CN108410729B - Primary cell extraction equipment and use method thereof - Google Patents
Primary cell extraction equipment and use method thereof Download PDFInfo
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- CN108410729B CN108410729B CN201710073993.9A CN201710073993A CN108410729B CN 108410729 B CN108410729 B CN 108410729B CN 201710073993 A CN201710073993 A CN 201710073993A CN 108410729 B CN108410729 B CN 108410729B
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- 238000000605 extraction Methods 0.000 title claims abstract description 60
- 238000000034 method Methods 0.000 title claims abstract description 16
- 239000007788 liquid Substances 0.000 claims abstract description 90
- 238000010438 heat treatment Methods 0.000 claims abstract description 26
- 239000000243 solution Substances 0.000 claims description 60
- 239000002244 precipitate Substances 0.000 claims description 37
- 239000006228 supernatant Substances 0.000 claims description 31
- 238000004140 cleaning Methods 0.000 claims description 25
- 102000004190 Enzymes Human genes 0.000 claims description 20
- 108090000790 Enzymes Proteins 0.000 claims description 20
- 238000005119 centrifugation Methods 0.000 claims description 17
- 238000006911 enzymatic reaction Methods 0.000 claims description 11
- 238000011084 recovery Methods 0.000 claims description 11
- 239000002699 waste material Substances 0.000 claims description 11
- 238000007599 discharging Methods 0.000 claims description 5
- 239000007853 buffer solution Substances 0.000 claims description 4
- 238000004113 cell culture Methods 0.000 claims description 4
- 238000011010 flushing procedure Methods 0.000 claims description 4
- 239000013049 sediment Substances 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 2
- 238000005580 one pot reaction Methods 0.000 claims description 2
- 239000001963 growth medium Substances 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 3
- 238000011109 contamination Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 230000004075 alteration Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/18—Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/52—Mobile; Means for transporting the apparatus
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
- C12M41/18—Heat exchange systems, e.g. heat jackets or outer envelopes
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/48—Automatic or computerized control
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M45/00—Means for pre-treatment of biological substances
- C12M45/02—Means for pre-treatment of biological substances by mechanical forces; Stirring; Trituration; Comminuting
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M45/00—Means for pre-treatment of biological substances
- C12M45/05—Means for pre-treatment of biological substances by centrifugation
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M45/00—Means for pre-treatment of biological substances
- C12M45/09—Means for pre-treatment of biological substances by enzymatic treatment
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- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
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- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
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- C12N2509/00—Methods for the dissociation of cells, e.g. specific use of enzymes
- C12N2509/10—Mechanical dissociation
Abstract
The invention discloses primary cell extraction equipment and a using method thereof. The sample mincing zone comprises a tissue mincing device. The operation area comprises a first gate, a heating temperature control device, a rotary fixture, a centrifugal device, a cover opening and closing device, a test tube placing frame and a liquid transferring device; the first gate is adjacent to the sample chopping zone; the heating temperature control device is adjacent to the first gate and is sequentially adjacent to the rotary fixture and the centrifugal device; the switch cover device is adjacent to the first gate, opposite to the heating temperature control device, and is adjacent to the test tube placing frame and the liquid transferring device in sequence. The extraction area comprises a second gate adjacent to the working area. The slide rail device is provided with a trolley which can slide on the slide rail device. The invention can achieve the effects of saving manpower and reducing pollution.
Description
Technical Field
The invention relates to an extraction device, in particular to a cell extraction device. The present invention further relates to a method of using, and more particularly to a method of performing the primary cell extraction apparatus described above.
Background
In recent years, the technology of extracting primary cell culture has been developed quite well, and the continuous culture of primary cells has been applied to various fields, so that the extraction of primary cells is now urgently required to be manufactured in large quantities.
In the prior art, since the extraction of primary cells involves multiple steps, multiple technicians are required to be employed to start with the tissue obtaining and to perform multiple steps of cutting, centrifuging, enzyme reaction, washing, culturing and the like in a laboratory, not only the tissue cut by each technician is different in size, but also the technicians are required to move back and forth between the steps to perform the steps repeatedly. The culture usually takes more than half a working day from the time of obtaining the tissue to the time of starting the culture, and once a large amount of primary cells are extracted, a plurality of technicians are required to complete the extraction.
However, since there are many processes in the primary cell extraction step, and it is not necessary to operate in a sterile room or a laminar flow cabinet (lab chamber), the probability of contamination of the primary cells is greatly increased, and the efficiency is also low.
In view of the above, there is a need in the art for an extraction apparatus that can save manpower, reduce errors between samples due to different extraction methods by different technicians, and reduce sample contamination.
Disclosure of Invention
In order to overcome the disadvantages of the prior art, the present invention provides a primary cell extraction apparatus and a method for using the same, so as to achieve the effects of saving manpower, achieving the same extraction efficiency, and reducing pollution.
To achieve the above objects, the present invention provides a primary cell extraction apparatus, which comprises a sample cutting area, an operation area, an extraction area, and a slide rail device. The sample mincing zone comprises a tissue mincing device. The operation area comprises a first gate, a heating temperature control device, a rotary fixture, a centrifugal device, a cover opening and closing device, a test tube placing frame and a liquid transferring device; the first gate is adjacent to the sample chopping area, so that the operation area is adjacent to the sample chopping area; the heating temperature control device is adjacent to the first gate; the rotary fixture is adjacent to one side of the heating temperature control device opposite to the first gate; the centrifugal device is adjacent to one side of the rotary fixture relative to the heating temperature control device; the switch cover device is adjacent to the first gate and opposite to the heating temperature control device; the test tube placing frame is adjacent to one side of the switch cover device relative to the first gate; the pipetting device is adjacent to one side of the test tube placement frame opposite to the switch cover device. The extraction area comprises a second gate which is adjacent to the operation area, so that the extraction area is adjacent to the operation area. The slide rail device penetrates through the sample chopping area, the operation area and the extraction area and penetrates below the first gate of the operation area and the second gate of the extraction area; the slide rail device is provided with a trolley which can slide on the slide rail device.
Preferably, the sample cutting area is a sterile laminar flow cabinet (sterile laminar flow cabinet) and has a UV lamp tube with sterilization function, so as to achieve a sterile operation space.
Preferably, the rotary clamp is a robot arm.
Preferably, the pipetting device comprises a frame, at least one pipette, a pipette tip frame, a rotating disc and a recovery area. The frame body is square; the at least one liquid transfer device is arranged above the frame body; the pipette tip rack is movably arranged on one side of the rack body; the rotary disc can be rotatably arranged in the frame body, and a plurality of tank bodies are arranged on the rotary disc; the recovery area is adjacent to the frame body and is arranged below the at least one liquid transfer device.
More preferably, the at least one liquid transfer device is two liquid transfer devices, and the two liquid transfer devices are adjacent to each other.
Preferably, the heating temperature control device, the rotary fixture and the centrifugal device in the operation area are arranged on one side of the slide rail device, and the switch cover device, the test tube placing frame and the liquid transfer device are arranged on the other side of the slide rail device.
The present invention further provides a method for using the primary cell extraction apparatus, comprising the following steps: (1) a tissue mincing device for placing a tissue specimen in the sample mincing zone; (2) the rotary fixture in the operation area clamps the test tube positioned in the test tube placing rack and moves the test tube to the trolley of the slide rail device; the trolley carrying the test tube moves to the cover opening and closing device to open the cover of the test tube; after the first gate is opened, the trolley carrying the test tube with the cover removed moves from the operation area to the sample chopping area, and the first gate is closed; (3) the tissue mincing device is used for mincing the tissue specimen and then flushing the minced tissue specimen into the test tube by buffer solution, opening the first gate, moving the trolley carrying the test tube of the tissue specimen to the opening and closing cover device of the operation area, and closing the first gate; (4) the cover opening and closing device covers the cover of the test tube on the test tube with the tissue sample, and the rotary clamp moves the test tube with the tissue sample to the centrifugal device for centrifugation, so that a first supernatant and a first precipitate are formed in the test tube; the test tube after centrifugation is moved back to the trolley by the rotary fixture; (5) the trolley carrying the centrifuged test tube is moved to the liquid transferring device from the cover opening and closing device, first supernatant in the test tube is sucked to leave a first precipitate, and then enzyme liquid is added into the test tube and mixed with the first precipitate; the trolley carrying the test tube with the enzyme solution moves to the switch cover device, and the switch cover device covers the cover; (6) the rotary fixture clamps the test tube loaded with the enzyme solution to shake and move to the heating temperature control device for enzyme reaction; (7) the rotary fixture clamps the test tube after the enzyme reaction, and moves to the centrifugal device for centrifugation, so that a second supernatant and a second precipitate are formed in the test tube; the rotary fixture moves the centrifuged test tube to the trolley and the cover is opened by the cover opening and closing device; (8) the trolley carrying the centrifuged test tube moves from the switch cover device to the liquid-transferring device, sucks the second supernatant in the test tube to leave a second precipitate, and then adds a cleaning solution into the test tube and mixes the cleaning solution with the second precipitate; the trolley carrying the test tubes of the cleaning solution moves to the switch cover device, and the switch cover device covers the cover; (9) the rotary clamp clamps the test tube loaded with the cleaning solution to shake, and moves the test tube to the centrifugal device for centrifugation, so that a third supernatant and a third precipitate are formed in the test tube; the rotary fixture moves the centrifuged test tube to the trolley and the cover is opened by the cover opening and closing device; (10) moving the trolley carrying the centrifuged test tube from the cover opening and closing device to the liquid transferring device, sucking a third supernatant in the test tube to leave a third precipitate, adding a culture solution into the test tube and mixing the culture solution with the third precipitate, wherein the test tube containing the culture solution has primary cells; the trolley carrying the test tubes of the culture solution moves to the switch cover device, and the switch cover device covers the cover; (11) the rotary clamp clamps the test tube loaded with the culture solution to shake and move to the trolley; opening the second gate, and moving the trolley carrying the test tubes with the culture solution from the operation area to the extraction area; and (12) closing the second gate, and the tube containing the culture solution is subjected to cell culture.
Preferably, the steps (5), (8) and (10) further include moving a pipette tip rack of the pipetting device to a position below each pipette, combining a plurality of pipette tips in the pipette tip rack with each pipette, moving the pipette tip rack away from each pipette, and then performing pipetting operation with the two pipettes.
Preferably, in the steps (5), (8) and (10), one of the pipettes sucks the supernatant in the test tube to leave a precipitate; the trolley carrying the test tube moves to a position far away from the liquid transfer device, the turntable carrying the waste liquid collecting tank body rotates and moves to the position below a liquid transfer device for absorbing the upper liquid from the frame body, and the upper liquid is discharged to the waste liquid collecting tank body; another liquid transfer device sucks the liquid in one pot body on the turntable, the trolley carrying the test tube moves back to the position below the liquid transfer device sucking the liquid and discharges the liquid to the test tube, and the liquid is mixed with the sediment in the test tube; the trolley carrying the test tube with the liquid moves to the switch cover device, and the switch cover device covers the cover; the pipette tip rack is moved to the lower part of each liquid transfer device again to assist in discharging the pipette tips combined with each liquid transfer device to the recovery area; wherein in the step (5), one of the tank bodies on the turntable is an enzyme tank body, and the liquid is an enzyme liquid; wherein in the step (8), one of the tank bodies on the turntable is a cleaning liquid tank body, and the liquid is cleaning liquid; wherein in the step (10), one of the tanks on the turntable is a culture solution tank, and the liquid is a culture solution.
The invention has the advantages that the primary cell can be automatically extracted by the sample chopping area, the operation area, the extraction area and the slide rail device of the primary cell extraction equipment, and various different processes are executed, so that the effects of saving manpower, having the same extraction efficiency and reducing pollution are achieved.
The invention is described in detail below with reference to the drawings and specific examples, but the invention is not limited thereto.
Drawings
FIG. 1 is a perspective view of the primary cell extraction apparatus of the present invention.
FIG. 2 is a top view of the primary cell extraction apparatus of the present invention.
FIG. 3 is a perspective view of the sample chopping area of the primary cell extraction device of the present invention.
FIG. 4 is a perspective view of a partial operation area of the primary cell extraction apparatus of the present invention.
FIG. 5 is a perspective view of another partial operation area of the primary cell extraction apparatus of the present invention.
FIG. 6 is a perspective view of the extraction section of the primary cell extraction apparatus of the present invention.
FIG. 7 is a flow chart of steps (1) to (6) of the method of the present invention using a primary cell extraction apparatus; s1 to S6 represent steps (1) to (6).
FIG. 8 is a flow chart of steps (7) to (12) of the method of the present invention using a primary cell extraction apparatus; s7 to S12 represent steps (7) to (12).
Wherein, the reference numbers:
1 Primary cell extraction equipment
10 sample cutting zone
11 tissue morcellating device
20 working area
21 first gate
22 heating temperature control device
23 rotating type clamp
24 centrifugal device
25 opening and closing cover device
26 test tube placing rack
27 liquid transfer device
271 frame body
272 liquid transfer device
273 pipette tip rack
274 rotating disk
275 recovery zone
30 extraction zone
31 second gate
40 sliding rail device
41 trolley
Detailed Description
The technical means adopted by the invention to achieve the purpose are further described below by combining the drawings and the preferred embodiments of the invention.
Referring to fig. 1 and 2, a primary cell extraction apparatus 1 according to the present invention is a sterile laminar flow cabinet, and the primary cell extraction apparatus 1 includes a sample crushing region 10, an operation region 20, an extraction region 30, and a slide rail device 40. Wherein the sample chopping zone 10, the working zone 20 and the extraction zone 30 are adjacent in sequence; the slide rail device 40 is disposed through the sample shredding zone 10, the working zone 20 and the extraction zone 30.
Referring to fig. 2 and 3, the sample-mincing region 10 includes a tissue-mincing device 11, and the tissue-mincing device 11 is fixedly arranged in the sample-mincing region 10. The sample chopping block 10 is a sterile laminar flow cabinet with UV lamps for sterilization to achieve sterile operating space.
Referring to fig. 2, 4 and 5, the operation area 20 includes a first gate 21, a heating temperature control device 22, a rotary clamp 23, a centrifugal device 24, a cover opening and closing device 25, a test tube holder 26 and a pipetting device 27. The first gate 21 is adjacent to the sample comminution zone 10. The heating temperature control device 22 is adjacent to the first gate 21. The rotary fixture 23 is adjacent to one side of the heating temperature control device 22 opposite to the first gate 21; in a preferred embodiment, the rotary gripper 23 is a robotic arm. The centrifugal device 24 is adjacent to the side of the rotary fixture 23 opposite to the heating and temperature control device 22. The switch cover device 25 is adjacent to the first gate 21 and opposite to the heating temperature control device 22. The test tube rack 26 is adjacent to a side of the opening and closing cover device 25 opposite to the first shutter 21. The pipetting device 27 is adjacent to the side of the test tube rack 26 opposite to the switch cover device 25; the pipetting device 27 comprises a frame 271, at least one pipette 272, a pipette tip frame 273, a turntable 274, and a recovery zone 275. The frame body 271 is square. The at least one pipette 272 is disposed above the frame body 271; in a preferred embodiment, the at least one pipette 272 is a micropipette (pipette); in another preferred embodiment, the at least one pipette 272 is two pipettes 272, and the pipettes are adjacent to each other. The pipette tip holder 273 is movably disposed at one side of the holder body 271. The turntable 274 is rotatably disposed in the frame body 271, and the turntable 274 has a plurality of tanks, wherein the tanks can be used as a waste liquid collecting tank, a ferment tank, a cleaning liquid tank, or a culture liquid tank. The recovery area 275 is adjacent to the frame 271 and is disposed below the at least one pipette 272. In a further preferred embodiment, the heating temperature control device 22, the rotary fixture 23 and the centrifugal device 24 of the working area 20 are disposed on one side of the slide rail device 40, and the lid opening and closing device 25, the test tube rack 26 and the pipetting device 27 are disposed on the other side of the slide rail device 40.
Referring to fig. 6, the extraction area 30 includes a second gate 31, and the second gate 31 is adjacent to the operation area 20.
Referring to fig. 2 and 6, the slide rail device 40 is disposed below the first gate 21 of the working area 20 and the second gate 31 of the extraction area 30; the slide rail device 40 has a trolley 41 sliding on the slide rail device 40.
Referring to fig. 7 and 8, the present invention further provides a method using the primary cell extraction apparatus 1, comprising the following steps:
(1) a tissue mincing means 11 for placing a tissue specimen in the sample mincing zone 10;
(2) the rotary gripper 23 of the working area 20 grips the test tube in the test tube rack 26 and moves the test tube to the trolley 41 of the slide rail device 40; the trolley 41 carrying the test tube moves to the cover opening and closing device 25 to open the cover of the test tube; after opening the first shutter 21, the trolley 41 carrying the test tube with the removed lid is moved from the working area 20 to the sample cutting area 10, and the first shutter 21 is closed;
(3) the tissue mincing device 11 is used for mincing the tissue specimen, then flushing the minced tissue specimen into the test tube by buffer solution, opening the first gate 21, moving the trolley 41 carrying the test tube of the tissue specimen to the cover opening and closing device 25 of the operation area 20, and closing the first gate 21;
(4) the cover opening and closing device 25 covers the cover of the test tube and closes the test tube with the tissue sample, and the rotary clamp 23 moves the test tube with the tissue sample to the centrifugal device 24 for centrifugation; centrifuging at 200x g deg.C for 4 min to form a first supernatant and a first precipitate; the rotary clamp 23 moves the centrifuged test tube back to the cover opening and closing device 25 to open the cover; the rotary gripper 23 then moves the centrifuged test tube back onto the trolley 41;
(5) a pipette tip rack 273 of the pipette device 27 is moved below each pipette 272, a plurality of pipette tips in the pipette tip rack 273 are combined with each pipette 272, and the pipette tip rack 273 is moved away from each pipette 272; the trolley 41 carrying the centrifuged test tube is moved from the switch cover device 25 to the pipetting device 27, and a first supernatant in the test tube is sucked by one of the pipettes 272 to leave a first precipitate; the trolley 41 carrying the test tube moves away from the pipetting device 27, the turntable 274 carrying the waste liquid collection tank rotates and moves from the frame 271 to the position below the pipetter 272 which sucks the first supernatant liquid and discharges the first supernatant liquid to the waste liquid collection tank; another pipette 272 sucks the ferment liquid in the ferment pot on the rotating disc 274, the trolley 41 carrying the test tube moves back to the position below the pipette 272 sucking the ferment liquid and discharges the ferment liquid to the test tube, and the ferment liquid is mixed with the first precipitate in the test tube; the trolley 41 carrying the test tube with the enzyme solution moves to the cover opening and closing device 25, and the cover is closed by the cover opening and closing device 25; the pipette tip rack 273 is moved again below each pipette 272 to assist in discharging the pipette tip 273, which is combined with each pipette 272, to the recovery area 275, the pipette tips in the pipette tip rack 273 are combined with each pipette 272, and the pipette tip rack 273 is moved away from each pipette 272;
(6) the rotary fixture 23 clamps the test tube loaded with the enzyme solution to shake, and move to the heating temperature control device 22 for enzyme reaction; the enzyme reaction condition is that the enzyme is shaken every 10 minutes at 37 ℃ for 6 times in total;
(7) the rotary clamp 23 clamps the test tube after the enzyme reaction and moves to the centrifugal device 24 for centrifugation; centrifuging at 200x g deg.C for 4 min to form a second supernatant and a second precipitate; the rotary clamp 23 moves the centrifuged test tube to the trolley 41, and the cover is opened by the cover opening and closing device 25;
(8) the difference between this step and step (5) is that the enzyme tank on the turntable 274 is replaced by a cleaning liquid tank. The detailed steps are as follows:
the trolley 41 carrying the test tube after enzyme reaction is moved from the switch cover device 25 to the pipetting device 27, and a second upper liquid in the test tube is sucked by one of the pipettes 272 to leave a second precipitate; the trolley 41 carrying the test tube is moved away from the pipetting device 27, the turntable 274 carrying the waste liquid collection tank is rotated from the frame body 271 to a position below the pipetter 272 which sucks the second supernatant liquid and discharges the second supernatant liquid to the waste liquid collection tank; another pipettor 272 sucks the cleaning liquid in the cleaning liquid tank on the turntable 274, the trolley 41 carrying the test tube moves back to the position below the pipettor 272 sucking the cleaning liquid and discharges the cleaning liquid to the test tube, and the cleaning liquid is mixed with the second sediment in the test tube; the trolley 41 carrying the test tubes of the cleaning solution moves to the cover opening and closing device 25, and the cover opening and closing device 25 covers the test tubes; the pipette tip rack 273 is moved again below each pipette 272 to assist in discharging the pipette tip 273, which is combined with each pipette 272, to the recovery area 275, the pipette tips in the pipette tip rack 273 are combined with each pipette 272, and the pipette tip rack 273 is moved away from each pipette 272;
(9) the rotary clamp 23 clamps the test tube loaded with the cleaning solution to shake, and moves to the centrifugal device 24 for centrifugation; centrifuging at 200x g deg.C for 4 min to form a third supernatant and a third precipitate; the rotary clamp 23 moves the centrifuged test tube to the trolley 41, and the cover is opened by the cover opening and closing device 25;
(10) the process is the same as step (5), except that the enzyme tank on the turntable 274 is replaced by a culture tank. The detailed steps are as follows:
the trolley 41 carrying the centrifuged test tube is moved from the switch cover device 25 to the pipetting device 27, and a third supernatant in the test tube is sucked by one of the pipettes 272 to leave a third precipitate; the trolley 41 carrying the test tube is moved away from the pipetting device 27, the turntable 274 carrying the waste liquid collection tank is rotationally moved from the frame body 271 to the position below the pipetter 272 which sucks the third supernatant liquid and discharges the third supernatant liquid to the waste liquid collection tank; another liquid transfer device 272 sucks the culture solution in the culture solution tank on the rotating disc 274, the trolley 41 carrying the test tube moves back to the position below the liquid transfer device 272 for sucking the culture solution and discharges the culture solution to the test tube, and the culture solution is mixed with the third precipitate in the test tube, and the test tube containing the culture solution has primary cells; the trolley 41 carrying the test tubes of the culture solution moves to the cover opening and closing device 25, and the cover is closed by the cover opening and closing device 25; the pipette tip rack 273 is moved again below each pipette 272 to assist in discharging the pipette tip 273, which is combined with each pipette 272, to the recovery area 275, the pipette tips in the pipette tip rack 273 are combined with each pipette 272, and the pipette tip rack 273 is moved away from each pipette 272;
(11) the rotary gripper 23 grips the test tube loaded with the culture solution, and moves the test tube to the carriage 41; the second gate 31 is opened, and the trolley 41 carrying the test tubes of the culture solution is moved from the operation area 20 to the extraction area 30;
(12) the second shutter 31 is closed and the tube containing the culture solution is subjected to cell culture.
Various modifications and alterations of this invention will become apparent to those skilled in the art without departing from the scope and spirit of this invention. Although the invention has been described in connection with specific preferred embodiments, it is to be understood that the invention is not to be unduly limited to such specific embodiments.
The present invention is capable of other embodiments, and various changes and modifications may be made by one skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.
Claims (9)
1. A primary cell extraction apparatus, comprising:
a sample chopping zone comprising:
a tissue mincing device disposed in the sample mincing region;
an operation area, comprising:
a first gate adjacent to the sample shredding zone, such that the working zone is adjacent to the sample shredding zone;
a heating temperature control device adjacent to the first gate;
the rotary fixture is adjacent to one side of the heating temperature control device, which is opposite to the first gate;
the centrifugal device is adjacent to one side of the rotary fixture, which is opposite to the heating temperature control device;
a switch cover device adjacent to the first gate and opposite to the heating temperature control device;
the test tube placing frame is adjacent to one side, opposite to the first gate, of the switch cover device; and
a pipetting device adjacent to one side of the test tube placing rack opposite to the switch cover device;
an extraction zone, comprising:
a second gate adjacent to the operation area to make the extraction area adjacent to the operation area; and
the sliding rail device penetrates through the sample chopping area, the operation area and the extraction area and penetrates below the first gate of the operation area and the second gate of the extraction area; the slide rail device is provided with a trolley which can slide on the slide rail device.
2. The primary cell extraction apparatus of claim 1, wherein the rotary gripper is a robotic arm.
3. The primary cell extraction apparatus of claim 1, wherein the pipetting device comprises:
a frame body which is square;
at least one liquid transfer device arranged above the frame body;
a pipette tip holder movably provided at one side of the holder body;
the rotary disc is rotatably arranged in the frame body and is provided with a plurality of tank bodies; and the number of the first and second groups,
and the recovery area is adjacent to the frame body and is arranged below the at least one liquid transfer device.
4. The primary cell extraction apparatus of claim 3, wherein the at least one pipette is two pipettors and is adjacent to each other.
5. The primary cell extraction apparatus according to claim 1 or 2, wherein the heating temperature control device, the rotary clamp and the centrifugation device of the operation area are disposed on one side of the slide rail device, and the lid opening and closing device, the test tube rack and the liquid transfer device are disposed on the other side of the slide rail device.
6. The primary cell extraction apparatus according to claim 3 or 4, wherein the heating temperature control device, the rotary fixture and the centrifugation device of the operation area are disposed on one side of the slide rail device, and the lid opening and closing device, the test tube rack and the liquid transfer device are disposed on the other side of the slide rail device.
7. A method of using the primary cell extraction apparatus of claim 1, 2 or 5, comprising the steps of:
(1) a tissue mincing device for placing a tissue specimen in the sample mincing zone;
(2) the rotary fixture in the operation area clamps the test tube positioned in the test tube placing rack and moves the test tube to the trolley of the slide rail device; the trolley carrying the test tube moves to the cover opening and closing device to open the cover of the test tube; after the first gate is opened, the trolley carrying the test tube with the cover removed moves from the operation area to the sample chopping area, and the first gate is closed;
(3) the tissue mincing device is used for mincing the tissue specimen and then flushing the minced tissue specimen into the test tube by buffer solution, opening the first gate, moving the trolley carrying the test tube of the tissue specimen to the cover opening and closing device of the operation area, and closing the first gate;
(4) the cover opening and closing device covers the cover of the test tube on the test tube with the tissue sample, and the rotary clamp moves the test tube with the tissue sample to the centrifugal device for centrifugation, so that a first supernatant and a first precipitate are formed in the test tube; the test tube after centrifugation is moved back to the trolley by the rotary fixture;
(5) moving the trolley carrying the centrifuged test tube from the cover opening and closing device to the liquid transferring device, sucking the first supernatant in the test tube to leave the first precipitate, adding an enzyme solution into the test tube, and mixing the enzyme solution with the first precipitate; the trolley carrying the test tube with the enzyme solution moves to the switch cover device, and the switch cover device covers the cover;
(6) the rotary fixture clamps the test tube loaded with the enzyme solution to shake and move to the heating temperature control device for enzyme reaction;
(7) the rotary fixture clamps the test tube after the enzyme reaction, and moves to the centrifugal device for centrifugation, so that a second supernatant and a second precipitate are formed in the test tube; the rotary fixture moves the centrifuged test tube to the trolley and the cover is opened by the cover opening and closing device;
(8) the trolley carrying the centrifuged test tube moves from the switch cover device to the liquid-transferring device, sucks the second supernatant in the test tube to leave the second precipitate, and then adds a cleaning solution into the test tube and mixes the cleaning solution with the second precipitate; the trolley carrying the test tubes of the cleaning solution moves to the switch cover device, and the switch cover device covers the cover;
(9) the rotary clamp clamps the test tube loaded with the cleaning solution to shake, and moves the test tube to the centrifugal device for centrifugation, so that a third supernatant and a third precipitate are formed in the test tube; the rotary fixture moves the centrifuged test tube to the trolley and the cover is opened by the cover opening and closing device;
(10) moving the trolley carrying the centrifuged test tube from the cover opening and closing device to the pipetting device, sucking the third supernatant in the test tube to leave the third precipitate, adding the culture solution into the test tube and mixing the culture solution with the third precipitate, wherein the test tube containing the culture solution has primary cells; the trolley carrying the test tubes of the culture solution moves to the switch cover device, and the switch cover device covers the cover;
(11) the rotary clamp clamps the test tube loaded with the culture solution to shake and move to the trolley; opening the second gate, and moving the trolley carrying the test tubes with the culture solution from the operation area to the extraction area; and the number of the first and second groups,
(12) the second gate is closed and the tube containing the culture medium will be subjected to cell culture.
8. A method of using the primary cell extraction apparatus of claim 3, 4 or 6, comprising the steps of:
(1) a tissue mincing device for placing a tissue specimen in the sample mincing zone;
(2) the rotary fixture in the operation area clamps the test tube positioned in the test tube placing rack and moves the test tube to the trolley of the slide rail device; the trolley carrying the test tube moves to the cover opening and closing device to open the cover of the test tube; after the first gate is opened, the trolley carrying the test tube with the cover removed moves from the operation area to the sample chopping area, and the first gate is closed;
(3) the tissue mincing device is used for mincing the tissue specimen and then flushing the minced tissue specimen into the test tube by buffer solution, opening the first gate, moving the trolley carrying the test tube of the tissue specimen to the cover opening and closing device of the operation area, and closing the first gate;
(4) the cover opening and closing device covers the cover of the test tube on the test tube with the tissue sample, and the rotary clamp moves the test tube with the tissue sample to the centrifugal device for centrifugation, so that a first supernatant and a first precipitate are formed in the test tube; the test tube after centrifugation is moved back to the trolley by the rotary fixture;
(5) moving the trolley carrying the centrifuged test tube from the cover opening and closing device to the liquid transferring device, sucking the first supernatant in the test tube to leave the first precipitate, adding an enzyme solution into the test tube, and mixing the enzyme solution with the first precipitate; the trolley carrying the test tube with the enzyme solution moves to the switch cover device, and the switch cover device covers the cover;
(6) the rotary fixture clamps the test tube loaded with the enzyme solution to shake and move to the heating temperature control device for enzyme reaction;
(7) the rotary fixture clamps the test tube after the enzyme reaction, and moves to the centrifugal device for centrifugation, so that a second supernatant and a second precipitate are formed in the test tube; the rotary fixture moves the centrifuged test tube to the trolley and the cover is opened by the cover opening and closing device;
(8) the trolley carrying the centrifuged test tube moves from the switch cover device to the liquid-transferring device, sucks the second supernatant in the test tube to leave the second precipitate, and then adds a cleaning solution into the test tube and mixes the cleaning solution with the second precipitate; the trolley carrying the test tubes of the cleaning solution moves to the switch cover device, and the switch cover device covers the cover;
(9) the rotary clamp clamps the test tube loaded with the cleaning solution to shake, and moves the test tube to the centrifugal device for centrifugation, so that a third supernatant and a third precipitate are formed in the test tube; the rotary fixture moves the centrifuged test tube to the trolley and the cover is opened by the cover opening and closing device;
(10) moving the trolley carrying the centrifuged test tube from the cover opening and closing device to the pipetting device, sucking the third supernatant in the test tube to leave the third precipitate, adding the culture solution into the test tube and mixing the culture solution with the third precipitate, wherein the test tube containing the culture solution has primary cells; the trolley carrying the test tubes of the culture solution moves to the switch cover device, and the switch cover device covers the cover;
(11) the rotary clamp clamps the test tube loaded with the culture solution to shake and move to the trolley; opening the second gate, and moving the trolley carrying the test tubes with the culture solution from the operation area to the extraction area; and the number of the first and second groups,
(12) closing the second gate, and culturing the cells in the test tube containing the culture solution;
wherein the steps (5), (8) and (10) further comprise moving a pipette tip rack of the pipette device to the lower part of each pipette, combining a plurality of pipette tips in the pipette tip rack with each pipette, moving the pipette tip rack away from each pipette, and then performing pipetting action by using two pipettes.
9. The method of claim 8, wherein steps (5), (8) and (10) comprise sucking supernatant from the test tube by one of the pipettes to leave a precipitate; the trolley carrying the test tube moves to a position far away from the liquid transfer device, the turntable carrying the waste liquid collecting tank body rotates and moves to the position below the liquid transfer device for absorbing the upper liquid from the frame body, and the upper liquid is discharged to the waste liquid collecting tank body; another liquid transfer device sucks the liquid in one pot body on the turntable, the trolley carrying the test tube moves back to the position below the liquid transfer device sucking the liquid and discharges the liquid to the test tube, and the liquid is mixed with the sediment in the test tube; the trolley carrying the test tube with the liquid moves to the switch cover device, and the switch cover device covers the cover; the pipette tip rack is moved to the lower part of each liquid transfer device again to assist in discharging the pipette tips combined with each liquid transfer device to the recovery area;
wherein in the step (5), one of the tank bodies on the turntable is an enzyme tank body, and the liquid is an enzyme liquid;
wherein in the step (8), one of the tank bodies on the turntable is a cleaning liquid tank body, and the liquid is cleaning liquid;
wherein in the step (10), one of the tanks on the turntable is a culture solution tank, and the liquid is a culture solution.
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| EP1506413B1 (en) * | 2002-05-17 | 2016-07-06 | Becton Dickinson and Company | Automated system for isolating, amplyifying and detecting a target nucleic acid sequence |
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| JP2009291104A (en) * | 2008-06-04 | 2009-12-17 | Kawasaki Heavy Ind Ltd | Automatic cell culture apparatus |
| CN101538612A (en) * | 2009-04-16 | 2009-09-23 | 上海科华生物工程股份有限公司 | Integrated blood nucleic acid screening platform |
| CN104661737A (en) * | 2012-07-18 | 2015-05-27 | 莱伯曼兹有限公司 | Automated solution dispenser |
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Effective date of registration: 20221118 Address after: 6th Floor, No. 250, Section 1, Neihu Road, Neihu District, Taipei, Taiwan, China, China Patentee after: DRSIGNAL BIOTECHNOLOGY Co.,Ltd. Address before: 1225 mailbox, Samoa Patentee before: SCL BIOTECH Ltd. |