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WO2019086018A1 - 微液滴生成装置 - Google Patents

微液滴生成装置 Download PDF

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Publication number
WO2019086018A1
WO2019086018A1 PCT/CN2018/113851 CN2018113851W WO2019086018A1 WO 2019086018 A1 WO2019086018 A1 WO 2019086018A1 CN 2018113851 W CN2018113851 W CN 2018113851W WO 2019086018 A1 WO2019086018 A1 WO 2019086018A1
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WO
WIPO (PCT)
Prior art keywords
micro
phase sample
droplet
microdroplet
component
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/CN2018/113851
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English (en)
French (fr)
Chinese (zh)
Inventor
荆高山
郭永
王博
苏世圣
白宇
朱修锐
付明珠
祝令香
刘宝霞
杨文军
王勇斗
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Targetingone Corp
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Targetingone Corp
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 claimed from CN201711074985.2A external-priority patent/CN109746062B/zh
Priority claimed from CN201711074977.8A external-priority patent/CN109746060B/zh
Priority claimed from CN201711074976.3A external-priority patent/CN109746059B/zh
Application filed by Targetingone Corp filed Critical Targetingone Corp
Priority to JP2020543685A priority Critical patent/JP7030361B2/ja
Publication of WO2019086018A1 publication Critical patent/WO2019086018A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/12—Apparatus for enzymology or microbiology with sterilisation, filtration or dialysis means

Definitions

  • the invention relates to the field of micro-droplet digital PCR technology, and in particular to a micro-droplet generating device.
  • Droplet digital PCR is a single-molecule PCR-based nucleic acid absolute quantitative analysis technique.
  • Microdroplet digital PCR technology is becoming the next revolutionary technology in the industry with its high sensitivity and high accuracy.
  • micro-droplet digital PCR technology has encountered the best opportunity to break through the technical bottleneck.
  • the technology uses a microfluidic chip to generate droplets having a diameter of several micrometers to hundreds of micrometers; the microdroplets encapsulate single molecules or single cells, and the reaction and detection are fully enclosed and fully integrated.
  • micro-droplet digital PCR device The working principle of the micro-droplet digital PCR device is: firstly, the sample to be tested is evenly distributed into a large number of "water-in-oil" micro-droplets (in the range of several micrometers to hundreds of micrometers) by a special micro-droplet generator. The number of droplets is on the order of a million.
  • microdroplets Since the number of microdroplets is sufficient, the microdroplets are separated from each other by the oil layer, so each microdroplet is equivalent to a "microreactor", and the microdroplet contains only the DNA single molecule of the sample to be tested; These microdroplets are respectively subjected to a PCR amplification reaction, and the fluorescence signals of the droplets are detected one by one by a microdroplet analyzer, the droplets of the fluorescent signal are interpreted as 1, and the droplets without the fluorescent signal are interpreted as 0. Finally, according to the Poisson distribution principle and the number and proportion of positive droplets, the number of target DNA molecules of the sample to be tested can be obtained, and the absolute quantification of the nucleic acid sample can be realized.
  • a key step in microdroplet digital PCR technology is the rapid, reliable, parallel generation of uniform micron-sized "water-in-oil” microdroplets.
  • One of the core technologies for generating microdroplets is the design and processing of microfluidic generation devices based on microfluidic technology.
  • Microdroplet generation devices are widely used in clinical testing and need to have the following principles: (1) fast, reliable, parallel generation of uniform micron-sized "water-in-oil” / "oil-in-water” microdroplets, (2) based on micro The flow control technology of the microfluidic chip has low material and processing cost, (3) easy operation, and (4) no cross-contamination during droplet formation and collection.
  • PDMS polydimethylsiloxane
  • the researchers used a soft lithography process (manually) to process PDMS microdroplets with micron levels.
  • the sample inlet and the micro-droplet generation outlet are punched by a mechanical processing process, and the sample introduction tube and the sample introduction tube are assembled.
  • the "oil phase” sample, the “aqueous phase” sample is manually inhaled into the syringe.
  • the "oil phase” sample, “water phase” sample is then injected into the PDMS microdroplet chip through a sample tube through an external syringe pump.
  • the generated microdroplets are collected through a sample tube into a conventional laboratory consumable such as an EP tube.
  • PDMS is a thermoelastic polymer material, which is not suitable for industrial grade injection molding and packaging processes. Manually processed PDMS microdroplet chips have poor reliability.
  • the present invention provides a microdroplet generating device based on a polymer material.
  • the microdroplet generating device is characterized by: (1) rapid, reliable, parallel generation of uniform micron-sized "water-in-oil” or “oil-in-water” microdroplets, and (2) microdroplet chip using thermoplastic material (eg polycarbonate material (PC), cyclic olefin copolymer (COP) or polymethyl methacrylate (PMMA), polypropylene (PP)), low cost in materials and batch processing, (3) by external pressure Source, using the micro-droplet generating device chip, sample injection, droplet collection process becomes convenient, (4) integrated micro-droplet generation device design, the whole process is not easy to produce cross-contamination.
  • thermoplastic material eg polycarbonate material (PC), cyclic olefin copolymer (COP) or polymethyl methacrylate (PMMA), polypropylene (PP)
  • PC polycarbonate material
  • COP cyclic olefin copolymer
  • the present invention proposes a microdroplet generating chip based on a polymer material.
  • the microdroplet chip combines the mature optical disc preparation process in the industry, and is characterized by: (1) rapidly and reliably generating uniform micron-sized "water-in-oil” or “oil-in-water” microdroplets, (2) pair The structure of the conventional circular ring disc is corrected, and the space of the optical disc is utilized to the maximum extent, and the micro-droplet generation flow path is arranged in parallel.
  • the present invention provides a micro-droplet generating device comprising a first component and a second component, the first component and the second component being fixedly coupled; the first component being a microdroplet generating chip for generating microdroplets; and the second component is a microdroplet sample loading and generating microdroplet collecting device for adding the oil phase sample and the water phase sample of the first component And the collection of generated microdroplets.
  • the microdroplet generating chip is circular or polygonal, and the polygon is preferably a hexagonal, octagonal or quadrilateral shape.
  • the microdroplet generating chip is a thermoplastic material, preferably a polycarbonate, a cyclic olefin copolymer, polymethyl methacrylate, and polypropylene.
  • the first component and the second component are sealed and fixedly connected by dispensing or ultrasonic welding.
  • first component and the second component are each integrally molded.
  • first component and the second component are thermoplastic materials, preferably polycarbonate materials, cyclic olefin copolymers or polymethyl methacrylate, polypropylene.
  • the first component is provided with at least one microdroplet generating unit, preferably four, eight and twelve microdroplet generating units;
  • the second component is provided with at least one microfluidic Drop loading and collecting unit, preferably four, eight and twelve micro-droplet generating units; each micro-droplet generating unit cooperates with its corresponding micro-droplet loading and collecting unit to perform micro-droplet addition Sample, generate, and collect.
  • each of the distances is equal to the distance between the standard eight-channel pipette tip.
  • the micro-droplet generating chip includes a central hole for injecting a plastic injection during the preparation of the micro-droplet generating chip and a micro-droplet generating chip during the mass production process. Transmitting; one or more micro-droplet generating units are disposed on both sides of the central hole centered on the central hole, and each micro-droplet generating unit independently generates micro-droplets; preferably, the central holes are equally spaced on both sides Four micro-droplet generating units are arranged.
  • the microdroplet generating unit includes an oil phase sample inlet, an oil phase sample conduit, an aqueous phase sample inlet, a water phase sample conduit, and a microdroplet outlet; the oil phase sample conduit And/or the aqueous phase sample line is an arcuate conduit structure remote from the central bore.
  • the microdroplet generating unit comprises two oil phase sample lines and one aqueous phase sample line, or one oil phase sample line and two aqueous phase sample lines; the oil phase The sample line and the aqueous phase sample line form a crisscross structure for forming microdroplets.
  • a return flow resistance zone is disposed in the pipeline after the oil phase sample inlet, and the oil phase sample flows through the return flow resistance zone, and then enters the oil phase sample pipeline separately into two channels.
  • a "water-in-oil" micro-droplet is formed; or a return-type flow-resistance zone is disposed in the pipeline after the inlet of the aqueous phase sample, and the aqueous phase sample flows through the return-type flow-resistance zone, and is divided into two paths respectively
  • the aqueous phase sample line produces "water-in-oil" microdroplets.
  • the oil phase sample line is provided with an oil phase sample filtration zone and/or the aqueous phase sample line is provided with an aqueous phase sample filtration zone; preferably, the oil phase sample filtration zone And/or the aqueous phase sample filtration zone is a set of columnar array structures, respectively.
  • an oil phase sample loading tank, an oil phase loading through hole, an aqueous phase sample loading tank, and a water phase loading through hole are disposed above the second component, wherein the oil phase loading a through hole and the aqueous phase loading through hole are respectively disposed in the bottom of the oil phase sample loading tank and the water phase sample loading tank, and the oil phase sample and the water phase sample respectively enter through the same a first component; and a sample collection device disposed below the second component.
  • the volume of the oil phase sample loading tank and the aqueous phase sample loading tank are respectively from 1 to 900 microliters, preferably from 5 to 500 microliters, more preferably from 100 to 200 microliters. .
  • the sample collection device includes a generated microdroplet receiving port and a microdroplet collecting port respectively disposed at both ends thereof, and a pre-storage cavity; the micro-droplet generated by the first component passes through the The microdroplet receiving port enters the second component, then passes through the pre-storage chamber, and finally collects from the micro-droplet collection port.
  • the microdroplet collection port is provided as an outlet having a slanted sidewall structure, and a connecting rod is disposed at an end thereof, and the microdroplets are dropped into the microdroplet collecting container through the connecting rod.
  • the micro-droplet collection container is a centrifuge tube
  • the connecting rod has a curved side wall; the connecting rod penetrates inside the centrifuge tube to facilitate micro-droplet collection.
  • the micro-droplet generation viewing area is disposed on the first component and the viewing window is disposed on the second component for monitoring the generated micro-droplets in real time in conjunction with the optical device.
  • the microdroplet generating device further includes a third component that seals the oil phase sample and the aqueous phase sample of the second component and applies external pressure therethrough.
  • the first component and the second component are respectively provided with positioning holes for facilitating the positioning of the two.
  • FIG. 1 is a schematic view showing the overall structure of a micro-droplet generating device of the present invention
  • Figure 2 is a schematic view showing the structure of a first component of the micro-droplet generating device of the present invention
  • Figure 3 is a schematic view showing the "cross structure" of the micro-droplet generation of the first component of the micro-droplet generating device of the present invention
  • Figure 4 is a schematic view showing the structure of a second component of the micro-droplet generating device of the present invention.
  • Figure 5 is a schematic view showing the structure of a collecting device of a second component of the present invention.
  • FIG. 6 is a schematic structural view of a micro-droplet generating chip of the present invention.
  • FIG. 7 is a structural view of a return flow resistance region of a micro-droplet-forming chip oil phase sample injection according to the present invention.
  • Figure 8 is a schematic view showing the structure of a filtration zone for injecting an oil phase sample of a micro-droplet generating chip of the present invention
  • Figure 9 is a schematic view showing the structure of the micro-droplet generating chip cross generation of the present invention.
  • Figure 10 is a schematic view showing the structure of an observation area of a micro-droplet generating chip of the present invention.
  • Embodiment 1 A micro-droplet generating device of the present invention
  • the micro-droplet generating device of the present invention comprises: a first component 1 which is a micro-droplet generating chip, and a second component 2 which is a sample and generation of a micro-droplet sample. Droplet collection device.
  • the first component 1 is fixedly connected to the second component 2.
  • the microdroplet generating device of the present invention further comprises a third component 3, the third component being a microdroplet oil phase sample loading bath and an aqueous phase sample loading slot for sealing the second component 2.
  • the third component 3 is, for example, a water-tight gasket, for example made of silica gel, which may be provided with air holes for applying pressure therethrough, which serves to ensure crimping between the external pressure source and the second component. It is airtight and easy to apply external pressure. By means of external pressure, two samples flow from the second component into the first component into the microchannel of the first component. Finally, the generated microdroplets are collected into a microdroplet collection container, such as a centrifuge tube (EP tube).
  • a microdroplet collection container such as a centrifuge tube (EP tube).
  • each micro-droplet generating unit is disposed in parallel on the first member 1, and from left to right, each micro-droplet generating unit is equally arranged in parallel at the first member. On the 1st, each micro-droplet generating unit independently generates micro-droplets.
  • the number of microdroplet generating units in the first component 1 depends somewhat on the size of the microdroplet generating chip and the need to generate droplets.
  • the currently used microdroplet generating structure is a "cross" structure that generates microdroplets, for example, to produce microdroplets having a diameter of 100 microns, and the "cross" structure has a width and depth of 70 micrometers.
  • each microdroplet generating unit includes an oil phase sample inlet 11, two oil phase sample lines 12, a water phase sample inlet 13, an aqueous phase sample line 14, and a microdroplet outlet 15.
  • oil phase sample enters the phase sample receiving port 11 of the first component 1, it is branched to the two oil phase sample lines 12.
  • aqueous phase sample enters the aqueous phase sample inlet 13 of the first component 1, it enters an aqueous phase sample line 14.
  • the two phase liquids meet in the cross structure to form microdroplets which then flow out of the microdroplet outlet 15.
  • the second component 2 is an injection that achieves an "oil phase” sample, a "aqueous phase” sample, and microdroplet collection.
  • eight micro-droplet loading and collecting units are disposed in parallel on the second member 2, from left to right.
  • Each microdroplet loading and collecting unit is arranged in parallel on the second component 2, and each microdroplet loading and collecting unit cooperates with each microdroplet generating unit for the microdroplet generating unit.
  • the second component 2 is provided with an oil phase sample loading tank 21, an oil phase loading through hole 22, an aqueous phase sample loading tank 23, and a water phase loading through hole 24, wherein the oil phase loading through hole 22 and the water phase
  • the sample insertion through holes 24 are respectively disposed at the center of the bottom of the oil phase sample loading tank 21 and the aqueous phase sample loading tank 23; a sample collecting device 25 is disposed below the second member 2, and micro generation is provided at both ends of the sample collecting device 25, respectively.
  • the droplet receiving port 26 and the microdroplet collecting outlet 27 is provided with an oil phase sample loading tank 21, an oil phase loading through hole 22, an aqueous phase sample loading tank 23, and a water phase loading through hole 24, wherein the oil phase loading through hole 22 and the water phase
  • the sample insertion through holes 24 are respectively disposed at the center of the bottom of the oil phase sample loading tank 21 and the aqueous phase sample loading tank 23; a sample collecting device 25 is disposed below the second member 2, and micro generation is provided at both ends of the sample collecting device 25,
  • the second component 2 is provided with a viewing window 4 for monitoring the generated microdroplets in real time in conjunction with the optical device.
  • the observation window 4 is a hollow design, and the observation window 4 is located before the micro droplet receiving port 26 is formed, and the micro droplets generated by the first member 1 can be observed.
  • the generated microdroplet receiving port 26 of the sample collecting device 25 is a through hole corresponding to the microdroplet outlet 15 in the first member 1, and a pre-storage chamber 28 is provided after the microdroplet receiving port 26 is formed.
  • the micro-droplet collection outlet 27 behind the pre-storage chamber 28 is provided as an outlet having a slanted side wall structure, and at its end is provided a connecting rod 29 for connection to the EP tube, the connecting rod 29 having curved side walls.
  • the connecting rod 29 penetrates the inside of the EP tube to facilitate micro-droplet collection.
  • the micro-droplets generated in the first member 1 pass through the micro-droplet outlet 15 and the droplet-forming through-hole 26 of the second member under pressure to enter the pre-storage chamber 28, and the density of the micro-droplets Less than the oil phase sample will float above the oil, and as the microdroplets and oil continue to flow, the microdroplets will slide into the EP tube along the oblique side walls of the microdroplet collection outlet 27.
  • the first component 1 and the second component 2 are in the oil phase sample inlet 11 and the oil phase loading via 22, the aqueous phase sample inlet 13 and the aqueous phase loading via 24, and the microdroplet outlet 15 is open.
  • the aperture and the resulting droplet outlet 26 are each surrounded by a dot seal.
  • the entire workflow is divided into three steps: (1) sample injection step, (2) microdroplet generation step, and (3) microdroplet collection step.
  • the "oil phase” sample and the "aqueous phase” sample are separately added to the oil phase sample loading tank 21 and the aqueous phase sample loading tank 23 of the second member 2.
  • the two samples pass through the oil phase loading through hole 22 and the water phase loading through hole 24, respectively, into the oil phase sample inlet 11 and the water phase sample receiving hole 13 of the first member 1, respectively; Entering the two oil phase sample line 12, the water phase enters one of the aqueous phase sample lines 14.
  • the oil phase and the aqueous phase form a uniform microdroplet at the cross structure of the first component 1.
  • the generated microdroplets pass through the microdroplet outlet 15 of the first component 1 and the resulting droplet exit 26 of the second component into the pre-storage cavity 28 of the second component, and the microdroplets of the pre-storage cavity 28 pass through the collection outlet 27
  • An EP tube connected to the sample collection device completes the generation and collection of microdroplets.
  • the oil phase sample and the aqueous phase sample are previously placed in the oil phase sample loading tank 21 and the aqueous phase sample loading tank 23 of the second member 2. Under the action of external pressure, the two samples pass through the oil phase loading through hole 22 and the water phase loading through hole 24, respectively, and enter the oil phase sample inlet 11 and the water phase sample receiving hole 13 of the first member 1, respectively; The phase enters two oil phase sample lines 12 and the water phase enters one of the aqueous phase sample lines 14.
  • FIG. 3 A typical "water-in-oil” micro-droplet generation process is shown in Figure 3:
  • the "oil phase” sample flows from the horizontal direction into the micron-scale pipeline under external air pressure; the "aqueous phase” sample is under the action of external air pressure. Flows into the micron-scale pipe in the vertical direction. Two immiscible liquids meet at the "cross” microfluidic structure. Due to the difference in liquid surface tension between the "oil phase” sample and the "aqueous phase” sample and the shear force generated by the applied pressure, the “aqueous phase” sample is separated from the continuous phase into discrete micro-phases by the "oil phase” sample at the cross structure. Droplet.
  • the microdroplets are in the form of "water-in-oil” and the outside is the "oil phase” sample.
  • the generated microdroplets pass through the microdroplet outlet 15 of the first component 1 and the resulting droplet exit 26 of the second component into the pre-storage cavity 28 of the second component, and the microdroplets of the pre-storage cavity 28 pass through the collection outlet 27
  • An EP tube connected to the sample collection device completes the generation and collection of microdroplets.
  • the micro droplets are generated in the first component microchannel and then flow to the microdroplet outlet 15 of the first component 1 and the droplet outlet 26 of the second component, and the droplet floats up under pressure.
  • the pre-storage chamber 28 of the two component 2 the density of the micro-droplets is smaller than that of the oil phase sample, and will float above the oil.
  • the micro-droplets and oil continue to flow, the micro-droplets will follow the oblique sidewall of the collection outlet 27. Slide into the EP tube and connect the rod 29 deep inside the EP tube to facilitate micro-droplet collection.
  • Embodiment 2 A microdroplet generating chip of the present invention
  • FIG. 6 is a schematic diagram showing the structure of a micro-droplet generating chip. From left to right, eight micro-droplet generating units 41 are designed on an octagonal chip, and a central hole 42 is formed in the center of the chip, and the central hole 42 is from a disc processing process for injecting plastic injection and batch The transport of substrates during the production process. The traditional circular disc structure is not easy to position, the chip is processed into an octagonal structure, and two positioning holes 43 are processed to facilitate the positioning and cooperation of the micro-droplet chip and related equipment.
  • Four identical micro-droplet generating units 41 are arranged at equal intervals on both sides of the center hole for generating micro-droplets in parallel.
  • each of the droplet generating units 41 includes an oil phase sample inlet 4111, a return flow resistance region 4112, two oil phase sample tubes 4113, two oil phase filtration regions 4114, and water from top to bottom.
  • the micro-droplet generation chip shown in FIG. 6 corrects the standard structure of the conventional optical disc, and can maximize the space of the optical disc and arrange the micro-droplet generation flow paths in parallel.
  • the chip processed by the precision injection molding process, combined with the return flow resistance zone and the filter zone design quickly and reliably generate uniform micron-sized "water-in-oil" microdroplets.
  • the oil phase sample is first injected into the oil phase sample inlet 4111 using an external air pump or a peristaltic pump.
  • a loop-shaped flow resistance region 4112 is designed after the oil phase sample inlet 4111, which is composed of a plurality of U-shaped tubes.
  • the oil phase sample will be infiltrated on the surface of the polymer material, and under capillary pressure, the oil phase sample will automatically flow into the microchannel under capillary pressure. In extreme cases, the oil phase sample continues to flow under capillary action.
  • the purpose of designing the flow resistance zone 4112 is to accurately control the injection volume of the oil phase sample, and to minimize the continuous flow of the oil phase sample in the microchannel under capillary action, so that the oil phase sample injection amount is only by the external air pump or the peristaltic pump. control.
  • the oil phase sample is passed through an oil phase split inlet and split into two oil phase sample lines 4113 of the same design, the oil phase sample line 4113 being a curved structural line away from the center hole 42.
  • each of the two oil phase sample lines 4113 enters an oil phase filtration zone 4114.
  • the oil phase filtration zone 4114 is a set of columnar array structures.
  • the columnar array structure has a plurality of rows of columnar arrays interlaced. Impurities (particles, silk fibers, etc.) present in the oil phase are blocked at the set of columnar structures and do not affect the formation of microdroplets.
  • the aqueous phase sample is first injected into the aqueous phase sample inlet 4121 using an external air pump or a peristaltic pump. Similar to the oil phase sample injection design, the aqueous phase sample enters an aqueous phase sample filtration zone 4122 and an aqueous phase sample line 4123.
  • the filtration zone is a set of columnar array structures designed to filter impurities in the aqueous phase sample and to eliminate the effects of impurities on droplet formation.
  • two oil phase sample lines 4113 and one aqueous phase sample line 4123 form a "cross" structure in the microdroplet formation region 413 for the generation of microdroplets.
  • Two “oil phase” samples flow through the two oil phase sample lines 4113 from the horizontal direction into the micron-scale pipeline under the action of external air pressure; one "aqueous phase” sample passes through the aqueous phase sample line 4123 under the action of external air pressure. Flows into the micron-scale pipe in the vertical direction. Two immiscible liquids meet at the "cross" microfluidic structure.
  • the "aqueous phase” sample Due to the difference in liquid surface tension between the "oil phase” sample and the “aqueous phase” sample and the shear force generated by the applied pressure, the "aqueous phase” sample is separated from the continuous phase into discrete micro-phases by the "oil phase” sample at the cross structure. Droplet.
  • the microdroplet generation viewing zone 414 is intended to facilitate real-time monitoring of microdroplets in conjunction with an optical device.
  • the characteristic structure of the microdroplet generation observation zone 414 is to the left of the micropipeline, and the closed microstructure is designed. Since the oil-phase/aqueous phase liquid and micro-droplets do not flow through the closed structure, the optical detecting device can collect a stable static image and focus on the micro-pipe plane, thereby obtaining clear detection results. The generated microdroplets flow out through the resulting microdroplet outlet 415.
  • the standard structure of the conventional optical disc is corrected to maximize the space of the optical disc and facilitate the positioning of the chip during use.
  • the central aperture 42 is derived from a disc processing process for injecting injection molding and for transporting the substrate during mass production.
  • the traditional circular disc structure is not easy to position. Therefore, the chip is processed into an octagonal structure, and two positioning holes 43 are machined to facilitate the positioning and cooperation of the micro-droplet chip and the related device.

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PCT/CN2018/113851 2017-11-06 2018-11-03 微液滴生成装置 Ceased WO2019086018A1 (zh)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2020543685A JP7030361B2 (ja) 2017-11-06 2018-11-03 微小液滴生成装置

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
CN201711074976.3 2017-11-06
CN201711074985.2A CN109746062B (zh) 2017-11-06 2017-11-06 微液滴生成装置
CN201711074977.8A CN109746060B (zh) 2017-11-06 2017-11-06 微液滴生成芯片
CN201711074976.3A CN109746059B (zh) 2017-11-06 2017-11-06 微液滴生成系统
CN201711074977.8 2017-11-06
CN201711074985.2 2017-11-06

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CN110756233A (zh) * 2019-10-28 2020-02-07 广东顺德工业设计研究院(广东顺德创新设计研究院) 微滴制备系统、微流控芯片及微滴制备方法
CN113413934A (zh) * 2021-07-16 2021-09-21 安徽骆华生物科技有限公司 十字流动聚焦型液滴微流控芯片的制备方法及应用

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