CN109735437B - Vessel and method for collecting and processing cells after ejection sorting of cells - Google Patents
Vessel and method for collecting and processing cells after ejection sorting of cells Download PDFInfo
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- CN109735437B CN109735437B CN201910078929.9A CN201910078929A CN109735437B CN 109735437 B CN109735437 B CN 109735437B CN 201910078929 A CN201910078929 A CN 201910078929A CN 109735437 B CN109735437 B CN 109735437B
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Abstract
A vessel and method for cell collection and processing after cell ejection sorting, comprising: the receiving cover is used for receiving the ejection sorted cells and containing cell lysis solution or other reagents; a reaction tube for holding a mixture for polymerase chain reaction and for performing polymerase chain reaction; the receiving cover and the reaction tube can be assembled and disassembled; the invention can meet the limitation of the cell ejection sorter on the receiving height and skillfully solves the requirement of the polymerase chain reaction on the volume; for cells subjected to catapulting sorting, the follow-up researches such as whole genome amplification and whole transcriptome amplification can be directly carried out in a cell receiver, the whole genome amplification success rate and the coverage rate are increased, the structure is simple, the method is simple, and the working result is efficient and reliable.
Description
Technical Field
The invention relates to the technical field of cell collectors and operation methods thereof, in particular to a vessel and a method for collecting and processing cells after ejection sorting of the cells.
Technical Field
Despite the rapid development of modern medicine, cancer, infectious diseases, metabolic diseases and other diseases still threaten the life and health of human beings, and the prevention and treatment of the diseases need to deeply explore the life process; in 2003, the human genome project completes the sequencing work of 30 hundred million base pairs of human bodies, but how to decipher the gene codes and analyze the complex relationship among genes, phenotypes and environments is still an unsolved problem; the intensive research on the cell phenotype interacting with the microenvironment is needed, the phenotype information relative to the genotype is supplemented, the connection between the genotype and the phenotype is established on the single cell level, the real systematic scientific decoding of the life is realized, and the work is beneficial to the accurate medical treatment and disease control and supports the national important requirement of population health;
the single cell ejection sorting technology is an effective cell separation means and has the following principle: the principle of interaction between laser and substances is used for accurately separating target single cells on the chip into a receiver, and the method has the characteristics of no mark, no contact, accuracy, wide application and the like; however, the existing cell collector adopted based on the technology is unreasonable in design, and the essential reasons are that the movement track of the cells in the sorting process is complex, the collection pore volume of the cell collector is small, and subsequent researches such as genome amplification and the like cannot be directly carried out in the cell collector, so that the receiving efficiency of cell receiving consumables is low, and the deep research of correlation between cell phenotype and genotype is directly influenced.
The invention content is as follows:
the invention aims to provide a vessel and a method for collecting and treating cells after cell ejection sorting, wherein the vessel adopts an assembled and disassembled sectional structure design, comprises a receiving cover used for being placed in a cell ejection sorter as a cell collecting container and a reaction tube used for subsequent experimental operations such as gene amplification and the like, and adopts an optimized structure design, so that the cell collector simultaneously meets the requirements of ejection sorting and subsequent research; the transfer steps and the operation flow when the sorted cells are subjected to subsequent treatment are reduced, the probability of exogenous pollution is effectively reduced, and the time cost is saved; meanwhile, the success rate of cell sorting and receiving and genome amplification is effectively improved by adopting a scientific method and application design.
A vessel and method for cell collection and processing after cell ejection sorting, wherein:
a vessel for cell collection and processing after cell catapulting sorting, comprising: a receiving cover which meets the receiving height limitation of the single cell ejection sorter and is used for receiving the single cells or cell groups obtained by sorting; the reaction tube is used for containing reagents for subsequent experimental operations; the receiving cover and the reaction tube can be assembled and disassembled;
by way of illustration, the subsequent experimental manipulation reagents are, for example, polymerase chain reaction mixtures and the like;
further, the receiving cover structure includes: a cell receiving structure for receiving catapulting sorted cells; the connecting structure is used for being connected, positioned and sealed with the reaction tube; the fixing structure is used for fixing the single cell ejection sorter receiving device;
wherein the cell receiving structure is disposed inside the connecting structure; the connecting structure is arranged on the upper part of the fixed structure;
as an example, the vessel is integrated with coupling structures of other consumables, instruments and tools, so that the universality of the vessel is ensured;
as an illustration, the cell receiving structure may also be used to hold cell lysates, culture media, or other solid and liquid reagents;
by way of illustration, the design of the connecting structure includes, but is not limited to, a cylinder, a cone, a cube, a sphere, a polyhedron, or one of a plurality of shape complexes;
as a preferable example, the upper part of the connecting structure is provided with a sealing ring structure around, and the sealing ring structure is used for forming a closed chamber with the reaction tube;
as a preferred illustration, the cell receiving structure has a volume preferably ranging between: 0.1-100 μ l; the volume optimization interval is designed through scientific mathematical modeling, is combined with actual experimental data, and is finally obtained through computer data summarization and analysis, the design of the interval greatly enhances the working efficiency of the vessel, and the requirement of the polymerase chain reaction on the volume is ingeniously solved; the ejection sorted cells can be directly subjected to follow-up researches such as whole genome and whole transcriptome amplification in a cell receiver;
as an illustration, the upper end of the cell receiving structure is provided with an opening, and the cross section of the opening is one of a circle, a square, a triangle or a composite figure of a plurality of figures;
as a preferred illustration, the cross-sectional area of the opening cross-section preferably ranges from: 0.002-80 square millimeters;
further, the fixing structure is used for being reversibly connected and fixed with a cell receiving device of the ejection sorter;
as an application example, the fixing structure may be designed with a clamping structure for facilitating clamping by a transfer device such as tweezers, which can effectively avoid temperature and pressure caused by touching the chamber and data interference generated by external flora;
as a preferred example, the overall height interval of the receiving cover is selected from: 0.1-50 mm;
as an illustration, the cell receiving structures, connecting structures, and securing structures can be arranged in a row-by-row array.
By way of illustration, the shape of the reaction tube includes, but is not limited to: one or a combination of cylinders, cones, cubes, spheres or a complex of multiple shapes.
As an illustration, the preferred range of the volume of the reaction tube is: 10-2000 μ l;
as an example of an application, the material of the vessel is common raw materials, including but not limited to: polypropylene (PP), polymethyl methacrylate (PMMA), Polydimethylsiloxane (PDMS), Polystyrene (PS), Polyethylene (PE), or a combination thereof.
An operation method of a vessel for collecting and processing cells after cell ejection sorting is used for performing polymerase chain reaction on the sorted cells in the vessel, and comprises the following specific steps:
step one, collecting the cells subjected to ejection sorting by using the receiving cover: placing the receiving cover into a receiving device of a single cell ejection sorter, and operating the single cell ejection sorter to sequentially sort target cells in a sample to be detected into one or more receiving covers;
further, the receiving covers are used as cell collecting vessels, and each receiving cover can collect a single cell or a cell group consisting of a plurality of cells;
further, the distance between the receiving cover and the sample chip is 0-5 mm;
furthermore, cell lysate, culture solution or other reagents can be added into the receiving cover in advance, so that a soft landing surface is formed in the chamber, and impact damage caused when the receiving cover contacts the inner wall of the receiving cover in the sorting process is reduced;
by way of illustration, the volume of the cell lysate, culture fluid, or other reagents is: 0.1-100 μ l;
further, the receiving cap may be pre-sterilized, including but not limited to: one or a combination of sterilization methods such as high-pressure sterilization, strong oxidant soaking, radionuclide irradiation and the like;
as an example of a preferred embodiment, the strong oxidant soak is: soaking with sodium hypochlorite;
as an example of a preferred embodiment, the radionuclide irradiation is: cobalt-60 irradiation;
combining the receiving cover containing the sorted cells and the reaction tube into a vessel to form a cell collecting and reacting device with an integrally closed chamber;
as an example, if no lysis solution, culture solution or other reagent or other solution is added to the receiving cap at step one, the solution or reagent may be added to the receiving cap before the receiving cap is combined with the reaction tube;
transferring the cells or cell lysate in the receiving cover to a reaction tube;
transferring the solution containing the cells or cell contents in the receiving cover to the bottom of the reaction tube by turning the combined vessel up and down and applying an acting force to the cells or cell lysate in the receiving cover;
by way of illustration, the forces include, but are not limited to: one or a combination of centrifugal force, gravity, electrostatic force, magnetic field force.
Step four, carrying out subsequent experimental treatment on the cells or cell lysate in the reaction tube;
by way of illustration, the subsequent experimental treatments include, but are not limited to: polymerase chain reaction, culture and biochemical analysis;
as an illustration, the polymerase chain reaction comprises: experiments relating to polymerase chain reaction, such as whole genome amplification, whole transcriptome amplification, target gene fragment amplification, and the like;
to better explain the design principle of the present invention, the following design basis and design concept will be briefly introduced:
firstly, a sectional cell collector is designed for convenient disassembly and assembly, and in the ejection sorting process, because the height of a receiving space of an ejection sorter is limited, a common polymerase chain reaction tube (PCR tube) cannot be put in the ejection sorter as a cell collecting container, the sectional cell collector is designed in advance for receiving the sorted single cells or cell groups and conveniently carrying out subsequent experiments such as polymerase chain reaction and the like in the ejection sorter;
secondly, after sorting is finished, the receiving cover is moved out of the receiving device of the sorter and assembled with the reaction tube to form a segmented cell collector, and after the segmented cell collector is turned over up and down, the cells in the cell collecting cover are transferred into the reaction tube by methods such as centrifugation;
thirdly, because the reaction tube has large enough volume and the size and the shape are matched with the thermal cycler, the subsequent researches such as single-cell whole gene amplification, single-cell whole transcriptome amplification and the like can be conveniently and directly carried out in the reaction tube, the operation flow is simplified, the transfer steps are reduced, and the introduction of exogenous pollution can be effectively prevented;
finally, the invention improves the cell collection efficiency and increases the whole genome amplification success rate and coverage rate by optimizing the single-cell whole gene amplification reaction system and conditions.
The invention has the advantages of
1. The optimized structural design adopted by the invention greatly enhances the use effect of the sectional cell collector, can meet the limitation of the cell ejection sorter on the receiving height, and skillfully solves the requirement of the polymerase chain reaction on the volume; the subsequent researches such as polymerase chain reaction and the like can be directly carried out on the cells subjected to ejection sorting in a cell receiver;
2. the operation flow is simplified, the transfer steps are reduced, and the introduction of exogenous pollution can be effectively prevented;
3. according to the invention, by optimizing the cell collection and treatment method of cell catapulting and sorting, the cell collection efficiency is improved, the whole genome amplification success rate and the coverage rate are increased, the structure is simple, the method is simple, and the working result is efficient and reliable.
Drawings
FIG. 1 is a schematic view of the structure of a vessel for cell collection and treatment after cell ejection and sorting according to the present invention
FIG. 2 is a schematic view of a receiving cover of a vessel for cell collection and processing after cell ejection and sorting according to the present invention
FIG. 3 is a diagram of the whole genome amplification electrophoresis of the segmented cell collector for cell receiving, transferring and polymerase chain reaction and the method using the same of example 1
FIG. 4 is a sectional view of a cell collector for receiving, transferring and PCR of the present invention and a method of using the same, as shown in example 2, which is a full genome amplification and 16S rRNA amplification electrophoretogram
Detailed Description
Preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings.
Detailed description of the drawings referring to fig. 1 and 2, a vessel and method for cell collection and processing after cell catapulting sorting, wherein:
a vessel for cell collection and processing after cell catapulting sorting, comprising: a receiving cover 101 which satisfies the receiving height limitation of the single cell ejection sorter and is used for receiving the single cells or cell groups obtained by sorting; a reaction tube 102 for holding reagents for subsequent experimental operations; the receiving cover 101 and the reaction tube 102 can be assembled and disassembled;
by way of illustration, the subsequent experimental manipulation reagents are, for example, polymerase chain reaction mixtures and the like;
further, the receiving cover 101 structure includes: a cell receiving structure 201 for receiving catapulting sorted cells; a connecting structure 202 for interconnecting, positioning and sealing with the reaction tube 102; a fixing structure 203 for fixing on the receiving device of the single cell ejection sorter;
wherein the cell receiving structure 201 is arranged inside the connecting structure 202; the connecting structure 202 is arranged on the upper part of the fixing structure 203;
as an example, the vessel 100 integrates a coupling structure with other consumables, instruments and tools, so that the universality of the vessel is ensured;
as an illustration, the cell receiving structure 201 may also be used to hold cell lysates, culture media, or other solid and liquid reagents;
by way of illustration, the design of the connecting structure 202 includes, but is not limited to, a cylinder, a cone, a cube, a sphere, a polyhedron, or one of a plurality of shape composites;
as a preferred illustration, the upper portion of the connecting structure 202 is provided with a sealing ring structure around for better forming a closed chamber with the reaction tube 102;
as a preferred illustration, the cell receiving structure 201 has a volume preferably set between: 0.1-100 μ l; the volume optimization interval is designed through scientific mathematical modeling, is combined with actual experimental data, and is finally obtained through computer data summarization and analysis, the design of the interval greatly enhances the working efficiency of the vessel, and the requirement of the polymerase chain reaction on the volume is ingeniously solved; the ejection sorted cells can be directly subjected to follow-up researches such as whole genome and whole transcriptome amplification in a cell receiver;
as an illustration, the upper end of the cell receiving structure 201 is provided with an opening, and the cross section of the opening is circular, square, triangular or one of a plurality of complex figures;
as a preferred illustration, the cross-sectional area of the opening cross-section preferably ranges from: 0.002-80 square millimeters;
further, the fixing structure 203 is used for reversibly connecting and fixing with a cell receiving device of the ejection sorter;
as an application example, the fixing structure 203 may be designed with a clamping structure for facilitating clamping by a transferring device such as tweezers, which can effectively avoid temperature and pressure caused by touching the chamber and data interference generated by external flora;
as a preferred example, the overall height of the receiving cover 101 is selected from: 0.1-50 mm;
as an illustration, the cell receiving structure 201, the connecting structure 202, and the fixing structure 203 may be arranged in a row array.
By way of illustration, the shape of the reaction tube 102 includes, but is not limited to: one or a combination of cylinders, cones, cubes, spheres or a complex of multiple shapes.
As an illustration, the volume of the reaction tube 102 is preferably in the range of: 10 to 2000. mu.l.
As an example of an application, the vessel 100 is made of common raw materials, including but not limited to: polypropylene (PP), polymethyl methacrylate (PMMA), Polydimethylsiloxane (PDMS), Polystyrene (PS), Polyethylene (PE), or a combination thereof.
An operation method of a vessel for collecting and processing cells after cell ejection sorting is used for performing polymerase chain reaction on the sorted cells in the vessel 100, and comprises the following specific steps:
step one, collecting the cells subjected to ejection sorting by using the receiving cover 101: putting the receiving cover 101 into a receiving device of a single cell ejection sorter, and operating the single cell ejection sorter to sequentially sort target cells in a sample to be detected into one or more receiving covers 101;
further, the receiving lids 101 serve as cell collecting vessels, and each receiving lid 101 can collect a single cell or a cell group composed of a plurality of cells;
further, the distance between the receiving cover 101 and the sample chip is 0-5 mm;
furthermore, cell lysate, culture solution or other reagents can be added into the receiving cover 101 in advance, so that a soft landing surface is formed in the chamber, and impact damage caused when the receiving cover 101 contacts the inner wall in the sorting process is reduced;
by way of illustration, the volume of the cell lysate, culture fluid, or other reagents is: 0.1-100 μ l;
further, the receiving cap 101 may be pre-sterilized, including but not limited to: one or a combination of sterilization methods such as high-pressure sterilization, strong oxidant soaking, radionuclide irradiation and the like;
as an example of a preferred embodiment, the strong oxidant soak is: soaking with sodium hypochlorite;
as an example of a preferred embodiment, the radionuclide irradiation is: cobalt-60 irradiation;
step two, combining the receiving cover 101 containing the sorted cells and the reaction tube 102 into a vessel 100 to form a cell collecting and reacting device with an integrally closed chamber;
as an example, if no lysis solution, culture solution or other reagents or other solutions are added to the receiving cap at step one, the above solutions or reagents may be added to the receiving cap 101 before the receiving cap 101 is combined with the reaction tube 102;
step three, transferring the cells or cell lysate in the receiving cover 101 to the reaction tube 102;
transferring the solution containing the cells or cell contents in the receiving cover 101 to the bottom of the reaction tube 102 by turning the combined vessel 100 up and down and applying a force to the cells or cell lysate in the receiving cover 101;
by way of illustration, the forces include, but are not limited to: one or a combination of centrifugal force, gravity, electrostatic force, magnetic field force.
Step four, carrying out subsequent experimental treatment on the cells or cell lysate in the reaction tube 102;
by way of illustration, the subsequent experimental treatments include, but are not limited to: polymerase chain reaction, culture and biochemical analysis;
the fourth step is a specific process taking polymerase chain reaction as an example: lysing the cells; adding a polymerase chain reaction mixture; performing a polymerase chain reaction; and detecting the reaction product.
As an illustration, the polymerase chain reaction comprises: experiments relating to polymerase chain reaction, such as whole genome amplification, whole transcriptome amplification, target gene fragment amplification, and the like;
to better explain the design principle of the present invention, the following design basis and design concept will be briefly introduced:
firstly, a sectional cell collector is designed for convenient disassembly and assembly, and in the ejection sorting process, because the height of a receiving space of an ejection sorter is limited, a common polymerase chain reaction tube (PCR tube) cannot be put in the ejection sorter as a cell collecting container, the sectional cell collector is designed in advance for receiving the sorted single cells or cell groups and conveniently carrying out subsequent experiments such as polymerase chain reaction and the like in the ejection sorter;
secondly, after sorting is finished, the receiving cover 101 is moved out of the receiving device of the sorter, the receiving device and the reaction tube 102 are assembled into a segmented cell collector, and after the segmented cell collector is turned over up and down, cells in the cell collecting cover are transferred into the reaction tube 102 by methods such as centrifugation;
thirdly, because the reaction tube 102 has a large enough volume and the size and the shape are matched with the thermal cycler, the subsequent researches such as single-cell whole gene amplification, single-cell whole transcriptome amplification and the like can be conveniently and directly carried out in the reaction tube, the operation flow is simplified, the transfer steps are reduced, and the introduction of exogenous pollution can be effectively prevented;
finally, the invention improves the cell collection efficiency and increases the whole genome amplification success rate and coverage rate by optimizing the single-cell whole gene amplification reaction system and conditions.
To better illustrate the application of the invention, the following example was carried out:
example 1: using the vessel 100 to receive cells and perform a polymerase chain reaction;
the vessel 100 of the present invention is subjected to aseptic processing; adding a suitable volume of lysis solution to the receiving lid 101 in a laminar flow cabinet; the receiving cover 101 is clamped by tweezers and is placed in a receiving device of the single-cell ejection sorter, and the limit of the single-cell ejection sorter on the height of cell collecting consumables can be met due to the lower height of the receiving cover; operating the single cell ejection sorter to sort one or more target cells into the receiving lid 101; then the receiving cover 101 is removed from the sorter, the reaction tube 102 is assembled with the receiving cover 101 in a reversed way to form a closed vessel 100, so that pollution can be avoided in the cell transfer process; the assembled vessel 100 is turned upside down, and the cell sap in the receiving cover 101 is transferred to the bottom of the reaction tube 102 by the centrifugal principle (at this time, the receiving cover 101 is positioned at the top of the reaction tube 102, and is in an inverted buckle shape), so that the efficient and pollution-free transfer of the sorted cells is realized; then placing the vessel 100 into a thermal cycler, and incubating at high temperature to thoroughly lyse cells; moving the vessel 100 back to the laminar flow cabinet, separating the receiving cover 101 from the reaction tube 102, adding a neutralization solution and a polymerase chain reaction mixed solution into the reaction tube 102, assembling the receiving cover 101 and the reaction tube 102 into a whole again, and then putting the segmented cell collector 102 into a thermal cycler for corresponding polymerase chain reaction; after the reaction is finished, the products are detected by agarose gel electrophoresis, and the data are shown in figure 3, wherein 1 is Marker, 2-8 are whole genome amplification products of sorted cells, and 9 is negative control.
Example 2: a method for whole genome amplification of cells after catapulting sorting based on a segmented cell collector;
carrying out sterile treatment on the vessel 100, adding 2 mu L of lysate in advance into a cavity of the receiving cover 101, clamping the receiving cover 101 by using tweezers, and placing the receiving cover into a receiving device of a single cell ejection sorter; operating a single cell ejection sorter to sort the target single cells into the receiving covers 101 in sequence; removing the receiving cap 101 from the sorter and combining with the reaction tube 102 to form the vessel 100; turning the vessel 100 upside down, centrifuging to transfer the lysate containing cells or cell contents in the receiving cover 101 to the bottom of the reaction tube 102; putting the vessel 100 into a thermal cycler, incubating at 65 ℃ for 10min, and setting the temperature of a hot cover to 70 ℃ to thoroughly lyse cells or microorganisms; adding a neutralizing solution into the reaction tube 102, uniformly mixing the tube walls of the reactor, centrifuging the mixture for a short time, and adjusting the pH value of the system to be neutral; then adding a whole genome amplification reaction mixture comprising random primers at two ends, dNTP, DNA polymerase buffer solution, non-nucleic acid water and the like into the reaction tube 102; then the receiving cover 101 and the reaction tube 201 are recombined into the vessel 100, and the flicked tube walls are uniformly mixed and centrifuged for a short time; putting the vessel 100 into a thermal cycler, incubating at 30 ℃ for 8 hours to complete the whole genome amplification reaction, and then adjusting the temperature to 65 ℃ for incubation for 3min to inactivate DNA polymerase; after the reaction is finished, detecting the product by using agarose gel electrophoresis, wherein a data chart is shown in detail in figure 4;
the optimized structural design adopted by the invention greatly enhances the use effect of the cell collecting vessel after the cell ejection sorting, can meet the limitation of the cell ejection sorting instrument on the receiving height, and skillfully solves the requirement of the polymerase chain reaction on the volume; the ejection sorted cells can be directly subjected to follow-up researches such as whole genome and whole transcriptome amplification in a cell receiver; the operation flow is simplified, the transfer steps are reduced, and the introduction of exogenous pollution can be effectively prevented; by optimizing a single-cell whole-gene amplification reaction system and conditions, the invention improves the cell collection efficiency, increases the whole-genome amplification success rate and the coverage rate, and has simple structure, simple method and high-efficiency and reliable working results.
The above embodiments are only preferred embodiments of the present invention, and it should be understood that the above embodiments are only for assisting understanding of the method and the core idea of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalents and the like made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (9)
1. A household utensils that is used for cell collection and processing after cell ejection is selected separately which characterized in that: the method comprises the following steps: the receiving cover meets the receiving height limit of the single cell ejection sorter, the distance between the receiving cover and the sample chip is 0-5mm, the receiving cover and the reaction tube are used for containing a polymerase chain reaction mixture, and the receiving cover and the reaction tube can be assembled and disassembled; the receiving cover structure includes: a cell receiving structure for receiving catapulting sorted cells; the connecting structure is used for being connected, positioned and sealed with the reaction tube; the fixing structure is used for fixing the single cell ejection sorter receiving device; the cell receiving structure is disposed inside the connecting structure; the connecting structure is arranged on the upper part of the fixed structure;
the fixing structure is used for being in reversible connection and fixation with a cell receiving device of the ejection sorter; the fixed structure is provided with a clamping structure.
2. The vessel for cell collection and processing after cell catapulting sorting according to claim 1, wherein the connecting structure is: one of a cylinder, a cone, a cube, a sphere, a polyhedron or a complex of multiple shapes; the reaction tube is in the shape of one or a combination of a cylinder, a cone, a cube, a sphere or a complex of multiple shapes.
3. The vessel for cell collection and processing after cell catapulting sorting according to claim 1, wherein the receiving cap volume is 0.1-100 μ l; the volume of the reaction tube is 10. mu.l-2000. mu.l.
4. The method for applying the vessel for cell collection and treatment after cell catapulting sorting according to claim 1, which is used for performing polymerase chain reaction on the sorted cells in the vessel, and is characterized by comprising the following specific steps:
step one, collecting the cells subjected to ejection sorting by using the receiving cover: placing the receiving cover into a receiving device of a single cell ejection sorter, and operating the single cell ejection sorter to sequentially sort target cells in a sample to be detected into one or more receiving covers;
combining the receiving cover containing the sorted cells and the reaction tube into a vessel to form a cell collecting and reacting device with an integrally closed chamber;
transferring the cells or cell lysate in the receiving cover to a reaction tube;
and step four, carrying out subsequent experimental treatment on the cells or cell lysate in the reaction tube.
5. The method for applying the vessel for cell collection and treatment after cell catapulting and sorting according to claim 4, wherein the specific process of the first step is as follows: and placing the receiving cover into a receiving device of a single cell ejection sorter, wherein the distance between the receiving cover and a sorting chip is 0-5mm, operating the single cell ejection sorter to sequentially sort single cells or single microorganisms in a sample to be detected into one or more receiving covers, and each receiving cover can receive a single cell or a cell group consisting of a plurality of cells.
6. The method for applying the vessel for cell collection and treatment after cell catapulting and sorting according to claim 4, wherein the specific process of the second step is as follows: and moving the receiving cover out of the single cell ejection sorter, and combining the receiving cover with the reaction tube to form the sectional cell collector.
7. The method for applying the vessel for cell collection and treatment after cell catapulting and sorting according to claim 4, wherein the specific process of the third step is as follows: the cells or cell lysate in the receiving lid are transferred to the bottom of the reaction tube using a centrifugation method.
8. The method for applying the vessel for collecting and processing the cells after the ejection sorting of the cells as claimed in claim 4, wherein the specific process of performing the subsequent experiment processing on the cells or cell lysates in the reaction tube in the fourth step is as follows: lysing the cells; adding a polymerase chain reaction mixture; performing a polymerase chain reaction; and detecting the reaction product.
9. The method of claim 4, wherein the subsequent experimental treatment comprises: polymerase chain reaction, culture and biochemical analysis.
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| CN106065391A (en) * | 2016-07-20 | 2016-11-02 | 国家纳米科学中心 | For unicellular sorting and the micro-fluidic chip of unicellular whole genome amplification |
| CN108130270A (en) * | 2016-12-01 | 2018-06-08 | 李治国 | Instant genetic test packet |
| CN108315389B (en) * | 2016-12-27 | 2021-07-02 | 中国科学院微生物研究所 | Micro-volume cell nucleic acid amplification method |
| CN106978488A (en) * | 2017-04-02 | 2017-07-25 | 北京军秀咨询有限公司 | A kind of gene tester, detection kit and its application |
| CN106906130A (en) * | 2017-04-19 | 2017-06-30 | 北京倍肯创新诊断技术研究院有限责任公司 | A kind of flat PCR reaction tubes of printing opacity high |
| CN106967592A (en) * | 2017-06-05 | 2017-07-21 | 无锡国盛生物工程有限公司 | A kind of PCR pipe |
| CN206887110U (en) * | 2017-06-07 | 2018-01-16 | 浙江省医学科学院 | A kind of PCR pipe, PCR pipe bar |
| CN107299049B (en) * | 2017-07-06 | 2020-08-25 | 浙江晟景生物技术有限公司 | Kit for extracting DNA from oral cells of infant |
| CN206970629U (en) * | 2017-07-20 | 2018-02-06 | 温和心 | A kind of paraffin cellular-type ring mediated responses pipe device |
| CN107490545A (en) * | 2017-07-21 | 2017-12-19 | 中国科学院青岛生物能源与过程研究所 | A kind of unicellular automation of high-flux microorganism sorts and reception system |
| CN107589105B (en) * | 2017-07-21 | 2021-08-24 | 中国科学院青岛生物能源与过程研究所 | Integrated device for single cell rapid Raman measurement and laser ejection sorting |
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2019
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