CN117448151B - Sorting device and sorting method for on-chip liquid flow path cells or particles - Google Patents
Sorting device and sorting method for on-chip liquid flow path cells or particles Download PDFInfo
- Publication number
- CN117448151B CN117448151B CN202311800730.5A CN202311800730A CN117448151B CN 117448151 B CN117448151 B CN 117448151B CN 202311800730 A CN202311800730 A CN 202311800730A CN 117448151 B CN117448151 B CN 117448151B
- Authority
- CN
- China
- Prior art keywords
- micro
- sorting
- flow
- liquid
- channel
- 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.)
- Active
Links
- 239000007788 liquid Substances 0.000 title claims abstract description 136
- 239000002245 particle Substances 0.000 title claims abstract description 80
- 238000000034 method Methods 0.000 title claims abstract description 25
- 238000002347 injection Methods 0.000 claims abstract description 121
- 239000007924 injection Substances 0.000 claims abstract description 121
- 230000003287 optical effect Effects 0.000 claims abstract description 66
- 238000010897 surface acoustic wave method Methods 0.000 claims abstract description 37
- 238000000926 separation method Methods 0.000 claims abstract description 22
- 238000001514 detection method Methods 0.000 claims abstract description 21
- 239000002699 waste material Substances 0.000 claims description 16
- 238000012545 processing Methods 0.000 claims description 14
- 238000011084 recovery Methods 0.000 claims description 12
- 238000003860 storage Methods 0.000 claims description 8
- 230000001154 acute effect Effects 0.000 claims description 3
- 108010076504 Protein Sorting Signals Proteins 0.000 claims description 2
- 238000005516 engineering process Methods 0.000 description 17
- 239000012530 fluid Substances 0.000 description 16
- 230000004907 flux Effects 0.000 description 14
- 229960000074 biopharmaceutical Drugs 0.000 description 7
- 230000008569 process Effects 0.000 description 5
- 230000009471 action Effects 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 230000007547 defect Effects 0.000 description 4
- 238000013461 design Methods 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 230000001360 synchronised effect Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 238000011010 flushing procedure Methods 0.000 description 2
- 238000009652 hydrodynamic focusing Methods 0.000 description 2
- 230000001678 irradiating effect Effects 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000010008 shearing Methods 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 238000001069 Raman spectroscopy Methods 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 230000005686 electrostatic field Effects 0.000 description 1
- 238000000684 flow cytometry Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 238000003754 machining Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000005855 radiation Effects 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000009987 spinning Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
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
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/04—Cell isolation or sorting
-
- 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
- C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1404—Handling flow, e.g. hydrodynamic focusing
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N2015/0019—Means for transferring or separating particles prior to analysis, e.g. hoppers or particle conveyors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N2015/1006—Investigating individual particles for cytology
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Sustainable Development (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Analytical Chemistry (AREA)
- Dispersion Chemistry (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Molecular Biology (AREA)
- Clinical Laboratory Science (AREA)
- Cell Biology (AREA)
- Automatic Analysis And Handling Materials Therefor (AREA)
Abstract
The invention provides a sorting device and a sorting method for on-chip liquid flow path cells or particles. The sorting apparatus includes: the device comprises a sheath liquid pump, a sample injection pump, a rotary disc slide, an optical system, a surface acoustic wave generator and a collecting device; n micro-channels are arranged on the rotary disc slide from the center to the edge in an extending mode, and M sorting branch channels are respectively arranged on each micro-channel. The invention adopts a rotary disk slide micro-channel network to realize high-precision flow control and micro-droplet generation by multi-time slot diversion, adopts a surface acoustic wave generator and combines a detection feedback control algorithm to realize accurate manipulation and separation of cells or particles, and realizes high-flux flow separation.
Description
Technical Field
The invention relates to the technical field of microfluidics, in particular to a sorting device and a sorting method for on-chip liquid flow path cells or particles.
Background
When single cells or particles flow through single or multiple lasers in the full spectrum flow cytometer, the detector of the flow cytometer can detect scattered light information and multiple fluorescent signals of each cell or particle, analyze information on the cells or particles, and rapidly identify and characterize the cells and particles. However, how cells or particles in a fluid suspension move in a random and unorganized manner, precisely manipulate and separate, is a liquid flow path core technique of flow cytometry.
CN104204768A discloses a sort flow cytometer comprising: fluid nozzles, lasers, computing devices, sorting control electronics, deflection plates. Wherein the computing device: receiving event data from the acquisition electronics and generating a primary sort decision assigned to a segment of the fluid stream; an undesired sequence of droplet charges allocated to at least two adjacent segments of the fluid stream is identified and a final sort decision is generated by modifying the primary sort decision. The sorting flow cytometer adopts a traditional sheath fluid hydrodynamic focusing and superimposed electrostatic field auxiliary separation method; but the instrument has more high-voltage components and higher overall cost.
CN101971247a discloses a system and method for acoustically focusing hardware and implementations that provides a capillary and at least one vibration source; machining a groove in the vibration source; and coupling the at least one vibration source to the capillary at the recess. The method adopts capillary hydrodynamic focusing and conventional acoustic wave assisted focusing methods, can realize fixed-point positioning, but cannot realize separation.
CN103331185a discloses a microfluidic device comprising: a substrate, a reagent receiving slot, a reagent structure, a microfluidic flow channel formed in the substrate, wherein the flow channel is fluidly coupled to the reagent receiving slot. The device adopts a method of on-chip micro-flow channel and magnetic field separation, can realize flow separation, but has lower flux.
In view of this, the present invention has been made.
Disclosure of Invention
The invention aims to provide a sorting device for on-chip liquid flow path cells or particles, which can obtain larger liquid flow, realize high-precision flow control and micro-droplet generation by multi-channel time slot diversion, and realize accurate manipulation and separation of cells or particles by combining a detection feedback control algorithm so as to realize flow sorting.
The second object of the present invention is to provide an application of the sorting device for on-chip liquid flow cell or particle in sorting of on-chip liquid flow cell or particle.
A third object of the present invention is to provide a method for sorting on-chip liquid flow cell or particle using the above-mentioned apparatus for sorting on-chip liquid flow cell or particle.
In order to achieve the above object of the present invention, the following technical solutions are specifically adopted:
in a first aspect, the present invention provides a sorting apparatus for on-chip liquid flow path cells or particles, the sorting apparatus comprising: sheath pump, sample injection pump, rotating disk slide, optical system, surface acoustic wave generator and collection system.
N micro-channels are arranged on the rotary disc slide from the center to the edge in an extending mode, and M sorting branch channels are respectively arranged on each micro-channel.
And an injection port of the sheath liquid pump is connected with a liquid inlet at the center of the rotary disc slide.
And an injection port of the sample injection pump is connected with a liquid inlet of the micro-channel.
The optical system comprises a light source and an optical detector, and the light source and the optical detector are arranged at one end close to the liquid inlet of the micro-channel.
The surface acoustic wave generator is arranged at the liquid inlet end of the separation branch flow passage.
The acquisition device is arranged at the liquid outlet end of the sorting branch flow passage.
Because various application scenes such as biopharmaceuticals need low-cost and high-flux flow separation technology, the invention adopts a large-flux injection pump to replace a high-precision small-flux high-pressure injection pump in a conventional scheme to obtain larger liquid flux, and simultaneously adopts a rotary disk slide micro-channel network to realize high-precision flow control and micro-droplet generation by multi-channel time slot diversion, and adopts a pair of interdigital electrode dipole source surface acoustic wave generators and a detection feedback control algorithm to realize accurate manipulation and separation of cells or particles, thereby realizing high-flux flow separation.
More specifically, the sorting device specifically includes the following structure:
sheath liquid pump
In the invention, the sheath liquid pump is used for injecting sheath liquid into the center of the rotary disc slide; i.e. the injection port of the sheath pump may be connected to the inlet port in the center of the rotating disc slide.
In alternative embodiments of the invention, the sheath pump may be a fixed position electrically driven high flux current carrying syringe pump; this is due to the low cost, high throughput flow-through sorting technology required in various application scenarios such as biopharmaceuticals; the invention adopts a large-flux injection pump to replace a small-flux high-pressure injection pump in the conventional scheme, so that larger liquid flow flux can be obtained.
II. Injection pump
In the invention, the sample injection pump is used for injecting a sample into any micro-channel arranged on the rotary disc slide, namely, an injection port of the sample injection pump can be connected with a liquid inlet of the micro-channel.
In preferred embodiments of the invention, the sample injection pump may be a fixed position electrically driven large throughput sample injection pump; this is due to the low cost, high throughput flow-through sorting technology required in various application scenarios such as biopharmaceuticals; the sorting device can adopt a large-flux injection pump to replace a small-flux high-pressure injection pump in the conventional scheme, so that larger liquid flow flux can be obtained.
It should be noted that the sample injected by the sample injection pump is usually a biological cell, but may also be a specific particle. Wherein the specific particles include, but are not limited to: various cells, microorganisms and synthetic microspheres.
III Microchannel network on rotating disc slide
In the invention, N micro-channels are arranged on the rotary disc slide from the center to the edge in an extending way, M sorting branch channels are respectively arranged on each micro-channel, and a micro-channel network arranged on the rotary disc slide is adopted to perform multi-channel time slot diversion to realize high-precision flow control and micro-droplet generation.
In a preferred embodiment of the present invention, N.gtoreq.10, for example, may be 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 150, 200, 250, 300, 400, 500, etc.; m.gtoreq.3, for example, may be 3, 4, 5, 6, 7, 8, 9, 10, etc. The sheath pump is used for injecting sheath liquid into the center of the rotary disc slide, and the sheath liquid flows from the center of the disc to the outer edge along the N paths of micro-channels under pressure. Thus, in principle, the larger N is, the better. Under the support of modern micron-level processing technology, the distribution density of micro-channels with N more than or equal to 10 on a single disc can be easily realized; and each micro-channel is respectively provided with M sorting branch channels, so that M is more than or equal to 3 can be easily realized. The method can realize NxM paths of parallel sorting paths, remove waste liquid paths and combine adjacent similar paths, can also realize Nx (M-1)/2 paths of parallel sorting paths, and can effectively support high-end special applications such as high-throughput cell subtype sorting.
IV. Optical System
In the invention, the optical system comprises a light source and an optical detector, wherein the light source and the optical detector are arranged at one end close to the liquid inlet of the micro-channel; the light source is used for irradiating the flow type micro-droplets in the current micro-flow channel and generating optical characteristic signals such as scattering, raman, fluorescence and the like; the optical detector is used for detecting the optical characteristic signal.
V. surface acoustic wave generator
In the invention, the surface acoustic wave generator is arranged at the liquid inlet end of the separation branch flow passage.
In a preferred embodiment of the present invention, the surface acoustic wave generator is a dipole source surface acoustic wave generator with a pair of interdigital electrodes, and the dipole source surface acoustic wave generator is disposed at two sides of the liquid inlet end of the sorting branch flow channel, and is used for controlling accurate flow of cells or particles into the selected sorting branch flow channel. The surface acoustic wave generator adopts a pair of interdigital electrode dipole source surface acoustic wave generator, and can combine with a detection feedback control algorithm to realize accurate manipulation and separation of cells or particles and realize high-flux flow separation.
VI, collecting device
In the invention, the acquisition device is arranged at the liquid outlet end of the separation branch flow passage.
In a preferred embodiment of the invention, the collection device may be a collection bottle, or other enrichment storage vessel optionally capable of collecting cells and/or particles. The device can be mounted under the vertical through holes at the tail ends of the branches of the micro-channels, can also carry out classification coding on each container, and is combined with the micro-channel branch network and the detection and sorting instruction to realize the advanced addressing function of enriching the appointed single cells or particles into the specific coding container.
In a preferred embodiment of the present invention, a microbump is disposed at the bottom of the microchannel just below the injection port of the sample injection pump.
It should be noted that a microbump is processed in the micro channel directly under the sample injection port, so as to change the hydrodynamic characteristics of the current-carrying sheath fluid, so that the sample is subjected to the action of a flexible shear force accelerating the current carrying, a large number of flow-type micro droplets containing single cells or particles are generated, and the flow-type micro droplets continue to flow to the edge of the disk under the driving of the sheath fluid.
In a preferred embodiment of the present invention, the micro bump is hemispherical in shape.
In a preferred embodiment of the present invention, the height of the microbump is 30 to 60% of the depth of the microchannel, for example, 30%, 32%, 35%, 39%, 40%, 43%, 45%, 50%, 54%, 58%, 60% or the like may be used.
In a preferred embodiment of the present invention, the light source and the optical detector are both disposed on one side of the rotating disc slide, and detect the reflected signal of the streaming micro-droplet in the current micro-channel.
In a preferred embodiment of the invention, the light source and the optical detector are respectively arranged at two sides of the rotary disk slide, and detect the reflection and/or refraction signals of the flow micro-droplets in the current light-transmitting and/or transparent micro-flow channels.
It should be noted that the light source and the detector may be placed on the same side of the disc to detect the reflected signal, or may be placed on both sides of the disc to detect the transmitted signal, and after the detected optical signal is subjected to photoelectric conversion, signal processing, and feature recognition, single cells or particles in the flowing micro-droplet are accurately detected, and then a sorting instruction is generated according to a preset standard, where the instruction will determine the final flow direction of the current micro-droplet.
In a preferred embodiment of the present invention, the sorting diversion flow path includes at least 1 waste liquid recovery flow path and at least 2 sorting collection flow paths for cells or particles.
In a preferred embodiment of the present invention, the sorting and collecting channel is disposed at an acute angle to the waste liquid recovery channel, and the selected cells or particles flow along the selected sorting and collecting channel into a collecting device disposed at the liquid outlet end of the sorting and branching channel.
In a preferred embodiment of the invention, the liquid outlet ends of adjacent sorting branch channels of adjacent micro-channels are connected with the same collecting device.
In a preferred embodiment of the present invention, the waste liquid recovery flow channel is arranged along the original micro flow channel direction, and unselected cells or particles continue to flow to the waste liquid recovery flow channel along the original direction, and are discharged from the edge of the rotating disc slide.
In a preferred embodiment of the present invention, the rotation of the rotating disc slide, and the injection of the sheath pump and the sample injection pump, are required to satisfy at least the following formula i and/or formula ii:
ti is less than or equal to Lc×Sc/Vs (formula I)
Wherein Vs represents the injection speed of the sample injection pump, ti represents the hovering injection time of the sample injection pump in the micro flow channel, sc represents the sectional area of the micro flow channel, and Lc represents the length of the micro flow channel.
Ti is larger than or equal to Vx/Vs (formula II)
Wherein Ti represents the hovering injection time of the sample injection pump in the micro flow channel, vx represents the minimum volume of micro liquid drops of the sample, and Vc represents the sheath liquid injection speed of the sheath liquid pump.
In a preferred embodiment of the present invention, the optical system detection and signal processing time and the rotational speed of the rotating disc slide are required to satisfy the following formula iii and/or formula iv:
Ti≥Ts×Lc 1 XSc/Vx (formula III)
Wherein Ti represents the hovering injection time of the sample injection pump in the micro flow channel, ts represents the time required by the optical system to detect cells or particles in micro liquid drops, lc 1 Represents the distance from the injection port of the sample injection pump to the optical detector on the microchannel, sc represents the cross-sectional area of the microchannel, and Vx represents the minimum volume of the microdroplet of the sample.
Tr≤N×Lc 2 XSc/Vc (formula IV)
Wherein Tr represents time required for identifying and sorting signal processing characteristics of an optical system, N represents the number of micro-channels and Lc 2 Representing the distance between the optical detector and the sorting intersection, sc represents the sectional area of the micro-flow channel, and Vc represents the sheath liquid injection speed of the sheath liquid pump.
In a preferred embodiment of the present invention, the saw generator is configured to control the flow direction of the micro-droplet, and at least the following formula v is satisfied:
Fs≥m×(M-1)×A×Vc 2 /(Ws×N 2 ×Sc 2 ) (formula V)
Wherein Fs represents the surface acoustic wave acting force exerted on cells or particles in the micro-droplets, M represents the mass of the micro-droplets, M represents the number of the sorting branch flow passages, a represents the included angle between adjacent sorting branch flow passages of each single micro-flow passage, vc represents the sheath liquid injection speed of the sheath liquid pump, ws represents the surface acoustic wave width, N represents the number of the micro-flow passages, and Sc represents the sectional area of the micro-flow passage.
In a preferred embodiment of the invention, the collection device is an enrichment storage container mounted under the vertical through hole at the tail end of each sorting and collecting flow channel;
in the preferred embodiment of the invention, the enrichment storage container is classified and encoded, and the addressing function of enriching the appointed single cells or particles into the specific encoding container is realized by combining with a micro-channel branch network and a detection and sorting instruction.
In a second aspect, the present invention provides the use of a sorting device for on-chip liquid flow path cells or particles as described in the first aspect in the sorting of on-chip liquid flow path cells or particles.
In a third aspect, the present invention provides a method for sorting on-chip liquid flow path cells or particles using the sorting apparatus for on-chip liquid flow path cells or particles according to the first aspect, comprising the steps of:
injecting sheath liquid into a liquid inlet at the center of the rotary disc slide by a sheath liquid pump, wherein the sheath liquid flows from the center of the disc slide to the outer edge along the N micro-channels under pressure;
injecting a sample into a micro-channel in the rotary disc slide by a sample injection pump, dispersing the sample into micro-droplets at the micro-bumps, and flowing along the N paths of micro-channels to the outer edge under the drive of sheath liquid;
illuminating the flow type micro liquid drops in the current micro flow channel by adopting a light source to generate an optical characteristic signal, and detecting the optical characteristic signal by an optical detector;
according to the sorting signal, the single cells or particles are driven to enter a selected sorting collecting flow passage by the directional sound wave generated by the surface acoustic wave generator, and finally flow into the collecting device.
Compared with the prior art, the invention has the following beneficial effects:
(1) The sorting device realizes the high-flux random multi-channel sorting function, and the defect that the capillary flow type technology is easy to block is avoided by utilizing the on-chip micro-channel injection flow type technology;
(2) The sorting device utilizes the design of a branching network of the large-flux injection multipath parallel micro-channels, so that the barrier of small flux and high cost of a common micro-fluidic chip is avoided;
(3) The sorting device utilizes a sample injection time slice interval current-carrying flushing method, thereby avoiding the inherent pollution-prone defect of the common sample injection technology;
(4) The sorting device utilizes the synchronous technology of the disk slide glass and sample injection and photoelectric detection, so that the detection speed and the recognition flux are improved;
(5) The sorting device disclosed by the invention utilizes a sound surface wave sound pressure driving technology to realize the branching flow sorting function at any angle, and combines the design of an on-chip mounted coding container to realize the addressing enrichment function of samples at any path;
(6) The sorting device can be widely applied to complex detection and sorting scenes, can doubly improve the flow type sorting flux of specific applications such as biopharmaceutical fast screening and the like, and has higher application value.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the description of the embodiments or the prior art will be briefly described, and it is obvious that the drawings in the description below are some embodiments of the present invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
Fig. 1 is a schematic structural diagram of a sorting apparatus for on-chip liquid flow path cells or particles according to the present invention.
The device comprises a sheath pump 1, a sample injection pump 2, a rotary disc slide 3, a micro-channel 4, a light source 5, an optical detector 6, a surface acoustic wave generator 7 and a collecting device 8.
Fig. 2 is a schematic diagram of a process in which a sample provided by the invention flows along an N-channel microchannel toward an outer edge under the drive of a sheath fluid.
Wherein 4 is a micro-channel, 5 is a light source, and 6 is an optical detector.
Fig. 3 is a schematic diagram of a process of driving single cells or particles into a sorting branch flow channel accurately by the surface acoustic wave generator provided by the invention.
Wherein 7 is a surface acoustic wave generator.
Detailed Description
Unless defined otherwise herein, scientific and technical terms used in connection with the present invention shall have the meanings commonly understood by one of ordinary skill in the art. The meaning and scope of terms should be clear, however, in the event of any potential ambiguity, the definitions provided herein take precedence over any dictionary or extraneous definition. In this application, the use of "or" means "and/or" unless stated otherwise. Furthermore, the use of the term "include" and other forms is not limiting.
In the description of the present invention, it should be noted that the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", "clockwise", "counterclockwise", and the like indicate an azimuth or a positional relationship based on that shown in the drawings, or an azimuth or a positional relationship in which the inventive product is conventionally put in use, are merely for convenience of describing the present invention and for simplifying the description, and do not indicate or imply that the apparatus or element to be referred to must have a specific azimuth, be constructed and operated in a specific azimuth, and thus should not be construed as limiting the present invention. Furthermore, the terms "first," "second," "third," and the like are used merely to distinguish between descriptions and should not be construed as indicating or implying relative importance.
Furthermore, the terms "horizontal," "vertical," "overhang," and the like do not denote a requirement that the component be absolutely horizontal or overhang, but rather may be slightly inclined. As "horizontal" merely means that its direction is more horizontal than "vertical", and does not mean that the structure must be perfectly horizontal, but may be slightly inclined.
In the description of the present invention, it should also be noted that, unless explicitly specified and limited otherwise, the terms "disposed," "mounted," "connected," and "connected" are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. The specific meaning of the above terms in the present invention will be understood in specific cases by those of ordinary skill in the art.
It is noted that in the following description, specific details are set forth in order to provide a thorough understanding of the present invention. The present invention may be embodied in many other forms than those herein described, and those skilled in the art may readily devise numerous other arrangements that do not depart from the spirit of the invention. Therefore, the present invention is not limited by the specific embodiments disclosed below.
The technical solutions of the present invention will be clearly and completely described in connection with the embodiments, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The invention is further illustrated by the following examples. The materials in the examples were prepared according to the existing methods or were directly commercially available unless otherwise specified.
Example 1
As shown in fig. 1, the present embodiment provides a sorting apparatus for on-chip liquid flow path cells or particles, the sorting apparatus comprising: a sheath liquid pump 1, a sample injection pump 2, a rotary disk slide 3, an optical system, a surface acoustic wave generator 7 and a collection device 8.
The injection port of the sheath liquid pump 1 is connected with the liquid inlet at the center of the rotary disc slide 3; i.e. the sheath pump 1 is used to inject sheath fluid into the centre of the spinning disc slide 3.
In alternative embodiments of the invention, the sheath pump 1 may be a fixed position electrically driven high flux current carrying syringe pump; this is due to the low cost, high throughput flow-through sorting technology required in various application scenarios such as biopharmaceuticals; the invention adopts a large-flux injection pump to replace a small-flux high-pressure injection pump in the conventional scheme, so that larger liquid flow flux can be obtained.
II, an injection port of the sample injection pump 2 is connected with a liquid inlet of the micro-channel; i.e. the sample injection pump 2 is used to inject a sample into a certain micro channel provided on the rotating disc slide 3.
In preferred embodiments of the invention, the sample injection pump may be a fixed position electrically driven large throughput sample injection pump; this is due to the low cost, high throughput flow-through sorting technology required in various application scenarios such as biopharmaceuticals; the sorting device can adopt a large-flux injection pump to replace a small-flux high-pressure injection pump in the conventional scheme, so that larger liquid flow flux can be obtained.
It should be noted that the sample injected by the sample injection pump is usually a biological cell, but may also be a specific particle.
And III, N micro-channels 4 are arranged on the rotary disc slide 3 from the center to the edge in an extending manner, M sorting branch channels are respectively arranged on each micro-channel, and multichannel time slot diversion is carried out by adopting a micro-channel network arranged on the rotary disc slide 3 to realize high-precision flow control and micro-droplet generation.
In a preferred embodiment of the present invention, N.gtoreq.10, for example, may be 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 120, 150, 200, 250, 300, 400, 500, etc.; m.gtoreq.3, for example, may be 3, 4, 5, 6, 7, 8, 9, 10, etc. The sheath pump is used for injecting sheath liquid into the center of the rotary disc slide, and the sheath liquid flows from the center of the disc to the outer edge along the N paths of micro-channels under pressure. Thus, in principle, the larger N is, the better. Under the support of modern micron-level processing technology, the distribution density of micro-channels with N more than or equal to 10 on a single disc can be easily realized; and each micro-channel is respectively provided with M sorting branch channels, so that M is more than or equal to 3 can be easily realized. The method can realize NxM paths of parallel sorting paths, remove waste liquid paths and combine adjacent similar paths, can also realize Nx (M-1)/2 paths of parallel sorting paths, and can effectively support high-end special applications such as high-throughput cell subtype sorting.
The optical system comprises a light source 5 and an optical detector 6, wherein the light source 5 and the optical detector 6 are arranged at one end close to the liquid inlet of the micro-channel; the light source 5 is used for irradiating the flow type micro-droplets in the current micro-flow channel to generate an optical characteristic signal; the optical detector 6 is used for detecting the optical characteristic signal.
And V, arranging the surface acoustic wave generators 7 on two sides of the liquid inlet end of the separation branch flow passage.
In the preferred embodiment of the invention, the surface acoustic wave generator adopts a pair of interdigital electrode dipole source surface acoustic wave generator, and the accurate control and separation of cells or particles are realized by combining a detection feedback control algorithm, so that the high-flux flow separation is realized.
And VI, the acquisition device 8 is arranged at the liquid outlet end of the separation branch flow passage.
Further, as shown in fig. 2, a micro bump is disposed at the bottom of the micro flow channel right below the injection port of the sample injection pump.
It should be noted that a microbump is processed in the micro channel directly under the sample injection port, so as to change the hydrodynamic characteristics of the current-carrying sheath fluid, so that the sample is subjected to the action of a flexible shear force accelerating the current carrying, a large number of flow-type micro droplets containing single cells or particles are generated, and the flow-type micro droplets continue to flow to the edge of the disk under the driving of the sheath fluid.
Further, the micro bump is hemispherical in shape.
Further, the height of the micro bump is 30-60% of the depth of the micro flow channel, for example, 30%, 32%, 35%, 39%, 40%, 43%, 45%, 50%, 54%, 58%, 60% or the like.
Further, the light source 5 and the optical detector 6 are both arranged on one side of the rotary disc slide, and detect the reflected signal of the flow type micro-droplet in the current micro-channel; or, the light source 5 and the optical detector 6 are respectively arranged at two sides of the rotary disc slide, and detect the reflection or/and refraction signals of the flow micro-droplets in the current light-transmitting and/or transparent micro-flow channels.
It should be noted that, the light source 5 irradiates the flow type micro-droplet in the current micro-channel to generate an optical characteristic signal, and the optical characteristic signal is detected by the optical detector 6, the light source 5 and the detector 6 can be placed on the same side of the disc to detect a reflected signal, or can be placed on the upper side and the lower side of the disc to detect a transmitted signal, after photoelectric conversion, signal processing and characteristic recognition are performed on the detected optical signal, single cells or particles in the flow type micro-droplet are accurately detected, and then a sorting instruction is generated according to a preset standard, and the instruction determines the final flow direction of the current micro-droplet.
Still further, the sort-by-pass flow path includes at least 1 waste-liquid recovery flow path and at least 2 sort-collection flow paths for cells or particles.
Further, the sorting and collecting channel is disposed at an acute angle to the waste liquid recovery channel, and the selected cells or particles flow into the collecting device 8 disposed at the liquid outlet end of the sorting and branching channel along the selected sorting and collecting channel.
Further, the liquid outlet ends of the adjacent sorting branch flow channels of the adjacent micro flow channels are connected with the same collecting device 8.
Further, the waste liquid recovery flow channel is arranged along the original micro flow channel direction, unselected cells or particles continue to flow to the waste liquid recovery flow channel along the original direction, and are discharged out of the disc from the edge of the rotary disc slide.
It should be noted that when the flow type micro-droplet just flows through the multi-branch of the micro-channel, the interdigital electrode surface acoustic wave generator 7 is driven according to the sorting signal to generate directional sound waves to drive single cells or particles to precisely enter the selected branch, and finally the single cells or particles flow into the collection device 8 mounted below the through hole of the branch sorting branch channel to realize the flow type sorting and enrichment function, and the unselected single cells or particles continuously flow to the waste liquid micro-channel along the original direction to be discharged out of the disc.
Furthermore, the device of the invention also comprises a set of parameter settings which ensure that the rotation of the disc for reasonably generating micro-droplets is coordinated with the injection of the sample, and the parameter settings are as follows:
and setting Vc to represent the sheath liquid injection speed of the sheath liquid pump, lc to represent the length of the micro flow channel, N to represent the number of channels of the micro flow channel, and Sc to represent the sectional area of the micro flow channel, wherein the flow velocity of the sheath liquid in the micro flow channel is Vc/(N×Sc).
Let Vs represent the sample injection speed of the sample injection pump, and the cross-sectional area of the injection port of the sample injection pump is Ss.
And setting Ti to represent the hovering injection time of the sample injection pump in the micro flow channels, and setting Tn to represent the cruising time of the sample injection pump between two adjacent micro flow channels, wherein the rotation period of the rotary disc slide is N× (Ti+Tn).
The length of the single injection sample fluid stream is Vs xTi/Sc.
Therefore, in order to make each flow channel work in parallel and time multiplexing, the length of the single injection sample liquid flow is smaller than or equal to the length of the micro flow channel, that is, vs×Ti/Sc is smaller than or equal to Lc, that is, the condition of the formula I needs to be satisfied:
ti is less than or equal to Lc×Sc/Vs (formula I)
Wherein Vs represents the injection speed of the sample injection pump, ti represents the hovering injection time of the sample injection pump in the micro flow channel, sc represents the sectional area of the micro flow channel, and Lc represents the length of the micro flow channel.
Furthermore, if Vx is set to represent the minimum volume of the micro-droplet of the sample, then in order to make the minimum micro-droplet be smoothly generated, the condition of formula ii must also be satisfied:
ti is larger than or equal to Vx/Vs (formula II)
Wherein Ti represents the hovering injection time of the sample injection pump in the micro flow channel, vx represents the minimum volume of micro liquid drops of the sample, and Vs represents the injection speed of the sample injection pump.
Furthermore, the device of the invention also comprises a set of parameter settings of optical detection and signal processing time to the rotating speed of the disc, and the parameter settings are specifically as follows:
setting Ts to represent the time required by the optical system to detect cells or particles in the micro-droplets, tr to represent the time required by the optical system to process the characteristic identification and the sorting instruction, and setting the flow rate of sheath liquid in the micro-flow channel to be Vc/(NxSc) as described above; lc (Lc) 1 Representing the distance Lc from the injection port of the sample injection pump to the optical detector on the microchannel 2 Representing the distance between the optical detector and the sorting intersection, the maximum number of micro-droplets to be detected at each time is Lc 1 ×Sc/Vx。
Based on this, to ensure that the injection, detection and sorting can be synchronized and coordinated with the detection and signal processing times of the optical system, and the rotational speed of the rotating disc slide, the following formulas iii and iv are satisfied:
ti is less than or equal to Lc×Sc/Vs (formula III)
Wherein Vs represents the injection speed of the sample injection pump, ti represents the hovering injection time of the sample injection pump in the micro flow channel, sc represents the sectional area of the micro flow channel, and Lc represents the length of the micro flow channel.
Ti is larger than or equal to Vx/Vs (formula IV)
Wherein Ti represents the hovering injection time of the sample injection pump in the micro flow channel, vx represents the minimum volume of micro liquid drops of the sample, and Vc represents the sheath liquid injection speed of the sheath liquid pump.
Furthermore, the device of the invention also comprises a set of parameter settings for controlling the accurate flow direction of the acoustic surface wave manipulation micro-droplet, and the parameter settings are specifically as follows:
setting Fs to represent the surface acoustic wave acting force of cells or particles in the micro-droplets, wherein the magnitude of Fs depends on the radius of the micro-droplets, the mass m of the micro-droplets, the relative density of the liquid, the relative density of the solid, the surface acoustic wave width Ws, the wave number and the amplitude.
Setting M to represent the number of the branch channels, wherein A represents the included angle between adjacent branch channels of each single micro channel, namely assuming that each single micro channel has M branch channels and the included angle between adjacent branches is A degrees, the maximum (M-1) x A/2 angle is required to deflect, and under the condition of moderate disc rotation speed, the centrifugal force is ignored, and only the condition of the following formula V needs to be ensured:
Fs≥m×(M-1)×A×Vc 2 /(Ws×N 2 ×Sc 2 ) (formula V)
Wherein Fs represents the surface acoustic wave acting force exerted on cells or particles in the micro-droplets, M represents the mass of the micro-droplets, M represents the number of sorting branches, a represents the included angle between adjacent sorting branches of each single micro-channel, vc represents the sheath liquid injection speed of the sheath liquid pump, ws represents the dipole source surface acoustic wave width, N represents the number of micro-channels, and Sc represents the cross-sectional area of the micro-channels.
It should be noted that when the rotation speed of the disc is high and the centrifugal force cannot be ignored, fs needs to be further increased, that is, the radiation power of the surface acoustic wave is increased. On the basis of meeting the constraint, the dipole source surface acoustic wave generator can be switched on and off according to the sorting instruction to perform voice control flow path manipulation on single cells or particles in micro-droplets, so that the flow sorting function is realized.
Further, the collection device is an enrichment storage container mounted under the vertical through holes at the tail ends of the sorting collection flow channels.
Furthermore, the enrichment storage container is classified and encoded, and the addressing function of enriching the appointed single cells or particles into the specific encoding container is realized by combining with a micro-channel branch network and a detection and sorting instruction.
It should be noted that the invention also comprises a set of sorted sample enrichment storage containers mounted below the vertical through holes at the tail ends of the branches of the micro-channels, and the containers can be classified and encoded, and combined with the micro-channel branch network and the detection sorting instruction, the invention realizes the advanced addressing function of enriching the appointed single cells or particles into the specific encoding container.
Example 2
The present embodiment provides a sorting method for on-chip liquid flow path cells, using the sorting apparatus provided in embodiment 1, specifically comprising the steps of:
s1, injecting sheath liquid into a liquid inlet at the center of a rotary disc slide 3 through a sheath liquid pump 1, wherein the sheath liquid flows from the center of the disc slide to the outer edge along N micro-channels under pressure;
s2, injecting a sample into a micro-channel of a rotating disc right below the sample injection pump 2, wherein the sample is a biological cell, and as shown in FIG. 2, processing a micro-bump in the micro-channel right below the sample injection port, wherein the micro-bump changes the hydrodynamic characteristics of a current carrying sheath fluid, so that the sample is subjected to the action of a flexible shearing force for accelerating the current carrying, a large number of flow micro-droplets containing single cells are generated, and flow to the edge of the disc continuously under the drive of the sheath fluid;
s3, in the continuous flowing process, the light source 5 irradiates the flow type micro-droplets in the current micro-flow channel to generate an optical characteristic signal, and the optical characteristic signal is detected by the optical detector 6; after photoelectric conversion, signal processing and feature recognition are carried out on the detected optical signals, single cells or particles in the flow type micro-droplets are accurately detected, and then a sorting instruction is generated according to a preset standard, wherein the instruction determines the final flow direction of the current micro-droplets;
s4, when the flow type micro-droplets just flow through the multi-branch of the micro-channel, as shown in fig. 3, the interdigital electrode surface acoustic wave generator 7 is driven according to the sorting signal to generate directional sound waves to drive single cells to precisely enter the selected branch, and finally the single cells flow into the collection device 8 mounted below the through hole of the sorting branch channel to realize the flow type sorting and enrichment function, and the unselected single cells continuously flow to the waste liquid micro-channel along the original direction and are discharged out of the disc.
Example 3
The present embodiment provides a sorting method of on-chip liquid flow path particles, using the sorting apparatus provided in embodiment 1, specifically including the steps of:
s1, injecting sheath liquid into a liquid inlet at the center of a rotary disc slide 3 through a sheath liquid pump 1, wherein the sheath liquid flows from the center of the disc slide to the outer edge along N micro-channels under pressure;
s2, injecting a sample into a micro-channel of a rotating disc right below the sample injection pump 2, wherein the sample is particles, and as shown in FIG. 2, processing a micro-bump in the micro-channel right below the sample injection port, wherein the micro-bump changes the hydrodynamic characteristics of a current carrying sheath fluid, so that the sample is subjected to the action of a flexible shearing force for accelerating the current carrying, a large number of flow micro-droplets containing the particles are generated, and flow towards the edge of the disc continuously under the drive of the sheath fluid;
s3, in the continuous flowing process, the light source 5 irradiates the flow type micro-droplets in the current micro-flow channel to generate an optical characteristic signal, and the optical characteristic signal is detected by the optical detector 6; after photoelectric conversion, signal processing and feature recognition are carried out on the detected optical signals, particles or granules in the flow type micro-droplets are accurately detected, and then a sorting instruction is generated according to a preset standard, wherein the instruction determines the final flow direction of the current micro-droplets;
s4, when the flow type micro-droplets just flow through the multi-branch of the micro-channel, as shown in fig. 3, the interdigital electrode surface acoustic wave generator 7 is driven according to the sorting signal to generate directional sound waves to drive particles to precisely enter the selected branch, and finally the particles flow into the collecting device 8 mounted below the through hole of the sorting branch channel to realize the flow type sorting and enrichment function, and the unselected particles continuously flow to the waste liquid micro-channel along the original direction and are discharged out of the disc.
The invention can realize the high-flux random multi-channel sorting function, utilizes the on-chip micro-channel injection flow type technology, avoids the defect of easy blockage of capillary flow type technology, utilizes the design of a large-flux injection multi-channel parallel micro-channel bifurcation network, avoids the obstacle of small flux and high cost of a common micro-fluidic chip, utilizes a sample injection time slice interval current-carrying flushing method, avoids the inherent easy pollution defect of the common sample injection technology, utilizes the disc slide and sample injection and photoelectric detection synchronous technology, improves the detection speed and the recognition flux, utilizes the acoustic surface wave sound pressure driving technology, realizes the random angle bifurcation flow type sorting function, combines the chip downloading and mounting of a coding container design, and realizes the random path sample addressing enrichment function. The invention can be widely applied to complex detection and separation scenes, can multiply improve the flow type separation flux of specific applications such as biopharmaceutical rapid screening and the like, and has higher application value.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit of the invention.
Claims (9)
1. A sorting apparatus for on-chip liquid flow path cells or particles, the sorting apparatus comprising: the device comprises a sheath liquid pump, a sample injection pump, a rotary disc slide, an optical system, a surface acoustic wave generator and a collecting device;
n micro-channels are arranged on the rotary disc slide from the center to the edge in an extending manner, and M sorting branch channels are respectively arranged on each micro-channel; wherein N is more than or equal to 10, M is more than or equal to 3;
the injection port of the sheath liquid pump is connected with the liquid inlet at the center of the rotary disc slide;
the injection port of the sample injection pump is connected with the liquid inlet of the micro-channel;
a micro-bump is arranged at the bottom of the micro-flow channel right below the injection port of the sample injection pump; the shape of the micro-bump is hemispherical; the height of the micro-bump is 30-60% of the depth of the micro-channel;
the optical system comprises a light source and an optical detector, and the light source and the optical detector are arranged at one end close to the liquid inlet of the micro-channel;
the surface acoustic wave generator is arranged at the liquid inlet end of the separation branch flow passage;
the acquisition device is arranged at the liquid outlet end of the sorting branch flow passage.
2. The on-chip liquid flow path cell or particle sorting apparatus according to claim 1, wherein the light source and the optical detector are both disposed on one side of the rotating disc slide, and detect a reflected signal of a flow type micro droplet in a current micro flow path;
or the light source and the optical detector are respectively arranged at two sides of the rotary disc slide, and detect the reflection and/or refraction signals of the flow micro-droplets in the current light-transmitting and/or transparent micro-flow channels.
3. The on-chip liquid flow path cell or particle sorting apparatus of claim 1, wherein the sorting divergent flow path includes at least 1 waste liquid recovery flow path and at least 2 sorting collection flow paths for cells or particles;
and/or the sorting and collecting flow channel and the waste liquid recovery flow channel are arranged at an acute angle, and the selected cells or particles flow into a collecting device arranged at the liquid outlet end of the sorting branch flow channel along the selected sorting and collecting flow channel;
and/or the liquid outlet ends of the adjacent sorting branch flow channels of the adjacent micro flow channels are connected with the same collecting device;
and/or the waste liquid recovery flow channel is arranged along the original micro flow channel direction, unselected cells or particles continue to flow to the waste liquid recovery flow channel along the original direction, and are discharged out of the disc from the edge of the rotary disc slide.
4. The on-chip liquid flow path cell or particle sorting apparatus according to claim 1, wherein the rotation of the rotary disc slide and the injection of the sheath pump and the sample injection pump satisfy at least the following conditions of formula i and/or formula ii:
ti is less than or equal to Lc×Sc/Vs (formula I)
Wherein Vs represents the injection speed of the sample injection pump, ti represents the hovering injection time of the sample injection pump in the micro flow channel, sc represents the sectional area of the micro flow channel, and Lc represents the length of the micro flow channel;
ti is larger than or equal to Vx/Vs (formula II)
Wherein Ti represents the hovering injection time of the sample injection pump in the micro flow channel, vx represents the minimum volume of micro liquid drops of the sample, and Vs represents the injection speed of the sample injection pump.
5. The apparatus according to claim 1, wherein the optical system detection and signal processing time and the rotational speed of the rotating disc slide are required to satisfy the following formula iii and/or formula iv:
Ti≥Ts×Lc 1 XSc/Vx (formula III)
Wherein Ti represents the hovering injection time of the sample injection pump in the micro flow channel, ts represents the time required by the optical system to detect cells or particles in micro liquid drops, lc 1 Represents the distance from the injection port of the sample injection pump to the optical detector on the micro flow channel, sc represents the sectional area of the micro flow channel,vx represents the minimum volume of the microdroplet of the sample;
Tr≤N×Lc 2 XSc/Vc (formula IV)
Wherein Tr represents time required for identifying and sorting signal processing characteristics of an optical system, N represents the number of micro-channels and Lc 2 Represents the distance between the optical detector and the sorting intersection, sc represents the sectional area of the micro-flow channel, and Vc represents the sheath liquid injection speed of the sheath liquid pump.
6. The device according to claim 1, wherein the surface acoustic wave generator is configured to control the flow direction of the micro-droplets under the condition of at least the following formula v:
Fs≥m×(M-1)×A×Vc 2 /(Ws×N 2 ×Sc 2 ) (formula V)
Wherein Fs represents the surface acoustic wave acting force exerted on cells or particles in the micro-droplets, M represents the mass of the micro-droplets, M represents the number of the sorting branch flow passages, a represents the included angle between adjacent sorting branch flow passages of each single micro-flow passage, vc represents the sheath liquid injection speed of the sheath liquid pump, ws represents the surface acoustic wave width, N represents the number of the micro-flow passages, and Sc represents the sectional area of the micro-flow passage.
7. The on-chip liquid flow path cell or particle sorting device according to claim 4, wherein the collection device is an enrichment storage container mounted under the end vertical through hole of each sorting collection flow path;
and/or classifying and encoding the enrichment storage containers, and combining with a micro-channel branch network and a detection and sorting instruction to realize the addressing function of enriching the appointed single cells or particles into the specific encoding containers.
8. Use of a sorting device of liquid flow path cells or particles on a chip according to any one of claims 1 to 7 in the sorting of liquid flow path cells or particles on a chip.
9. A method for sorting on-chip liquid flow path cells or particles, characterized in that the sorting method uses the sorting device for on-chip liquid flow path cells or particles according to any one of claims 1 to 7, specifically comprising the steps of:
injecting sheath liquid into a liquid inlet at the center of the rotary disc slide by a sheath liquid pump, wherein the sheath liquid flows from the center of the disc slide to the outer edge along the N micro-channels under pressure;
injecting a sample into a micro-channel in the rotary disc slide by a sample injection pump, dispersing the sample into micro-droplets at the micro-bumps, and flowing along the N paths of micro-channels to the outer edge under the drive of sheath liquid;
illuminating micro liquid drops in the current micro flow channel by adopting a light source to generate an optical characteristic signal, and detecting the optical characteristic signal by an optical detector;
according to the sorting signal, the single cells or particles are driven to enter a selected sorting collecting flow passage by the directional sound wave generated by the surface acoustic wave generator, and finally flow into the collecting device.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311800730.5A CN117448151B (en) | 2023-12-26 | 2023-12-26 | Sorting device and sorting method for on-chip liquid flow path cells or particles |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311800730.5A CN117448151B (en) | 2023-12-26 | 2023-12-26 | Sorting device and sorting method for on-chip liquid flow path cells or particles |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117448151A CN117448151A (en) | 2024-01-26 |
| CN117448151B true CN117448151B (en) | 2024-03-29 |
Family
ID=89582268
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311800730.5A Active CN117448151B (en) | 2023-12-26 | 2023-12-26 | Sorting device and sorting method for on-chip liquid flow path cells or particles |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117448151B (en) |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005108963A1 (en) * | 2004-05-06 | 2005-11-17 | Nanyang Technological University | Microfluidic cell sorter system |
| CN101653768A (en) * | 2002-09-16 | 2010-02-24 | 塞通诺米公司 | Method and apparatus for sorting particles |
| CN102879377A (en) * | 2012-10-29 | 2013-01-16 | 重庆科技学院 | Microfluidic chip for detecting early leukemia |
| CN108823065A (en) * | 2018-07-05 | 2018-11-16 | 中国科学院苏州生物医学工程技术研究所 | Microparticle sorting unit based on intermittent inclination surface acoustic wave |
| CN110793905A (en) * | 2019-11-06 | 2020-02-14 | 中国科学院苏州生物医学工程技术研究所 | Microfluid chip and microfluid cell sorting system |
| CN111889153A (en) * | 2020-08-04 | 2020-11-06 | 桂林电子科技大学 | Flow cytometer based on optical fiber integrated microfluidic chip |
| CN112111385A (en) * | 2020-08-19 | 2020-12-22 | 东南大学 | Cancer cell sorting and detecting device |
| CN114574357A (en) * | 2022-02-14 | 2022-06-03 | 大连交通大学 | Microalgae and bacteria sorting device based on combination of deterministic lateral displacement and dielectrophoresis technologies |
| CN115791576A (en) * | 2022-11-25 | 2023-03-14 | 中国科学院苏州生物医学工程技术研究所 | Cell flow type sorting device |
| CN116165123A (en) * | 2023-02-03 | 2023-05-26 | 杭州谱康医学科技有限公司 | Method for detecting antibody by mass spectrometry and application thereof |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7232687B2 (en) * | 2004-04-07 | 2007-06-19 | Beckman Coulter, Inc. | Multiple sorter monitor and control subsystem for flow cytometer |
-
2023
- 2023-12-26 CN CN202311800730.5A patent/CN117448151B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101653768A (en) * | 2002-09-16 | 2010-02-24 | 塞通诺米公司 | Method and apparatus for sorting particles |
| WO2005108963A1 (en) * | 2004-05-06 | 2005-11-17 | Nanyang Technological University | Microfluidic cell sorter system |
| CN102879377A (en) * | 2012-10-29 | 2013-01-16 | 重庆科技学院 | Microfluidic chip for detecting early leukemia |
| CN108823065A (en) * | 2018-07-05 | 2018-11-16 | 中国科学院苏州生物医学工程技术研究所 | Microparticle sorting unit based on intermittent inclination surface acoustic wave |
| CN110793905A (en) * | 2019-11-06 | 2020-02-14 | 中国科学院苏州生物医学工程技术研究所 | Microfluid chip and microfluid cell sorting system |
| CN111889153A (en) * | 2020-08-04 | 2020-11-06 | 桂林电子科技大学 | Flow cytometer based on optical fiber integrated microfluidic chip |
| CN112111385A (en) * | 2020-08-19 | 2020-12-22 | 东南大学 | Cancer cell sorting and detecting device |
| CN114574357A (en) * | 2022-02-14 | 2022-06-03 | 大连交通大学 | Microalgae and bacteria sorting device based on combination of deterministic lateral displacement and dielectrophoresis technologies |
| CN115791576A (en) * | 2022-11-25 | 2023-03-14 | 中国科学院苏州生物医学工程技术研究所 | Cell flow type sorting device |
| CN116165123A (en) * | 2023-02-03 | 2023-05-26 | 杭州谱康医学科技有限公司 | Method for detecting antibody by mass spectrometry and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 流式细胞分选仪研究进展与展望;孙福齐等;《中国医学物理学杂志》;20230731;第40卷(第7期);第890-898页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117448151A (en) | 2024-01-26 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11446665B2 (en) | Hydrodynamic focusing apparatus and methods | |
| CN107505249B (en) | Microfluidic chip system for rare cell screening | |
| Chung et al. | Recent advances in miniaturized microfluidic flow cytometry for clinical use | |
| US8387803B2 (en) | Particle sorting | |
| CN108432132B (en) | Microfluidic particle manipulation | |
| US9550215B2 (en) | Method and apparatus for sorting particles | |
| US20160231223A1 (en) | Fluidic chip for flow cytometry and methods of use | |
| US20110005978A1 (en) | Method and apparatus for sorting particles | |
| US11686665B2 (en) | Microfluidic system with combined electrical and optical detection for high accuracy particle sorting and methods thereof | |
| WO2011097032A1 (en) | Multiple flow channel particle analysis system | |
| ZA200408705B (en) | Method and apparatus for sorting particles | |
| CN201548547U (en) | Flow cell analysis device based on microfluidic chip | |
| CN101726585A (en) | Flow cytometry based on microfluidic chip | |
| US20070131554A1 (en) | Multi-sample microfluidic dielectrophoresis separating device and method thereof | |
| Frenzel et al. | Microfluidic train station: highly robust and multiplexable sorting of droplets on electric rails | |
| CN117448151B (en) | Sorting device and sorting method for on-chip liquid flow path cells or particles | |
| CN107297334B (en) | Cell sorting device and method based on micro-spark cavitation | |
| EP3834942A1 (en) | Microchip and microparticle fractionating device | |
| CN210215391U (en) | Cell sorting device | |
| CN110093271B (en) | Cell sorting device | |
| US20230249182A1 (en) | Microfluidic chip device based on magnetic field-controlled fluorescently-labeled cell sorting method and use method | |
| CN115791576A (en) | Cell flow type sorting device | |
| CN212975166U (en) | Microfluidic chip for particle manipulation | |
| WO2013044109A1 (en) | Microfluidic device for separating cells from a fluid | |
| JP2020519305A (en) | MicroFACS for detection and isolation of target cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |