CN111521549A - Particle sorting device and method - Google Patents
Particle sorting device and method Download PDFInfo
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- CN111521549A CN111521549A CN202010406155.0A CN202010406155A CN111521549A CN 111521549 A CN111521549 A CN 111521549A CN 202010406155 A CN202010406155 A CN 202010406155A CN 111521549 A CN111521549 A CN 111521549A
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- 239000002245 particle Substances 0.000 title claims abstract description 106
- 238000000034 method Methods 0.000 title claims abstract description 17
- 238000001514 detection method Methods 0.000 claims description 11
- 125000006850 spacer group Chemical group 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 3
- 238000005192 partition Methods 0.000 claims description 2
- 238000004513 sizing Methods 0.000 claims description 2
- 230000003287 optical effect Effects 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 6
- 239000012530 fluid Substances 0.000 description 5
- 239000008187 granular material Substances 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 239000013618 particulate matter Substances 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 108091005461 Nucleic proteins Proteins 0.000 description 1
- 239000004793 Polystyrene Substances 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 241000723873 Tobacco mosaic virus Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- -1 and the like Substances 0.000 description 1
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- 238000004113 cell culture Methods 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
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- 229910010272 inorganic material Inorganic materials 0.000 description 1
- 239000011147 inorganic material Substances 0.000 description 1
- 239000004816 latex Substances 0.000 description 1
- 229920000126 latex Polymers 0.000 description 1
- 239000000696 magnetic material Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 239000007769 metal material Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920002223 polystyrene Polymers 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000001846 repelling effect Effects 0.000 description 1
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- 238000006748 scratching Methods 0.000 description 1
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- 239000000377 silicon dioxide Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
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- 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
- G01N15/1434—Optical arrangements
- G01N15/1436—Optical arrangements the optical arrangement forming an integrated apparatus with the sample container, e.g. a flow cell
-
- 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
- G01N15/149—Optical investigation techniques, e.g. flow cytometry specially adapted for sorting particles, e.g. by their size or optical properties
-
- 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
- G01N2015/1477—Multiparameters
- G01N2015/1479—Using diffuse illumination or excitation
-
- 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
- G01N2015/1493—Particle size
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- Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The embodiment of the application provides a particle sorting device and a method, wherein the device comprises a flow chamber and a nozzle, the flow chamber is used for limiting the flow of particles, and the nozzle is used for adjusting the size according to the characteristics of the particles; the nozzle includes a third adjustment member provided at a lower portion of the flow chamber, the third adjustment member being in a ring shape having a variable inner diameter. The particle sorting device and the particle sorting method can continuously adjust the diameter of the nozzle.
Description
Technical Field
The present application relates to the field of optical detection, and in particular, to a particle sorting apparatus and method.
Background
The current flow cytometer on the market needs to manually replace and install nozzles with different sizes according to different particle sizes, the size of the nozzle is fixed, the operation of replacing and installing different nozzles is complex, the nozzle needs to be continuously adjusted after the nozzle is replaced every time in order to ensure the accuracy of particle sorting after the nozzle is replaced, the nozzle is frequently replaced and is easily damaged, and the nozzle is easily blocked. The operation efficiency of the sorting device is low due to the fact that the flow of replacing the nozzle is troublesome and the sorting device cannot automatically adapt to the characteristics of the sorted particles.
Disclosure of Invention
The application provides a particle sorting device and method, the size of a nozzle is variable, and the stability and the sorting efficiency of particle sorting can be improved.
A particle sorting apparatus is provided, the apparatus comprising a flow chamber for restricting the flow of particles and a nozzle for sizing the particles according to their characteristics;
the nozzle includes a third adjustment member provided at a lower portion of the flow chamber, the third adjustment member being in a ring shape having a variable inner diameter.
Preferably, the third adjustment member is an iris.
Preferably, the nozzle further comprises a first adjustment member;
the first adjusting piece is arranged at the lower part of the flow chamber, and the third adjusting piece is arranged inside the first adjusting piece; the third adjusting piece is an elastic element;
the first adjusting piece is used for adjusting the transverse size of the third adjusting piece.
Preferably, the nozzle further comprises a second adjustment member;
the second adjusting piece is used for adjusting the longitudinal size of the third adjusting piece; the second adjusting member is disposed at a lower portion of the third adjusting member.
Preferably, the device further comprises a spacer;
the partition is disposed between the flow chamber and the third trim.
Preferably, the device further comprises a detection module and a control module;
the detection module is used for acquiring the size of the particles;
the control module is used for adjusting the nozzle according to the size of the particles.
The present application also provides a method of sorting particles, the method comprising:
controlling a laser to emit at least one laser, the laser being irradiated onto the particles in the flow cell; the particles generate fluorescence signals and scattered light signals under the irradiation of the laser in sequence;
receiving the fluorescent signal and/or the scattered light signal with a detection module, and acquiring one or more of the following characteristics of the particle: the size of the particle, the internal composition of the particle, the structure of the particle, the complexity of the particle, the fluorescence characteristic information of the particle, the morphology image of the particle, the fluorescence characteristic information of the particle;
and adjusting the inner diameter of the third adjusting member according to the characteristics of the particles, and enlarging or reducing the diameter of the nozzle.
Preferably, the third adjustment member is an iris.
Preferably, the adjusting the inner diameter of the third adjusting member specifically includes moving the first adjusting member left and right to change the lateral size of the third adjusting member.
Preferably, the adjusting the inner diameter of the third adjusting member specifically includes moving the second adjusting member up and down to change the longitudinal size of the third adjusting member.
The particle sorting device and method provided by the embodiment of the application can realize the following beneficial effects:
the diameter size of the nozzle can be continuously adjusted, and compared with the traditional nozzle with a fixed through hole, the nozzle has the advantages of wide application range, simple and convenient operation for adjusting the diameter of the nozzle, high efficiency and high precision.
Drawings
FIG. 1 is a schematic view of a first configuration of a particle sorting apparatus according to an embodiment of the present application;
FIG. 2 is a schematic view of a second configuration of a particle sorting apparatus according to an embodiment of the present application;
FIG. 3 is a schematic view of a third configuration of a particle sorting apparatus according to an embodiment of the present application;
FIG. 4 is a schematic view of a fourth configuration of a particle sorting apparatus according to an embodiment of the present application;
FIG. 5 is a schematic view of a fifth configuration of a particle sorting apparatus according to an embodiment of the present application;
fig. 6 is a sixth configuration diagram of a particle sorting apparatus according to an embodiment of the present application.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application, and it is obvious that the described embodiments are some embodiments of the present application, but not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.
The singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise. As used herein, the terms "first" and "second" are used interchangeably to distinguish one element or class of elements from another element or class of elements, respectively, and are not intended to denote the position or importance of the individual elements.
The term "and/or" herein is merely an association describing an associated object, meaning that three relationships may exist, e.g., a and/or B, may mean: a exists alone, A and B exist simultaneously, and B exists alone. In addition, the character "/" herein generally indicates that the former and latter related objects are in an "or" relationship.
Reference herein to "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment can be included in at least one embodiment of the application. The appearances of the phrase in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments. It is explicitly and implicitly understood by one skilled in the art that the embodiments described herein can be combined with other embodiments.
In order to make the technical solutions better understood by those skilled in the art, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the accompanying drawings.
As shown in fig. 1-3, embodiments of the present application provide a particle sorting apparatus that includes a flow chamber and a nozzle. The flow chamber is used to restrict the flow of particles and the nozzle is used to adjust the size according to the characteristics of each particle, for example, the size of the nozzle is adjusted according to the size of the particle. For example, the nozzle includes at least one of a first trim and a second trim, and a third trim. The third adjustment member may be a resilient member. The third adjustment member is disposed at a lower portion of the flow chamber. For example, the third adjustment member may be annular, or the third adjustment member may be disposed in a dotted pattern around the elastic member. The first adjusting piece is used for adjusting the inner diameter of the elastic element. The second adjusting member may also be annular, and the second adjusting member is used for adjusting the inner diameter of the elastic element. For example, the first adjusting member is used for adjusting the transverse size of the third adjusting member, and the second adjusting member is used for adjusting the longitudinal size of the third adjusting member. For example, the third adjustment member is disposed inside the first adjustment member; the second adjusting member is disposed at a lower portion of the third adjusting member. The first adjusting piece is fixed, and the second adjusting piece moves up and down to change the inner diameter of the third adjusting piece. Or the second adjusting piece is fixed, and the first adjusting piece moves left and right to change the inner diameter of the third adjusting piece.
The particles may be cells, bacteria, etc., including but not limited to, biological particulate matter, such as microorganisms including bacteria such as E.coli, viruses such as tobacco mosaic virus, fungi such as yeast, ribosomes, chromosomes, mitochondria, organelles, etc., and biologically-relevant polymers such as nucleic acids, proteins, and complexes thereof; the particles may also be artificial particles such as latex particles, gel particles, industrial particles, and the like, including but not limited to particles formed of organic polymeric materials including polystyrene and the like, inorganic materials including glass, silica, magnetic materials, and the like, and metallic materials including metal colloids and the like, and the like. Although the particulate matter is generally spherical in shape, the particles may have a non-spherical shape. Further, the size, mass, etc. of the particles are also not limited. For example, the particles are encapsulated by the sample fluid and can flow in a flow cell.
In an alternative embodiment of the present application, as shown in fig. 4, the device further comprises a spacer. The spacer is between the flow chamber and the third trim and/or the first trim. The spacer is used to prevent the third trim and/or the first trim from scratching the flow cell.
In another embodiment of the present application, as shown in fig. 5 and 6, the third adjusting member is disposed at a lower portion of the flow chamber, and an inner diameter of the third adjusting member is variable, for example, the third adjusting member is an inelastic element, and the third adjusting member may be an iris diaphragm (iris diaphragm).
In another embodiment of the present application, the apparatus further includes a detection module and a control module, the detection module is configured to obtain a size of the particle, for example, the detection module includes a first detector, the particle is irradiated by the laser to generate an optical signal, the optical signal includes a fluorescent signal and a scattered light signal, the first detector is configured to receive a first portion of the optical signal, the first portion of the optical signal includes the scattered light signal, a direction of the first portion of the optical signal is parallel to a direction of the laser, and the first portion of the optical signal may be referred to as Forward Scatter (FSC). The first detector is used for acquiring the size of the particles. The control module is configured to adjust the nozzle based on the size or other characteristic of the particles such that the particles have a diameter of approximately 1/5 to 1/2 of the diameter of the nozzle. The application provides a particle sorting unit can continuous adjustment nozzle's diameter size, compares with the fixed nozzle of traditional through-hole size, and applicable scope is extensive, adjusts nozzle diameter's easy and simple to handle, efficient, the precision is high. The application provides a granule sorting unit is applicable to the different waiting of characteristic, flow velocity difference and selects separately the granule, and the automatic characteristics, the velocity of flow etc. of surveying the granule automatically adjust the nozzle size.
In another embodiment of the present application, the apparatus further comprises a sorting module for sorting the particles into different sample tubes/cell culture dishes/cell wells according to the classification of the particles by the control module. For example, the sorting module is used to control the movement of particles so that they fall into a specified location. The method of controlling the movement of particles is not limited, for example, the sorting module further comprises a charging plate for charging the droplets containing particles when the droplets containing particles are about to be separated from the stream of sample fluid, a deflecting plate, such that the droplets containing particles are charged after exiting the stream of fluid; the deflection plate is used for attracting or repelling the charged droplets containing the particles, so that the charged droplets containing the particles are deflected and move according to the speed, direction and trajectory predicted by the control module, or the uncharged droplets containing the particles are not deflected, so that each droplet containing the particles falls into a designated position respectively. The sample fluid may be a sheath fluid. In another embodiment of the present application, the sorting module is further configured to obtain an interval time between each droplet, and control the charging plate to charge and discharge the deflection plate and each droplet by using the interval time between the droplets, so that each droplet falls into a specific position.
Based on the particle sorting device, the particle sorting method comprises the following steps:
controlling a laser to emit at least one laser, the laser being irradiated onto the particles in the flow cell; the particles sequentially generate optical signals under the irradiation of the laser, and the optical signals comprise fluorescence signals and scattered light signals;
receiving the fluorescent signal and/or the scattered light signal with a detection module, and acquiring one or more of the following characteristics of the particle: the size of the particle, the internal composition of the particle, the structure of the particle, the complexity of the particle, the fluorescence characteristic information of the particle, the morphology image of the particle, the fluorescence characteristic information of the particle;
and adjusting the inner diameter of the third adjusting member according to the characteristics of the particles, and enlarging or reducing the diameter of the nozzle.
In another embodiment of the present application, the adjusting the inner diameter of the third adjusting member according to the characteristics of the particles specifically includes moving the first adjusting member left and right to change the lateral size of the third adjusting member. Or, moving the second adjusting piece up and down changes the longitudinal size of the third adjusting piece.
In another embodiment of the present application, the method further comprises placing the particles in a designated location based on the characteristics of the particles. For example, charging the droplet containing the particles such that the droplet containing the particles is charged after exiting the flow chamber; the charged particle-containing droplets are attracted or repelled such that the charged particle-containing droplets are deflected, or the uncharged particle-containing droplets are not deflected such that each particle-containing droplet falls into a designated location, respectively.
The above embodiments are only used for illustrating the technical solutions of the present application, and not for limiting the same; although the present application has been described in detail with reference to the foregoing embodiments, it should be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions in the embodiments of the present application.
Claims (10)
1. A particle sorting apparatus, comprising a flow chamber for restricting the flow of particles and a nozzle for sizing the particles according to their characteristics;
the nozzle includes a third adjustment member provided at a lower portion of the flow chamber, the third adjustment member being in a ring shape having a variable inner diameter.
2. The apparatus of claim 1, wherein the third adjustment member is an iris diaphragm.
3. The apparatus of claim 1, wherein the nozzle further comprises a first adjustment member;
the first adjusting piece is arranged at the lower part of the flow chamber, and the third adjusting piece is arranged inside the first adjusting piece; the third adjusting piece is an elastic element;
the first adjusting piece is used for adjusting the transverse size of the third adjusting piece.
4. The apparatus of claim 1, wherein the nozzle further comprises a second adjustment member;
the second adjusting piece is used for adjusting the longitudinal size of the third adjusting piece; the second adjusting member is disposed at a lower portion of the third adjusting member.
5. The device of claims 1-4, further comprising a spacer;
the partition is disposed between the flow chamber and the third trim.
6. The apparatus of claims 1 to 4, further comprising a detection module and a control module;
the detection module is used for acquiring the size of the particles;
the control module is used for adjusting the nozzle according to the size of the particles.
7. A method of sorting particles, the method comprising:
controlling a laser to emit at least one laser, the laser being irradiated onto the particles in the flow cell; the particles generate fluorescence signals and scattered light signals under the irradiation of the laser in sequence;
receiving the fluorescent signal and/or the scattered light signal with a detection module, and acquiring one or more of the following characteristics of the particle: the size of the particle, the internal composition of the particle, the structure of the particle, the complexity of the particle, the fluorescence characteristic information of the particle, the morphology image of the particle, the fluorescence characteristic information of the particle;
and adjusting the inner diameter of the third adjusting member according to the characteristics of the particles, and enlarging or reducing the diameter of the nozzle.
8. The method of claim 7, wherein the third adjustment member is an iris diaphragm.
9. The method of claim 7, wherein adjusting the inner diameter of the third adjustment member comprises moving the first adjustment member left or right to change the lateral dimension of the third adjustment member.
10. The method of claim 7, wherein adjusting the inner diameter of the third adjustment member comprises moving the second adjustment member up and down to change the longitudinal dimension of the third adjustment member.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010406155.0A CN111521549B (en) | 2020-05-13 | 2020-05-13 | Particle sorting device and method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010406155.0A CN111521549B (en) | 2020-05-13 | 2020-05-13 | Particle sorting device and method |
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| Publication Number | Publication Date |
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| CN111521549A true CN111521549A (en) | 2020-08-11 |
| CN111521549B CN111521549B (en) | 2021-01-01 |
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| US20030138967A1 (en) * | 2002-01-22 | 2003-07-24 | Dakocytomation Denmark A/S | Environmental containment system for a flow cytometer |
| US20050153458A1 (en) * | 1994-10-14 | 2005-07-14 | University Of Washington | Droplet formation systems for flow cytometers |
| US7056430B1 (en) * | 2002-01-09 | 2006-06-06 | Axon Instruments, Inc. | Detachable cell-delivery system for patch-clamp unit |
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| CN107449716A (en) * | 2017-07-25 | 2017-12-08 | 深圳市孔雀生物科技有限公司 | A kind of sorting unit of flow cytometer |
| US20180180526A1 (en) * | 2004-07-27 | 2018-06-28 | Beckman Coulter, Inc. | Enhancing Flow Cytometry Discrimination with Geometric Transformation |
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2020
- 2020-05-13 CN CN202010406155.0A patent/CN111521549B/en active Active
Patent Citations (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS63172943A (en) * | 1987-01-12 | 1988-07-16 | Showa Denko Kk | Apparatus for analyzing and dispensing microparticle |
| JPH041568A (en) * | 1990-04-17 | 1992-01-07 | Fuji Electric Co Ltd | Cell sorter |
| US20050153458A1 (en) * | 1994-10-14 | 2005-07-14 | University Of Washington | Droplet formation systems for flow cytometers |
| CN1265195A (en) * | 1997-07-24 | 2000-08-30 | 美国政府农业部 | Flow cytometry nozzle for high efficiency cell sorting |
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| US7056430B1 (en) * | 2002-01-09 | 2006-06-06 | Axon Instruments, Inc. | Detachable cell-delivery system for patch-clamp unit |
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