WO2023236760A1 - Appareil d'impression couplé à une puce microfluidique et procédé d'impression - Google Patents

Appareil d'impression couplé à une puce microfluidique et procédé d'impression Download PDF

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Publication number
WO2023236760A1
WO2023236760A1 PCT/CN2023/095303 CN2023095303W WO2023236760A1 WO 2023236760 A1 WO2023236760 A1 WO 2023236760A1 CN 2023095303 W CN2023095303 W CN 2023095303W WO 2023236760 A1 WO2023236760 A1 WO 2023236760A1
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WO
WIPO (PCT)
Prior art keywords
printing
channel
auxiliary
liquid
droplets
Prior art date
Application number
PCT/CN2023/095303
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English (en)
Chinese (zh)
Inventor
陈华英
余恩
赵文涛
Original Assignee
珠海大略科技有限公司
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
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Application filed by 珠海大略科技有限公司 filed Critical 珠海大略科技有限公司
Publication of WO2023236760A1 publication Critical patent/WO2023236760A1/fr

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Classifications

    • 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
    • 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
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06F—ELECTRIC DIGITAL DATA PROCESSING
    • G06F3/00—Input arrangements for transferring data to be processed into a form capable of being handled by the computer; Output arrangements for transferring data from processing unit to output unit, e.g. interface arrangements
    • G06F3/12—Digital output to print unit, e.g. line printer, chain printer
    • G06F3/1201—Dedicated interfaces to print systems
    • G06F3/1202—Dedicated interfaces to print systems specifically adapted to achieve a particular effect
    • G06F3/1211—Improving printing performance
    • G—PHYSICS
    • G06—COMPUTING; CALCULATING OR COUNTING
    • G06T—IMAGE DATA PROCESSING OR GENERATION, IN GENERAL
    • G06T7/00—Image analysis
    • G06T7/0002—Inspection of images, e.g. flaw detection
    • G06T7/0012—Biomedical image inspection

Definitions

  • hSCP Hand-Held Single-Cell Pipettes
  • the piezoelectric single-cell printing of Peter Koltay et al. has high requirements on equipment, is complex to manufacture, expensive, and requires other precision equipment to provide electrical signals.
  • the method of Feng, Lin et al. and the pipette-type single-cell printing of Lidong Qin et al. cannot ensure gentle fluid shear force through pressurized printing, which is more harmful to cells. The most important thing is that pressurization will cause It greatly affects the hydrodynamic pressure distribution of the flow channel inside the chip, causing fluid backflow inside the chip, which is not suitable for high-throughput cell printing. List
  • the upward punching and drainage method of Huaying et al. can easily cause cells to settle at the outlet due to gravity.
  • the purpose of the invention is to develop a printing outlet that assists a single cell printing system to realize printing inoculation.
  • the outlet includes one or more auxiliary printing channels parallel to the main printing channel, which can control the flow rate at will, in addition to the main working channel.
  • the auxiliary printing channel will flow out liquid to form large droplets that can drip quickly and merge with the microdroplets at the printing outlet.
  • the flow rate of the auxiliary printing channel can be increased to accelerate the formation of droplets, which greatly improves the efficiency of printing cells or particles.
  • This method does not require additional equipment, is low cost, simple to manufacture, and has flexible control methods.
  • this method can also control the time interval of droplet formation through image recognition, so that a falling droplet contains a desired number of cells or particles. Ultimately, high-precision printing of single or multiple cells/particles can be achieved.
  • the purposes of the invention include:
  • the printing outlet is formed in one step without secondary processing, so that no other factors block the printed cells or microspheres in the channel.
  • the auxiliary printing channel is not directly connected to the printing outlet and has no impact on the hydrodynamic characteristics of the fluid in the chip;
  • the present invention provides a printing device and a printing method that are matched with the microfluidic chip; dynamically adjusting the flow rate of the auxiliary printing channel through image recognition of cells and particles to control droplet formation, achieving efficient and continuous
  • the printing of single or multiple cells or particles can also be achieved with the printing of water-in-oil or oil-in-water droplets.
  • a printing device that matches the microfluidic chip including a single-sided auxiliary printing channel, a double-sided auxiliary printing channel and a four-sided surrounding auxiliary channel, are all used to help particles print quickly.
  • the single-sided auxiliary printing channel includes: a particle sample inlet; a printing channel is provided at the bottom of the particle sample inlet, an auxiliary printing channel is provided on one side of the printing channel, and an auxiliary liquid supply inlet is provided at the top of the auxiliary printing channel.
  • the double-sided auxiliary printing channel includes a particle sample inlet, a printing channel is set at the bottom of the particle sample inlet, auxiliary printing channels are set on both sides of the printing channel, an auxiliary liquid supply inlet is set at the bottom of the auxiliary printing channel, and the printing outlet is set at the printing The size of the bottom end of the channel, the tip of the print outlet, is used to determine the size of the final droplet.
  • the four-sided surrounding auxiliary channel includes an auxiliary liquid supply outlet, an auxiliary printing channel and an auxiliary printing channel annular liquid; an auxiliary liquid supply outlet is arranged at the top of the auxiliary printing channel, and an auxiliary printing channel annular liquid is arranged at the bottom of the auxiliary liquid supply outlet.
  • the present invention can also form oil-in-water or water-in-oil droplets by controlling the flow rate.
  • the printing method of a printing device matching the microfluidic chip includes the following steps:
  • Step 1 Before the system works, the printing system needs to be placed vertically;
  • Step 2 The printing channel is open, the auxiliary printing channel is closed, and the image recognition system is used to determine that single or multiple particles flow out with the liquid to the outside of the printing port to form tiny droplets;
  • Step 3 Open the auxiliary printing channel, inject a certain amount of auxiliary printing liquid into the outlet at a specific time according to the required droplet size, so that it merges with the tiny droplets in step 2 to form large droplets and drips immediately;
  • Step 4 Close the auxiliary printing channel
  • Step 5 Repeat the above steps 2 to 4 to achieve loop printing.
  • the printing channel in step 2 can be liquid water without any particles.
  • the auxiliary channel is oil
  • this method can be used to produce water-in-oil droplets. The amounts of water and oil in the droplets can be adjusted;
  • the printing channel is oil and the auxiliary channel is water, producing oil-in-water droplets.
  • the size of the droplets is related to the tip size of the printing outlet. The smaller the tip of the chip, the smaller the droplets that can be dropped;
  • the auxiliary channel is oil. At this time, water-in-oil is printed, with single or multiple particles or cells in the water;
  • the auxiliary channel can be the same water or cell culture medium as the liquid in the printing channel. At this time, large droplets will be printed and drip directly.
  • this inventive principle is applied to the chip through the matching printing principle of the microfluidic chip, including the following forms: one-time molding and manufacturing, combined bonding of the print head and chip, and wraparound liquid printing.
  • the number of particles is identified based on the optical signal image, and then the flow rate of the auxiliary printing channel is controlled by electrical signals to control the size and dripping time of the droplets to achieve the printing of multiple droplets.
  • Different wrapped droplets are formed according to the flow rate, enabling printing of water-in-oil or oil-in-water droplets.
  • Figure 1 is a schematic structural diagram of the method of the present invention.
  • A) a schematic diagram of the auxiliary printing channel on one side;
  • B) a schematic diagram of the auxiliary printing channel on both sides.
  • FIG. 2 Schematic diagram of the principle of printing single particles in the printing structure;
  • FIG. 3 Schematic diagram of the printing structure printing single particle process
  • A) Schematic diagram of image recognition dynamic control-assisted printing. After the particles arrive at the exit of the printing channel and are recognized by the camera image and receive a high-frequency pulse, the program controls to increase the flow rate of the auxiliary printing channel to accelerate the formation of large droplets at the exit end;
  • Figure 5 is a schematic diagram of photolithography of the printing channel, auxiliary printing channel and upstream sorting microfluidic chip designed by the present invention, including a printing channel and an auxiliary printing channel.
  • Figure 6 is a schematic diagram of the bonding between the print head and the upstream microfluidic chip.
  • the print head includes an auxiliary printing channel and a printing channel
  • the microfluidic chip includes an auxiliary printing liquid supply channel and a particle channel.
  • the present invention is mainly a printing device that can be matched with a microfluidic chip and help particles to be printed quickly.
  • the printing design is shown in Figure 1.
  • Figure 1A is a single-sided auxiliary printing channel design, including a Particle sample inlet, an auxiliary liquid supply inlet, an auxiliary printing channel and a printing channel.
  • Figure 1B shows the design of the double-sided auxiliary printing channel, which includes a particle sample inlet, an auxiliary liquid supply inlet, two auxiliary printing channels and a printing channel. aisle.
  • Step 1 Before the system works, the printing system needs to be placed vertically;
  • Step 3 Open the auxiliary printing channel, inject a certain amount of auxiliary printing liquid into the outlet at a specific time according to the required droplet size, so that it merges with the tiny droplets in step 2 to form large droplets and drips immediately;
  • Step 5 Repeat the above steps 2 to 4 to achieve loop printing.
  • this fast printing method can also be combined with an image recognition system to realize automated printing and save operating steps.
  • the printing channel can be liquid without any particles, such as water. If the auxiliary channel is oil, this method can be used to produce water-in-oil droplets. The amounts of water and oil are adjustable. Conversely, oil-in-water droplets can be produced.
  • the auxiliary channel can be the same as the liquid in the printing channel, such as water or cell culture medium. At this time, the printed liquid will be large droplets that drip directly.
  • the function of the auxiliary channel is: (1) Provide a different liquid from the printing channel to form water-in-oil or oil-in-water, and make it drip quickly by forming larger droplets (2) Provide the same liquid as the printing channel , mainly to increase the volume of liquid and make it drip quickly.
  • Step 1 Before the system works, the printing system needs to be placed vertically;
  • Step 2 The printing channel is open, the auxiliary printing channel is closed, and the image recognition system is used to determine that single or multiple particles flow out with the liquid to the outside of the printing port to form tiny droplets;
  • Step 3 Open the auxiliary printing channel, inject a certain amount of auxiliary printing liquid into the outlet at a specific time according to the required droplet size, so that it merges with the tiny droplets in step 2 to form large droplets and drips immediately;
  • Step 4 Close the auxiliary printing channel
  • Step 5 Repeat the above steps 2 to 4 to achieve loop printing.
  • This printing method is a structural design downstream of the particle sorting system. After the upstream sorting system completes the particle sorting, it is quickly printed through this printing structure.
  • the method of the present invention can be divided into four types when applied to the upstream microfluidic system.
  • the print head contains an auxiliary printing channel
  • the microfluidic chip contains a printing channel.
  • the liquid flowing out of the last auxiliary printing channel flows out in an annular shape.
  • the annular channel surrounds the particle printing channel, so that the liquid in the auxiliary printing channel can better wrap the particle sample flowing out of the printing channel.
  • the print head contains an auxiliary liquid supply inlet, an auxiliary printing channel and an auxiliary printing channel annular liquid outlet, and the microfluidic chip contains a particle channel.
  • the present invention can also be applied to droplet microfluidics to form water pockets by controlling the flow rate.
  • Oil or water-in-oil droplets As shown in Figure 9, water is introduced into the printing channel, and oil is introduced into the auxiliary printing channel.
  • the steps to form water-in-oil droplets are as follows:
  • Step 1 Before the system works, the printing system needs to be placed vertically.
  • Step 2 Keep the auxiliary printing channel closed, open the printing channel, and allow a certain amount of printing liquid (water) to flow out of the printing port to form small water droplets;
  • Combination bonding of print head and chip means that the print head is made separately and then assembled with the chip.

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  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Sustainable Development (AREA)
  • Theoretical Computer Science (AREA)
  • General Physics & Mathematics (AREA)
  • Physics & Mathematics (AREA)
  • Dispersion Chemistry (AREA)
  • Molecular Biology (AREA)
  • Analytical Chemistry (AREA)
  • Human Computer Interaction (AREA)
  • Clinical Laboratory Science (AREA)
  • Cell Biology (AREA)
  • Medical Informatics (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Radiology & Medical Imaging (AREA)
  • Quality & Reliability (AREA)
  • Computer Vision & Pattern Recognition (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

La présente invention concerne un appareil d'impression couplé à une puce microfluidique et un procédé d'impression. La tête d'impression comprend : un canal d'impression conçu pour produire des particules et un liquide spécifique, et un canal auxiliaire situé sur un ou deux côtés du canal d'impression ou entourant le canal d'impression. Lorsqu'un système de reconnaissance d'image détermine qu'un certain nombre de particules ou un certain volume de liquide est délivré par l'intermédiaire d'un orifice d'impression, un liquide d'impression auxiliaire est rapidement délivré à travers le canal auxiliaire et mélangé avec les gouttelettes autour de l'orifice d'impression, facilitant ainsi la chute naturelle des gouttelettes sous l'action de la gravité. La présente invention favorise la chute naturelle des gouttelettes de liquide contenant le liquide d'impression ou les particules en augmentant le volume des gouttelettes, ce qui permet d'éviter efficacement l'influence des procédés d'impression conventionnels reposant sur la pulvérisation rapide sur les particules à l'intérieur du liquide, notamment sur l'activité cellulaire.
PCT/CN2023/095303 2022-06-10 2023-05-19 Appareil d'impression couplé à une puce microfluidique et procédé d'impression WO2023236760A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202210655537.6A CN114958566A (zh) 2022-06-10 2022-06-10 一款和微流控芯片匹配的打印装置及打印方法
CN202210655537.6 2022-06-10

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WO2023236760A1 true WO2023236760A1 (fr) 2023-12-14

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114958566A (zh) * 2022-06-10 2022-08-30 珠海大略科技有限公司 一款和微流控芯片匹配的打印装置及打印方法

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110016429A (zh) * 2018-01-09 2019-07-16 四川大学华西医院 一种含细胞的双重液滴及其制备方法
CN110295109A (zh) * 2019-07-08 2019-10-01 中国科学院深圳先进技术研究院 基于微流控液滴打印系统的数字pcr检测方法及应用
CN112342137A (zh) * 2020-11-25 2021-02-09 中国科学技术大学 一种基于图像处理和微流控打印的单细胞分选装置及方法
CN113477282A (zh) * 2021-04-25 2021-10-08 深圳大学 一种基于液滴微流控的单细胞分离系统及方法
CN114958566A (zh) * 2022-06-10 2022-08-30 珠海大略科技有限公司 一款和微流控芯片匹配的打印装置及打印方法

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110016429A (zh) * 2018-01-09 2019-07-16 四川大学华西医院 一种含细胞的双重液滴及其制备方法
CN110295109A (zh) * 2019-07-08 2019-10-01 中国科学院深圳先进技术研究院 基于微流控液滴打印系统的数字pcr检测方法及应用
CN112342137A (zh) * 2020-11-25 2021-02-09 中国科学技术大学 一种基于图像处理和微流控打印的单细胞分选装置及方法
CN113477282A (zh) * 2021-04-25 2021-10-08 深圳大学 一种基于液滴微流控的单细胞分离系统及方法
CN114958566A (zh) * 2022-06-10 2022-08-30 珠海大略科技有限公司 一款和微流控芯片匹配的打印装置及打印方法

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