CN111307695B - Calibration device and method of laser scattering classification device for cell counting - Google Patents
Calibration device and method of laser scattering classification device for cell counting Download PDFInfo
- Publication number
- CN111307695B CN111307695B CN202010164965.XA CN202010164965A CN111307695B CN 111307695 B CN111307695 B CN 111307695B CN 202010164965 A CN202010164965 A CN 202010164965A CN 111307695 B CN111307695 B CN 111307695B
- Authority
- CN
- China
- Prior art keywords
- injector
- valve
- stepping motor
- cell
- solution
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000000034 method Methods 0.000 title claims abstract description 22
- 238000001514 detection method Methods 0.000 claims abstract description 40
- 230000003287 optical effect Effects 0.000 claims abstract description 24
- 238000005086 pumping Methods 0.000 claims abstract description 6
- 239000000243 solution Substances 0.000 claims description 142
- 239000007788 liquid Substances 0.000 claims description 44
- 239000002699 waste material Substances 0.000 claims description 37
- 238000012795 verification Methods 0.000 claims description 23
- 230000008676 import Effects 0.000 claims description 13
- 238000002356 laser light scattering Methods 0.000 claims description 12
- 238000002347 injection Methods 0.000 claims description 7
- 239000007924 injection Substances 0.000 claims description 7
- 238000000386 microscopy Methods 0.000 claims description 6
- 238000011010 flushing procedure Methods 0.000 claims description 2
- 239000011521 glass Substances 0.000 claims 1
- 239000006059 cover glass Substances 0.000 abstract description 4
- 239000012530 fluid Substances 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 81
- 230000008878 coupling Effects 0.000 description 4
- 238000010168 coupling process Methods 0.000 description 4
- 238000005859 coupling reaction Methods 0.000 description 4
- 238000010586 diagram Methods 0.000 description 3
- 230000000903 blocking effect Effects 0.000 description 2
- 238000007635 classification algorithm Methods 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 230000003139 buffering effect Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1434—Optical arrangements
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/1012—Calibrating particle analysers; References therefor
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N2015/1006—Investigating individual particles for cytology
Landscapes
- Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Materials By Optical Means (AREA)
Abstract
The invention discloses a calibration device of a laser scattering classification device for cell counting, which comprises a bearing main body and a cover glass arranged on the bearing main body, wherein a laser scattering area and an optical detection area are arranged on the bearing main body; a fluid pumping system is also included. The invention also discloses a checking control method. Compared with the prior art, the same cell sample in the checking device is classified in the laser scattering classification device and then is conveyed to the microscopic examination device through the checking device for reclassification, and whether the classification result of the laser scattering classification device is correct or not is verified by taking the microscopic examination result as reference; the adjustment is carried out before delivery, and the accuracy of the device is ensured.
Description
Technical Field
The invention relates to a detection technology in the medical field, in particular to a verification device and a verification control method of a laser scattering classification device for cell counting.
Background
At present, there are two methods for verifying whether the classification result of a flow cytometer or a leukocyte analyzer is correct. The first method is to analyze and judge whether the classification of the laser scattering classification device (classification device) for cell counting is correct or not by comparing the known parameters of the quality control material with the inspection result of the instrument. The second method is to take a part of a sample to perform microscopic observation, record the number of related cells, and calculate the result of the proportion of each cell. And comparing the microscopic examination result with the classification device detection result, and analyzing and judging whether the instrument classification is correct or not.
The above two methods have a similar disadvantage, and both methods are used for analyzing and judging the performance of the classification device in a statistical sense. Both methods are based on the whole sample, and do not depend on single cells, so that the methods cannot judge whether the single cells are classified correctly or not, and can only judge whether a large number of cells are roughly classified correctly on the whole; there is therefore a need for a device that can assess the accuracy of the classification of the device.
Disclosure of Invention
The invention aims to provide a verification device and a verification method for a laser scattering classification device for cell counting, aiming at solving the technical problem of verifying the classification result of the laser scattering classification device so as to ensure the classification accuracy of the device when the device leaves a factory.
In order to solve the problems, the invention adopts the following technical scheme: a calibration device of a laser scattering classification device for cell counting comprises a bearing main body for bearing a cell sample and a cover glass arranged on the bearing main body, wherein a laser scattering area and an optical detection area are arranged on the bearing main body, at least the laser scattering area and the optical detection area on the bearing main body are transparent, a cell solution inlet, an oil inlet, a first solution inlet, a second solution inlet and an outflow port are arranged on the bearing main body, a microscopic examination groove is arranged on the bearing main body at the optical detection area, runners respectively communicated with the cell solution inlet, the oil inlet, the first solution inlet, the second solution inlet and the microscopic examination groove are arranged on the bearing main body, the cell solution inlet, the oil inlet and the first solution inlet are arranged at one end of the bearing main body far away from the optical detection area, and the second solution inlet is arranged at the optical detection area, the outflow port is arranged in one end of the microscopic examination groove far away from the laser scattering area;
the cell culture device also comprises a liquid pump system, so that oil, cell solution and cell-free solution are injected into the flow channel through the cell solution inlet, the oil inlet, the first solution inlet and the second solution inlet by the liquid pump system.
Further, the liquid pumping system comprises a first valve, a second valve, a third valve, a fourth valve, a fifth valve, a sixth valve, a seventh valve, an eighth valve, a first injector, a second injector, a third injector, a fourth injector, a waste liquid pump, a first container, a second container, a third container, a waste liquid pool and an air bag, wherein the first injector, the second injector, the third injector and the fourth injector are respectively driven by a first stepping motor, a second stepping motor, a third stepping motor and a fourth stepping motor, an oil inlet is inserted into the first container through a pipeline, the third valve and the fourth valve are arranged on a pipeline between the oil inlet and the first container, and the second injector is connected between the third valve and the fourth valve through a pipeline; the cell solution inlet is inserted into the second container through a pipeline, the first valve and the second valve are arranged on the pipeline between the cell solution inlet and the second container, and the first injector is connected between the first valve and the second valve through a pipeline; the first solution inlet is inserted into the third container through a pipeline, the fifth valve and the sixth valve are arranged on the pipeline between the first solution inlet and the third container, and the third injector is connected between the fifth valve and the sixth valve through a pipeline; the second solution inlet is inserted into the third container through a pipeline, and the seventh valve and the eighth valve are arranged on the pipeline between the second solution inlet and the third container; the outflow port is inserted into the waste liquid pool through a pipeline, and the air bag and the waste liquid pump are arranged on the pipeline between the outflow port and the waste liquid pool.
Further, the runner includes trunk and a plurality of branches, the trunk is located on the central line that bears the weight of the main part, and from cell solution import through laser scattering district straight line extend to the optics detection zone and with the mirror examination recess intercommunication, oil import, first solution import, second solution import then through branch and trunk intercommunication.
Further, the distance of the branch between the oil inlet and the trunk is equal to the distance of the branch between the first solution inlet and the trunk.
Further, the flow passage is a slit groove.
Further, the second solution inlets are two and are symmetrically arranged on two sides of the microscopic examination groove respectively.
Furthermore, the first syringe, the second syringe, the third syringe and the fourth syringe are respectively fixed on the respective corresponding syringe supports, the first stepping motor, the second stepping motor, the third stepping motor and the fourth stepping motor are respectively fixed on the corresponding syringe supports through the motor supports, the axis of the output shaft of the first stepping motor is parallel to the axis of the piston rod of the first syringe, the output shafts of the first stepping motor, the second stepping motor, the third stepping motor and the fourth stepping motor are respectively connected with a lead screw through a coupler, one end of the lead screw, far away from the coupler, is fixed on the corresponding syringe support through a bearing so that the lead screw can axially rotate, a nut in threaded connection is sleeved on the lead screw, and the nut is respectively connected with the piston rod of the first syringe, the piston rod of the second syringe and the piston rod of the third syringe through the connecting supports, The piston rod of the fourth syringe is connected.
Furthermore, the first stepping motor and the first syringe, the second stepping motor and the second syringe, the third stepping motor and the third syringe, and the fourth stepping motor and the fourth syringe are respectively arranged on two side surfaces which are deviated from the corresponding syringe supports, guide grooves are arranged on the syringe supports, and the connecting support penetrates through the guide grooves and then is fixedly connected with the piston rod of the corresponding first syringe, the piston rod of the second syringe, the piston rod of the third syringe and the piston rod of the fourth syringe.
The invention also discloses a verification control method of the laser scattering classification device for cell counting, which comprises the following steps:
step S1, setting a checking device of the laser scattering classification device for cell counting;
step S2, opening the first valve, the third valve, the fifth valve and the seventh valve, respectively driving the first injector, the second injector, the third injector and the fourth injector to pump the oil, the cell solution and the cell-free solution in the first container, the second container and the third container into the first injector, the second injector, the third injector and the fourth injector through the first stepping motor, the second stepping motor, the third stepping motor and the fourth stepping motor, and then closing the first injector, the second injector, the third injector and the fourth injector;
step S3, opening a second valve, and injecting the cell solution in the first injector into the trunk from the cell solution inlet;
step S4, the laser scattering classification device detects whether cells exist in the cell solution in the laser scattering area through laser; if yes, the first injector stops injecting, the second valve is closed, and the step S5 is executed; otherwise, continuing laser detection;
step S5, the second valve is closed, and the first stepping motor is stopped to stop the injection of the first injector; opening a fourth valve, and pushing the oil in the second injector from the branch to the trunk through the oil inlet; blocking the cell solution in the trunk to divide the cell solution, forming a cell solution drop on the trunk close to the laser scattering area, and stopping the second stepping motor to stop the injection of the second injector and close the fourth valve;
step S6, opening a sixth valve, and driving a third injector to push the cell-free solution into the trunk from the branch through a first solution inlet by a third stepping motor;
step S7, the laser scattering classification device carries out laser detection on the cell solution drop in the step S4 again, and whether a second cell exists in the cell solution drop in the laser scattering area is detected; if yes, go to step S11; otherwise, go to step S8;
step S8, the laser scattering classification device detects whether an oil drop signal exists in the laser scattering area through laser; if yes, go to step S9; otherwise, returning to the step S6;
step S9, driving a third injector to continuously inject cell-free solution by a third stepping motor, so that the cell solution drops in the laser scattering area are pushed into the microscopy groove;
step S10, the third step motor stops and the sixth valve is closed; the microscopic examination device photographs the cell solution drops in the optical detection area through an objective lens; comparing the photographing result with the laser detection result;
step S11, starting a waste liquid pump, and dripping the cell solution in the microscopic examination groove into a waste liquid pool; and then opening a sixth valve and an eighth valve, flushing the trunk and the microscopic examination groove through a third injector and a fourth injector, pumping the waste liquid into the waste liquid pool through the waste liquid pump again, and finishing.
Compared with the prior art, the cell sample sorting device has the advantages that the checking device is arranged, the same cell sample in the checking device is sorted in the laser scattering sorting device and then is conveyed to the microscopic examination device through the checking device for reclassification, and the microscopic examination result is used as a reference, so that whether the sorting result of the laser scattering sorting device is correct or not is verified; the laser scattering classification device can be adjusted before leaving the factory, and the accuracy of the device is ensured.
Drawings
Fig. 1 is a schematic structural diagram of the verification apparatus of the present invention.
Fig. 2 is a schematic structural diagram of the load bearing body of the present invention.
FIG. 3 is a schematic view of the structure of the carrier body and the cover glass of the present invention.
Fig. 4 is a sectional view taken along a-a in fig. 3.
Fig. 5 is a state diagram of the use of the present invention.
FIG. 6 is a flow chart of verification control of the present invention.
Fig. 7 is a schematic view of the connection of the stepper motor of the present invention to the injector.
Fig. 8 is a sectional view taken along the direction B-B in fig. 7.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings and examples.
As shown in fig. 1, 2, 3 and 4, the present invention discloses a verification apparatus for a laser light scattering classification apparatus (classification apparatus) for cell counting, comprising: a bearing main body 1 for bearing cell samples, a cover glass 2 arranged on the bearing main body 1, wherein the bearing main body 1 is provided with a laser scattering area 100 and an optical detection area 200, at least the laser scattering area 100 and the optical detection area 200 on the bearing main body 1 are transparent, the bearing main body 1 is provided with a cell solution inlet 3, an oil inlet 4, a first solution inlet 5, a second solution inlet 6 and an outflow opening 7, a microscopic examination groove 8 is arranged on the bearing main body 1 at the optical detection area 200, a flow channel 9 respectively communicating the cell solution inlet 3, the oil inlet 4, the first solution inlet 5, the second solution inlet 6 and the microscopic examination groove 8 is arranged on the bearing main body 1, the cell solution inlet 3, the oil inlet 4 and the first solution inlet 5 are arranged at one end of the bearing main body 1 far away from the optical detection area 200, the second solution inlet 6 is arranged at the optical detection area 200, the outflow port 7 is arranged in one end of the microscopic examination groove 8 far away from the laser scattering area 100;
and a liquid pumping system including a first valve 39, a second valve 10, a third valve 11, a fourth valve 12, a fifth valve 13, a sixth valve 14, a seventh valve 15, an eighth valve 16, a first syringe 17, a second syringe 18, a third syringe 19, a fourth syringe 20, a waste liquid pump 21, a first container 22 for containing oil, a second container 23 for containing a cell solution, a third container 24 for containing a solution (without cells), a waste liquid tank 25, and a bladder 26, wherein the first syringe 17, the second syringe 18, the third syringe 19, and the fourth syringe 20 are driven by a first stepping motor 27, a second stepping motor 28, a third stepping motor 29, and a fourth stepping motor 30, respectively, to inject oil, a cell solution, or a cell-free solution, an oil inlet 4 is inserted into the first container 22 through a pipe, the third valve 11, the fourth valve 12 are provided on a pipe between the oil inlet 4 and the first container 22, the second injector 18 is connected between the third valve 11 and the fourth valve 12 through a pipeline; the cell solution inlet 3 is inserted into the second container 23 through a pipe, the first valve 39 and the second valve 10 are provided on the pipe between the cell solution inlet 3 and the second container 23, and the first syringe 17 is connected between the first valve 39 and the second valve 10 through a pipe; the first solution inlet 5 is inserted into the third container 24 through a pipeline, the fifth valve 13 and the sixth valve 14 are arranged on the pipeline between the first solution inlet 5 and the third container 24, and the third injector 19 is connected between the fifth valve 13 and the sixth valve 14 through a pipeline; the second solution inlet 6 is inserted into the third container 24 through a pipeline, and the seventh valve 15 and the eighth valve 16 are arranged on the pipeline between the second solution inlet 6 and the third container 24; the outflow port 7 is inserted into the waste liquid tank 25 through a pipeline, and the air bag 26 and the waste liquid pump 21 are arranged on the pipeline between the outflow port 7 and the waste liquid tank 25; the air bag 26 is used for temporarily storing the waste liquid so as to solve the problem of unsmooth outflow of the waste liquid.
The oil in the first container 22 is opened through the third valve 11, the fourth valve 12 is closed, and the oil is pumped into the second injector 18 under the driving of the second stepping motor 28; the fourth valve 12 is opened and the third valve 11 is closed, and the oil in the second syringe 18 is pushed into the flow channel 9 from the oil inlet 4 by the driving of the second stepping motor 28 to isolate the cell solution in the flow channel 9, so that the cell solution in the laser scattering area 100 forms a cell solution droplet.
The cell solution in the second container 23 is pumped into the first syringe 17 by the driving of the first stepping motor 27 through the opening of the first valve 39 and the closing of the second valve 10; the cell solution in the first syringe 17 is pushed from the cell solution inlet 3 into the flow channel 9 by the driving of the first stepping motor 27 with the second valve 10 opened and the first valve 39 closed to send the cell solution into the flow channel 9.
The cell-free solution in the third container 24 is opened by the fifth valve 13, closed by the sixth valve 14, and is pumped into the third syringe 19 by the driving of the third step motor 29; the sixth valve 14 is opened, the fifth valve 13 is closed, and the solution in the third syringe 19 is pushed into the flow channel 9 from the first solution inlet 5 by the driving of the third stepping motor 29, so as to push the oil in the flow channel 9 towards the optical detection area 200, so as to drive the droplets of the cell solution to move towards the optical detection area 200 and clean the flow channel.
The cell-free solution in the third container 24 is also opened by the seventh valve 15, closed by the eighth valve 16, and pumped into the fourth syringe 20 by the driving of the fourth stepping motor 30; the eighth valve 16 is opened, the seventh valve 15 is closed, and the cell-free solution in the fourth syringe 20 is pushed into the flow channel 9 from the second solution inlet 6 by the driving of the fourth stepping motor 30, so as to wash the cell solution droplets located in the optical detection zone 200.
The waste liquid pump 21 is used for sucking waste liquid in the microscopic examination groove 8, buffering the generated waste liquid through the air bag 26, and pumping the waste liquid into the waste liquid pool 25.
As shown in fig. 1, the flow channel 9 includes a main trunk 91 and a plurality of branches 92, the main trunk 91 is disposed on a central line of the main carrying body 1, and extends linearly from the cell solution inlet 3 to the optical detection region 200 through the laser scattering region 100 and communicates with the microscopy groove 8, and the oil inlet 4, the first solution inlet 5, and the second solution inlet 6 communicate with the main trunk 91 through the branches 92.
As shown in fig. 1, as a preferred embodiment of the present invention, the carrying body 1 has a plate-like structure, and the distance between the oil inlet 4 and the branch 92 of the trunk 91 is equal to the distance between the first solution inlet 5 and the branch 92 of the trunk 91; the second solution inlets 6 are provided with two, which are respectively symmetrically arranged on two sides of the microscopic examination groove 8, so that the cell-free solution can be well flowed towards the microscopic examination groove 8.
As shown in fig. 2, the flow passage 9 is a slit groove; the microscopic examination groove 8 comprises a circular main groove 81 and an extension groove 82, the main groove 81 is arranged at the optical detection region 200, the extension groove 82 is arranged at one end of the main groove 81 far away from the laser scattering region 100, and the outflow port 7 is arranged at one end of the extension groove 82 far away from the main groove 81.
As shown in FIG. 5, in use, the inspection device 600 is placed on the stage 300 and the laser light scattering classification device 400 of the microscopy device 500; the optical detection area 200 is located below the objective lens 501 of the microscopy device 500 and is opposite to the light gathering device 502 and the light source 503; the general laser scattering classification device 400 mainly includes a laser 401, a reflective mirror 402, a beam splitter 403, a first condenser 404, a second condenser 405, a first photosensor 406, and a second photosensor 407, as can be seen from the figure, the laser 401 is opposite to the laser scattering area 100 and is located at the lower end of the verification device 600, and the reflective mirror 402 is arranged opposite to the laser 401 and is located above the verification device 600; the beam splitter 403 is opposite to the reflective mirror 402 to guide the light to the first and second photosensors 406 and 407 through the first and second condensing lenses 403 and 405, respectively.
In the present invention, the first syringe 17, the second syringe 18, the third syringe 19 and the fourth syringe 20 are respectively fixed on the respective corresponding syringe supports 31, the first stepping motor 27, the second stepping motor 28, the third stepping motor 29 and the fourth stepping motor 30 are respectively fixed on the corresponding syringe supports 31 through the motor supports 38, that is, each syringe and each stepping motor are provided with one syringe support, as shown in fig. 7 and 8, the axis of the output shaft of the first stepping motor 27 is parallel to the axis of the piston rod of the first syringe 17, the output shafts of the first stepping motor 27, the second stepping motor 28, the third stepping motor 29 and the fourth stepping motor 30 are respectively connected with the lead screw 33 through the coupling 32, one end of the lead screw 33 far away from the coupling 32 is fixed on the corresponding syringe support 31 through the bearing 37 so that the lead screw 33 can axially rotate, the screw rod 33 is sleeved with a nut 34 in threaded connection, and the nut 34 is respectively connected with the piston rod of the first syringe 17, the piston rod of the second syringe 18, the piston rod of the third syringe 19 and the piston rod of the fourth syringe 20 through a connecting bracket 35. The first stepping motor 27 and the first syringe 17, the second stepping motor 28 and the second syringe 18, the third stepping motor 29 and the third syringe 19, and the fourth stepping motor 30 and the fourth syringe 20 are respectively arranged on two side surfaces of the syringe bracket 31 which are deviated from each other, a guide groove 36 is arranged on the syringe bracket 31, and the connecting bracket 35 is fixedly connected with the piston rod of the corresponding first syringe 17, the piston rod of the second syringe 18, the piston rod of the third syringe 19, and the piston rod of the fourth syringe 20 after penetrating through the guide groove 36.
Since the first syringe 17, the second syringe 18, the third syringe 19, the fourth syringe 20 and the first stepping motor 27, the second stepping motor 28, the third stepping motor 29 and the fourth stepping motor 30 have the same connection structure, the connection between the first syringe 17 and the first stepping motor 27 is further described herein by taking as an example the connection between the first syringe 17 and the first stepping motor 27, as shown in fig. 7 and 8, the first syringe 17 is fixed on a syringe bracket 31, the first stepping motor 27 is fixed on the syringe bracket 31 through a motor bracket 38, the axis of the output shaft of the first stepping motor 27 is parallel to the axis of the piston rod of the first syringe 17 and can be arranged on the same side or opposite sides of the syringe bracket 31, the output shaft of the first stepping motor 27 is connected with a lead screw 33 through a coupling 32, and the end of the lead screw 33 far away from the coupling 32 is fixed on the syringe bracket 31 through a bearing 37, so that the screw 33 can axially rotate, a nut 34 in threaded connection is sleeved on the screw 33, the nut 34 is connected with the piston rod 171 of the first injector 17 through a connecting bracket 35, so that when the first stepping motor 27 drives the screw 33 to rotate, the nut 34 moves on the screw 33, thereby driving the piston rod of the first injector 17 to move; specifically, the first stepping motor 27 and the first syringe 17 are disposed on two side surfaces of the syringe bracket 31 facing away from each other, a guide groove 36 is disposed on the syringe bracket 31, and the connecting bracket 35 is fixedly connected to the piston rod of the first syringe 17 after passing through the guide groove 36.
As shown in fig. 1 and 6, the verification control method of the present invention includes the steps of:
step S1, setting the checking device of the laser scattering classification device for cell counting;
step S2, opening the first valve 39, the third valve 11, the fifth valve 13 and the seventh valve 15, driving the first injector 17, the second injector 18, the third injector 19 and the fourth injector 20 to pump the oil, the cell solution and the cell-free solution in the first container 22, the second container 23 and the third container 24 into the first injector 17, the second injector 18, the third injector 19 and the fourth injector 20 respectively through the first stepping motor 27, the second stepping motor 28, the third stepping motor 29 and the fourth stepping motor 30, and then closing the first injector, the second injector, the third injector and the fourth injectors;
step S3, opening the second valve 10, and injecting the cell solution in the first syringe 17 into the stem 91 from the cell solution inlet 3;
step S4, detecting whether there is a cell in the cell solution in the laser scattering area 100 by the laser light scattering classification device (classification device); if yes, the first injector 17 stops injecting, the second valve 10 is closed, and the process goes to step S5; otherwise, continuing laser detection;
step S5, the second valve 10 is closed, and the first stepping motor 27 is stopped to stop the injection of the first syringe 17; opening the fourth valve 12 to push the oil in the second syringe 18 from the branch 92 into the trunk 91 through the oil inlet 4; thereby blocking the cell solution in the trunk 91 to divide the cell solution, so that a droplet of the cell solution can be formed on the trunk 91 near the laser light scattering area 100, and then the second stepping motor 28 is stopped to stop the injection of the second injector 18 and close the fourth valve 12;
step S6, opening the sixth valve 14, and the third step motor 29 driving the third injector 19 to push the cell-free solution from the branch 92 to the trunk 91 through the first solution inlet 5, so as to push the oil and the blocked droplets of the cell solution to move continuously toward the optical detection area 200;
step S7, the sorting device performs laser detection on the droplet of cell solution in step S4 again, and detects whether a second cell exists in the droplet of cell solution in the laser scattering area 100; if yes, go to step S11; otherwise, go to step S8;
step S8, the sorting device detects whether an oil drop signal exists in the laser scattering area 100 through laser; if yes, go to step S9; otherwise, returning to the step S6; detecting the oil drop signal ensures that the laser detection scan has been completed in the laser scattering region and that no second cell is detected;
step S9, driving the third injector 19 to continue injecting the cell-free solution by the third stepping motor 29, so that the cell solution droplets in the laser scattering area 100 are pushed into the microscopy groove 8;
step S10, the third stepper motor 29 is stopped to stop the injection of the third injector 19 and close the sixth valve 14; the microscopic examination device photographs the cell solution drops in the optical detection area 200 through the objective lens; comparing the photographing result with the laser detection result; therefore, whether the classification method of the classification device is correct or not can be judged; if the classification of the classification device is wrong, the parameters of the corresponding classification algorithm can be adjusted according to the reason of the wrong image analysis in the microscopic examination; thereby optimizing and improving the accuracy of the classification device;
step S11, starting the waste liquid pump 21, and dripping the cell solution in the microscopic examination groove 8 into the waste liquid pool; then, the sixth valve 14 and the eighth valve 16 are opened, the trunk 91 and the microscopic examination groove 8 are washed by the third syringe 19 and the fourth syringe 20, and then the waste liquid is pumped into the waste liquid pool 25 by the waste liquid pump 21 again, and the process is finished.
By the verification device and the verification control method, cell solution droplets of the same cell sample can pass through the laser scattering area first, laser scattering signals when the cell solution flows through the droplet are obtained, and if the laser scattering signals judge that the droplets only contain one cell, the droplets are conveyed to a microscope detection area for microscopic examination and photographing; if it is judged that one of the droplets contains no cell, the droplet of the cell solution is directly discharged. And then comparing the cell classification result of the laser scattering signal with the microscopic examination classification result to judge whether the laser scattering classification method is accurate or not, so as to verify the classification result of the laser scattering classification device (classification device) and timely adjust the parameters of the corresponding classification algorithm of the laser scattering classification device (classification device), thereby optimizing and improving the accuracy of the classification device.
Claims (9)
1. A verification device for a laser light scattering classification device for cell counting, characterized in that: including a main part (1) that bears that is used for bearing the weight of the cell sample, locate glass cover (2) on bearing the weight of main part (1), be equipped with laser scattering area (100), optics detection zone (200) on bearing the weight of main part (1), it is transparent to bear laser scattering area (100) and optics detection zone (200) at least on main part (1), it imports (3), oil import (4), first solution import (5), second solution import (6), egress opening (7) to be equipped with cell solution on bearing the weight of main part (1), lie in optics detection zone (200) on bearing the weight of main part (1) and locate to establish mirror examination recess (8), be equipped with runner (9) that communicate cell solution import (3), oil import (4), first solution import (5), second solution import (6) and mirror examination recess (8) respectively on bearing the weight of main part (1), cell solution import (3), The oil inlet (4) and the first solution inlet (5) are positioned at one end, far away from the optical detection area (200), of the bearing main body (1), the second solution inlet (6) is arranged at the optical detection area (200), and the outflow port (7) is arranged in one end, far away from the laser scattering area (100), of the microscopic examination groove (8);
the device also comprises a liquid pump system, so that oil, cell solution and cell-free solution are injected into the flow channel (9) through the cell solution inlet (3), the oil inlet (4), the first solution inlet (5) and the second solution inlet (6) by the liquid pump system.
2. The verification device for a laser light scattering classification apparatus for cell counting as claimed in claim 1, wherein: the liquid pump system comprises a first valve (39), a second valve (10), a third valve (11), a fourth valve (12), a fifth valve (13), a sixth valve (14), a seventh valve (15), an eighth valve (16), a first injector (17), a second injector (18), a third injector (19), a fourth injector (20), a waste liquid pump (21), a first container (22), a second container (23), a third container (24), a waste liquid pool (25) and an air bag (26), wherein the first injector (17), the second injector (18), the third injector (19) and the fourth injector (20) are respectively driven by a first stepping motor (27), a second stepping motor (28), a third stepping motor (29) and a fourth stepping motor (30), an oil inlet (4) is inserted into the first container (22) through a pipeline, the third valve (11) and the fourth valve (12) are arranged on a pipeline between the oil inlet (4) and the first container (22), and the second injector (18) is connected between the third valve (11) and the fourth valve (12) through a pipeline; the cell solution inlet (3) is inserted into the second container (23) through a pipeline, the first valve (39) and the second valve (10) are arranged on the pipeline between the cell solution inlet (3) and the second container (23), and the first injector (17) is connected between the first valve (39) and the second valve (10) through a pipeline; the first solution inlet (5) is inserted into the third container (24) through a pipeline, the fifth valve (13) and the sixth valve (14) are arranged on the pipeline between the first solution inlet (5) and the third container (24), and the third injector (19) is connected between the fifth valve (13) and the sixth valve (14) through a pipeline; the second solution inlet (6) is inserted into the third container (24) through a pipeline, and the seventh valve (15) and the eighth valve (16) are arranged on the pipeline between the second solution inlet (6) and the third container (24); the outflow port (7) is inserted into the waste liquid tank (25) through a pipe, and the air bag (26) and the waste liquid pump (21) are arranged on the pipe between the outflow port (7) and the waste liquid tank (25).
3. The verification device for a laser light scattering classification apparatus for cell counting as claimed in claim 1, wherein: the runner (9) comprises a main body (91) and a plurality of branches (92), wherein the main body (91) is arranged on a central line of the bearing main body (1), the main body extends to the optical detection area (200) from the cell solution inlet (3) through the laser scattering area (100) and is communicated with the microscopic examination groove (8), and the oil inlet (4), the first solution inlet (5) and the second solution inlet (6) are communicated with the main body (91) through the branches (92).
4. The verification device for a laser light scattering classification apparatus for cell counting as claimed in claim 3, wherein: the distance of the branch (92) between the oil inlet (4) and the trunk (91) is equal to the distance of the branch (92) between the first solution inlet (5) and the trunk (91).
5. The verification device for a laser light scattering classification apparatus for cell counting as claimed in any one of claims 1 to 4, wherein: the flow channel (9) is a slit groove.
6. The verification device for a laser light scattering classification apparatus for cell counting as claimed in claim 1, wherein: the two second solution inlets (6) are symmetrically arranged on two sides of the microscopic examination groove (8).
7. The verification device for a laser light scattering classification apparatus for cell counting as claimed in claim 2, wherein: the first injector (17), the second injector (18), the third injector (19) and the fourth injector (20) are respectively fixed on the corresponding injector supports (31), the first stepping motor (27), the second stepping motor (28), the third stepping motor (29) and the fourth stepping motor (30) are respectively fixed on the corresponding injector supports (31) through motor supports (38), the axis of the output shaft of the first stepping motor (27) is parallel to the axis of the piston rod of the first injector (17), the output shafts of the first stepping motor (27), the second stepping motor (28), the third stepping motor (29) and the fourth stepping motor (30) are respectively connected with a screw rod (33) through a coupler (32), and one end of the screw rod (33) far away from the coupler (32) is fixed on the corresponding injector support (31) through a bearing (37), so that the screw rod (33) can axially rotate, a nut (34) in threaded connection is sleeved on the screw rod (33), and the nut (34) is respectively connected with a piston rod of the first syringe (17), a piston rod of the second syringe (18), a piston rod of the third syringe (19) and a piston rod of the fourth syringe (20) through a connecting bracket (35).
8. The verification device for a laser light scattering classification apparatus for cell counting as claimed in claim 7, wherein: the first stepping motor (27) and the first injector (17), the second stepping motor (28) and the second injector (18), the third stepping motor (29) and the third injector (19), and the fourth stepping motor (30) and the fourth injector (20) are respectively arranged on two side surfaces which are deviated from the corresponding injector supports (31), guide grooves (36) are formed in the injector supports (31), and the connecting support (35) penetrates through the guide grooves (36) and then is connected and fixed with the piston rod of the corresponding first injector (17), the piston rod of the corresponding second injector (18), the piston rod of the corresponding third injector (19), and the piston rod of the corresponding fourth injector (20).
9. A verification control method of a laser scattering classification device for cell counting is characterized in that: the method comprises the following steps:
step S1, setting up the verification device of the laser scattering classification device for cell counting as claimed in claim 2, 7 or 8;
step S2, opening a first valve (39), a third valve (11), a fifth valve (13) and a seventh valve (15), and respectively driving a first injector (17), a second injector (18), a third injector (19) and a fourth injector (20) to pump oil, cell solution and cell-free solution in a first container (22), a second container (23) and a third container (24) into the first injector, the second injector (18), the third injector (19) and the fourth injector (20) through a first stepping motor (27), a second stepping motor (28), a third stepping motor (29) and a fourth stepping motor (30) and then closing the first injector, the second injector, the third injector and the fourth injector;
step S3, opening a second valve (10), and injecting the cell solution in the first injector (17) into the trunk (91) from the cell solution inlet (3);
step S4, the laser scattering classification device detects whether cells exist in the cell solution in the laser scattering area (100) through laser; if yes, the first injector (17) stops injecting, the second valve (10) is closed, and the step S5 is carried out; otherwise, continuing laser detection;
step S5, the second valve (10) is closed, and the first stepping motor (27) is stopped to stop the injection of the first injector (17); opening the fourth valve (12) to push the oil in the second syringe (18) from the branch (92) into the trunk (91) through the oil inlet (4); so as to block the cell solution in the trunk (91) to divide the cell solution, form a drop of the cell solution on the trunk (91) near the laser scattering area (100), and then stop the second stepping motor (28) to stop the injection of the second injector (18) and close the fourth valve (12);
step S6, opening a sixth valve (14), driving a third injector (19) by a third stepping motor (29) to push the cell-free solution from the branch (92) to the trunk (91) through a first solution inlet (5);
step S7, the laser scattering classification device carries out laser detection on the cell solution drop in the step S4 again, and detects whether a second cell exists in the cell solution drop in the laser scattering area (100); if yes, go to step S11; otherwise, go to step S8;
step S8, the laser scattering classification device detects whether an oil drop signal exists in the laser scattering area (100) through laser; if yes, go to step S9; otherwise, returning to the step S6;
step S9, driving a third injector (19) to continue injecting the cell-free solution by a third stepping motor (29) so that the cell solution drops in the laser scattering area (100) are pushed into the microscopy groove (8);
step S10, stopping the third stepping motor (29) and closing the sixth valve (14); the microscopic examination device photographs the cell solution drops in the optical detection area (200) through an objective lens;
step S11, starting a waste liquid pump (21), and dripping the cell solution in the microscopic examination groove (8) into a waste liquid pool; and then, opening a sixth valve (14) and an eighth valve (16), flushing the main stem (91) and the microscopic examination groove (8) through a third injector (19) and a fourth injector (20), and pumping the waste liquid into a waste liquid pool (25) through a waste liquid pump (21) again to finish the process.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010164965.XA CN111307695B (en) | 2020-03-11 | 2020-03-11 | Calibration device and method of laser scattering classification device for cell counting |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010164965.XA CN111307695B (en) | 2020-03-11 | 2020-03-11 | Calibration device and method of laser scattering classification device for cell counting |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111307695A CN111307695A (en) | 2020-06-19 |
| CN111307695B true CN111307695B (en) | 2022-06-14 |
Family
ID=71157112
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010164965.XA Active CN111307695B (en) | 2020-03-11 | 2020-03-11 | Calibration device and method of laser scattering classification device for cell counting |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111307695B (en) |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1608963B1 (en) * | 2003-03-28 | 2009-12-30 | Inguran, LLC | Apparatus and methods for providing sex-sorted animal sperm |
| CA2614319A1 (en) * | 2005-07-15 | 2007-01-25 | Chemimage Corporation | Digitizing biology |
| US8691164B2 (en) * | 2007-04-20 | 2014-04-08 | Celula, Inc. | Cell sorting system and methods |
| CA2761630A1 (en) * | 2008-04-25 | 2009-10-25 | James W. Winkelman | Method of determining a complete blood count and a white blood cell differential count |
| CN103267838B (en) * | 2013-05-15 | 2013-08-28 | 中山市滔略生物科技有限公司 | Quality control material and calibration material for verifying hematology analyzer and preparation method thereof |
| CN109269964A (en) * | 2018-10-15 | 2019-01-25 | 深圳博大博聚科技有限公司 | A kind of artificial counting and calibration equipment based on image recognition method cell counter |
| CN112795463A (en) * | 2020-12-28 | 2021-05-14 | 山东大学 | Automatic single cell separation system and method |
-
2020
- 2020-03-11 CN CN202010164965.XA patent/CN111307695B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN111307695A (en) | 2020-06-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US20230075298A1 (en) | Autofocus systems and methods for particle analysis in blood samples | |
| CN101185001B (en) | For blood sample analysis method and implement the equipment of the method and reagent | |
| CN101189503B (en) | Optical device for blood analysis, analyzing apparatus equipped with same | |
| CN105074420B (en) | Hematology system and method | |
| US8715573B2 (en) | Fluidic system for a flow cytometer with temporal processing | |
| EP3207358B1 (en) | Systems and methods for imaging fluid samples | |
| CN101184984B (en) | Hydraulic device for blood analyzing apparatus, related method and analyzing apparatus equipped with such a device | |
| CN102393705B (en) | Sample formed component analyzer automatic detection control apparatus and control method thereof | |
| US20160011098A1 (en) | Compound optical flow cells and method of manufacture and use | |
| CN104458544A (en) | Liquid flow system of flow cytometer | |
| CN111307695B (en) | Calibration device and method of laser scattering classification device for cell counting | |
| CN104422646A (en) | Sample analyzing method and sample analyzer | |
| CN111808737A (en) | Cell sorting system | |
| US20030110826A1 (en) | Sheath liquid supplying apparatus, sheath liquid supplying method, and evaluating method of sheath liquid supplying condition | |
| WO2017167361A1 (en) | Aligning a non-spherical biological entity in a sample flow using ambient viscoelastic fluid flows | |
| CN101184985B (en) | Vessel for blood analyzing optical device, analyzing apparatus equipped with same | |
| CN114166609A (en) | Full-automatic sample pretreatment device | |
| CN209485972U (en) | A kind of on-line checking bacteria concentration device | |
| CN114594040A (en) | Flow cytometer and working method thereof | |
| CN211061420U (en) | Automatic zooming fluorescence detection device based on cell nondestructive detection | |
| CA3192517A1 (en) | H-type filter device for analysing a component | |
| CN111289447A (en) | On-line measuring fungus concentration device | |
| US20240024867A1 (en) | Flowcell holder and biological analysis systems and methods | |
| CN116593722A (en) | Single-molecule detection liquid path system and single-molecule detection method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |