536W WO 2020063864 A1 WO2020063864 A1 WO 2020063864A1
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WO
WIPO (PCT)
Prior art keywords
generating device
droplet
microfluidic chip
collection bottle
chip system
Prior art date
Application number
PCT/CN2019/108536
Other languages
English (en)
French (fr)
Inventor
陈宜真
伍涛
将肯迈克尔
Original Assignee
青岛华大智造普惠科技有限公司
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority to SG11202103193SA priority Critical patent/SG11202103193SA/en
Priority to CN201980062474.XA priority patent/CN112969779B/zh
Priority to EP19866082.1A priority patent/EP3858972A4/en
Priority to KR1020237010587A priority patent/KR20230045109A/ko
Priority to AU2019351502A priority patent/AU2019351502B2/en
Priority to JP2021542250A priority patent/JP7197717B2/ja
Priority to CA3114261A priority patent/CA3114261A1/en
Priority to KR1020217012700A priority patent/KR102626171B1/ko
Application filed by 青岛华大智造普惠科技有限公司 filed Critical 青岛华大智造普惠科技有限公司
Priority to CN202310133635.8A priority patent/CN116139953A/zh
Priority to EP23165514.3A priority patent/EP4219678A1/en
Publication of WO2020063864A1 publication Critical patent/WO2020063864A1/zh
Priority to US17/209,463 priority patent/US20210205806A1/en
Priority to JP2022200239A priority patent/JP2023051960A/ja
Priority to US18/196,422 priority patent/US20230278034A1/en

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502769—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
    • 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
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    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
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    • 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
    • B01L3/50273—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 the means or forces applied to move the fluids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01F—MIXING, e.g. DISSOLVING, EMULSIFYING OR DISPERSING
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    • B01F25/42—Static mixers in which the mixing is affected by moving the components jointly in changing directions, e.g. in tubes provided with baffles or obstructions
    • B01F25/43—Mixing tubes, e.g. wherein the material is moved in a radial or partly reversed direction
    • B01F25/433—Mixing tubes wherein the shape of the tube influences the mixing, e.g. mixing tubes with varying cross-section or provided with inwardly extending profiles
    • B01F25/4331—Mixers with bended, curved, coiled, wounded mixing tubes or comprising elements for bending the flow
    • 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
    • B01F33/3011—Micromixers using specific means for arranging the streams to be mixed, e.g. channel geometries or dispositions using a sheathing stream of a fluid surrounding a central stream of a different fluid, e.g. for reducing the cross-section of the central stream or to produce droplets from the central stream
    • 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
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
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    • 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
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    • B01L3/502753—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation
    • 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
    • C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
    • C12M25/01—Drops
    • 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
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/025—Align devices or objects to ensure defined positions relative to each other
    • B—PERFORMING OPERATIONS; TRANSPORTING
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    • 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
    • B—PERFORMING OPERATIONS; TRANSPORTING
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    • B01L2200/06—Fluid handling related problems
    • B01L2200/0673—Handling of plugs of fluid surrounded by immiscible fluid
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    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
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    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0864—Configuration of multiple channels and/or chambers in a single devices comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
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    • B01L2400/049—Moving fluids with specific forces or mechanical means specific mechanical means and fluid pressure fluid pressure, pneumatics vacuum

Definitions

  • the present application relates to the field of single-cell sequencing, and in particular, to a microfluidic chip system and a method for preparing a droplet.
  • single-cell sequencing is often performed based on a microfluidic chip technology.
  • a single-cell sample preparation platform is required to prepare a droplet sample.
  • These platforms have been able to perform droplet sample preparation well and can be automated, but the structure of these platforms is relatively complex, not only including at least three pumps as a source of power for generating flow and generating droplets, but also other Device or system.
  • These platforms require synchronous operation of each pump to ensure the consistent flow of the three reagents, otherwise the formation of droplets or the uniformity of droplet size will be affected, and the pressure drop, continuous phase and dispersion on the droplet generation device will be affected. Phase flow rate and droplet generation frequency are not predictable, programmable, and reproducible.
  • the single-cell sample preparation platform in the prior art has a large volume, is not easy to carry, and has high requirements on the environment.
  • the present application provides a microfluidic chip system capable of solving the above problems.
  • the present application also provides a method for preparing a droplet capable of solving the above problems.
  • a microfluidic chip system is used for generating droplets.
  • the microfluidic chip system includes a droplet generating device for generating droplets; a power generating device for providing the droplet generating device with generated droplets A collection bottle for collecting droplets flowing from the droplet generation device; and a connecting device for communicating with the droplet generation device, the power generation device, and the collection bottle; a preparation platform for The liquid droplet generating device, the power generating device, and the collection bottle are fixed.
  • the droplet generation device includes a chip body, and a continuous phase inlet and a dispersed phase inlet are formed in the chip body, and the continuous phase and the dispersed phase enter the chip from the continuous phase inlet and the dispersed phase inlet. main body.
  • microfluidic chip system further includes a continuous phase reservoir and a dispersed phase reservoir, and the continuous phase reservoir and the dispersed phase reservoir are respectively fixed on the chip body or formed on the chip body. Outside the chip body.
  • the preparation platform includes a base and a droplet generating device card slot and a power generating device card slot formed on the base, and the droplet generating device and the power generating device are sequentially housed in the droplet generating device.
  • the base includes a first portion and a second portion connected to the first portion, the first portion is lower than the second portion, and the droplet generating device card slot is formed in the first portion On the part, the power generating device slot is formed on the second part.
  • the collection bottle is located outside the droplet generating device, and a collection bottle clamping slot is formed on the base, and the collection bottle is received and fixed in the collection bottle clamping slot.
  • the collection bottle is housed and fixed in the chip body, and the chip body is housed and fixed in a card slot of the droplet generation device.
  • the microfluidic chip system further includes at least one oscillating device, the oscillating device is disposed in a slot of the droplet generating device and corresponds to the inlet of the dispersed phase, and is used to apply vibration to the dispersed phase.
  • the microfluidic chip system further includes at least one oscillating device, the oscillating device is disposed on the chip body and corresponds to the dispersed phase inlet for applying vibration to the dispersed phase.
  • the power generating device includes an operation portion and a first stopper formed at an end of the operation portion
  • the preparation platform further includes a first stopper slot
  • the first stopper slot is used for The first limiting member is received and fixed to maintain the operation portion at a predetermined position.
  • the power generating device is an injection device.
  • the power generating device is a negative pressure generating device, and the negative pressure generating device is hermetically connected to the collection bottle through the connecting device.
  • the preparation platform further includes a fixed base and a dust cover reversibly connected to the fixed base, the dust cover is located on one side of the droplet generating device, and the dust cover is used for Prevent dust from entering the droplet generating device.
  • the power generating device is a positive pressure generating device, and the positive pressure generating device is sealedly connected to the continuous phase inlet and the dispersed phase inlet through the connecting device, respectively.
  • a method for preparing liquid droplets implemented by using the microfluidic chip system as described above includes the steps of measuring a predetermined amount of a continuous phase and a dispersed phase by using a pipette or a pipette, and The continuous phase is introduced into the continuous phase inlet, and the measured dispersed phase is introduced into the dispersed phase inlet; and a power generating device is started to cause a pressure difference in the collection bottle, the connecting device, and the chip body, and the pressure difference promotes The dispersed phase and the continuous phase converge and flow into a collection bottle in the form of droplets.
  • the method further includes the step of applying vibration to the dispersed phase by using at least one oscillating device.
  • the microfluidic chip system provided in this application firstly: 1) fixes the droplet generating device, the power generating device and the collection bottle to a preparation platform to ensure the stability of the droplet generated by the microfluidic chip system; 2) A single power source is applied to the droplet outlet of the droplet generating device, so that a vacuum is formed in the droplet generating device, and the cell fluid, the microbead solution and the oil phase flow synchronously in the droplet generating device, which has a high communication efficiency.
  • the first limiting groove keeps the first limiting piece of the injection device in a predetermined position to maintain its vacuum, which is low cost and easy to operate;
  • the pressure in the power generating device changes with the injection and flow of the reagent, and the reagent is filled to In the collection device, the pressure change in the power generation device also follows the ideal gas law; 5) the pressure drop on the droplet generation device, the flow rates of all continuous and dispersed phases, the size of the droplets and the generation frequency All are predictable, programmable, and reproducible; 6)
  • the microfluidic chip system is small in size and can be self-assembled. Therefore, the microfluidic chip system has lower requirements for the use environment, is easy to carry, and Can be scaled up.
  • FIG. 1 is a schematic perspective view of a microfluidic chip system according to a first embodiment of the present application.
  • FIG. 2 is a schematic diagram of an internal structure of a chip body of the microfluidic chip shown in FIG. 1.
  • FIG. 3 is a schematic diagram of a microfluidic chip, a collection bottle, and a power generating device in the microfluidic chip system shown in FIG. 1, wherein the power generating device and the chip body (a cross-sectional view taken along the line III-III described in FIG. 2) ) Is a sectional view.
  • FIG. 4 is a schematic diagram after adding the oil phase to the first reservoir of the microfluidic chip shown in FIG. 3, the cell fluid to the second reservoir, and the microbead solution to the third reservoir.
  • FIG. 5 is a schematic diagram of the microfluidic chip system shown in FIG. 4 after the power generating device is evacuated.
  • FIG. 6 is a schematic diagram of a microfluidic chip system including an oscillation device according to an embodiment of the present application.
  • FIG. 7 is a schematic diagram of droplets (excluding microbeads and cells) generated by using the microfluidic chip system provided by the present invention.
  • FIG. 8 is a schematic diagram of droplets (including microbeads and cells) generated by using the microfluidic chip system provided by the present invention.
  • FIG. 9 is a schematic diagram of an internal structure of a chip body according to a second embodiment of the present application.
  • FIG. 10 is a schematic diagram of an internal structure of a chip body according to a third embodiment of the present application.
  • FIG. 11 is a schematic diagram of a connection relationship between a microfluidic chip, a collection bottle, and a power generating device according to a fourth embodiment of the present application, where the power generating device and the chip body are in a cross-sectional state.
  • FIG. 12 is a schematic diagram of a connection relationship between a microfluidic chip, a collection bottle, and a power generating device according to a fifth embodiment of the present application, in which the power generating device and the chip body are in a cross-sectional state.
  • FIG. 13 is a schematic diagram of a connection relationship between a microfluidic chip, a collection bottle, and a power generating device according to a sixth embodiment of the present application, where the power generating device and the chip body are in a cross-sectional state.
  • FIG. 14 is a schematic diagram of a connection relationship between a microfluidic chip, a collection bottle, and a power generating device according to a seventh embodiment of the present application, in which the power generating device and the chip body are in a cross-sectional state.
  • FIG. 15 is a schematic diagram of a connection relationship between a microfluidic chip, a collection bottle, and a power generating device according to an eighth embodiment of the present application, where the power generating device and the chip body are in a cross-sectional state.
  • FIG. 16 is a schematic diagram of a connection relationship between a microfluidic chip, a collection bottle, and a power generating device according to a ninth embodiment of the present application, in which the power generating device and the chip body are in a sectional view.
  • FIG. 17 is a schematic diagram of a connection relationship between a microfluidic chip, a collection bottle, and a power generating device according to a tenth embodiment of the present application, where the power generating device and the chip body are in a cross-sectional state.
  • FIG. 18 is a schematic perspective view of a preparation platform provided by an eleventh embodiment of the present application.
  • FIG. 19 is a schematic perspective view of another state of the preparation platform shown in FIG. 18.
  • FIG. 20 is a flowchart of preparing a droplet using a microfluidic chip system without an oscillating device.
  • FIG. 21 is a flowchart of preparing a droplet using a microfluidic chip system with an oscillating device.
  • Microfluidic chip system 100 Microbead solution introduction channel 219 Preparation platform 10, 101 Confluence channel 210 Base 11 First reservoir twenty two First surface 111 Second reservoir twenty three Second surface 112 Third reservoir twenty four First side 113 First oscillating device 25 Second side 114 Second oscillating device 26 First 115 Collection bottle 30 Second part 116 Cover 31 Drop slot of the droplet generating device 12 Connecting device 40 Collection bottle slot 13 First connecting tube 41 Power generating device card slot 14 Second connecting tube 42 First Containment Department 141 Power generating device 50 Second Containment Department 142 Main body 51 First limit slot 15 Containment space 511 Second limit slot 18 Operation Department 52 Fixed seat 16 Gas inlet and outlet 53 Dust cover 17 First stop 54 Liquid droplet generating device 20 Second stop 55 Chip body 21, 110, 120 Cell fluid 60 Third surface 211 Microbead solution 70 Fourth surface 212 Oil phase 80 Oil phase inlet 213 Droplet 200 Cell fluid inlet 214 Third connecting tube 43 Microbead solution inlet 215 Fourth connection tube 44 Droplet outlet 216 Fifth connecting pipe 45 Oil phase
  • a first embodiment of the present application provides a microfluidic chip system 100, which is used to prepare a droplet 200.
  • the microfluidic chip system 100 includes a preparation platform 10, a droplet generation device 20, a collection bottle 30, a connection device 40, and a power generation device 50.
  • the preparation platform 10 is used to fix the liquid droplet generating device 20, the collection bottle 30, the connection device 40, and the power generating device 50
  • the liquid droplet generating device 20 is used to generate the liquid droplet 200
  • the collection bottle 30 The liquid droplet generating device 20 is configured to collect the liquid droplets 200 flowing out of the liquid droplet generating device 20.
  • the connecting device 40 is configured to connect the liquid droplet generating device 20 and the collection bottle 30 and / or the liquid droplet generating device 20 and the power generation device 50 and / or the power generation device 50 and the collection bottle 30.
  • “fixed” includes two cases: the droplet generation device 20, the collection bottle 30, and the power generation device 50 cannot be detached from the preparation platform 10; the droplet generation device 20, the collection bottle 30, and power The generating device 50 can be detached from the preparation platform 10. In this embodiment, the droplet generation device 20, the collection bottle 30, and the power generation device 50 can be detached from the preparation platform 10.
  • the droplet generation device 20, the collection bottle 30, and the power generation device 50 are all fixed on the preparation platform 10.
  • the connection device 40 connects the droplet generation device 20 and the collection bottle 30, and connects the collection bottle 30 and the power generation device 50.
  • the preparation platform 10 includes a base 11 and a droplet generating device card slot 12, a collection bottle card slot 13 and a power generating device card slot 14 that are opened on the base 11.
  • the droplet generating device 20, the collection bottle 30, and the power generating device 50 are sequentially housed and fixed in the droplet generating device holding slot 12, the collecting bottle holding slot 13, and the power generating device holding slot 14.
  • the droplet generating device 20, the collection bottle 30, and the power generating device 50 may also be provided or installed on the base 11 by other means, for example, a buckle or The locking device fixes the droplet generating device 20, the collection bottle 30, and the power generating device 50 on the base 11 by means of snapping or locking.
  • the base 11 includes a first surface 111, a second surface 112 opposite to the first surface 111, a first side surface 113 connecting the first surface 111 and the second surface 112, and a side opposite to the first side surface 113, respectively. ⁇ ⁇ ⁇ 114 ⁇ The second side 114.
  • the droplet generating device slot 12 and the power generating device slot 14 are formed from the first surface 111 and the first side 113 to the second surface 112 and the second side 114, respectively, and the collection bottle slot 13 It is recessed from the first surface 111 to the second surface 112.
  • the droplet generating device latching slot 12 and the power generating device latching slot 14 both penetrate the first surface 111 and the first side surface 113, and the collection bottle latching slot 13 only penetrates the first surface 111.
  • the droplet generating device latching slot 12 and the power generating device latching slot 14 both penetrate only the first surface 111.
  • the preparation platform 10 can be made by, but not limited to, a 3D printing method.
  • the preparation platform 10 is manufactured by a 3D printing method, and the cost is low and the size can be adjusted according to actual needs.
  • the power generating device slot 14 includes a first receiving portion 141 and a second receiving portion 142.
  • the first receiving portion 141 is used to receive the main body portion 51 of the power generating device 50 (see FIG. 1 and the following), and the second receiving portion 142 is used to receive the operation portion 52 of the power generating device 50 (see FIG. 1). And below).
  • the preparation platform 10 further includes a first limiting slot 15 and a second limiting slot 18.
  • the first limiting slot 15 is used for receiving and fixing the first limiting member 54 (see FIG. 1 and the following) of the power generating device 50 to maintain the operation portion 52 at a predetermined position.
  • the second limiting slot 18 is in communication with the power generating device clamping slot 14 and is used for receiving and fixing the second limiting member 55 of the power generating device 50 (see FIG. 1 and the following).
  • the first limiting groove 15 is a recess formed from the first surface 111 and the first side surface 113 to the second surface 112 and the second side surface 114 respectively.
  • the first side surface 113 may also serve as a limiting function of the first limiting groove 15, or the first limiting groove 15 is a through hole that only penetrates the first surface 111 and does not pass through. A groove on the first side 113.
  • the droplet generation device 20 includes a chip body 21.
  • the chip body 21 is provided with a continuous phase inlet and a dispersed phase inlet.
  • the chip body 21 is provided with a continuous phase introduction channel, a dispersed phase introduction channel, and a confluence channel.
  • the continuous phase inlet is in communication with the continuous phase introduction channel.
  • the dispersed phase inlet is in communication with the dispersed phase introduction channel, the continuous phase introduction channel and the dispersed phase introduction channel intersect and communicate with one end of the confluence channel.
  • a continuous phase enters the continuous phase introduction channel from the continuous phase inlet
  • a dispersed phase enters the dispersed phase introduction channel from the dispersed phase inlet
  • the continuous phase and the dispersed phase are in the continuous phase introduction channel and the dispersed phase introduction channel.
  • the dispersed phase is introduced into a confluence at the intersection of the channels and enters the confluence channel.
  • the continuous phase may be equal to oil, and the dispersed phase may be cell fluid, microbead fluid, water, and the like.
  • the continuous phase is an oil phase
  • the dispersed phase is a cell fluid and a microbead fluid.
  • the chip body 21 includes a third surface 211 and a fourth surface 212 opposite to the third surface 211.
  • An oil phase inlet 213 continuous phase inlet
  • a cell liquid inlet 214 disersed phase inlet
  • a microbead solution inlet 215 disersed phase inlet
  • a droplet outlet 216 are recessed from the third surface 211 to the fourth surface 212, respectively.
  • the oil phase inlet 213, the cell liquid inlet 214, the microbead solution inlet 215, and the droplet outlet 216 only penetrate the third surface 211. It can be understood that, in other embodiments, the droplet outlet 216 may also be disposed on one of the sides connecting the third surface 211 and the fourth surface 212.
  • the chip main body 21 also has an oil phase introduction channel 217 (continuous phase introduction channel), a cell fluid introduction channel 218 (dispersed phase introduction channel), and a microbead solution introduction channel 219 (dispersed phase introduction channel).
  • the oil phase inlet 213 is in communication with the droplet outlet 216 through the oil phase introduction channel 217
  • the cell liquid inlet 214 is in communication with the droplet outlet 216 through the cell liquid introduction channel 218.
  • the bead solution inlet 215 is in communication with the droplet outlet 216 through the bead solution introduction channel 219.
  • the chip body 21 is also formed with a confluence channel 210, the oil phase introduction channel 217, cell fluid introduction channel 218, and microbead solution introduction channel 219 meet at one end of the confluence channel 210, and the confluence channel 210 The other end is in sealed communication with the droplet outlet 216.
  • the chip body 21 is further formed with two oil phase introduction channels 217, two cell fluid introduction channels 218, and a microbead solution introduction channel 219.
  • two of the oil phase introduction channels 217 surround the cell fluid inlet 214 and two of the cell fluid introduction channels 218, and two of the cell fluid introduction channels 218 surround the microbead solution inlet 215 and all The microbead solution introduction channel 219 is provided.
  • the two oil phase introduction channels 217 and the two cell fluid introduction channels 218 are symmetrically distributed at The microbead solution is introduced on both sides of the channel 219.
  • the oil phase inlet 213, the cell fluid inlet 214, and the microbead solution inlet 215 are distributed on the same straight line.
  • the oil phase inlet 213, the cell liquid inlet 214, the microbead solution inlet 215, and the droplet outlet 216 are not distributed on the same line.
  • the microfluidic chip system 100 may further include a continuous phase storage tank and a dispersed phase storage tank.
  • the continuous phase storage tank is in communication with the continuous phase inlet
  • the dispersed phase storage tank is in communication with the continuous phase inlet.
  • the inlet of the dispersed phase is in communication.
  • the continuous phase reservoir and the dispersed phase reservoir may be located on the chip body 21 or outside the chip body 21.
  • the microfluidic chip system 100 further includes a first storage tank 22 (continuous phase storage tank) and a second storage tank 23 (dispersed phase storage tank). ) And the third storage tank 24 (dispersed phase storage tank).
  • the first storage tank 22 is in communication with the oil phase inlet 213, the second storage tank 23 is in communication with the cellular liquid inlet 214, and the third storage tank 24 is in communication with the microbead solution inlet 215.
  • the first liquid storage tank 22, the second liquid storage tank 23, and the third liquid storage tank 24 are located on the chip body 21.
  • the chip body 21 is integrally formed with the first liquid storage tank 22, the second liquid storage tank 23, and the third liquid storage tank 24.
  • the first storage tank 22, the second storage tank 23, and the third storage tank 24 may also be directly connected to the oil phase inlet 213, the cell liquid inlet 214, and the microbead solution inlet.
  • the first liquid storage tank 22 is used to temporarily store the oil phase 80.
  • the storage volume of the first liquid storage tank 22 may be, but is not limited to, 100 microliters, 200 microliters, 500 microliters, and 1 milliliter.
  • the second liquid storage tank 23 is used to temporarily store the cell liquid 60.
  • the storage volume of the second liquid storage tank 23 may be, but is not limited to, 100 microliters, 200 microliters, 500 microliters, and 1 milliliter.
  • the third liquid storage tank 24 is used to temporarily store the microbead solution 70.
  • the storage volume of the third liquid storage tank 24 may be, but is not limited to, 100 microliters, 200 microliters, 500 microliters, and 1 milliliter.
  • the microbead solution 70 may be one of a polystyrene microbead solution, a resin microbead solution, or a magnetic bead solution.
  • an opening (not shown) is formed in each of the first storage tank 22, the second storage tank 23, and the third storage tank 24, so as to facilitate the oil phase 80, the cell liquid 60, and the microbeads.
  • the solution 70 is transferred into the first liquid storage tank 22, the second liquid storage tank 23, and the third liquid storage tank 24.
  • the volume ratio of the oil phase 80, the cell liquid 60, and the microbead solution 70 in the first storage tank 22, the second storage tank 23, and the third storage tank 24 may be, but is not limited to, 2 : 1: 1.
  • the cell liquid 60 is 200 microliters
  • the microbead solution 70 is 200 microliters
  • the oil phase 80 is 400 microliters.
  • the liquid droplet generating device 20 may not include the first liquid storage tank 22, the second liquid storage tank 23, and the third liquid storage tank 24.
  • the oil phase 80 can be directly added to the oil phase inlet 213, the cell liquid 60 can be directly added to the cell liquid inlet 214, and the microbead solution 70 can be directly added to the microbead solution. Entrance 215.
  • the collection bottle 30 is used to collect the liquid droplets 200.
  • the collection bottle 30 includes a cover body 31, and two through holes (not shown) are formed in the cover body 31.
  • the lid body 31 is detachably connected to the body of the collection bottle 30 or is inseparable.
  • the volume of the collection bottle 30 may be, but is not limited to, 1 ml and 5 ml. In this embodiment, the volume of the collection bottle 30 is 1 ml.
  • connection device 40 includes a first connection pipe 41 and a second connection pipe 42.
  • One end of the first connection pipe 41 is in communication with the power generating device 50 and the other end is in communication with the collection bottle 30.
  • One end of the second connection pipe 42 is connected to the chip body 21 and is connected to the chip body 21.
  • the droplet outlet 216 is in communication, and the other end is in communication with the collection bottle 30.
  • one end of the second connection pipe 42 is connected to the third surface 211, and the other end is connected to a through hole of the cover 31 of the collection bottle 30 and communicates with the collection bottle. 30 connected.
  • One end of the first connection pipe 41 is connected to the power generating device 50, and the other end is connected to another through hole of the cover 31 and communicates with the collection bottle 30.
  • first connection pipe 41 and the second connection pipe 42 are both 10 cm long hoses.
  • the power generating device 50 is configured to provide the liquid droplet generating device 20 with power for generating the liquid droplets 200 so as to be located in the first liquid storage tank 22, the second liquid storage tank 23, and the third liquid storage tank 24.
  • the oil phase 80, the cell liquid 60, and the microbead solution 70 flow in the oil phase introduction channel 217, the cell liquid introduction channel 218, and the microbead solution introduction channel 219 of the droplet generation device 20, respectively, and then, in the confluence channel 210, Confluence of the two forms a droplet 200 that flows out of the droplet outlet 216 and further flows into the collection bottle 30.
  • the power generating device 50 is a negative pressure generating device.
  • the power generating device 50 includes a main body portion 51, an operation portion 52, and a gas inlet and outlet end 53.
  • the main body portion 51 includes a receiving space 511, one end of the operation portion 52 is received in the receiving space 511, and the other end protrudes out of the main body portion 51.
  • One end of the operation portion 52 accommodated in the accommodation space 511 is closely adhered to the inner wall of the accommodation space 511.
  • the gas inlet / outlet end 53 is fixed to an end of the main body portion 51 away from the exposed operation portion 52 and communicates with the accommodation space 511.
  • An end of the second connection pipe 42 remote from the collection bottle 30 is connected to the gas inlet and outlet end 53.
  • the power generating device 50 is an injection device.
  • the power generating device 50 further includes a first limiter 54 and a second limiter 55.
  • the first limiter 54 is disposed on the operation portion 52 and is exposed on the main body portion 51. The end.
  • the first limiting member 54 cooperates with the first limiting slot 15 to pull the operation portion 52 to a fixed position to maintain a vacuum inside the power generating device 50.
  • the second stopper 55 is fixed on the main body portion 51, and the second stopper 55 cooperates with the second stopper groove 18 to limit the position of the second stopper 55 during the process of pulling the operation portion 52. The movement of the main body portion 51 will be described.
  • P1 and P2 are the pressures corresponding to the initial volume V1 and the volume V2, respectively, at this time, the second connection pipe 42, the collection bottle 30, the first connection pipe 41, and the chip body
  • the pressure in the cell 24 is lower than the atmospheric pressure,
  • the power of the power generating device 50 can be generated in the following ways: 1) it is generated in a closed container before being connected to the chip body 21; 2) the power generating device 50 is driven by a pump; 3) through This is done by pulling or pushing the plunger of a syringe (or similar device).
  • the microfluidic chip system 100 may further include at least one oscillating device, and the oscillating device is corresponding to the third liquid storage tank and / or the second liquid storage tank, and is configured to apply vibration to the liquid storage device.
  • the microfluidic chip system 100 further includes at least one oscillating device, and the oscillating device is used for applying vibration to the dispersed phase.
  • the microfluidic chip system 100 further includes a first oscillation device 25 and a second oscillation device 26.
  • the first oscillating device 25 and the second oscillating device 26 are fixed in the card slot 12 of the droplet generating device and correspond to the cell liquid inlet 214 and the microbead solution inlet 215, respectively.
  • the first oscillating device 25 and the second oscillating device 26 may both be ceramic oscillating plates.
  • the first oscillating device 25 and the second oscillating device 26 generate vibrations after startup, and the vibrations are applied to the cell fluid 60 and the microbead solution 70, which can effectively improve the cell fluid 60 and the microbead solution 70.
  • the problem of sedimentation of cells or microbeads during the experiment of the bead solution 70 can ensure the consistency of the concentration of the cell solution 60 and the microbead solution 70.
  • the first oscillating device 25 and the second oscillating device 26 may be located on the chip body 21.
  • the first oscillating device 25 and the second oscillating device 26 are located on the fourth surface of the chip body 21. 212 and corresponding to the cell fluid inlet 214 and the microbead solution inlet 215, respectively, or located on the side wall connecting the third surface 211 and the fourth surface 212 of the chip body 21 and corresponding to the cell fluid inlet 214 and the microbead solution inlet 215 location. It is only necessary that the vibration energy generated after the first oscillation device 25 and the second oscillation device 26 is activated is applied to the cell fluid 60 and the microbead solution 70 to improve the cell fluid 60 and the microbead solution 70. Cell or bead sedimentation problem.
  • the microfluidic chip system 100 may include only one oscillating device, for example, only the second oscillating device 26, and the second oscillating device 26 may be fixed in a slot of the droplet generating device.
  • the second oscillating device 26 may also be fixed on the fourth surface 212 of the chip body 21 and corresponds to the microbead solution inlet 215. Therefore, the bead sedimentation problem generated during the experiment of the bead solution 70 can be effectively improved, and the consistency of the concentration of the bead solution 70 can be ensured.
  • the microfluidic chip system 100 includes only one oscillating device.
  • the oscillating device is provided corresponding to the cell fluid inlet 214 and the microbead solution inlet 215, and applies vibration to the cell fluid 60 and the microbeads at the same time. On solution 70.
  • a vacuum is formed inside the power generating device 50.
  • the vacuum will cause the dispersed phase and the continuous phase to flow in the chip body 10, because an injection device is used as
  • the power generating device can generate a vacuum with a simple action.
  • reagents dispensersed phase and continuous phase
  • the volume of air in the power generation device will expand or compress in a positive or negative pressure system, which will cause the chip inlet (continuous phase)
  • the pressure drop at the inlet and the dispersed phase inlet and the outlet (droplet outlet) decreases, which causes the flow rate of the reagent to gradually decrease, and changes in the flow rate of the reagent will cause changes in droplet size. Therefore, the pressure drop in the chip body 10, the flow rates of the dispersed and continuous phases, the droplet size, and the generation frequency are all predictable, programmable, and reproducible.
  • FIG. 7 is a schematic diagram of droplets (excluding microbeads and cells) generated by using the microfluidic chip system provided by the present invention.
  • the droplet size variation of the droplet generated by using the microfluidic chip system 100 provided by the present invention is small, and the uniformity is good.
  • the size of the droplet is 55.7um, and the size change of the droplet is 2%.
  • FIG. 8 is a schematic diagram of a droplet (including microbeads and cells) generated by using the microfluidic chip system 100 provided by the present invention.
  • the microfluidic chip system provided by the present invention can encapsulate the microbeads and cells in the oil phase.
  • FIG. 9 is a schematic diagram of an internal structure of a chip body 110 according to a second embodiment of the present application.
  • the internal structure of the chip body 110 is basically the same as the internal structure of the chip body 21 provided in the first embodiment of the present application, and the only difference is that two oil phase introduction channels 217 are formed in the chip body 110, One cell fluid is introduced into the channel 218 and two microbead solutions are introduced into the channel 219.
  • two of the oil phase introduction channels 217 surround the microbead solution inlet 215 and two of the microbead solution introduction channels 219, and two of the microbead solution introduction channels 219 surround the cell fluid inlet 214.
  • one cell fluid introduction channel 218 is provided.
  • the two oil phase introduction channels 217 and the two microbead solution introduction channels 219 are symmetrically distributed in all places.
  • the microbead solution is introduced to both sides of the channel 219.
  • the oil phase inlet 213, the cell liquid inlet 214, the microbead solution inlet 215, and the droplet outlet 216 are distributed on the same straight line.
  • FIG. 10 is a schematic diagram of an internal structure of a chip body 120 according to a third embodiment of the present application.
  • the internal structure of the chip body 120 is basically the same as the internal structure of the chip body 21 provided in the first embodiment of the present application. The only difference is that two oil phase introduction channels 217 are formed in the chip body 120. A cell fluid is introduced into the channel 218 and a microbead solution is introduced into the channel 219. Specifically, two of the oil phase introduction channels 217 surround the microbead solution inlet 215, one of the microbead solution introduction channel 219, the cell fluid inlet 214, and one cell fluid introduction channel 218.
  • One of the cell fluid introduction channels 218 and one of the microbead solution introduction channels 219 are symmetrically distributed, and the two oil phase introduction channels 217 are distributed in one of the cell fluid introduction channels 218 and one of the microbead solution introduction channels 219 On both sides.
  • the oil phase inlet 213 and the droplet outlet 216 are distributed on the same straight line, and the oil phase inlet 213, the cell liquid inlet 214, the microbead solution inlet 215, and the droplet outlet 216 are not distributed. On the same line.
  • FIG. 11 is a schematic diagram of a connection relationship between a chip body 21, a collection bottle 30, and a power generating device 50 according to a fourth embodiment of the present application, where the power generating device 50 is in a cross-sectional state.
  • the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 is basically the same as the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 shown in FIG.
  • first liquid storage tank 22, the second liquid storage tank 23, and the third liquid storage tank 24 are formed outside the chip body 21 and pass through a third connection pipe 43 and a fourth connection pipe 44
  • a fifth connecting tube 45 is in sealed communication with the oil phase inlet 213, the cell fluid inlet 214, and the microbead solution inlet 215, respectively.
  • FIG. 12 is a schematic diagram of a connection relationship between a chip body 21, a collection bottle 30, and a power generating device 50 according to a fifth embodiment of the present application, where the power generating device 50 is a cross-sectional state.
  • the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 is basically the same as the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 shown in FIG.
  • the droplet generating device 20 further includes a convex portion 27 formed on the chip body 21, the collection bottle 30 is placed in the convex portion 27, the convergence channel 210 and the collection
  • the bottle 30 is in sealed communication, and the collection bottle 30 is in sealed communication with the power generating device 50 through the first connection pipe 41.
  • FIG. 13 is a schematic diagram of a connection relationship between a chip body 21, a collection bottle 30, and a power generating device 50 according to a sixth embodiment of the present application, where the power generating device 50 is a cross-sectional state.
  • the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 is basically the same as the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 shown in FIG.
  • the only difference is that the first reservoir 22, the second reservoir 23, and the third reservoir 24 are formed on the chip body 21, and the first reservoir 22 is in sealed communication with the oil phase inlet 213
  • the second storage tank 23 is in sealed communication with the cell fluid inlet 214, and the third storage tank 24 is in sealed communication with the bead solution inlet 215.
  • FIG. 14 is a schematic diagram of a connection relationship between the chip body 21, the collection bottle 30, and the power generating device 50 according to the seventh embodiment of the present application, where the power generating device 50 is a cross-sectional state.
  • the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 is basically the same as the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 shown in FIG.
  • the power generating device 50 is a positive pressure generating device.
  • the power generating device 50 is an injection device.
  • the power generating device 50 is in communication with the first liquid storage tank 22, the second liquid storage tank 23, and the third liquid storage tank 24 through the first connection pipe 41, respectively.
  • the first connecting pipe 41 is a multi-joint bifurcated connecting pipe.
  • the first connection pipe 41 includes a main connection section 411, a first branch connection section 412, a second branch connection section 413, and a third branch connection section 414.
  • the main connection section 411 One end of the power generating device 50 is connected to the gas inlet and outlet end 53 of the power generating device 50 and is in sealed communication, and the other end of the main connection section 411 is respectively connected to the first branched connection section 412 and the second branched connection section 413 and the third bifurcated connecting section 414 are connected and hermetically communicated.
  • FIG. 15 is a schematic diagram of the connection relationship between the chip body 21, the collection bottle 30, and the power generating device 50 according to the eighth embodiment of the present application, where the power generating device 50 is a cross-sectional state.
  • the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 is basically the same as the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 shown in FIG.
  • the only difference is that the first liquid storage tank 22, the second liquid storage tank 23, and the third liquid storage tank 24 are formed outside the chip body 21 and pass through a third connection pipe 43 and a fourth connection pipe 44
  • a fifth connecting tube 45 is in sealed communication with the oil phase inlet 213, the cell fluid inlet 214, and the microbead solution inlet 215, respectively.
  • FIG. 16 is a schematic diagram of the connection relationship between the chip body 21, the collection bottle 30, and the power generating device 50 according to the ninth embodiment of the present application, where the power generating device 50 is a cross-sectional state.
  • the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 is basically the same as the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 shown in FIG.
  • the droplet generating device 20 further includes a convex portion 27 formed on the chip body 21, the collection bottle 30 is placed in the convex portion 27, the convergence channel 210 and the collection Bottle 30 is hermetically communicated.
  • FIG. 17 is a schematic diagram of the connection relationship between the chip body 21, the collection bottle 30, and the power generating device 50 according to the tenth embodiment of the present application, where the power generating device 50 is a cross-sectional state.
  • the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 is basically the same as the connection relationship between the chip body 21, the collection bottle 30, and the power generation device 50 shown in FIG.
  • the only difference is that the first reservoir 22, the second reservoir 23, and the third reservoir 24 are formed on the chip body 21, and the first reservoir 22 is in sealed communication with the oil phase inlet 213
  • the second storage tank 23 is in sealed communication with the cell fluid inlet 214, and the third storage tank 24 is in sealed communication with the bead solution inlet 215.
  • FIGS. 18-19 are schematic perspective views of the preparation platform 101 provided by the eleventh embodiment of the present application.
  • the structure of the preparation platform 101 is similar to the structure of the preparation platform 10 in the first embodiment, and the only difference is that when the pressure generating device is a negative pressure generating device, the preparation
  • the base 11 of the platform 101 includes a first portion 115 and a second portion 116 connected to the first portion 115.
  • the first portion 115 is lower than the second portion 116, and the droplet generating device is stuck in a slot.
  • 12 is formed on the first portion 115, and the power generating device slot 14 is formed on the second portion 116.
  • the droplet generating device locking slot 12 may be omitted, and the droplet generating device 20 may be directly fixed to the first portion 115.
  • the collection bottle clamping groove 13 is also formed on the second portion 116.
  • the preparation platform 101 further includes a fixing base 16 fixed on the first part 115 and a dust cover 17 reversibly connected to the fixing base 16.
  • the dust cover 17 is located on one side of the chip body 21.
  • the dust cover 17 is used to prevent dust from entering the chip body 21. Specifically, the dust cover 17 is located above the continuous phase inlet and / or the dispersed phase inlet, or is located above the continuous phase reservoir and / or the dispersed phase reservoir.
  • the preferred embodiment of the present application further provides a method for preparing droplets.
  • the method can be performed in combination with the microfluidic chip system 100 or a similar system described above.
  • the method includes the following steps: step:
  • step S801 a predetermined amount of a continuous phase and a dispersed phase are respectively measured by a pipette or a pipette, the measured continuous phase is introduced into the continuous phase inlet, and the measured dispersed phase is introduced into the Said dispersed phase inlet.
  • the step S801 may be expressed as: using a pipette or a pipette to measure a predetermined amount of the oil phase 80, the cell liquid 60, and the microbead solution 70, respectively, and measure the measured oil phase 80 is introduced into the oil phase inlet 213 of the microfluidic chip system 100, the measured cell fluid 60 is introduced into the cell fluid inlet 214, and the measured bead solution 70 is introduced into the bead solution inlet 215.
  • the step S801 can be expressed as: measuring a predetermined amount of continuous phase and For the dispersed phase, the measured continuous phase is introduced into the continuous phase storage tank, and the measured dispersed phase is introduced into the dispersed phase storage tank.
  • step S801 is specifically: using a pipette
  • a predetermined amount of the oil phase 80, the cell liquid 60, and the microbead solution 70 are respectively measured by a gun or a pipette, and the measured oil phase 80 is introduced into the first storage tank 22, and the measured cell liquid 60 is introduced into the second In the liquid storage tank 23, the measured bead solution 70 is introduced into the third liquid storage tank 24, and the oil phase 80 is introduced into the oil phase inlet 213 through the first liquid storage tank 22, and the cell liquid 60 is introduced into the cell fluid inlet 214 through the second storage tank 23, and the microbead solution 70 is introduced into the microbead solution inlet 215 through the third storage tank 24.
  • the cell liquid 60 is 200 microliters
  • the microbead solution 70 is 200 microliters
  • the oil phase 80 is 400 microliters.
  • the order of adding the cell liquid 60, the microbead solution 70, and the oil phase 80 may be: firstly add the cell liquid 60, then the microbead solution 70, and finally the oil phase 80; or it may be: first add the microbead solution 70. Add cell liquid 60, and finally add oil phase 80; it is also possible to add the cell liquid 60, microbead solution 70, and oil phase 80 at the same time.
  • step S802 the power generating device 50 is started, so that a pressure difference is formed between the pressure in the collection bottle 30, the connection device 40, the chip body 21 and the atmospheric pressure, and the pressure difference causes the dispersed phase and the continuous phase to converge and form droplets.
  • the form flows into the collection bottle 30.
  • step S802 may be: activating the power generating device 50 to enable the collection bottle 30, the second connection tube 42 and the droplet outlet 216 in the chip body 21, the convergence channel 210, at least one dispersed phase introduction channel, and the dispersed phase
  • a pressure difference is formed between the pressure in the inlet and the continuous phase inlet and the atmospheric pressure, and the pressure difference causes the continuous phase and the dispersed phase to flow toward the convergence channel 210, and merges to form a droplet 200 at the convergence channel 210.
  • the droplet 200 is under the effect of the pressure difference Further, it flows into the collection bottle 30 through the droplet outlet 216 and the second connection pipe 42.
  • the step S802 can be expressed as: activating the power generating device 50, so that the collection bottle 30, the second connection tube 42, and the droplet outlet 216, the confluence channel 210, and the microbead solution introduction channel in the chip body 21 219.
  • the cell fluid introduction channel 218 and the oil phase introduction channel 217, the microbead solution inlet 215, the cell liquid inlet 214, and the oil phase inlet 213 form a pressure difference between atmospheric pressure and atmospheric pressure, and the pressure difference promotes the cell liquid 60,
  • the microbead solution 70 and the oil phase 80 flow to the confluence channel 210, and merge at the confluence channel 210 to form a droplet 200.
  • the droplet 200 further flows into the collection bottle 30 through the droplet outlet 216 and the second connection pipe 42 under the effect of the pressure difference.
  • step S802 may For: starting the power generating device 50 to enable the collection bottle 30, the second connection pipe 42, and the droplet outlet 216, the confluence channel 210, the microbead solution introduction channel 219, the cell fluid introduction channel 218, and the oil phase introduction in the chip body 21
  • the pressure difference between the pressure in the channel 217, the third storage tank 24, the second storage tank 23, and the first storage tank 22 and the atmospheric pressure forms a pressure difference, which promotes the cell fluid 60, the microbead solution 70, and the oil.
  • the phase 80 flows to the confluence channel 210, and merges to form a droplet 200 at the confluence channel 210.
  • the droplet 200 further flows into the collection bottle 30 through the droplet outlet 216 and the second connection pipe 42 under the effect of the pressure difference.
  • the power generating device 50 is a negative pressure generating device, so step S802 is specifically: activating the power generating device 50 to make the liquid droplets in the collection bottle 30, the second connecting pipe 42 and the chip body 21
  • step S802 is specifically: activating the power generating device 50 to make the liquid droplets in the collection bottle 30, the second connecting pipe 42 and the chip body 21
  • the outlet 216, the microbead solution introduction channel 219, the cell fluid introduction channel 218, and the oil phase introduction channel 217 generate a negative pressure, which promotes the cell fluid 60, the microbead solution 70, and the oil phase 80 to flow to the confluence channel 210.
  • a droplet 200 is formed at the confluence channel 210, and the droplet 200 further flows into the collection bottle 30 through the droplet outlet 216 and the second connection pipe 42 under the effect of the pressure difference.
  • the specific operation of generating the negative pressure is as follows: the operating portion 52 of the power generating device 50 is pulled to a preset position and the first limiting member 54 is fixed
  • step S802 is specifically: activating the power generating device 50 so that the dispersed phase introduction channel, the continuous phase introduction channel, and the liquid in the chip body 21 are introduced.
  • a positive pressure is generated in the drip outlet 216 and the collection bottle 30.
  • the positive pressure causes the dispersed phase and the continuous phase to flow toward the confluence channel 210, and merges at the confluence channel 210 to form a droplet 200.
  • the droplet 200 is further affected by the positive pressure. It flows into the collection bottle 30 through the droplet outlet 216.
  • the preparation method may further include step S900 before starting the power generating device 50: using the first oscillation device 25 and the second oscillation device 26 to apply vibration to the microbead solution 70 and the cell fluid 60, for example, turning on a power switch (not shown) provided on the base 11 of the first oscillating device 25 and the second oscillating device 26, the first oscillating device 25 and the first The two oscillating devices 26 start to vibrate to apply vibration to the microbead solution 70 and the cell solution 60.
  • step S900 before starting the power generating device 50: using the first oscillation device 25 and the second oscillation device 26 to apply vibration to the microbead solution 70 and the cell fluid 60, for example, turning on a power switch (not shown) provided on the base 11 of the first oscillating device 25 and the second oscillating device 26, the first oscillating device 25 and the first The two oscillating devices 26 start to vibrate to apply vibration to the microbead solution 70 and the cell solution 60.
  • step S900 can also be expressed as: using at least one oscillating device to apply vibration to the microbead solution 70 and / or cell fluid 60 on.
  • the step S900 may also be expressed as: applying vibration to the dispersed phase by using the first oscillation device 25 and the second oscillation device 26, for example, turning on the first oscillation device 25 and the first oscillation device 25;
  • a power switch (not shown) is provided on the two oscillating devices 26 on the base 11. The first oscillating device 25 and the second oscillating device 26 start to vibrate to apply vibration to the dispersed phase.
  • the microfluidic chip system provided in this application firstly: 1) fixes the droplet generating device, the power generating device and the collection bottle to a preparation platform to ensure the stability of the droplet generated by the microfluidic chip system; 2) A single power source is applied to the droplet outlet of the droplet generating device, so that a vacuum is formed in the droplet generating device, and the cell fluid, the microbead solution and the oil phase flow synchronously in the droplet generating device, which has a high communication efficiency.
  • the first limit groove of the first position of the injection device maintains the first limit of the injection device to maintain its vacuum, low cost and easy operation;
  • the pressure in the power generating device changes with the injection and flow of the reagent, the reagent Filled into the collection device, the pressure change in the power generation device also follows the ideal gas law; 5) pressure drop on the droplet generation device, the flow rates of all continuous and dispersed phases, the size and generation of droplets Frequency is predictable, programmable and reproducible; 6) Oscillating device configured in the microfluidic chip system can effectively improve cells and / or microbeads generated during the experiment The sedimentation of the beads can ensure the consistency of the concentration of the cell fluid and / or the pico bead solution.
  • the microfluidic chip system provided in this application can prepare 250,000 droplets in 2 minutes, which means that 10,000 single-cell target products can be obtained, that is, one droplet contains one magnetic bead and one cell. And the droplet generation time can be adjusted according to actual needs to control the target product amount of the reagent.
  • the volume of the entire microfluidic chip system is only the size of an iPad mini produced by Apple, and users can greatly expand their use space.

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Abstract

本申请公开了一种微流控芯片系统，其用于产生液滴，所述微流控芯片系统包括液滴生成装置，其用于生成液滴；动力产生装置，其用于给所述液滴生成装置提供生成液滴的动力；收集瓶，其用于收集从所述液滴生成装置流出的液滴；连接装置，其用于连通所述液滴生成装置、所述动力产生装置及所述收集瓶；及制备平台，其用于固定所述液滴生成装置、所述动力产生装置及所述收集瓶。本申请还公开了一种液滴的制备方法。

Description

微流控芯片系统及液滴的制备方法 技术领域
本申请涉及单细胞测序领域，尤其涉及一种微流控芯片系统及液滴的制备方法。
背景技术
单细胞基因组学领域近年来发展得非常迅速，为人们揭示了复杂生物学体系的许多重要线索，包括微生物群落的生态多样性和人类癌症的基因组。
现有技术中常采用基于微流控芯片技术进行单细胞测序，基于微流控芯片技术需要通过配套单细胞样品制备平台进行液滴样品的制备。这些平台已经能够很好的进行液滴样品的制备且能够实现自动化，但这些平台的结构比较复杂，不仅包括至少三个泵作为产生流动和产生液滴的动力来源，还包括与泵配合的其他的装置或系统。这些平台需要各个泵之间同步作动，才能保证三种试剂的流动一致，否则将会影响液滴的形成或影响液滴的尺寸均一性，液滴生成装置上的压降、连续相与分散相的流速及液滴的生成频率可预测性、可编程性和可重现性不高。
另外，现有技术中的单细胞样品制备平台的体积较大，不便于携带，对环境的要求较高。
发明内容
有鉴于此，本申请提供一种能够解决上述问题的微流控芯片系统。
另外，本申请还提供一种能够解决上述问题的液滴的制备方法。
一种微流控芯片系统，用于产生液滴，所述微流控芯片系统包括液滴生成装置，用于生成液滴；动力产生装置，用于给所述液滴生成装置提供生成液滴的动力；收集瓶，用于收集从所述液滴生成装置流出的液滴；及连接装置，用于连通所述液滴生成装置、所述动力产生装置及所述收集瓶；制备平台，用于固定所述液滴生成装置、所述动力产生装置及所述收集瓶。
进一步地，所述液滴生成装置包括一芯片主体，所述芯片主体内形成有连续相入口及分散相入口，所述连续相及分散相从所述连续相入口及分散相入口进入所述芯片主体。
进一步地，所述微流控芯片系统还包括连续相储液池及分散相储液池，所述连续相储液池及分散相储液池分别固定在所述芯片主体上或形成在所述芯片主体外。
进一步地，所述制备平台包括底座及形成在所述底座上的液滴生成装置卡槽及动力产生装置卡槽，所述液滴生成装置及所述动力产生装置依次收容在所述液滴生成装置卡槽及所述动力产生装置卡槽内。
进一步地，所述底座包括第一部及与所述第一部连接的第二部，所述第一部低于所述第二部，所述液滴生成装置卡槽形成在所述第一部上，所述动力产生装置卡槽形成在所述第二部上。
进一步地，所述收集瓶位于所述液滴生成装置外，所述底座上还形成有一收集瓶卡槽，所述收集瓶收容并固定在所述收集瓶卡槽内。
进一步地，所述收集瓶收容并固定在所述芯片主体内，所述芯片主体收容并固定在所述液滴生成装置卡槽内。
进一步地，所述微流控芯片系统还包括至少一振荡装置，所述振荡装置设置在所述液滴生成装置卡槽内且对应所述分散相入口，用于施加振动于分散相。
进一步地，所述微流控芯片系统还包括至少一振荡装置，所述振荡装置设置在所述芯片主体上且对应所述分散相入口，用于施加振动于所述分散相。
进一步地，所述动力产生装置包括一操作部及一形成在所述操作部末端的第一限位件，所述制备平台还包括一第一限位槽，所述第一限位槽用于收容并固定所述第一限位件，以将所述操作部保持在预定位置处。
进一步地，所述动力产生装置为注射装置。
进一步地，所述动力产生装置为负压产生装置，所述负压产生装置通过所述连接装置与所述收集瓶密封连接。
进一步地，所述制备平台还包括一固定座及一可翻转连接在所述固定座上的防尘盖，所述防尘盖位于所述液滴生成装置一侧，所述防尘盖用于防止灰尘进入所述液滴生成装置内。
进一步地，所述动力产生装置为正压产生装置，所述正压产生装置通过所述连接装置分别与所述连续相入口及所述分散相入口密封连接。
一种利用如上所述的微流控芯片系统来实施的液滴的制备方法，包括步骤：通过移液枪或移液管分别量取一预定量的连续相及分散相，将量取的所述连续相导入所述连续相入口内，将量取的所述分散相导入所述分散相入口内；及启动动力产生装置，使收集瓶、连接装置及芯片主体内产生压强差，压强差促使所述分散相及所述连续相汇流并以液滴的形式流入收集瓶内。
进一步地，在启动所述动力产生装置之前，还包括步骤：利用至少一振荡装置施加振动于所述分散相上。
本申请提供的微流控芯片系统，首先，1)将液滴生成装置、动力产生装置及收集瓶均固定到一制备平台上，以保证所述微流控芯片系统生成液滴的稳定性；2)将单一动力源施加至液滴生成装置的液滴出口处，使液滴生成装置中形成真空，促使细胞液、微珠溶液与油相在液滴生成装置中同步流动，具有较高通量，生成液滴的尺寸均一性较好；3)进一步以注射装置作为单一动力源并根据理想气体定律P 1V 1＝P 2V 2来控制注射装置操作部前后的位置且以底座上的第一限位槽将注射装置的第一限位件保持在预定位置以保持其真空，成本低且易于操作；4)动力 产生装置中的压力随着试剂的注入和流动发生变化，试剂填充到收集装置中，动力产生装置中压力的变化也遵循理想气体定律；5)液滴生成装置上的压降，所有连续相与分散相的流速，液滴的尺寸和生成频率都是可预测的、可编程的和可重现的；6)所述微流控芯片系统的体积较小且可自行组装，因此，微流控芯片系统对使用环境要求较低、便于携带且可规模化生产。
附图说明
图1为本申请第一实施方式提供的一种微流控芯片系统的立体示意图。
图2是图1所示的微流控芯片的芯片主体的内部构造示意图。
图3是图1所示的微流控芯片系统中的微流控芯片、收集瓶及动力产生装置的示意图，其中动力产生装置及芯片本体(沿图2所述的剖线III-III的剖视图)为剖视状态。
图4是在图3所示的微流控芯片的第一储液池中加入油相、第二储液池中加入细胞液、第三储液池中加入微珠溶液后的示意图。
图5是图4所示的微流控芯片系统在动力产生装置抽真空后的示意图。
图6是本申请实施例提供的一种包含振荡装置的微流控芯片系统的示意图。
图7为采用本发明提供的微流控芯片系统生成的液滴(不包含微珠和细胞)的示意图。
图8为采用本发明提供的微流控芯片系统生成的液滴(包含微珠和细胞)的示意图。
图9是本申请第二实施方式提供的芯片主体的内部构造示意图。
图10是本申请第三实施方式提供的芯片主体的内部构造示意图。
图11是本申请第四实施方式提供的微流控芯片、收集瓶及动力产生装置的连接关系示意图，其中动力产生装置及芯片本体均为剖视状态。
图12是本申请第五实施方式提供的微流控芯片、收集瓶及动力产生装置的连接关系示意图，其中动力产生装置及芯片本体均为剖视状态。
图13是本申请第六实施方式提供的微流控芯片、收集瓶及动力产生装置的连接关系示意图，其中动力产生装置及芯片本体均为剖视状态。
图14是本申请第七实施方式提供的微流控芯片、收集瓶及动力产生装置的连接关系示意图，其中动力产生装置及芯片本体均为剖视状态。
图15是本申请第八实施方式提供的微流控芯片、收集瓶及动力产生装置的连接关系示意图，其中动力产生装置及芯片本体均为剖视状态。
图16是本申请第九实施方式提供的微流控芯片、收集瓶及动力产生装置的连接关系示意图，其中动力产生装置及芯片本体均为剖视状态。
图17是本申请第十实施方式提供的微流控芯片、收集瓶及动力产生装置的连接关系示意图，其中动力产生装置及芯片本体均为剖视状态。
图18本申请第十一实施方式提供的制备平台的立体示意图。
图19是图18所示的制备平台的另一种状态的立体示意图。
图20是采用不带振荡装置的微流控芯片系统制备液滴的流程图。
图21是采用带有振荡装置所示微流控芯片系统制备液滴的流程图。
主要元件符号说明
微流控芯片系统 100 微珠溶液引入通道 219
制备平台 10，101 汇流通道 210
底座 11 第一储液池 22
第一表面 111 第二储液池 23
第二表面 112 第三储液池 24
第一侧面 113 第一振荡装置 25
第二侧面 114 第二振荡装置 26
第一部 115 收集瓶 30
第二部 116 盖体 31
液滴生成装置卡槽 12 连接装置 40
收集瓶卡槽 13 第一连接管 41
动力产生装置卡槽 14 第二连接管 42
第一收容部 141 动力产生装置 50
第二收容部 142 主体部 51
第一限位槽 15 收容空间 511
第二限位槽 18 操作部 52
固定座 16 气体进出口端 53
防尘盖 17 第一限位件 54
液滴生成装置 20 第二限位件 55
芯片主体 21，110，120 细胞液 60
第三表面 211 微珠溶液 70
第四表面 212 油相 80
油相入口 213 液滴 200
细胞液入口 214 第三连接管 43
微珠溶液入口 215 第四连接管 44
液滴出口 216 第五连接管 45
油相引入通道 217 凸部 27
细胞液引入通道 218    
如下具体实施方式将结合上述附图进一步说明本申请。
具体实施方式
为能进一步阐述本申请达成预定申请目的所采取的技术手段及功效，以下结合附图1-21及较佳实施方式，对本申请提供的微流控芯片系统及液滴的制备方法的具体实施方式、结构、特征及其功效，作出如下详细说明。
请参阅图1-6，本申请第一实施方式提供一种微流控芯片系统100，所述微流控芯片系统100用于制备液滴200。
请参阅图1，所述微流控芯片系统100包括制备平台10、液滴生成装置20、收集瓶30、连接装置40及动力产生装置50。其中，所述制备平台10用于固定所述液滴生成装置20、收集瓶30、连接装置40及动力产生装置50，所述液滴生成装置20用于生成液滴200，所述收集瓶30用于收集从所述液滴生成装置20流出的所述液滴200，所述连接装置40用于连接所述液滴生成装置20与所述收集瓶30及/或连接所述液滴生成装置20与所述动力产生装置50及/或连接所述动力产生装置50与所述收集瓶30。其中，“固定”包括两种情形：所述液滴生成装置20、收集瓶30及动力产生装置50不可从所述制备平台10上拆卸下来；所述液滴生成装置20、收集瓶30及动力产生装置50可以从所述制备平台10上拆卸下来。在本实施方式中，所述液滴生成装置20、收集瓶30及动力产生装置50可以从所述制备平台10上拆卸下来。
在本实施方式中，所述液滴生成装置20、收集瓶30及动力产生装置50均固定在所述制备平台10上。所述连接装置40连接所述液滴生成装置20与所述收集瓶30，并连接所述收集瓶30与所述动力产生装置50。具体地，所述制备平台10包括底座11以及开设于底座11上的液滴生成装置卡槽12、收集瓶卡槽13及动力产生装置卡槽14。所述液滴生成装置20、收集瓶30及动力产生装置50依次收容并固定在所述液滴生成装置卡槽12、收集瓶卡槽13及动力产生装置卡槽14内。
可以理解，在其他实施方式中，所述液滴生成装置20、收集瓶30及动力产生装置50也可通过其他方式设置或安装于所述底座11上，例如，在底座11上设置卡扣或锁固装置，通过卡扣或锁固等方式将所述液滴生成装置20、收集瓶30及动力产生装置50固定于底座11上。
具体地，所述底座11包括第一表面111、与第一表面111相背的第二表面112、分别连接第一表面111及第二表面112的第一侧面113及与第一侧面113相背的第二侧面114。其中，所述液滴生成装置卡槽12及动力产生装置卡槽14均自第一表面111及第一侧面113分别向第二表面112及第二侧面114凹陷形成，所述收集瓶卡槽13自第一表面111向第二表面112凹陷形成。具体地，所述液滴生成装置卡槽12及动力产生装置卡槽14均贯穿第一表面111及第一侧面113，所述收集瓶卡槽13仅贯穿第一表面111。
在其他实施方式中，所述液滴生成装置卡槽12及动力产生装置卡槽14均仅贯穿第一表面111。
所述制备平台10可以但不局限于通过3D打印的方法制作而成。在本实施方式中，所述制备平台10通过3D打印方法制作而成，其成本较低且大小可以根据实际需要进行调节。
在本实施方式中，所述动力产生装置卡槽14包括第一收容部141及第二收容部142。其中，所述第一收容部141用于收容动力产生装置50的主体部51(见图1及下文)，所述第二收容部142用于收容动力产生装置50的操作部52(见图1及下文)。
所述制备平台10还包括第一限位槽15及一第二限位槽18。第一限位槽15用于收容并固定所述动力产生装置50的第一限位件54(见图1及下文)，以将所述操作部52保持在预定位置处。所述第二限位槽18与所述动力产生装置卡槽14连通，用于收容并固定所述动力产生装置50的第二限位件55(见图1及下文)。
在本实施方式中，所述第一限位槽15为分别自所述第一表面111及第一侧面113分别向第二表面112及第二侧面114凹陷形成的凹槽。在其他实施方式中，也可以是所述第一侧面113起到第一限位槽15具有的限位作用，或是第一限位槽15为一仅贯穿所述第一表面111、不贯穿第一侧面113的凹槽。
其中，所述液滴生成装置20包括芯片主体21。所述芯片主体21上开设有连续相入口及分散相入口，所述芯片主体21内部开设有连续相引入通道、分散相引入通道及汇流通道，所述连续相入口与所述连续相引入通道连通，所述分散相入口与所述分散相引入通道连通，所述连续相引入通道及所述分散相引入通道相交并与所述汇流通道的一端连通。连续相自所述连续相入口进入所述连续相引入通道，分散相自所述分散相入口进入所述分散相引入通道，所述连续相与所述分散相在所述连续相引入通道与所述分散相引入通道的相交处汇流并进入所述汇流通道内。
其中，所述连续相可以为油相等，所述分散相可以为细胞液、微珠液、水等。在本实施方式中，所述连续相为油相，所述分散相为细胞液及微珠液。
具体地，在本实施方式中，所述芯片主体21包括第三表面211及与第三表面211相背的第四表面212。分别自第三表面211向第四表面212凹陷形成油相入口213(连续相入口)、细胞液入口214(分散相入口)、微珠溶液入口215(分散相入口)及液滴出口216。所述油相入口213、细胞液入口214、微珠溶液入口215及液滴出口216仅贯穿所述第三表面211。可以理解，在其他实施方式中，所述液滴出口216也可以设置于连接第三表面211与第四表面212的其中一个侧面上。
其中，所述芯片主体21内还形成有油相引入通道217(连续相引入通道)、细胞液引入通道218(分散相引入通道)及微珠溶液引入通道219(分散相引入通道)。其中，所述油相入口213通过所述油相引入通道217与所述液滴出口216相连通，所述细胞液入口214通过所述细胞液引入通道218与所述液滴出口216相连通，所述微珠溶液入口215通过所述微珠溶液引入通道219与所述液滴出口216相连 通。
其中，所述芯片主体21内还形成有汇流通道210，所述油相引入通道217、细胞液引入通道218及微珠溶液引入通道219于所述汇流通道210的一端交汇，所述汇流通道210的另一端与所述液滴出口216相密封连通。
在本实施方式中，所述芯片主体21内还形成有两条油相引入通道217、两条细胞液引入通道218及一条微珠溶液引入通道219。具体地，两条所述油相引入通道217环绕所述细胞液入口214及两条所述细胞液引入通道218设置，两条所述细胞液引入通道218环绕所述微珠溶液入口215及所述微珠溶液引入通道219设置。在所述油相引入通道217、所述细胞液引入通道218及所述微珠溶液引入通道219汇流之前，两条所述油相引入通道217与两条所述细胞液引入通道218对称分布在所述微珠溶液引入通道219的两侧。
在本实施方式中，所述油相入口213、细胞液入口214及微珠溶液入口215分布在同一条直线上。
在本实施方式中，所述油相入口213、细胞液入口214、微珠溶液入口215及液滴出口216不分布在同一条直线上。
其中，所述微流控芯片系统100还可以包括连续相储液池及分散相储液池，所述连续相储液池与所述连续相入口连通，所述分散相储液池与所述分散相入口连通。其中，所述连续相储液池与所述分散相储液池可以位于所述芯片主体21上，也可以位于所述芯片主体21外。
具体地，请参阅图3，在本实施方式中，所述微流控芯片系统100还包括第一储液池22(连续相储液池)、第二储液池23(分散相储液池)及第三储液池24(分散相储液池)。所述第一储液池22与油相入口213连通，所述第二储液池23与细胞液入口214连通，所述第三储液池24与所述微珠溶液入口215连通。
在本实施方式中，所述第一储液池22、第二储液池23及第三储液池24位于所述芯片主体21上。具体地，所述芯片主体21与所述第一储液池22、第二储液池23及第三储液池24之间是一体成型的。在其他实施方式中，所述第一储液池22、第二储液池23及第三储液池24也可以直接插接在所述油相入口213、细胞液入口214及微珠溶液入口215内。
在本实施方式中，所述第一储液池22用于暂时存储油相80。所述第一储液池22的存储体积可以为但不局限于100微升、200微升、500微升及1毫升。
在本实施方式中，所述第二储液池23用于暂时存储细胞液60。所述第二储液池23的存储体积可以为但不局限于100微升、200微升、500微升及1毫升。
在本实施方式中，所述第三储液池24用于暂时存储微珠溶液70。所述第三储液池24的存储体积可以为但不局限于100微升、200微升、500微升及1毫升。
其中，所述微珠溶液70可以为聚苯乙烯微珠溶液、树脂微珠溶液或磁珠溶液等中的一种。
其中，所述第一储液池22、第二储液池23及第三储液池24上均形成有一开口(图未示)，以便于将所述油相80、细胞液60及微珠溶液70转移至所述第一储液池22、第二储液池23及第三储液池24内。
其中，所述第一储液池22、第二储液池23及第三储液池24中的所述油相80、细胞液60及微珠溶液70的体积比可以为但不局限于2:1:1。在本实施方式中，所述细胞液60为200微升，所述微珠溶液70为200微升，所述油相80为400微升。
在其他实施方式中，所述液滴生成装置20还可以不包括所述第一储液池22、第二储液池23及第三储液池24。此时，可以将所述油相80直接加入所述油相入口213中，将所述细胞液60直接加入所述细胞液入口214内，将所述微珠溶液70直接加入所述微珠溶液入口215内。
其中，所述收集瓶30用于收集液滴200。所述收集瓶30包括一盖体31，所述盖体31上形成有两个通孔(图未示)。所述盖体31与收集瓶30的主体为可分离式连接或者为不可分离式连接。
其中，所述收集瓶30的体积可以为但不局限于1毫升、5毫升。在本实施方式中，所述收集瓶30的体积为1毫升。
在本实施方式中，所述连接装置40包括第一连接管41及第二连接管42。所述第一连接管41的一端与所述动力产生装置50相连通，另一端与所述收集瓶30相连通，所述第二连接管42的一端连接在所述芯片主体21上且与所述液滴出口216相连通，另一端与所述收集瓶30相连通。
具体地，在本实施方式中，所述第二连接管42的一端连接在第三表面211上，另一端连接在所述收集瓶30的盖体31的一个通孔内并与所述收集瓶30相连通。所述第一连接管41的一端连接在所述动力产生装置50上，另一端连接在所述盖体31的另一个通孔内并与所述收集瓶30相连通。
在本实施方式中，所述第一连接管41及第二连接管42均为10厘米长的软管。
其中，所述动力产生装置50用于给所述液滴生成装置20提供生成液滴200的动力，以使得位于第一储液池22、第二储液池23及第三储液池24中的油相80、细胞液60及微珠溶液70分别在液滴生成装置20的油相引入通道217、细胞液引入通道218及微珠溶液引入通道219内流动，之后，在所述汇流通道210中汇合，形成液滴200，所述液滴200从所述液滴出口216流出，并进一步流入所述收集瓶30。
在本实施方式中，所述动力产生装置50为负压产生装置。所述动力产生装置50包括主体部51、操作部52及气体进出口端53。其中，所述主体部51包括收容空间511，所述操作部52的一端收容在所述收容空间511内，另一端凸出于所述主体部51外。所述操作部52收容在收容空间511内的一端与收容空间511 的内壁紧密贴合。当所述操作部52位于所述主体部51之外的一端受到外力作用并致动所述操作部52，使所述操作部52在所述收容空间511内滑动。所述气体进出口端53固定在所述主体部51上远离外露的所述操作部52的一端且与所述收容空间511相连通。所述第二连接管42远离收集瓶30的一端连接在所述气体进出口端53上。
具体地，在本实施方式中，所述动力产生装置50为一注射装置。
在本实施方式中，所述动力产生装置50还包括一第一限位件54及第二限位件55，所述第一限位件54设置在所述操作部52的外露于主体部51的一端。所述第一限位件54与所述第一限位槽15相配合，以将所述操作部52拉至固定的位置，以保持所述动力产生装置50内部真空。所述第二限位件55固定在所述主体部51上，所述第二限位件55与所述第二限位槽18相配合，以在拉动所述操作部52的过程中限制所述主体部51的移动。
所述操作部52在外力作用下移动，使位于其上的所述第一限位件54被移动并限位于所述第一限位槽15处时，所述第二连接管42、收集瓶30、第一连接管41及芯片主体21的所述液滴出口216、汇流通道210、微珠溶液引入通道219、细胞液引入通道218、油相引入通道217从初始体积V1增长到体积V2，根据理想气体状态方程P1V1＝P2V2，其中，P1与P2为分别对应初始体积V1和体积V2时的压强，此时，所述第二连接管42、收集瓶30、第一连接管41及芯片主体21的所述液滴出口216、汇流通道210、微珠溶液引入通道219、细胞液引入通道218、油相引入通道217、第一储液池22、第二储液池23、第三储液池24内的压强小于大气压强，产生的压强差使得所述液滴产生装置20内的所述分散相与所述连续相以液滴的形式流入收集瓶30内。
其中，所述动力产生装置50的动力可以通过以下方式产生：1)与所述芯片主体21连接之前在在密闭容器中预先产生；2)由泵驱动所述动力产生装置50产生；3)通过拉动或推动注射器(或类似装置)的柱塞产生。
在一些实施例中，所述微流控芯片系统100还可以包括至少一振荡装置，所述振荡装置对应所述第三储液池及/或第二储液池设置，用于施加振动于所述微珠溶液70及/或细胞液60。
其中，请参阅图6，所述微流控芯片系统100还包括至少一个振荡装置，所述振荡装置用于施加振动于分散相。
具体地，在本实施方式中，所述微流控芯片系统100还包括一第一振荡装置25及一第二振荡装置26。其中，所述第一振荡装置25及第二振荡装置26固定在所述液滴生成装置卡槽12内且分别对应所述细胞液入口214及微珠溶液入口215。
其中，所述第一振荡装置25及第二振荡装置26均可为陶瓷振荡片。
其中，所述第一振荡装置25及所述第二振荡装置26在启动后产生振动，振 动施加于所述细胞液60与所述微珠溶液70上，可以有效改善所述细胞液60及微珠溶液70在实验过程中产生的细胞或微珠沉降问题，从而能够保证所述细胞液60及微珠溶液70浓度的一致性。
在其他实施方式中，所述第一振荡装置25及第二振荡装置26也可位于芯片主体21上，例如，第一振荡装置25及第二振荡装置26位于所述芯片主体21的第四表面212上且分别对应所述细胞液入口214及微珠溶液入口215，或位于连接芯片主体21的第三表面211与第四表面212的侧壁上且对应细胞液入口214及微珠溶液入口215所在位置。仅需所述第一振荡装置25及所述第二振荡装置26启动后产生的振动能施加于所述细胞液60与微珠溶液70上即可，以改善细胞液60及微珠溶液70的细胞或微珠沉降问题。
在另一实施方式中，所述微流控芯片系统100可以仅包括一个振荡装置，例如，仅包括所述第二振荡装置26，第二振荡装置26可固定在所述液滴生成装置卡槽12内且对应微珠溶液入口215。第二振荡装置26也可以固定在芯片主体21的第四表面212上且对应所述微珠溶液入口215。从而，可以有效改善微珠溶液70在实验过程中产生的微珠沉降问题，从而能够保证所述微珠溶液70浓度的一致性。
在再一实施方式中，微流控芯片系统100仅包括一个振荡装置，所述振荡装置同时对应所述细胞液入口214与微珠溶液入口215设置，并同时施加振动于细胞液60与微珠溶液70上。
当施加一动力在所述动力产生装置50上时，所述动力产生装置50内部形成真空，真空会促使所述分散相与连续相在所述芯片本体10内流动，因采用的是注射装置作为动力产生装置，从而可以通过简单的动作即可产生真空。当试剂(分散相与连续相)开始在所述芯片本体内流动，会导致所述动力产生装置中的空气的体积在正压或负压系统中膨胀或压缩，从而会导致芯片入口(连续相入口及分散相入口)处和出口(液滴出口)处的压降减小，从而导致所述试剂的流速逐渐减慢，所述试剂的流速的变化会引起液滴尺寸的变化。因此，所述芯片本体10内的压降、所述分散相与连续相的流速、液滴的尺寸和生成频率都是可预测的、可编程的和可重现的。
请参阅图7，图7为采用本发明提供的微流控芯片系统生成的液滴(不包含微珠和细胞)的示意图。由图7可知，采用本发明提供的微流控芯片系统100生成的液滴的液滴尺寸变化较小，均一性较好。在本实施方式中，所述液滴的尺寸为55.7um，所述液滴的尺寸变化为2％。
请参阅图8，图8为采用本发明提供的微流控芯片系统100生成的液滴(包含微珠和细胞)的示意图。由图8可知，采用本发明提供的微流控芯片系统可以将所述微珠与细胞包裹在所述油相内。
请参阅图9，图9为本申请第二实施方式提供的一种芯片主体110的内部构 造示意图。所述芯片主体110的内部构造与本申请第一实施方式提供的所述芯片主体21的内部构造基本相同，其区别点仅在于：所述芯片主体110内形成有两条油相引入通道217、一条细胞液引入通道218及两条微珠溶液引入通道219。具体地，两条所述油相引入通道217环绕所述微珠溶液入口215及两条所述微珠溶液引入通道219设置，两条所述微珠溶液引入通道219环绕所述细胞液入口214及一条所述细胞液引入通道218设置。在所述油相引入通道217、所述细胞液引入通道218及所述微珠溶液引入通道219汇流之前，两条所述油相引入通道217与两条微珠溶液引入通道219对称分布在所述微珠溶液引入通道219的两侧。
在本实施方式中，所述油相入口213、细胞液入口214、微珠溶液入口215及液滴出口216分布在同一条直线上。
请参阅图10，图10为本申请第三实施方式提供的一种芯片主体120的内部构造示意图。所述芯片主体120的内部构造与本申请第一实施方式提供的所述芯片主体21的内部构造基本相同，其区别点仅在于：所述芯片主体120内形成有两条油相引入通道217、一条细胞液引入通道218及一条微珠溶液引入通道219。具体地，两条所述油相引入通道217环绕所述微珠溶液入口215、一条所述微珠溶液引入通道219、所述细胞液入口214及一条所述细胞液引入通道218设置。一条所述细胞液引入通道218及一条所述微珠溶液引入通道219对称分布，两条所述油相引入通道217分布在一条所述细胞液引入通道218及一条所述微珠溶液引入通道219的两侧。
在本实施方式中，所述油相入口213与所述液滴出口216分布在同一条直线上，所述油相入口213、细胞液入口214、微珠溶液入口215及液滴出口216不分布在同一条直线上。
请参阅图11，图11为本申请第四实施方式提供的芯片主体21、收集瓶30及动力产生装置50的连接关系示意图，其中动力产生装置50为剖视状态。在本实施方式中，所述芯片主体21、收集瓶30及动力产生装置50的连接关系与图3所示的所述芯片主体21、收集瓶30及动力产生装置50的连接关系基本相同，其区别仅在于：所述第一储液池22、第二储液池23及第三储液池24形成在所述芯片主体21外，且通过一第三连接管43、一第四连接管44及一第五连接管45分别与所述油相入口213、细胞液入口214及微珠溶液入口215密封连通。
请参阅图12，图12为本申请第五实施方式提供的芯片主体21、收集瓶30及动力产生装置50的连接关系示意图，其中动力产生装置50为剖视状态。在本实施方式中，所述芯片主体21、收集瓶30及动力产生装置50的连接关系与图9所示的所述芯片主体21、收集瓶30及动力产生装置50的连接关系基本相同，其区别仅在于：所述液滴生成装置20还包括一形成在所述芯片主体21上的凸部27，所述收集瓶30放置于所述凸部27内，所述汇流通道210与所述收集瓶30密封连通，所述收集瓶30通过所述第一连接管41与所述动力产生装置50密封连通。
请参阅图13，图13为本申请第六实施方式提供的芯片主体21、收集瓶30及动力产生装置50的连接关系示意图，其中动力产生装置50为剖视状态。在本实施方式中，所述芯片主体21、收集瓶30及动力产生装置50的连接关系与图12所示的所述芯片主体21、收集瓶30及动力产生装置50的连接关系基本相同，其区别仅在于：所述第一储液池22、第二储液池23及第三储液池24形成在所述芯片主体21上，所述第一储液池22与油相入口213密封连通，所述第二储液池23与细胞液入口214密封连通，所述第三储液池24与所述微珠溶液入口215密封连通。
请参阅图14，图14为本申请第七实施方式提供的芯片主体21、收集瓶30及动力产生装置50的连接关系示意图，其中动力产生装置50为剖视状态。在本实施方式中，所述芯片主体21、收集瓶30及动力产生装置50的连接关系与图3所示的所述芯片主体21、收集瓶30及动力产生装置50的连接关系基本相同，其区别仅在于：所述动力产生装置50为正压产生装置。具体地，所述动力产生装置50为一注射装置。所述动力产生装置50通过所述第一连接管41分别与所述第一储液池22、第二储液池23及第三储液池24相连通。在本实施方式中，所述第一连接管41为一多接头的分叉连接管。具体地，所述第一连接管41包括一主连接段411、一第一分叉连接段412、一第二分叉连接段413及一第三分叉连接段414，所述主连接段411的一端与所述动力产生装置50的所述气体进出口端53相连接并密封连通，所述主连接段411的另一端分别与所述第一分叉连接段412、第二分叉连接段413及第三分叉连接段414相连接并密封连通。
当然，本实施方式中的这种结构还适用于图9及图10所示的芯片本体的结构。
请参阅图15，图15为本申请第八实施方式提供的芯片主体21、收集瓶30及动力产生装置50的连接关系示意图，其中动力产生装置50为剖视状态。在本实施方式中，所述芯片主体21、收集瓶30及动力产生装置50的连接关系与图14所示的所述芯片主体21、收集瓶30及动力产生装置50的连接关系基本相同，其区别仅在于：所述第一储液池22、第二储液池23及第三储液池24形成在所述芯片主体21外，且通过一第三连接管43、一第四连接管44及一第五连接管45分别与所述油相入口213、细胞液入口214及微珠溶液入口215密封连通。
请参阅图16，图16为本申请第九实施方式提供的芯片主体21、收集瓶30及动力产生装置50的连接关系示意图，其中动力产生装置50为剖视状态。在本实施方式中，所述芯片主体21、收集瓶30及动力产生装置50的连接关系与图15所示的所述芯片主体21、收集瓶30及动力产生装置50的连接关系基本相同，其区别仅在于：所述液滴生成装置20还包括一形成在所述芯片主体21上的凸部27，所述收集瓶30放置于所述凸部27内，所述汇流通道210与所述收集瓶30 密封连通。
请参阅图17，图17为本申请第十实施方式提供的芯片主体21、收集瓶30及动力产生装置50的连接关系示意图，其中动力产生装置50为剖视状态。在本实施方式中，所述芯片主体21、收集瓶30及动力产生装置50的连接关系与图16所示的所述芯片主体21、收集瓶30及动力产生装置50的连接关系基本相同，其区别仅在于：所述第一储液池22、第二储液池23及第三储液池24形成在所述芯片主体21上，所述第一储液池22与油相入口213密封连通，所述第二储液池23与细胞液入口214密封连通，所述第三储液池24与所述微珠溶液入口215密封连通。
请参阅图18-19，图18-19为本申请第十一实施方式提供的制备平台101的立体示意图。在本实施方式中，所述制备平台101的结构与第一实施方式中的所述制备平台10的结构类似，其区别仅在于：当所述压力产生装置为负压产生装置时，所述制备平台101的所述底座11包括第一部115及与所述第一部115连接的第二部116，所述第一部115低于所述第二部116，所述液滴生成装置卡槽12形成在所述第一部115上，所述动力产生装置卡槽14形成在所述第二部116上。在其他实施方式中，也可以省略所述液滴生成装置卡槽12，而直接将所述液滴生成装置20固定在所述第一部115上。在本实施方式中，所述收集瓶卡槽13也形成在所述第二部116上。当然，若所述收集瓶30形成在所述芯片主体21内，则所述第二部116上则不设置所述收集瓶卡槽13。所述制备平台101还包括一固定在所述第一部115上的固定座16及一可翻转连接在所述固定座16上的防尘盖17。所述防尘盖17位于所述芯片主体21一侧，所述防尘盖17用于防止灰尘进入所述芯片主体21内。具体地，所述防尘盖17位于所述连续相入口及/或分散相入口的上方，或者是位于所述连续相储液池及/或分散相储液池的上方。
请参阅图20-21，本申请较佳实施方式还提供一种液滴的制备方法，所述制备方法可以结合上述介绍的微流控芯片系统100或类似系统来执行，所述制备方法包括如下步骤：
步骤S801，通过移液枪或移液管分别量取一预定量的连续相及分散相，将量取的所述连续相导入所述连续相入口内，将量取的所述分散相导入所述分散相入口内。
具体地，在本实施方式中，所述步骤S801可以表述为：使用移液枪或移液管分别量取预定量的油相80、细胞液60及微珠溶液70，将量取的油相80导入微流控芯片系统100的油相入口213内，将量取的细胞液60导入细胞液入口214内，将量取的微珠溶液70导入微珠溶液入口215内。
当所述微流控芯片系统100还包括连续相储液池及分散相储液池时，所述步骤S801可以表述为，通过移液枪或移液管分别量取一预定量的连续相及分散相，将量取的所述连续相导入所述连续相储液池内，将量取的所述分散相导入所述分 散相储液池内。
具体地，在本实施方式中，所述微流控芯片系统100还包括第一储液池22、第二储液池23及第三储液池24，因此，步骤S801具体为：使用移液枪或移液管分别量取预定量的油相80、细胞液60及微珠溶液70，将量取的油相80导入第一储液池22内，将量取的细胞液60导入第二储液池23内、将量取的微珠溶液70导入第三储液池24内，所述油相80通过所述第一储液池22导入所述油相入口213内，所述细胞液60通过所述第二储液池23导入细胞液入口214内，所述微珠溶液70通过所述第三储液池24导入所述微珠溶液入口215内。
在本实施方式中，所述细胞液60为200微升，所述微珠溶液70为200微升，所述油相80为400微升。
其中，所述细胞液60、微珠溶液70及油相80的加样顺序可以是：先加细胞液60、再加微珠溶液70、最后加油相80；也可以是：先加微珠溶液70、再加细胞液60、最后加油相80；还可以是同时添加所述细胞液60、微珠溶液70及油相80。
步骤S802，启动动力产生装置50，使收集瓶30、连接装置40、芯片主体21内的压强与大气压强之间形成压强差，压强差促使所述分散相及所述连续相汇流并以液滴的形式流入收集瓶30内。
具体地，步骤S802可以为：启动动力产生装置50，使收集瓶30、第二连接管42及芯片主体21内的液滴出口216、汇流通道210、至少一分散相引入通道、所述分散相入口及连续相入口内的压强与大气压强之间形成压强差，压强差促使连续相及分散相向汇流通道210流动，在汇流通道210处汇合形成液滴200，液滴200在压强差的作用下进一步通过液滴出口216与第二连接管42流入收集瓶30内。
在本实施方式中，所述步骤S802可以表述为：启动动力产生装置50，使收集瓶30、第二连接管42及芯片主体21内的液滴出口216、汇流通道210、微珠溶液引入通道219、细胞液引入通道218及油相引入通道217、所述微珠溶液入口215、细胞液入口214及油相入口213内的压强与大气压强之间形成压强差，压强差促使细胞液60、微珠溶液70及油相80向汇流通道210流动，在汇流通道210处汇合形成液滴200，液滴200在压强差的作用下进一步通过液滴出口216与第二连接管42流入收集瓶30内。
在本实施方式中，所述微流控芯片系统100的液滴生成装置20还包括所述第一储液池22、第二储液池23及第三储液池24，因此，步骤S802可以为：启动动力产生装置50，使收集瓶30、第二连接管42、及芯片主体21内的液滴出口216、汇流通道210、微珠溶液引入通道219、细胞液引入通道218、油相引入通道217、所述第三储液池24、第二储液池23及第一储液池22内的压强与大气压强之间形成压强差，压强差促使细胞液60、微珠溶液70及油相80向汇流通道 210流动，在汇流通道210处汇合形成液滴200，液滴200在压强差的作用下进一步通过液滴出口216与第二连接管42流入收集瓶30内。
在本实施方式中，所述动力产生装置50为一负压产生装置，因此步骤S802具体为：启动所述动力产生装置50使收集瓶30、第二连接管42及芯片主体21内的液滴出口216、微珠溶液引入通道219、细胞液引入通道218、油相引入通道217内产生负压，所述负压促使细胞液60、微珠溶液70及油相80向汇流通道210流动，在汇流通道210处汇合形成液滴200，液滴200在压强差的作用下进一步通过液滴出口216与第二连接管42流入收集瓶30内。进一步地，产生负压的具体操作为：拉动动力产生装置50的操作部52至一预设位置并利用底座11上的第一限位槽15将第一限位件54固定。
在其他实施方式中，所述动力产生装置50为一正压产生装置，因此步骤S802具体为：启动所述动力产生装置50，使芯片主体21内的分散相引入通道、连续相引入通道、液滴出口216及收集瓶30内产生正压，所述正压促使分散相与连续相向汇流通道210流动，在汇流通道210处汇合形成液滴200，液滴200在所述正压的作用下进一步通过液滴出口216流入收集瓶30内。
请参阅图19，在另一种实施方式中，所述制备方法在启动动力产生装置50前还可包括步骤S900：利用第一振荡装置25及第二振荡装置26施加振动于所述微珠溶液70与细胞液60上，例如，打开所述第一振荡装置25及所述第二振荡装置26设置于底座11上的电源开关(图未示)，所述第一振荡装置25及所述第二振荡装置26开始振动以施加振动于所述微珠溶液70与细胞液60上。
可以理解，由于微流控芯片系统100可以仅包括一个振荡装置，因此在另一实施方式中步骤S900也可表述为：利用至少一个振荡装置施加振动于所述微珠溶液70及/或细胞液60上。
在其他实施方式中，所述步骤S900还可以表述为：利用第一振荡装置25及第二振荡装置26施加振动于所述分散相上，例如，打开所述第一振荡装置25及所述第二振荡装置26设置于底座11上的电源开关(图未示)，所述第一振荡装置25及所述第二振荡装置26开始振动以施加振动于所述分散相上。
本申请提供的微流控芯片系统，首先，1)将液滴生成装置、动力产生装置及收集瓶均固定到一制备平台上，以保证所述微流控芯片系统生成液滴的稳定性；2)将单一动力源施加至液滴生成装置的液滴出口处，使液滴生成装置中形成真空，促使细胞液、微珠溶液与油相在液滴生成装置中同步流动，具有较高通量，生成液滴的尺寸均一性较好；；3)进一步以注射装置作为单一动力源并根据理想气体定律P 1V 1＝P 2V 2来控制注射装置操作部前后的位置且以底座上的第一限位槽将注射装置的第一限位件保持在预定位置以保持其真空，成本低且易于操作；；4)动力产生装置中的压力随着试剂的注入和流动发生变化，试剂填充到收集装置中，动力产生装置中压力的变化也遵循理想气体定律；5)液滴生成装置上的压降，所 有连续相与分散相的流速，液滴的尺寸和生成频率都是可预测的、可编程的和可重现的；6)微流控芯片系统中配置的振荡装置可以有效改善细胞液及/或微珠溶液在实验过程中产生的细胞及/或微珠的沉降问题，从而能够保证细胞液及/或微微珠溶液浓度的一致性。
另外，本申请提供的微流控芯片系统在2分钟内可以制备250,000个液滴，意味着能得到10000个单细胞目标产物，即一个液滴内包含一个磁珠和一个细胞。并且可以根据实际需求调节液滴产生时间，来控制试剂目标产物量。整个微流控芯片系统的体积仅有一个苹果公司生产的ipad mini大小，使用者可以极大的拓展其使用空间。
以上所述，仅是本申请的较佳实施方式而已，并非对本申请任何形式上的限制，虽然本申请已是较佳实施方式揭露如上，并非用以限定本申请，任何熟悉本专业的技术人员，在不脱离本申请技术方案范围内，当可利用上述揭示的技术内容做出些许更动或修饰为等同变化的等效实施方式，但凡是未脱离本申请技术方案内容，依据本申请的技术实质对以上实施方式所做的任何简单修改、等同变化与修饰，均仍属于本申请技术方案的范围内。

Claims (16)

  1. 一种微流控芯片系统，用于产生液滴，所述微流控芯片系统包括
    液滴生成装置，用于生成液滴；
    动力产生装置，用于给所述液滴生成装置提供生成液滴的动力；
    收集瓶，用于收集从所述液滴生成装置流出的液滴；
    连接装置，用于连通所述液滴生成装置、所述动力产生装置及所述收集瓶；及
    制备平台，用于固定所述液滴生成装置、所述动力产生装置及所述收集瓶。
  2. 如权利要求1所述的微流控芯片系统，其特征在于，所述液滴生成装置包括一芯片主体，所述芯片主体内形成有连续相入口及分散相入口，所述连续相及分散相从所述连续相入口及分散相入口进入所述芯片主体。
  3. 如权利要求2所述的微流控芯片系统，其特征在于，所述微流控芯片系统还包括连续相储液池及分散相储液池，所述连续相储液池及分散相储液池分别固定在所述芯片主体上或形成在所述芯片主体外。
  4. 如权利要求2所述的微流控芯片系统，其特征在于，所述制备平台包括底座及形成在所述底座上的液滴生成装置卡槽及动力产生装置卡槽，所述液滴生成装置及所述动力产生装置依次收容在所述液滴生成装置卡槽及所述动力产生装置卡槽内。
  5. 如权利要求4所述的微流控芯片系统，其特征在于，所述底座包括第一部及与所述第一部连接的第二部，所述第一部低于所述第二部，所述液滴生成装置卡槽形成在所述第一部上，所述动力产生装置卡槽形成在所述第二部上。
  6. 如权利要求4所述的微流控芯片系统，其特征在于，所述收集瓶位于所述液滴生成装置外，所述底座上还形成有一收集瓶卡槽，所述收集瓶收容并固定在所述收集瓶卡槽内。
  7. 如权利要求4所述的微流控芯片系统，其特征在于，所述收集瓶收容并固定在所述芯片主体内，所述芯片主体收容并固定在所述液滴生成装置卡槽内。
  8. 如权利要求4所述的微流控芯片系统，其特征在于，所述微流控芯片系统还包括至少一振荡装置，所述振荡装置设置在所述液滴生成装置卡槽内且对应所述分散相入口，用于施加振动于分散相。
  9. 如权利要求2所述的微流控芯片系统，其特征在于，所述微流控芯片系统还包括至少一振荡装置，所述振荡装置设置在所述芯片主体上且对应所述分散相入口，用于施加振动于所述分散相。
  10. 如权利要求1所述的微流控芯片系统，其特征在于，所述动力产生装置包括一操作部及一形成在所述操作部末端的第一限位件，所述制备平台还包括一第一限位槽，所述第一限位槽用于收容并固定所述第一限位件，以将所述操作部保持在预定位置处。
  11. 如权利要求10所述的微流控芯片系统，其特征在于，所述动力产生装置为注射装置。
  12. 如权利要求1-11任一项所述的微流控芯片系统，其特征在于，所述动力产生装置为负压产生装置，所述负压产生装置通过所述连接装置与所述收集瓶密封连接。
  13. 如权利要求12所述的微流控芯片系统，其特征在于，所述制备平台还包括一固定座及一可翻转连接在所述固定座上的防尘盖，所述防尘盖位于所述液滴生成装置一侧，所述防尘盖用于防止灰尘进入所述液滴生成装置内。
  14. 如权利要求2-11任一项所述的微流控芯片系统，其特征在于，所述动力产生装置为正压产生装置，所述正压产生装置通过所述连接装置分别与所述连续相入口及所述分散相入口密封连接。
  15. 一种利用权利要求2-11任一项所述的微流控芯片系统来实施的液滴的制备方法，包括步骤：
    通过移液枪或移液管分别量取一预定量的连续相及分散相，将量取的所述连续相导入所述连续相入口内，将量取的所述分散相导入所述分散相入口内；及
    启动动力产生装置，使收集瓶、连接装置及芯片主体内产生压强差，压强差促使所述分散相及所述连续相汇流并以液滴的形式流入收集瓶内。
  16. 如权利要求15所述的液滴的制备方法，其特征在于，在启动所述动力产生装置之前，还包括步骤：利用至少一振荡装置施加振动于所述分散相上。
PCT/CN2019/108536 2018-09-29 2019-09-27 微流控芯片系统及液滴的制备方法 WO2020063864A1 (zh)

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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101570776A (zh) * 2008-04-29 2009-11-04 中国科学院大连化学物理研究所 一种基于微流控芯片的高通量纳升级微液滴形成和固定方法、其专用芯片及应用
CN104877899A (zh) * 2014-02-28 2015-09-02 中国科学院青岛生物能源与过程研究所 一种基于液滴的微生物快速直接绝对定量检测系统和方法
CN104877898A (zh) * 2014-02-27 2015-09-02 中国科学院青岛生物能源与过程研究所 一种低成本、高效分离获取单细胞的系统和方法
CN107418872A (zh) * 2016-08-31 2017-12-01 四川蓝光英诺生物科技股份有限公司 制备微球的装置和制备微球的方法
CN108020490A (zh) * 2017-06-23 2018-05-11 中国科学院天津工业生物技术研究所 一种使用液滴微流控芯片的高通量筛选设备
CN209144161U (zh) * 2018-09-29 2019-07-23 深圳华大生命科学研究院 微流控芯片系统

Family Cites Families (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4190448A3 (en) 2006-05-11 2023-09-20 Bio-Rad Laboratories, Inc. Microfluidic devices
JP4572973B2 (ja) 2008-06-16 2010-11-04 ソニー株式会社 マイクロチップ及びマイクロチップにおける送流方法
US9156010B2 (en) * 2008-09-23 2015-10-13 Bio-Rad Laboratories, Inc. Droplet-based assay system
CA2745636A1 (en) * 2008-12-02 2010-06-10 California Institute Of Technology Self-powered microfluidic devices, methods and systems
GB201211342D0 (en) 2012-06-26 2012-08-08 Cambridge Entpr Ltd Microfluidic device
CN105498875A (zh) * 2016-01-27 2016-04-20 杭州霆科生物科技有限公司 一种用于制备液滴的离心式微流控芯片
CN106190779B (zh) * 2016-08-29 2018-08-24 苏州汶颢芯片科技有限公司 基于微流控芯片的单细胞分离和包封装置和方法
CN205990393U (zh) * 2016-08-29 2017-03-01 苏州汶颢芯片科技有限公司 基于微流控芯片的单细胞分离和包封装置
CN108499500A (zh) 2018-04-10 2018-09-07 中国计量大学 一种振动管路控制流动聚焦型微流控芯片生成微液滴的方法

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101570776A (zh) * 2008-04-29 2009-11-04 中国科学院大连化学物理研究所 一种基于微流控芯片的高通量纳升级微液滴形成和固定方法、其专用芯片及应用
CN104877898A (zh) * 2014-02-27 2015-09-02 中国科学院青岛生物能源与过程研究所 一种低成本、高效分离获取单细胞的系统和方法
CN104877899A (zh) * 2014-02-28 2015-09-02 中国科学院青岛生物能源与过程研究所 一种基于液滴的微生物快速直接绝对定量检测系统和方法
CN107418872A (zh) * 2016-08-31 2017-12-01 四川蓝光英诺生物科技股份有限公司 制备微球的装置和制备微球的方法
CN108020490A (zh) * 2017-06-23 2018-05-11 中国科学院天津工业生物技术研究所 一种使用液滴微流控芯片的高通量筛选设备
CN209144161U (zh) * 2018-09-29 2019-07-23 深圳华大生命科学研究院 微流控芯片系统

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