CN112083170A - Cell protein sample extraction element - Google Patents
Cell protein sample extraction element Download PDFInfo
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- CN112083170A CN112083170A CN202010781670.7A CN202010781670A CN112083170A CN 112083170 A CN112083170 A CN 112083170A CN 202010781670 A CN202010781670 A CN 202010781670A CN 112083170 A CN112083170 A CN 112083170A
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
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Abstract
The invention relates to a cell protein sample extraction device which comprises a three-axis motion platform, an electric pipetting gun, a gripper device, a refrigerator, a centrifuge, a PBS solution box, a supernatant centrifuge tube placing box, a lysate placing box, a PMSF solution placing box, a culture dish operation table, a cell scraping plate device, a waste liquid placing box, a gun head placing box, a waste placing box, a centrifuge tube pressing device, a centrifuge tube operation table, a control system and a centrifuge tube placing box. The invention can automatically extract the cell protein sample, has high repeatability precision, high efficiency, low requirement on operators and low labor intensity, and can effectively avoid the harm of toxic substances to the operators.
Description
Technical Field
The invention belongs to the technical field of preparation of protein samples, and relates to an auxiliary device for preparation of a cell protein sample in a biochemical experiment Western blotting (Western Blot) process, in particular to a cell protein sample extraction device.
Background
The Western Blot detection technology is more and more widely used in the fields of basic research such as protein molecule signal channels, proteomics and the like and research application of disease markers, and aims to qualitatively detect whether a certain protein is expressed or not or semi-quantitatively detect the expression level of the certain protein. It uses the marked antibody to detect the corresponding antigen in the tissue cell, and achieves the purpose of diagnosing the cause of infectious diseases, classifying and identifying tumors, and providing important reference basis for the prognosis and gene therapy of tumors. The scientific and normative operation has extremely important significance for ensuring the immunohistochemical flaking quality, improving the clinical pathological diagnosis level and promoting the development of pathological research work.
The first step of Western Blot is protein sample preparation, protein extraction is crucial in the Western Blot process, and protein extraction can be roughly divided into three steps: the first step is to obtain material, the second step is to separate and break the cell or tissue components of interest, and the third step is to perform lysis sample preparation.
The specific operation steps for preparing the cell protein sample comprise:
1. when the cell state is observed to be suitable for extracting protein, removing the culture solution;
2. adding 3ml of 4 ℃ precooled PBS (0.01M pH7.2-7.3) into each bottle of cells, wherein the Chinese name of the PBS is phosphate buffer salt solution, flatly placing and slightly shaking the cells for 1 min to wash the cells, then removing washing liquid, repeating the above operations twice, washing the cells for three times to wash out the culture solution, removing the PBS completely, and then placing the culture bottle on ice;
3. add 10. mu.l PMSF (100 mM) to 1ml lysate, the Chinese name of PMSF is: uniformly shaking phenylmethylsulfonyl fluoride, placing on ice, and mixing PMSF with a lysis solution when the PMSF is not crystallized;
4. adding 400 μ l of lysis solution containing PMSF into each bottle of cells, and lysing for 30min on ice, wherein the culture bottle is frequently shaken back and forth to fully lyse the cells;
5. after the lysis, scraping the cells on one side of the culture bottle by using a clean scraper, then moving cell fragments and lysate into a 1.5ml centrifuge tube by using a gun, and performing the whole operation on ice as much as possible;
6. centrifuging at 12000rpm at 4 deg.C for 5 min;
7. the centrifuged supernatant is separately packed and transferred into a centrifuge tube for 0.5 min and stored at the temperature of minus 20 ℃, and ten million of white flocculent precipitates cannot be absorbed to the lower part of the centrifuge tube of 1.5 ml.
At present, cell sample preparation is operated manually, the cell sample preparation process is complex, the operation time is long, the requirement on operators is high, the whole process relates to a toxic substance PMSF, the health of the operators is affected, and the manual operation efficiency is low, unstable and easy to make mistakes.
Disclosure of Invention
Aiming at the problems, the invention provides a cell protein sample extraction device which can automatically extract cell protein samples, has high repeatability precision, high efficiency, stability, low requirement on operators and low labor intensity, is not easy to make mistakes, and can effectively avoid the harm of toxic substances to the operators.
The technical scheme adopted by the invention for solving the technical problems is as follows: the utility model provides a cell protein sample extraction element, includes that three-axis motion platform, electronic pipetting gun, tongs, refrigerator, centrifuge, PBS solution box, supernatant centrifuge tube place the box, the box is placed to the lysate, PMSF solution places box, culture dish operation panel, cell scraper means, waste liquid place the box, the box is placed to the rifle head, the box is placed to the waste material, centrifuging tube compression fittings, centrifuging tube operation panel, control system, centrifuging tube place the box.
Preferably, the three-axis motion platform comprises a base and a motion platform, wherein the motion platform can move X, Y, Z in three directions relative to the base, and the structure of the motion platform is the prior art and is not described in detail herein.
Preferably, the electric pipetting gun is mounted on the three-axis motion platform, can move in X, Y, Z three directions along with the three-axis motion platform and can move relative to the three-axis motion platform along the Z direction, the electric pipetting gun is provided with a pipetting device and a gun head withdrawing device, the pipetting device is used for sucking and discharging solution, and the gun head withdrawing device is used for automatically withdrawing the gun head.
Preferably, the pipetting device is preferably a multi-pump head multi-channel electronic peristaltic pump, the multi-pump head multi-channel electronic peristaltic pump is suitable for sucking and discharging solutions with different pipetting quantities, and the pipetting speed is increased under the condition of ensuring the precision.
Preferably, the gun head withdrawing device comprises a pressing plate and a mobile power device, the mobile power device is installed on a gun body of the electric liquid-transfering gun, the pressing plate is installed at one end of the mobile power device, the pressing plate is driven by the mobile power device to withdraw the gun head from the electric liquid-transfering gun, and the mobile power device preferably selects an air cylinder.
Preferably, the gripping device comprises a centrifuge tube gripping hand and a culture dish gripping hand, the centrifuge tube gripping hand is arranged on the three-axis motion platform and used for gripping a centrifuge tube, and the culture dish gripping hand is used for gripping a culture dish.
Preferably, the centrifugal tube hand grip adopts a finger cylinder.
Preferably, the culture dish hand grip adopts a finger cylinder.
Preferably, the refrigerator is mounted on the base and is provided with three thermostats, namely a first temperature thermostat, a second temperature thermostat and a third temperature thermostat, and sealing covers which can be opened by the control system are arranged on the upper parts of the three thermostats respectively.
Preferably, the temperatures of the first temperature oven, the second temperature oven and the third temperature oven are set to 4 ℃, -5 ℃ and-20 ℃, respectively.
Preferably, the centrifuge is installed in the first temperature incubator for centrifugal stratification of the cellular fluid in the centrifuge tube, and the structure thereof is configured in the prior art and will not be described in detail herein.
Preferably, the PBS solution box is arranged in the first temperature incubator and is used for placing the PBS solution.
Preferably, the supernatant centrifuge tube placing box is arranged in a third temperature incubator and is used for storing centrifuge tubes separately filled with supernatant.
Preferably, the lysate containing box is installed in a third temperature incubator and used for storing the lysate.
Preferably, the PMSF solution storage box is installed in the third temperature incubator and used for storing PMSF solution.
Preferably, the culture dish operating platform is arranged in the second temperature constant temperature box and comprises a culture dish placing platform, a placing platform angle adjusting device and a rotating device, the angle adjusting device of the placing table comprises an upper plate, a base and a telescopic device, wherein two ends of the upper plate are provided with hinged supports, one end of the base is hinged with the hinged support at one end of the upper plate, the other end of the base is hinged with one end of the telescopic device, the other end of the telescopic device is hinged with the hinged support at the other end of the upper plate, the rotating device is arranged on the upper plate, the culture dish placing table is connected with one end of the rotating device and is used for placing a culture dish, the placing table angle adjusting device is used for adjusting the inclination angle of the culture dish placing table and the base, the rotating device is used for driving the culture dish placing platform to rotate, so that the culture dish rotates.
Preferably, the telescopic device adopts a cylinder.
Preferably, the rotating device adopts a stepping motor.
Preferably, the cell scraper device is installed in the second temperature incubator, and includes a rotating guide, a slider device, a scraper, and a scraper rotating device, wherein the rotating guide is rotatable, the slider device is movable up and down along the rotating guide, the scraper rotating device is installed on the slider device, the scraper is installed on the scraper rotating device, and the scraper is rotatable along a rotating shaft by driving of the scraper rotating device.
Preferably, the rotation axis coincides with the culture dish axis, and the length (i.e., the rotation radius) of the scraper is slightly smaller than the radius of the culture dish.
Preferably, the waste liquid housing box is used for housing various waste liquids such as a culture solution sucked from a culture dish and a used PBS solution.
Preferably, the lance tip placement box is used for placing a plurality of lance tips.
Preferably, the waste holding box is used for holding used gun tips and centrifuge tubes.
As preferred, centrifuging tube compression fittings places the chamber and fastens the device including pressfitting board, centrifuging tube, the centrifuging tube is placed the chamber and is equipped with a plurality of appearance chambeies and is used for placing the centrifuging tube, it installs to fasten the device the centrifuging tube places one side in chamber, the pressfitting board is fixed fasten on the device, the pressfitting board is in make the centrifuging tube detain rotatory to the centrifugation lumen under the drive of fastening the device soon, realize the pressfitting of centrifuging tube.
As preferred, the centrifuging tube operation panel includes that the centrifuging tube deposits chamber, centrifuging tube clamping device, laser emitter and laser receiver, the centrifuging tube is deposited the chamber top and is equipped with and holds the chamber and be used for placing the centrifuging tube, and relative both sides are respectively opened there is the bar hole, centrifuging tube clamping device one end is fixed the centrifuging tube is deposited the chamber bottom, and the other end is equipped with the tongs for press from both sides tight centrifuging tube, laser emitter with laser receiver fixes respectively the centrifuging tube is deposited the both sides that open that the chamber has the bar hole, laser emitter is used for transmitting laser signal, and laser signal arrives through the bar hole laser receiver, laser receiver is used for receiving laser emitter transmits the laser signal of coming.
Preferably, the centrifugal tube clamping device adopts a finger air cylinder.
Preferably, the control system is used for receiving signals of the sensors and sending control signals to control the on and off of power elements such as a motor and an air cylinder.
Preferably, the centrifuge tube placing box is used for placing a plurality of centrifuge tubes of various types.
Compared with the prior art, the invention has the following beneficial effects:
1. the cell protein sample can be automatically extracted, the repeatability precision is high, the efficiency is high, and the stability is not easy to make mistakes;
2. the extraction is automatic, operators do not need to contact toxic substances, and the harm of the toxic substances to the operators can be effectively avoided;
3. the control system is used for accurately controlling the operation, so that the influence of manual operation on the result accuracy is reduced;
4. the automatic control system is suitable for automation, has low requirements on operators, and reduces the labor intensity of the operators.
Drawings
FIG. 1 is a schematic structural view of the present invention;
FIG. 2 is a schematic diagram of the construction of an electric pipette 2 of the present invention;
FIG. 3 is a schematic view of the structure of the petri dish operating table 10;
FIG. 4 is a schematic view of the cell scraper 11 according to the present invention;
FIG. 5 is a schematic structural diagram of the centrifuge tube pressing device 15 of the present invention;
FIG. 6 is a schematic diagram of the centrifuge tube station 16 of the present invention;
figure 7 is a schematic cross-sectional view of the centrifuge tube station 16 of the present invention.
Detailed Description
The present invention will be described in detail with reference to fig. 1 to 7, and the exemplary embodiments and descriptions of the present invention are provided to explain the present invention, but not to limit the present invention.
The utility model provides a cell protein sample extraction element, include that triaxial motion platform 1, electronic pipetting gun 2, tongs 3, refrigerator 4, centrifuge 5, PBS solution box 6, the box 7 is placed to the centrifugal tube of supernatant, the box 8 is placed to the lysate, PMSF solution is placed box 9, culture dish operation panel 10, cell scraper means 11, box 12 is placed to the waste liquid, the box 13 is placed to the rifle head, the box 14 is placed to the waste matter, centrifuging tube compression fittings 15, centrifuging tube operation panel 16, control system 17, box 18 is placed to the centrifuging tube.
The three-axis motion platform 1 comprises a base and a motion platform, wherein the motion platform can move X, Y, Z in three directions relative to the base.
The electric liquid-transfering gun 2 is arranged on the three-axis motion platform 1, can move in X, Y, Z three directions along with the three-axis motion platform 1 and can move relative to the three-axis motion platform 1 along the Z direction, the electric liquid-transfering gun 2 is provided with a liquid-transfering device 21 and a gun head withdrawing device 22, the liquid-transfering device 21 is used for sucking and discharging solution, and the gun head withdrawing device 22 is used for automatically withdrawing the gun head.
The pipetting device 21 is preferably a multi-pump head multi-channel electronic peristaltic pump, and the multi-pump head multi-channel electronic peristaltic pump is suitable for sucking and discharging solutions with different pipetting quantities, so that the pipetting speed is increased under the condition of ensuring the precision.
When the gun head is installed at the interface of the large-displacement pump head, more solution is sucked by the gun head when the peristaltic pump rotates for one circle, when the gun head is installed at the interface of the small-displacement pump head, less solution is sucked by the gun head when the peristaltic pump rotates for one circle, when the capacity of the solution to be sucked is large, the interface of the large-displacement pump head is connected, and when the capacity of the solution to be sucked is small, the interface of the small-displacement pump head is connected, so that the liquid-moving speed is improved under the condition of ensuring the precision.
The gun head withdrawing device 22 comprises a pressure plate 22a and a moving power device 22b, the moving power device 22b is installed on the gun body of the electric pipette gun 2, the pressure plate 22a is installed at one end of the moving power device 22b, the pressure plate 22a is driven by the moving power device 22b to withdraw the gun head from the electric pipette gun 2, and the moving power device 22b is preferably an air cylinder.
The gripper device 3 comprises a centrifuge tube gripper and a culture dish gripper, and is arranged on the three-axis motion platform 1, the centrifuge tube gripper is used for gripping a centrifuge tube, the culture dish gripper is used for gripping a culture dish, a finger cylinder is preferably selected by the centrifuge tube gripper, and a finger cylinder is preferably selected by the culture dish gripper.
The refrigerator 4 is provided with three thermostats of a first temperature thermostat, a second temperature thermostat and a third temperature thermostat, the upper parts of the three thermostats are respectively provided with a sealing cover which can be opened by the control of a control system 17, and the temperatures of the three thermostats are preferably set to be 4 ℃, 5 ℃ and 20 ℃ below zero respectively.
And the centrifuge 5 is arranged in the first temperature incubator and is used for centrifugally layering the cell sap in the centrifuge tube.
The PBS solution box 6 is installed in the first temperature incubator and used for placing the PBS solution.
And the supernatant centrifuge tube placing box 7 is arranged in a third temperature thermostat and is used for storing the centrifuge tubes separately filled with the supernatant.
And the lysate containing box 8 is arranged in the third temperature incubator and is used for storing the lysate.
The PMSF solution placing box 9 is arranged in the third temperature constant temperature box and used for storing PMSF solution.
The culture dish operating platform 10 is arranged in a second temperature constant temperature box and comprises a culture dish placing platform 10a, a placing platform angle adjusting device 10b and a rotating device 10c, wherein the placing platform angle adjusting device 10b comprises an upper plate, a base and a telescopic device, two ends of the upper plate are provided with hinged supports, one end of the base is hinged with the hinged support at one end of the upper plate, the other end of the base is hinged with one end of the telescopic device, the other end of the telescopic device is hinged with the hinged support at the other end of the upper plate, the rotating device is arranged on the upper plate, the culture dish placing platform is connected with one end of the rotating device and is used for placing a culture dish, and when the telescopic device performs telescopic action, the angles of the upper plate and the base can be, thereby adjusting the inclination angle of the culture dish placing table with the base, the rotating device is used for driving the culture dish placing table to rotate, thereby rotating the culture dish, and the rotating device 10c is preferably a stepping motor.
The cell scraper 11 is installed in the second temperature incubator, and includes a rotating guide 11a, a slider 11b, a scraper 11c, and a scraper rotating device 11d, wherein the rotating guide 11a is rotatable, the slider 11b is movable up and down along the rotating guide 11a, the scraper rotating device 11d is installed on the slider 11b, the scraper 11c is installed on the scraper rotating device 11d, and the scraper 11c is rotatable along the rotating shaft by driving the scraper rotating device 11 d.
The rotation axis coincides with the culture dish axis, and the length of the scraper 11c, i.e. the rotation radius, is slightly smaller than the radius of the culture dish.
The waste liquid storage box 12 is used for storing various waste liquids such as a culture solution sucked from a culture dish and a used PBS solution.
The tip placement box 13 is used to place a plurality of tips.
The waste placement box 14 is used to place used tips, centrifuge tubes.
Centrifuging tube compression fittings 15 includes that pressfitting board 15a, centrifuging tube place chamber 15b and spiral shell device 15c, and centrifuging tube place chamber 15b is equipped with a plurality of appearance chambeies and is used for placing the centrifuging tube, and spiral shell device 15c installs in the centrifuging tube and places one side of chamber 15b, and pressfitting board 15a fixes on spiral shell device 15c, and pressfitting board 15a makes the centrifuging tube detain rotatory to the centrifugation tube chamber under spiral shell device 15 c's drive, realizes the pressfitting of centrifuging tube.
Centrifuge tube operation panel 16 includes that the centrifuging tube deposits chamber 16a, centrifuging tube clamping device 16b, laser emission device 16c and laser receiving device 16d, centrifuging tube deposits chamber 16a top and is equipped with the appearance chamber and is used for placing the centrifuging tube, relative both sides are respectively opened there is the bar hole, centrifuging tube clamping device 16b one end is fixed in the centrifuging tube and is deposited chamber 16a bottom, the other end is equipped with the tongs, be used for pressing from both sides tight centrifuging tube, laser emission device 16c and laser receiving device 16d fix respectively that the centrifuging tube deposits the both sides that open that chamber 16a has the bar hole, laser emission device 16c is used for transmitting laser signal, laser signal reaches laser receiving device 16d through the bar hole, laser receiving device 16d is used for receiving the laser signal that laser emission device 16c transmitted.
The control system 17 is used for receiving signals of the sensors and sending control signals to control the on and off of power elements such as motors and cylinders, and the specific control systems are all in the prior art.
The centrifuge tube placement box 18 is used to place a plurality of multi-type centrifuge tubes, preferably 1.5ml centrifuge tubes, 0.5 ml centrifuge tubes.
The step of installing the gun head is as follows: the control system 17 controls the three-axis motion platform 1 to drive the electric liquid-transferring gun 2 to the position of the gun head placing box 13, and then the electric liquid-transferring gun 2 moves downwards to sleeve the gun head on the electric liquid-transferring gun 2.
The step of withdrawing the gun head comprises the following steps: when the gun head needs to be withdrawn, the control system 17 controls the three-axis motion platform 1 to drive the electric pipetting gun 2 to the position of the waste placement box 14, the control system 17 controls the movable power device 22b to extend out, so that the pressing plate 22a extends out, the gun head is pushed out to withdraw from the electric pipetting gun 2, and after the gun head is withdrawn and falls into the waste placement box 14, the control system 17 controls the movable power device 22b to retract, so that the pressing plate 22a retracts.
The step of replacing the gun head is as follows: and after the gun head is withdrawn, a new gun head is installed again, and the process is to replace the gun head.
The specific implementation process comprises the following implementation steps:
1. when the cell state is observed to be suitable for protein extraction, the culture dish is placed on a culture dish operation platform 10;
2. the control system 17 controls the placing platform angle adjusting device 10b to adjust the inclination angle of the culture dish placing platform 10a and the base, and culture solution is concentrated at the lower position of the culture dish;
3. installing a gun head, controlling the three-axis motion platform 1 to move by the control system 17 to drive the electric liquid-transferring gun 2 to the position of the culture dish operation platform 10, sucking the culture liquid from the low position of the culture dish, sucking and taking the culture liquid to the waste liquid placing box 12, and withdrawing the gun head;
4. the control system 17 controls to open a sealing cover of a second temperature thermostat of the refrigerator 4, a preset amount of PBS solution in the first temperature thermostat is sucked to the culture dish, the control system 17 controls the placing table angle adjusting device 10b to adjust the inclination angle between the culture dish placing table 10a and the base, the rotating device 10c drives the culture dish placing table 10a to rotate, the culture dish rotates, the PBS solution is made to be fully contacted with cells in the culture dish, and after a preset time, washing liquor in the culture dish is sucked to the waste liquid placing box 12;
5. repeating the step 4 twice;
6. the control system 17 controls the three-axis motion platform 1 to move to drive the gripping device 3 to grip the centrifugal tube placed in the centrifugal tube placing box 18 to the centrifugal tube placing cavity 15 b;
7. replacing the gun head, controlling a sealing cover of a third temperature thermostat of the refrigerator 4 to be opened by the control system 17, sucking a preset amount of PMSF solution into a centrifugal tube placed in the centrifugal tube placing cavity 15b, driving the pressing plate 15a by the screwing device 15c to enable the centrifugal tube to be screwed to the centrifugal tube cavity, and pressing the centrifugal tube;
8. the control system 17 controls the three-axis motion platform 1 to move to drive the gripper device 3 to grip a centrifugal tube with PMSF solution and shake the centrifugal tube for a preset time;
9. the control system 17 controls the three-axis motion platform 1 to move to drive the gripper device 3 to the position of the centrifuge tube operating platform 16, the centrifuge tube with the PMSF solution placed therein is placed in the centrifuge tube storage cavity 16a, the control system 17 controls the centrifuge tube clamping device 16b to clamp the centrifuge tube, the gripper of the centrifuge tube in the gripper device 3 grasps the tube buckle of the centrifuge tube and moves upwards, and the tube buckle is separated from the tube cavity of the centrifuge tube;
10. the control system 17 controls the three-axis motion platform 1 to move to drive the gripping device 3 to grip another centrifugal tube placed in the centrifugal tube placing box 18 into the centrifugal tube placing cavity 15 b;
11. replacing the gun head, and controlling the sealing cover of a third temperature incubator of the refrigerator 4 to be opened by the control system 17, and absorbing a preset amount of lysate solution into a centrifuge tube placed in the centrifuge tube placing cavity 15 b;
12. changing a gun head, sucking and shaking uniformly, placing PMSF solution in a centrifuge tube storage cavity 16a into a centrifuge tube with lysate solution in a centrifuge tube placing cavity 15b, driving a centrifugal tube buckle of a pressing plate 15a to rotate to a centrifuge tube cavity by a screwing device 15c, and pressing the centrifuge tube;
13. the control system 17 controls the three-axis motion platform 1 to move to drive the gripping device 3 to grip the centrifuge tube placed in the centrifuge tube storage cavity 16a to the waste placing box 14;
14. the control system 17 controls the three-axis motion platform 1 to move to drive the gripper device 3 to grip a centrifuge tube with PMSF solution and lysate solution mixed liquid and shake the centrifuge tube for a preset time;
15. the control system 17 controls the three-axis motion platform 1 to move to drive the gripper device 3 to the position of the centrifuge tube operating platform 16, the centrifuge tube with the PMSF solution and lysate solution mixed solution is placed in the centrifuge tube storage cavity 16a, the control system 17 controls the centrifuge tube clamping device 16b to clamp the centrifuge tube, the gripper of the centrifuge tube in the gripper device 3 grasps the tube buckle of the centrifuge tube and moves upwards, and the tube buckle is separated from the tube cavity of the centrifuge tube;
16. sucking the uniformly shaken PMSF solution and lysate solution mixed solution into a culture dish, controlling a placing table angle adjusting device 10b by a control system 17 to adjust the inclination angle between a culture dish placing table 10a and the bottom plane, and driving the culture dish placing table 10a to rotate by a rotating device 10c to enable the culture dish to rotate, so that the PMSF solution and lysate solution mixed solution is in full contact with cells in the culture dish, and the cells are fully cracked;
17. after cell lysis is carried out for 30min, the rotating guide post 11a rotates, the sliding block device 11b moves downwards to enable the rotating center of the scraper 11c to be the center of the culture dish and the lower edge of the scraper to be in contact with the culture dish, and the scraper rotating device 11d drives the scraper 11c to rotate for a circle to enable the cells to be scraped and collected on one side;
18. the control system 17 controls the placing table angle adjusting device 10b to adjust the inclination angle of the culture dish placing table 10a and the base, and the inclination is made to make the side where the cells are collected be the lowest position;
19. the control system 17 controls the three-axis motion platform 1 to move to drive the gripping device 3 to grip the centrifuge tube placed in the centrifuge tube storage cavity 16a to the waste placing box 14;
20. the control system 17 controls the three-axis motion platform 1 to move to drive the gripping device 3 to grip 1.5ml of centrifuge tubes placed in the centrifuge tube placing box 18 to the centrifuge tube placing cavity 15 b;
21. changing the gun head, and transferring the cell fragments and the lysate to a 1.5ml centrifuge tube placed in a centrifuge tube placing box 15;
22. pressing a 1.5ml centrifuge tube which is placed in a centrifuge tube placing box 15 and contains cell fragments and lysis solution, placing the 1.5ml centrifuge tube which is placed in a centrifuge tube placing box 15 and contains the cell fragments and the lysis solution into a centrifuge by a gripping device 3 after pressing, and centrifuging for 5 min at 12000 rpm;
23. placing a centrifuged 1.5ml centrifuge tube with cell fragments and lysis solution in a centrifuge tube storage cavity 16a, controlling a centrifuge tube clamping device 16b by a control system 17 to clamp the centrifuge tube, grasping a centrifuge tube buckle by a centrifuge tube gripper in a gripper device 3 and moving upwards to separate the tube buckle from the tube cavity of the centrifuge tube;
24. the laser emitting device 16c emits laser signals, the laser receiving device 16d detects the laser signals emitted by the laser emitting device 16c, the laser signals can penetrate through the supernatant to reach the laser receiving device 16d, the white flocculent precipitate prevents the laser signals from reaching the laser receiving device 16d, therefore, the height of the supernatant can be detected through the laser signals detected on the laser receiving device 16d, and the control system 17 limits the maximum position of the gun head moving downwards according to the detected height;
25. the control system 17 controls the three-axis motion platform 1 to move to drive the gripping device 3 to grip 0.5 ml of centrifuge tube placed in the centrifuge tube placing box 18 to the centrifuge tube placing cavity 15 b;
26. replacing the gun head, sucking the centrifuged supernatant into a 0.5 ml centrifuge tube placed in the centrifuge tube placing cavity 15b, and pressing;
27. the 0.5 ml centrifuge tube containing the supernatant was transferred to a third temperature incubator of a refrigerator 4 and stored.
The technical solutions provided by the embodiments of the present invention are described in detail above, and the principles and embodiments of the present invention are explained herein by using specific examples, and the descriptions of the embodiments are only used to help understanding the principles of the embodiments of the present invention; meanwhile, for a person skilled in the art, according to the embodiments of the present invention, there may be variations in the specific implementation manners and application ranges, and in summary, the content of the present description should not be construed as a limitation to the present invention.
Claims (9)
1. A cell protein sample extraction device, characterized in that: the device comprises a three-axis motion platform (1), an electric pipette gun (2), a gripper device (3), a refrigerator (4), a centrifuge (5), a PBS solution box (6), a supernatant centrifuge tube placing box (7), a lysate placing box (8), a PMSF solution placing box (9), a culture dish operating platform (10), a cell scraping plate device (11), a waste liquid placing box (12), a gun head placing box (13), a waste placing box (14), a centrifuge tube pressing device (15), a centrifuge tube operating platform (16), a control system (17) and a centrifuge tube placing box (18);
the three-axis motion platform (1) comprises a base and a motion platform, wherein the motion platform can move X, Y, Z in three directions relative to the base;
the electric pipette gun (2) is arranged on the three-axis motion platform (1), can move in X, Y, Z three directions along with the three-axis motion platform (1) and can move relative to the three-axis motion platform (1) along the Z direction;
the gripping device (3) is arranged on the three-axis motion platform (1) and is used for gripping a centrifuge tube and a culture dish;
the refrigerator (4) is arranged on the base and is provided with a first temperature thermostat, a second temperature thermostat and a third temperature thermostat which have different temperatures, and the upper parts of the three thermostats are respectively provided with a sealing cover which can be opened by the control system (17);
the centrifuge (5) is arranged in the first temperature thermostat and is used for centrifuging and layering cell sap in a centrifuge tube;
the PBS solution box (6) is arranged in the first temperature incubator and is used for placing PBS solution;
the supernatant centrifuge tube placing box (7) is arranged in the third temperature thermostat and is used for storing centrifuge tubes which are separately filled with supernatant;
the lysate containing box (8) is arranged in the third temperature incubator and is used for storing the lysate;
the PMSF solution placing box (9) is installed in the third temperature thermostat and used for storing PMSF solution;
the culture dish operating platform (10) is arranged in the second temperature incubator;
the cell scraper device (11) is arranged in the second temperature incubator;
the waste liquid placing box (12) is arranged on the base and is used for storing various waste liquids such as culture solution sucked from a culture dish and used PBS solution;
the gun head placing box (13) is arranged on the base and used for storing gun heads;
the waste placing box (14) is arranged on the machine base and is used for placing used gun heads and centrifuge tubes;
the centrifugal tube pressing device (15) is arranged on the machine base and used for pressing the centrifugal tube;
the centrifuge tube operating platform (16) is arranged on the machine base;
the control system (17) is arranged on the base and used for receiving signals of the sensors and sending control signals to control the on-off of power elements such as a motor and an air cylinder;
the centrifuge tube placing box (18) is installed on the machine base and used for storing centrifuge tubes.
2. The cellular protein sample extraction device according to claim 1, wherein: the electric liquid transferring gun (2) is provided with a liquid transferring device (21) and a gun head withdrawing device (22).
3. The cellular protein sample extraction device according to claim 2, wherein: the liquid-transfering device (21) is a multi-pump head multi-channel electronic peristaltic pump.
4. The cellular protein sample extraction device according to claim 2, wherein: the gun head withdrawing device (22) comprises a pressing plate and a movable power device, the movable power device is installed on a gun body of the electric liquid-transferring gun (2), the pressing plate is installed at one end of the movable power device, and the pressing plate is pushed out under the driving of the movable power device so that the gun head is withdrawn from the electric liquid-transferring gun (2).
5. The cellular protein sample extraction device according to claim 1, wherein: the temperature of the first temperature thermostat is 4 ℃, the temperature of the second temperature thermostat is-5 ℃, and the temperature of the third temperature thermostat is-20 ℃.
6. The cellular protein sample extraction device according to claim 1, wherein: the culture dish operation platform (10) comprises a culture dish placing platform (10 a), a placing platform angle adjusting device (10 b) and a rotating device (10 c), the placing platform angle adjusting device (10 b) comprises an upper plate, a base and a telescopic device, hinged supports are arranged at two ends of the upper plate, one end of the base is hinged to one end of the upper plate, the other end of the base is hinged to one end of the telescopic device, the other end of the telescopic device is hinged to the hinged support at the other end of the upper plate, the rotating device (10 c) is installed on the upper plate, and the culture dish placing platform 10 a) is connected with one end of the rotating device (10 c).
7. The cellular protein sample extraction device according to claim 1, wherein: the cell scraper device (11) comprises a rotary guide post (11 a), a slider device (11 b), a scraper (11 c) and a scraper rotating device (11 d), wherein the rotary guide post (11 a) can rotate, the slider device (11 b) can move up and down along the rotary guide post (11 a), the scraper rotating device (11 d) is installed on the slider device (11 b), the scraper (11 c) is installed on the scraper rotating device (11 d), and the scraper (11 c) can rotate along a rotating shaft under the driving of the scraper rotating device (11 d).
8. The cellular protein sample extraction device according to claim 1, wherein: centrifuging tube compression fittings (15) place chamber (15 b) and spiral shell and put (15 c) including pressfitting board (15 a), centrifuging tube places chamber (15 b) and is used for placing the centrifuging tube, pressfitting board (15 a) are in make the centrifugation tube detain rotatory to the centrifugation lumen under the drive of spiral shell and put (15 c), realize the pressfitting of centrifuging tube.
9. The cellular protein sample extraction device according to claim 1, wherein: the centrifuge tube operating platform (16) comprises a centrifuge tube storage cavity (16 a), a centrifuge tube clamping device (16 b), a laser emitting device (16 c) and a laser receiving device (16 d), the top of the centrifuge tube storage cavity (16 a) is provided with a containing cavity for placing a centrifuge tube, two opposite sides are respectively provided with a strip-shaped hole, one end of the centrifugal tube clamping device (16 b) is fixed at the bottom of the centrifugal tube storage cavity (16 a), the other end is provided with a hand grip, used for clamping a centrifuge tube, the laser emitting device (16 c) and the laser receiving device (16 d) are respectively fixed at two sides of the centrifuge tube storage cavity (16 a) provided with strip-shaped holes, the laser emitting device (16 c) is used for emitting laser signals, the laser signals reach the laser receiving device (16 d) through the strip-shaped hole, the laser receiving device (16 d) is used for receiving the laser signals transmitted by the laser transmitting device (16 c).
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010781670.7A CN112083170A (en) | 2020-08-06 | 2020-08-06 | Cell protein sample extraction element |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010781670.7A CN112083170A (en) | 2020-08-06 | 2020-08-06 | Cell protein sample extraction element |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112083170A true CN112083170A (en) | 2020-12-15 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010781670.7A Withdrawn CN112083170A (en) | 2020-08-06 | 2020-08-06 | Cell protein sample extraction element |
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| Country | Link |
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| CN (1) | CN112083170A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116987579A (en) * | 2023-08-07 | 2023-11-03 | 四川省畜牧科学研究院 | Animal cell extraction device and method thereof |
-
2020
- 2020-08-06 CN CN202010781670.7A patent/CN112083170A/en not_active Withdrawn
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116987579A (en) * | 2023-08-07 | 2023-11-03 | 四川省畜牧科学研究院 | Animal cell extraction device and method thereof |
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