CN109735437A - It is collected and the vessel and method that handle after a kind of ejection sorting of cell for cell - Google Patents
It is collected and the vessel and method that handle after a kind of ejection sorting of cell for cell Download PDFInfo
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- CN109735437A CN109735437A CN201910078929.9A CN201910078929A CN109735437A CN 109735437 A CN109735437 A CN 109735437A CN 201910078929 A CN201910078929 A CN 201910078929A CN 109735437 A CN109735437 A CN 109735437A
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Abstract
It is collected and the vessel and method that handle after a kind of ejection sorting of cell for cell, comprising: for being received and being contained cell pyrolysis liquid or the reception lid of other reagents to ejection sorting cell；For containing polymerase chain reaction mixture and carrying out the reaction tube of polymerase chain reaction；Receiving lid can be assembled and split with reaction tube；The present invention can meet limitation of the cell ejection sorter to reception height, and cleverly solve demand of the polymerase chain reaction to volume；The cell of ejection sorting may be implemented in cell receiver and directly carry out the follow-up studies such as full-length genome, the amplification of full transcript profile, increase whole genome amplification success rate and coverage rate, and structure is simple, method is concise, working result high efficient and reliable.
Description
Technical field
The present invention relates to a kind of cell harvestor and its operating method technical field, after especially a kind of cell ejection sorting
The vessel and method with processing are collected for cell.
Technical background
Although modern medicine develops rapidly, the diseases such as cancer, infectious diseases, metabolic still threaten the life of the mankind
Life is with health, the prevention and treatment to these diseases, needs to explore life process deeper into ground；2003, " Human Genome Project " was complete
At the examining order of 3,000,000,000 base-pairs of human body, but these gene-codes how are decoded, between parsing gene, phenotype and environment
Complex relationship is still yet unresolved issue；This needs is furtherd investigate to the cell phenotype that microenvironment interacts, and is mended
The phenotypic information opposite with genotype is filled, contacting between genotype and phenotype is established in unicellular level, is realized to life
Real system sexology decoding, this work facilitate precisely medical treatment and disease prevention and cure, and support this country of population health is great
Demand；
Unicellular ejection sorting technology is a kind of effective cell separation means, principle are as follows: laser and substance phase interaction
Principle has unmarked, non-contact, accurate, suitable extensively by the unicellular accurate separation of the target on chip into receiver
The features such as using；But it is unreasonable based on existing cell harvestor design used by the technology, essential reason is that cell exists
Motion profile is complicated in assorting room, and the collection pore volume of cell harvestor is smaller, and gene can not be directly carried out in collector
The follow-up studies such as group amplification, the receiving efficiency for causing cell to receive consumptive material is low, directly affects cell phenotype and genotype correlation
Further investigation.
Summary of the invention:
The vessel and method collected and handled for cell after being sorted it is an object of the present invention to provide a kind of ejection of cell,
The vessel are designed using composable with segment type structure that is splitting, including are received for being put into cell ejection sorter as cell
Collect the reception lid of container, and for the reaction tube of the experimental implementations such as subsequent gene amplification, is designed using the structure of optimization, so that
The cell harvestor meets the needs of ejection sorting and follow-up study simultaneously；Reduce and turns when sorting cell carries out subsequent processing
Walk is rapid and operating process, effectively reduces external source pollution probability, and saved time cost；It is answered simultaneously using the method for science
With design, the success rate of cell sorting reception and genome amplification is effectively raised.
It is collected and the vessel and method that handle after a kind of ejection sorting of cell for cell, in which:
It is collected and the vessel that handle after a kind of ejection sorting of cell for cell, comprising: meet unicellular ejection sorter
Height limitation is received, for receiving unicellular or cell mass the reception lid that sorting obtains；For containing subsequent experimental operation examination
The reaction tube of agent；The reception lid and reaction tube can be assembled and be split；
Illustrate as an example, the subsequent experimental operation reagent is such as polymerase chain reaction mixture；
Further, the lid structure that receives includes: to receive structure for receiving the cell of ejection sorting cell；For with
The connection structure that reaction tube is connected with each other, positions, seals；Fixation for being fixed in unicellular ejection sorter reception device
Structure；
Wherein, the cell receives structure setting in the inside of connection structure；The connection structure is arranged in the fixation
Structure upper；
Illustrate as an example, the vessel are integrated with the coupled structure with other consumptive materials, instrument, tool, ensure that
The versatility of vessel；
Illustrate as an example, the cell receive structure can be also used for containing cell pyrolysis liquid, culture medium or its
His solid and liquid reagent；
Illustrate as an example, the design of the connection structure, including but not limited to cylinder, cone, cube, sphere,
One of polyhedron or various shapes complex；
Illustrate as a kind of preferable examples, connection structure top surrounding is provided with ring structure, for better
Closed chamber is formed with reaction tube；
Illustrate as a kind of preferable examples, the preferred section setting of volume that the cell receives structure exists: 0.1 μ l-100 μ
l；The preferred section of this volume combines actual experiment data by the mathematical modeling design of science, is summarized point by computer data
It is finally obtained after analysis, the design in the section greatly strengthens the working efficiency of vessel, and cleverly solves polymerase chain reaction
Cope with the demand of volume；The cell of ejection sorting may be implemented in cell receiver and directly carry out full-length genome, full transcription
The follow-up studies such as group amplification；
Illustrate as an example, the upper end that the cell receives structure is equipped with opening, opening section be it is round, rectangular,
One of triangle or the compound pattern of a variety of figures；
Illustrate as a kind of preferable examples, the preferred section of the sectional area of the opening section are as follows: 0.002~80 square of milli
Rice；
Further, be used to carry out with the cell reception device of ejection sorter can reverse connection and solid for the fixed structure
It is fixed；
Illustrate as a kind of applicating example, the transfer utensil clamping such as tweezers easy to use can have been designed on the fixed structure
Grasping structures, this measure can effectively avoid touching the data interference that chamber bring temperature, pressure and external flora generate；
Illustrate as a kind of preferable examples, its whole height section selection: 0.1~50mm is covered in the reception；
Illustrate as an example, the cell, which receives structure, connection structure, fixed structure, can be set to townhouse battle array
Column.
Illustrate as an example, the shape of the reaction tube includes but is not limited to: cylinder, cone, cube, sphere or more
One of complex of kind shape or combination.
Illustrate as an example, the volume preferred scope of the reaction tube: 10 μ l-2000 μ l；
Illustrate as a kind of applicating example, the material of the vessel is common raw materials, including but not limited to: polypropylene
(PP), polymethyl methacrylate (PMMA), dimethyl silicone polymer (PDMS), polystyrene (PS), in polyethylene (PE)
A kind of or combination.
The operating method with the vessel handled is collected for cell after a kind of ejection sorting of cell, for sorting in vessel
Cell carries out polymerase chain reaction, comprises the following specific steps that:
Step 1: being collected using the cell for covering and being sorted to ejection that receives: lid will be received and be put into unicellular ejection
In the reception device of sorter, operate unicellular ejection sorter the target cell in sample to be tested is successively sorted to one or
In multiple reception lids；
Further, the reception lid is used as cell harvestor ware, and each reception, which is covered, collects individual cells or by multiple
The cell mass of cell composition；
Further, it receives and covers at a distance between sample chip as 0-5mm；
Further, cell pyrolysis liquid, culture solution or other reagents can be previously added to receiving in lid, this measure is in chamber
Soft landing surface is formed, is reduced in assorting room, the impact injury when contact receives lid inner wall；
Illustrate as an example, the volume of the cell pyrolysis liquid, culture solution or other reagents are as follows: 0.1-100 μ l；
Further, the reception lid can carry out aseptic process in advance, including but not limited to: high pressure sterilization, strong oxidizer
One of method for disinfection such as immersion and radionuclide irradiation or combination；
It illustrates as a kind of preferred embodiment, the strong oxidizer impregnates are as follows: sodium hypochlorite impregnates；
It illustrates as a kind of preferred embodiment, the radionuclide irradiation are as follows: cobalt -60 irradiates；
Step 2: the reception lid for including sorting cell and reaction tube group are combined into vessel, being formed, there is one to seal
The cell of closed chamber room is collected and reactor；
Illustrate as an example, if lysate, culture solution or other examinations are not added into reception lid in step 1
Above-mentioned solution or reagent can be added to receiving in lid before receiving lid and combining with reaction tube in agent or other solution；
Step 3: the cell received in lid or cell pyrolysis liquid are transferred in reaction tube；
By by combination after the vessel spin upside down, then pass through to receive lid in cell or cell pyrolysis liquid apply
Described receive is included that the solution of cell or cellular content is transferred to reaction bottom of the tube in lid by the method for active force；
Illustrate as an example, the active force includes but is not limited to: centrifugal force, gravity, electrostatic force, in magnetic field force
A kind of or combination.
Step 4: in reaction tube cell or cell pyrolysis liquid carry out subsequent experimental processing；
Illustrate as an example, the subsequent experimental processing includes but is not limited to: polymerase chain reaction, culture, life
Change analysis；
Illustrate as an example, the polymerase chain reaction includes: whole genome amplification, the amplification of full transcript profile, mesh
Mark gene fragment amplification etc. is related to the experiment of polymerase chain reaction；
Design principle in order to better illustrate the present invention now briefly introduces its design considerations and mentality of designing is as follows:
Firstly, design segment type cell harvestor is to be able to easily split and assemble, in ejection assorting room,
Due to launching the reception space limited height system of sorter, common polymerase chain reaction pipe (PCR pipe) can not be put into wherein
As cell collection container, so the present invention designs a kind of segment type cell harvestor in advance, obtain for receiving sorting
Unicellular or cell mass, while the subsequent experimentals such as polymerase chain reaction are easily carried out wherein；
Secondly, covering the reception device for removing sorter after sorting for receiving, being assembled into segment type cell with reaction tube
Cell after spinning upside down, is collected the cell in lid by the methods of centrifugation and is transferred in reaction tube by collector；
Again, since reaction pipe volume is enough big, and size, shape and thermal cycler match, can be easily straight wherein
The follow-up studies such as the unicellular full genome amplification of row, unicellular full transcript profile amplification are tapped into, operating process is simplified, reduces transfer step
Suddenly, it can effectively prevent introducing external source pollution；
Finally, the present invention by optimizing unicellular full genome amplification reaction system and condition, is improving cell collection efficiency
While, increase whole genome amplification success rate and coverage rate.
Beneficial effects of the present invention
1, the optimizing structure design that the present invention uses, greatly strengthens the using effect of segment type cell harvestor, can expire
Sertoli cell launches limitation of the sorter to reception height, and cleverly solves demand of the polymerase chain reaction to volume；It is right
It may be implemented in the cell of ejection sorting and directly carry out the follow-up studies such as polymerase chain reaction in cell receiver；
2, operating process is simplified, transfer step is reduced, can effectively prevent introducing external source pollution；
3, the cell collection and processing method of the invention by optimization cell ejection sorting, is improving cell collection efficiency
Meanwhile whole genome amplification success rate and coverage rate being increased, and structure is simple, method is concise, working result high efficient and reliable.
Detailed description of the invention
Fig. 1 is the structural schematic diagram collected for cell after a kind of cell ejection of the present invention sorts with the vessel handled
Fig. 2 is that the structure after a kind of cell ejection of the present invention sorts for the reception lid of the vessel of cell collection and processing is shown
It is intended to
Fig. 3 be cell of the present invention receive, the segment type cell harvestor and its application method of transfer, polymerase chain reaction
1 whole genome amplification electrophoretogram of embodiment
Fig. 4 be cell of the present invention receive, the segment type cell harvestor and its application method of transfer, polymerase chain reaction
2 whole genome amplification of embodiment and 16S rRNA expand electrophoretogram
Specific embodiment
With reference to the accompanying drawing to a preferred embodiment of the present invention will be described in detail.
Be detailed in shown in Fig. 1 and Fig. 2, after a kind of ejection sorting of cell for cell collect with the vessel and method that handle,
In:
It is collected and the vessel that handle after a kind of ejection sorting of cell for cell, comprising: meet unicellular ejection sorter
Height limitation is received, for receiving unicellular or cell mass the reception lid 101 that sorting obtains；For containing subsequent experimental operation
The reaction tube 102 of reagent；The reception lid 101 and reaction tube 102 can be assembled and be split；
Illustrate as an example, the subsequent experimental operation reagent is such as polymerase chain reaction mixture；
Further, 101 structure of lid that receives includes: to receive structure 201 for receiving the cell of ejection sorting cell；
For being connected with each other, positioning, the connection structure 202 of sealing with reaction tube 102；It is received for being fixed on unicellular ejection sorter
Fixed structure 203 on device；
Wherein, the cell receives the inside that connection structure 202 is arranged in structure 201；The setting of connection structure 202 exists
203 top of fixed structure；
Illustrate as an example, the vessel 100 are integrated with the coupled structure with other consumptive materials, instrument, tool, guarantee
The versatilities of vessel；
Illustrate as an example, the cell receive structure 201 can be also used for containing cell pyrolysis liquid, culture medium or
Other solids and liquid reagent；
Illustrate as an example, the design of the connection structure 202, including but not limited to cylinder, cone, cube, ball
One of body, polyhedron or various shapes complex；
Illustrate as a kind of preferable examples, the 202 top surrounding of connection structure is provided with ring structure, for more preferable
With reaction tube 102 formed closed chamber；
Illustrate as a kind of preferable examples, the preferred section setting of volume that the cell receives structure 201 exists: 0.1 μ l-
100μl；The preferred section of this volume is designed by the mathematical modeling of science, and combines actual experiment data, is converged by computer data
It is finally obtained after bulk analysis, the design in the section greatly strengthens the working efficiency of vessel, and cleverly solves polymerase chain
Formula reacts the demand to volume；The cell of ejection sorting may be implemented in cell receiver and directly carry out full-length genome, complete
The follow-up studies such as transcript profile amplification；
Illustrate as an example, the upper end that the cell receives structure 201 is equipped with opening, and opening section is round, side
One of shape, triangle or compound pattern of a variety of figures；
Illustrate as a kind of preferable examples, the preferred section of the sectional area of the opening section are as follows: 0.002~80 square of milli
Rice；
Further, the fixed structure 203 be used for ejection sorter cell reception device carry out can reverse connection with
It is fixed；
Illustrate as a kind of applicating example, the transfer utensil such as tweezers easy to use can have been designed on the fixed structure 203
The grasping structures of clamping, this measure can effectively avoid touching the data that chamber bring temperature, pressure and external flora generate
Interference；
Illustrate as a kind of preferable examples, described its whole height section of reception lid 101 is chosen: 0.1~50mm；
Illustrate as an example, it is settable that the cell receives structure 201, connection structure 202, fixed structure 203
At townhouse array.
Illustrate as an example, the shape of the reaction tube 102 includes but is not limited to: cylinder, cone, cube, sphere
One of complex of various shapes or combination.
Illustrate as an example, the volume preferred scope of the reaction tube 102: 10 μ l-2000 μ l.
Illustrate as a kind of applicating example, the material of the vessel 100 is common raw materials, including but not limited to: poly- third
Alkene (PP), polymethyl methacrylate (PMMA), dimethyl silicone polymer (PDMS), polystyrene (PS), in polyethylene (PE)
One kind or combination.
The operating method with the vessel handled is collected for cell after a kind of ejection sorting of cell, for in vessel 100
It sorts cell and carries out polymerase chain reaction, comprise the following specific steps that:
Step 1: using it is described receive 101 pairs of lid ejection sorting cells be collected: will receive lid 101 be put into it is slender
Born of the same parents launch in the reception device of sorter, operate unicellular ejection sorter by the target cell in sample to be tested successively sort to
In one or more reception lids 101；
Further, the reception lid 101 is used as cell harvestor ware, each reception lid 101 collect individual cells or
The cell mass being made of multiple cells；
Further, receive lid 101 between sample chip at a distance from be 0-5mm；
Further, cell pyrolysis liquid, culture solution or other reagents can be previously added to receiving in lid 101, this measure is to chamber
Indoor formation soft landing surface reduces in assorting room, the impact injury when contact receives 101 inner wall of lid；
Illustrate as an example, the volume of the cell pyrolysis liquid, culture solution or other reagents are as follows: 0.1-100 μ l；
Further, the reception lid 101 can carry out aseptic process in advance, including but not limited to: high pressure sterilization, Strong oxdiative
One of method for disinfection such as agent immersion and radionuclide irradiation or combination；
It illustrates as a kind of preferred embodiment, the strong oxidizer impregnates are as follows: sodium hypochlorite impregnates；
It illustrates as a kind of preferred embodiment, the radionuclide irradiation are as follows: cobalt -60 irradiates；
Step 2: by it is described include to sort the reception lid 101 of cell to be combined into vessel 100 with 102 groups of reaction tube, form tool
There are cell collection and the reactor of integrated closed chamber；
Illustrate as an example, if lysate, culture solution or other examinations are not added into reception lid in step 1
Above-mentioned solution or examination can be added to receiving in lid 101 before receiving lid 101 and combining with reaction tube 102 in agent or other solution
Agent；
Step 3: the cell received in lid 101 or cell pyrolysis liquid are transferred in reaction tube 102；
By by combination after the vessel 100 spin upside down, then by receive lid 101 in cell or cell cracking
The method that liquid applies active force will include that the solution of cell or cellular content is transferred to reaction tube in the reception lid 101
102 bottoms；
Illustrate as an example, the active force includes but is not limited to: centrifugal force, gravity, electrostatic force, in magnetic field force
A kind of or combination.
Step 4: in reaction tube 102 cell or cell pyrolysis liquid carry out subsequent experimental processing；
Illustrate as an example, the subsequent experimental processing includes but is not limited to: polymerase chain reaction, culture, life
Change analysis；
Detailed process of the step 4 by taking polymerase chain reaction as an example are as follows: lytic cell；Polymerase chain reaction is added
Answer mixture；Carry out polymerase chain reaction；Reaction product is detected.
Illustrate as an example, the polymerase chain reaction includes: whole genome amplification, the amplification of full transcript profile, mesh
Mark gene fragment amplification etc. is related to the experiment of polymerase chain reaction；
Design principle in order to better illustrate the present invention now briefly introduces its design considerations and mentality of designing is as follows:
Firstly, design segment type cell harvestor is to be able to easily split and assemble, in ejection assorting room,
Due to launching the reception space limited height system of sorter, common polymerase chain reaction pipe (PCR pipe) can not be put into wherein
As cell collection container, so the present invention designs a kind of segment type cell harvestor in advance, obtain for receiving sorting
Unicellular or cell mass, while the subsequent experimentals such as polymerase chain reaction are easily carried out wherein；
Secondly, receiving the reception device of the removal sorter of lid 101 after sorting, being assembled into merogenesis with reaction tube 102
Cell after spinning upside down, is collected the cell in lid by the methods of centrifugation and is transferred in reaction tube 102 by formula cell harvestor；
Again, since 102 volume of reaction tube is enough big, and size, shape and thermal cycler match, can easily wherein
The follow-up studies such as unicellular full genome amplification, unicellular full transcript profile amplification are directly carried out, operating process is simplified, reduces transfer
Step can effectively prevent introducing external source pollution；
Finally, the present invention by optimizing unicellular full genome amplification reaction system and condition, is improving cell collection efficiency
While, increase whole genome amplification success rate and coverage rate.
Application method in order to better illustrate the present invention, citing operation embodiment are as follows:
Embodiment 1: cell is received using this vessel 100 and carries out polymerase chain reaction；
Aseptic process is carried out to the vessel 100 of the present invention；Splitting for proper volume is added into reception lid 101 in Streamline cabinet
Solve liquid；With tweezers clamp receive lid 101, be put into it is unicellular ejection sorter reception device in, due to this receptions lid highly compared with
It is low, it can satisfy the limitation that unicellular ejection sorter collects consumptive material height to cell；Unicellular ejection sorter is operated, by one
A or multiple target cells are sorted into reception lid 101；Lid 101 will then be received and remove sorter, reaction tube 102 is buckled to
It is assembled with lid 101 is received, forms closed vessel 100, to can be avoided pollution during transcellular；By group
Vessel 100 after dress turn upside down, and the cell liquid in the reception lid 101 is transferred to 102 bottom of reaction tube by centrifugal principle
(lid 101 that receives described at this time be in the top of reaction tube 102, itself is in back-off shape), realize sort cell it is efficient, without dirt
Dye transfer；Vessel 100 are put into thermal cycler again, high temperature incubation cracks cell thoroughly；Vessel 100 are moved back into laminar flow
In cabinet, lid 101 will be received and separated with reaction tube 102, neutralizer and polymerase chain reaction mixing are added into reaction tube 102
Liquid, then lid 101 will be received again and be assembled integrally with reaction tube 102, segment type cell harvestor 102 is then put into thermal cycle
Corresponding polymerase chain reaction is carried out in instrument；After reaction, product is detected using agarose gel electrophoresis, data
In detail as shown in Figure 3, wherein 1 is Marker, 2-8 is the whole genome amplification product for sorting cell, and 9 be negative control.
Embodiment 2: the method that whole genome amplification is carried out to the cell after ejection sorting based on segment type cell harvestor；
To vessel 100 carry out aseptic process, to receive lid 101 chamber in be previously added 2 μ L volume lysates, use tweezer
Sub-folder, which takes, receives lid 101, is put into the reception device of unicellular ejection sorter；Unicellular ejection sorter is operated, by target
It is unicellular successively to sort into each reception lid 101；Lid 101 will be received and remove sorter, and combined with reaction tube 102, formed
Vessel 100；Spin upside down vessel 100, it includes that the lysate of cell or cellular content is transferred in lid 101 that centrifugation, which will receive,
102 bottom of reaction tube；Vessel 100 are put into thermal cycler, carry out 65 DEG C of incubations, duration 10min, while hot Gai Wendu is set
It is 70 DEG C, cracks cell or microorganism thoroughly；Neutralizer is added into reaction tube 102, flicks tube wall mixing and of short duration centrifugation,
System pH is adjusted to neutrality；Then to addition whole genome amplification reaction mixture in reaction tube 102, including both ends are with power traction
The ingredients such as object, dNTP, archaeal dna polymerase, DNA polymerase buffer liquid, seedless sour water；Lid 101 and reaction tube 201 will be received again again
Vessel 100 are combined into, tube wall mixing and of short duration centrifugation are flicked；Vessel 100 are put into thermal cycler, 30 DEG C of incubations are carried out, when
Long 8h completes whole genome amplification reaction, then adjusts the temperature to 65 DEG C of incubation 3min, inactivates archaeal dna polymerase；Reaction terminates
Afterwards, product is detected using agarose gel electrophoresis, data drawing list is in detail as shown in Figure 4；
The optimizing structure design that the present invention uses greatly strengthens the use effect of cell harvestor ware after cell ejection sorting
Fruit can meet limitation of the cell ejection sorter to reception height, and cleverly solve polymerase chain reaction to volume
Demand；The cell of ejection sorting may be implemented in cell receiver after directly carrying out full-length genome, the amplification of full transcript profile etc.
Continuous research；Operating process is simplified, transfer step is reduced, can effectively prevent introducing external source pollution；The present invention is slender by optimizing
Born of the same parents' full genome amplification reaction system and condition increase whole genome amplification success rate while improving cell collection efficiency
And coverage rate, and structure is simple, method is concise, working result high efficient and reliable.
It is above-described to be merely a preferred embodiment of the present invention, it should be understood that the explanation of above embodiments is only used
In facilitating the understanding of the method and its core concept of the invention, it is not intended to limit the scope of protection of the present invention, it is all of the invention
Any modification for being made within thought and principle, equivalent replacement etc., should all be included in the protection scope of the present invention.
Claims (9)
1. the vessel collected and handled for cell after a kind of cell ejection sorting, it is characterised in that: include: to meet unicellular bullet
The reception lid that sorter receives height limitation, and the reaction tube for containing polymerase chain reaction mixture are penetrated, lid is received
It can assemble and split with reaction tube；The lid structure that receives includes: to receive structure for receiving the cell of ejection sorting cell；
Connection structure for being connected with each other, positioning, sealing with reaction tube；For being fixed on unicellular ejection sorter reception device
Fixed structure；The cell receives structure setting in the inside of connection structure；The connection structure is arranged in the fixed knot
Structure top.
2. the vessel collected and handled for cell after a kind of cell ejection sorting according to claim 1, feature exist
In the design of the connection structure, including but not limited to cylinder, cone, cube, sphere, polyhedron or various shapes complex
One of；The shape of the reaction tube is one of complex of cylinder, cone, cube, sphere or various shapes or group
It closes.
3. the vessel collected and handled for cell after a kind of cell ejection sorting according to claim 1, feature exist
In the reception cover volume is 0.1 μ l-100 μ l；The volume of reaction tube is 10 μ l-2000 μ l.
4. the application method with the vessel handled is collected for cell after a kind of cell ejection sorting, for thin to sorting in vessel
Born of the same parents carry out polymerase chain reaction, which is characterized in that comprise the following specific steps that:
Step 1: being collected using the cell for covering and being sorted to ejection that receives: lid will be received and be put into unicellular ejection sorting
In the reception device of instrument, operates unicellular ejection sorter and successively sort the target cell in sample to be tested to one or more
In the reception lid；
Step 2: the reception lid for including sorting cell and reaction tube group are combined into vessel, being formed has integrated closing chamber
The cell of room is collected and reactor；
Step 3: the cell received in lid or cell pyrolysis liquid are transferred in reaction tube；
Step 4: in reaction tube cell or cell pyrolysis liquid carry out subsequent experimental processing.
5. the application side with the vessel handled is collected after a kind of cell ejection sorting according to claim 2 for cell
Method, which is characterized in that the detailed process of step 1 are as follows: the reception lid of segment type cell harvestor is put into unicellular ejection and is sorted
It is 0-10mm at a distance from sorting chip, operating unicellular ejection sorter will be in sample to be tested in the reception device of instrument
Unicellular or single microbial is successively sorted into one or more reception lids, the receivable individual cells of each receptions lid or
The cell mass being made of multiple cells.
6. the application side with the vessel handled is collected after a kind of cell ejection sorting according to claim 2 for cell
Method, which is characterized in that the detailed process of the step 2 are as follows: reception lid is removed into unicellular ejection sorter, and with it is anti-
Guan Zuhe is answered, segment type cell harvestor is formed.
7. the application side with the vessel handled is collected after a kind of cell ejection sorting according to claim 2 for cell
Method, which is characterized in that the detailed process of the step 3 are as follows: the cell in lid or cell cracking will be received using the methods of centrifugation
Liquid is transferred to reaction bottom of the tube.
8. the application side with the vessel handled is collected after a kind of cell ejection sorting according to claim 2 for cell
Method, which is characterized in that detailed process of the step 4 by taking polymerase chain reaction as an example are as follows: lytic cell；Polymerase is added
Chain reaction mixture；Carry out polymerase chain reaction；Reaction product is detected.
9. the application side with the vessel handled is collected after a kind of cell ejection sorting according to claim 2 for cell
Method, which is characterized in that the subsequent experimental processing includes but is not limited to: polymerase chain reaction, culture, biochemical analysis.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111004706A (en) * | 2019-12-23 | 2020-04-14 | 长春长光辰英生物科学仪器有限公司 | High-flux receiving system for micro-size target sorting instrument |
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