CN110940568A - Integrated system for cell enrichment, separation, dyeing and flaking - Google Patents
Integrated system for cell enrichment, separation, dyeing and flaking Download PDFInfo
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- CN110940568A CN110940568A CN201911303162.1A CN201911303162A CN110940568A CN 110940568 A CN110940568 A CN 110940568A CN 201911303162 A CN201911303162 A CN 201911303162A CN 110940568 A CN110940568 A CN 110940568A
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- 238000004043 dyeing Methods 0.000 title claims abstract description 14
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- 238000010186 staining Methods 0.000 claims abstract description 28
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N1/31—Apparatus therefor
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/34—Purifying; Cleaning
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/40—Concentrating samples
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Abstract
The invention belongs to the field of biological sample analysis and detection, and particularly relates to an integrated system for cell enrichment, separation, staining and flaking. The integrated system for cell enrichment, separation, staining and slide production comprises an automatic control module, and a mechanical control device, a cell separation device and a specimen slide production device which can be controlled in a linkage manner under the control of the automatic control module. According to the system, after the CTCs tumor cells are extracted by the cell separation device in a filtering mode, the CTCs tumor cells are transferred to the specimen slide-making device for dyeing treatment, and the whole operation process has high automation degree and high flux. The system has reasonable layout and compact structure of each module, integrates the functions of sample transfer, sample fixation, sample filtration, cell staining and sample mounting into one system for operation, has extremely high integration degree, has the advantages of automation and high flux, can adapt to the requirement of individualized medical treatment, and is applied to various medical institution occasions, timely diagnosis and the like.
Description
Technical Field
The invention belongs to the field of biological sample analysis and detection, and particularly relates to an integrated system for cell enrichment, separation, staining and flaking.
Background
Circulating Tumor Cells (CTCs) are tumor cells that are released into the peripheral blood circulation from solid tumors or metastases, either spontaneously or as a result of medical procedures, and are important causes of postoperative recurrence and distant metastasis in malignant patients. In recent years, with the continuous improvement of detection technology, the application value of CTCs detection as a novel non-invasive diagnostic tool in the aspects of early detection of postoperative recurrence and distant metastasis of patients, evaluation of curative effect and prognosis and the like has become a hot spot of clinical research. However, since the number of CTCs in peripheral blood is very small, and the only few tumor cells are usually found in about 1 hundred million white blood cells and 500 hundred million red blood cells, the CTCs are usually enriched before detection, i.e., isolated from collected blood or other samples, in order to increase the detection rate of the CTCs.
Currently, the enrichment methods of CTCs are mainly classified into immunomagnetic separation methods and enrichment methods based on the physical properties of CTCs according to their principles. Among them, enrichment methods based on the physical properties of CTCs mainly include density gradient centrifugation and filtration. The filtration method is a method for separating tumor cells based on their sizes, and is mainly used for separating tumor cells according to their sizes from normal cells. The filtration method is a method for separating cells by a physical method, so that the cell morphology can be well and completely preserved, various antigens or molecular markers on the surface are not damaged, the characteristics of the cells are not influenced, good conditions are provided for subsequent detection, the technical requirement on equipment is not high, the process is easy to master, and the method becomes the most common CTCs enrichment method in practical operation.
However, in the actual CTCs separation and enrichment method, in the process of cell separation and staining, the separation efficiency is not high enough due to the slow separation speed of cells, and the filter is easily blocked due to the sedimentation of cells in the sample; in addition, in practical operation, a lot of non-tumor cells are easily remained on the back surface of the filtering membrane after cell separation is finished, and great interference is brought to the reading process after staining; moreover, in the dyeing process after the enrichment of the CTCs, the processes of drying, sealing and drying the filtering membrane sample are also needed manually, so that the whole CTCs detection process is relatively complicated, and the working efficiency is influenced.
Disclosure of Invention
Therefore, the technical problem to be solved by the present invention is to provide an integrated system for cell enrichment, separation, staining and slide production, which has the advantages of high sample processing throughput, fast sample processing speed and high automation degree.
In order to solve the technical problems, the integrated system for cell enrichment, separation, staining and slide production is characterized by comprising an automatic control module, and a mechanical control device, a cell separation device and a specimen slide production device which can be controlled in a linkage manner under the control of the automatic control module; wherein the content of the first and second substances,
the mechanical control device is used for picking up consumables and reagents required in the whole operation process and conveying the consumables and the reagents to a specified position for corresponding operation; the mechanical control device comprises a mechanical arm sliding rail assembly, a liquid suction assembly and a cover glass capping assembly, wherein the liquid suction assembly can move to a specified position along the mechanical arm sliding rail and is used for picking up required consumables and reagents, and the cover glass capping assembly is used for picking up required cover glass;
the cell separation device comprises a cell filtration component and a cleaning component which is adaptive to the cell filtration component, and is used for enriching target cells in a sample to be separated and separating the target cells to obtain target cells which can be used for flaking;
the specimen slide making device comprises a heating assembly arranged on a workbench and a cover glass extracting assembly for controlling automatic extraction of required cover glass, and is used for dyeing, making a slide and sealing the enriched target cells to obtain specimen samples which can be directly used for observation.
Specifically, the mechanical arm sliding rail assembly comprises a group of fixed rails arranged in parallel and a sliding rail which is vertically arranged between the fixed rails and can move along the vertical direction of the fixed rails;
the liquid suction assembly and the cover glass capping assembly are respectively arranged on two sides of the sliding rail and are respectively conveyed to a designated working position along with the movement of the sliding rail.
Specifically, the liquid suction assembly comprises a TIP head pick-up hole, the required TIP head is picked up and pushed out under the control of a linear motor, and the required reagent is extracted and filled at a specified position at a fixed point under the control of a reagent extraction injection pump communicated with the TIP head pick-up hole.
Specifically, the cover slip capping assembly comprises a cover slip picking suction cup, and fixed-point picking of the required cover slip is realized and capping is carried out on the corresponding glass slide under the control of a cover slip picking suction cup driving motor;
the cover glass picking sucker is respectively communicated with the diaphragm pump and the corresponding air release valve through a pipeline, and the suction and the placement of the required cover glass are realized by controlling the alternate work of the diaphragm pump and the corresponding air release valve.
Specifically, the cell separation assembly comprises a cell filter, wherein a filtering membrane is arranged in the cell filter, and after target cells are injected into the cell filter, negative pressure is formed under the action of a pipeline pinch valve to perform cell sedimentation filtration and enrich the target cells at the filtering membrane.
Specifically, wash the subassembly including set up in the under membrane of cell filter below washs the needle, can be right the back of filtration membrane enrichment face is washd, and the liquid after the washing is then kept in temporarily in with the waste liquid that the cell filter is linked together keeps in the intracavity.
Specifically, the cover glass extracting assembly comprises a cover glass placing frame arranged on a workbench, and a cover glass stacking area capable of accommodating the cover glass is formed in the cover glass placing frame;
a gap which can only be pushed out by a single cover glass is arranged between the cover glass placing frame and the plane of the workbench, and the cover glass push rod arranged on the workbench and corresponding to the gap pushes the required cover glass to move out of the cover glass placing frame to a fixed point;
be provided with in the cover glass rack and follow the gliding cover glass briquetting of cover glass rack inner wall, the cover glass briquetting is along with the continuous release of cover glass and the slip descends, works as when the cover glass briquetting descend to predetermineeing the height can trigger set up in the cover glass surplus alarm of cover glass rack side reminds the replenishment cover glass.
Specifically, the heating assembly comprises a heating table top arranged on the workbench, and a glass slide placing area is arranged on the heating table top;
and a waste liquid sucker slide rail is further arranged on the heating table top and can drive the waste liquid sucker to a specified glass slide position to pump and remove waste reagents generated in the dyeing process.
Specifically, the system further comprises a consumable placement assembly;
the consumable placing assembly comprises a TIP head placing frame, a reagent placing frame and/or a sample placing frame.
Specifically, the system further comprises a waste liquid storage assembly;
the waste storage assembly includes a TIP waste chamber and/or a waste reagent collection bucket.
The integrated system for cell enrichment, separation, staining and slide production comprises an automatic control module, and a mechanical control device, a cell separation device and a specimen slide production device which can be controlled in a linkage manner under the control of the automatic control module. According to the system, after the CTCs tumor cells are extracted by the cell separation device in a filtering mode, the CTCs tumor cells are transferred to the specimen slide-making device for dyeing treatment, and the whole operation process has high automation degree and high flux. The system has reasonable layout and compact structure of each module, integrates the functions of sample transfer, sample fixation, sample filtration, cell staining and sample mounting into one system for operation, has extremely high integration degree, has the advantages of automation and high flux, can adapt to the requirement of individualized medical treatment, and is applied to various medical institution occasions, timely diagnosis and the like.
In the integrated system for cell enrichment, separation, staining and slide production, the liquid adding of staining solution and the picking of the cover glass can be completed together by arranging the mechanical control device and the sample slide production device which can be controlled in a linkage manner, the automatic extraction of the required cover glass can be realized by the heating slide-sealing device, and the effect of sealing is further optimized by utilizing the heating table top arranged on the surface of the workbench. In the system, the mounting process can be completed automatically, the liquid adding of the dyeing liquid and the cover glass picking are completed based on the same device in the whole process, the complexity of operation is effectively reduced, and the mounting effect is improved by increasing the control of the heating table top.
Drawings
In order that the present disclosure may be more readily and clearly understood, the following detailed description of the present disclosure is provided in connection with specific embodiments thereof and the accompanying drawings, in which,
FIG. 1 is a schematic structural view of an integrated system according to the present invention;
FIG. 2 is a schematic view of the internal construction of the wicking assembly of the present invention;
FIG. 3 is a schematic view of the internal structure of the cover slip capping assembly of the present invention;
FIG. 4 is a schematic view of another angular internal configuration of the cover slip capping assembly of the present invention;
FIG. 5 is a schematic view of the structure of the cell separation apparatus according to the present invention;
FIG. 6 is a schematic view showing the structure of a specimen-producing apparatus according to the present invention;
FIG. 7 is a schematic view of the cover slip extraction assembly of the present invention;
FIG. 8 is a schematic view of a heating assembly according to the present invention;
the reference numbers in the figures denote: 1-liquid absorption component, 2-cover glass cover component, 3-cover glass extraction component, 4-heating component, 5-mechanical arm sliding rail component, 6-cell separation component, 7-cleaning component, 11-TIP head picking hole, 12-TIP head, 13-linear motor, 14-main control board, 15-position judgment sensor, 16-sucker air pressure sensor, 17-reagent extraction injection pump, 21-cover glass picking sucker, 22-cover glass picking sucker driving motor, 23-diaphragm pump, 31-cover glass placing rack, 32-cover glass pressing block, 33-cover glass stacking area, 34-cover glass allowance alarm sensor, 35-cover glass push rod, 36-cover glass push rod motor and 41-glass placing area, 42-heating table top, 43-waste liquid sucker slide rail, 44-waste liquid sucker, 51-fixed rail, 52-sliding rail, 61-cell filter, 62-cell filter interface, 62-pipeline pinch valve, 64-pipeline monitoring module, 71-cleaning needle under membrane, 71-waste liquid temporary storage cavity, 81-TIP head placing frame, 82-reagent placing frame, 83-sample placing frame, 91-TIP head waste cavity and 92-waste reagent collecting barrel.
Detailed Description
In the following embodiments of the present invention, the structure and function of the integrated device are described by taking the enrichment and separation of CTCs cells as an example.
The integrated system for cell enrichment, separation, staining and slide production comprises an automatic control module, and a mechanical control device, a cell separation device and a specimen slide production device which can be controlled in a linkage manner under the control of the automatic control module, as shown in the structure of FIG. 1. The automatic control module can be a control module or a control assembly known in the prior art, and the writing of a corresponding control program is carried out. The mechanical control device, the cell separation device and the specimen slide making device can be arranged in a reasonable space among the working supports 10 of the whole system. Preferably, in the configuration shown in fig. 1, the mechanical control device is provided in the circumferential direction of the top of the work frame 10, and the cell separation device and the specimen slide device are provided in parallel in the internal space of the work frame 10, thereby forming a system with a small space occupation.
With the structure shown in fig. 1-4, the mechanical control device is used for picking up consumables and reagents required in the whole operation process and conveying the consumables and the reagents to a designated position for corresponding operation. The mechanical control device comprises a mechanical arm slide rail assembly 5, a liquid suction assembly 1 which can move to a designated position along the mechanical arm slide rail assembly 5 and is used for picking up required consumables and reagents, and a cover glass capping assembly 2 used for picking up required cover glass. The washing liquid assembly 1 is mainly used for filling reagents such as a sample to be separated, a staining solution and the like to a fixed point position at a fixed point, sucking the required cover glass by utilizing the moving process of the cover glass capping assembly 2 while adding the staining solution, and capping the sucked cover glass on the glass slide at the fixed point after adding the staining solution. Because the liquid adding of the staining solution and the suction of the cover glass are completed based on the same device assembly, the suction of the cover glass is completed while the liquid adding is performed, and the working time of the step can be effectively saved.
In the structure shown in fig. 1, the robot sliding rail assembly 5 is disposed at the upper circumference of the working support 10, and specifically, the robot sliding rail assembly 5 includes a set of fixed rails 51 disposed in parallel, and a sliding rail 52 disposed vertically between the fixed rails 51 and capable of moving along the vertical direction of the fixed rails 51; the fixed rails 51 are fixed at both side end positions of the working bracket 10 in parallel, a sliding rail 52 capable of sliding along the fixed rails 51 in the transverse direction is arranged between the fixed rails 51, and both ends of the sliding rail 52 are respectively positioned in the sliding grooves of the fixed rails 51 and can slide along the fixed rails 51. The liquid suction assembly 1 and the cover glass capping assembly 2 are respectively arranged on two side surfaces of the sliding rail 52 and are respectively connected with sliding grooves on two sides of the sliding rail 52 in a sliding manner, so that the liquid suction assembly 1 and the cover glass capping assembly 2 can be respectively conveyed to a specified working position along with the movement of the sliding rail 52.
As shown in the structure of fig. 1-4, the washing liquid component 1 comprises a TIP head picking hole 11 arranged at the position of the opposite surface (i.e. the close surface) of the washing liquid component 1 and the working bracket 10, and is used for picking up a required TIP head 12 at a fixed point, and under the control of a reagent extraction injection pump 17 communicated with the TIP head picking hole 11, the liquid flow power during liquid suction and liquid discharge is provided, and the required sample, reagent and the like are extracted and filled at the corresponding position at the fixed point.
The washing solution assembly 1 slides along the mechanical arm slide rail assembly 5 under the driving and control of the linear motor 13 and the main control board 14, drops a sample and a reagent to be processed to a designated position, and can control the picking and pushing of the TIP head 12. The washing liquid component 1 further comprises a position judgment sensor 15 which comprises 1 group of optical fiber sensors and 1 group of color difference sensors and is used for detecting the placement of consumables, detecting the state of a sample in the filter and judging whether the sample and the consumables are placed in proportion. The suction cup pressure sensor 16 can be used to determine whether the negative pressure state of the whole module meets the requirement.
As a preferred configuration, in the integrated system of the present invention, the TIP head pick-up holes 11 may be arranged in at least 2, preferably 6 as shown in fig. 1 to 4, regularly arranged, and more preferably arranged corresponding to the number and positions of the slides at the position of the slide placing region 41 of the heating assembly 4. The dyeing liquid can be contained in an independent liquid storage tank to be convenient to take and use, and the liquid storage tank can also be arranged at a fixed position of the whole device.
In the structure shown in fig. 1-4, the cover slip capping assembly 2 comprises a cover slip picking suction cup 21 disposed at a position opposite to (i.e. proximate to) the working support 10 of the cover slip capping assembly 2, the cover slip picking suction cup 21 is controlled by a cover slip picking suction cup driving motor 22 to realize fixed-point picking of a desired cover slip and to cap the corresponding slide, and the cover slip picking suction cup 21 is provided with a negative pressure by a diaphragm pump 23.
In order to control the suction and placement of the glass slide, in the system, the cover glass picking suction disc 21 is connected with a tee joint through a pipeline and is respectively communicated with the diaphragm pump 23, the air release valve and the barometer, the suction and placement of the required cover glass are realized by controlling the alternate work of the diaphragm pump 23 and the air release valve, and whether the effective cover glass is picked by the cover glass picking suction disc 21 is monitored by reading the air pressure data in the pipeline. Specifically, when the cover glass picking suction disc 21 needs to pick up a required cover glass, the diaphragm pump 23 is started to extract negative pressure in the pipeline, so that the cover glass is picked up; when the cover glass is transferred to a preset position, the diaphragm pump 23 is controlled to stop working and the air escape valve is opened, so that the pressure in the pipeline is recovered, and the sucked cover glass can be separated from the cover glass pickup suction cup 21 and placed at the position of the slide glass; meanwhile, when the cover glass picking suction disc 21 sucks the cover glass, the barometer can monitor the pressure in the pipeline, and whether the cover glass picking suction disc 21 takes the effective cover glass is further monitored by observing the read air pressure value. Preferably, the surface of the cover glass picking suction disc 21 for sucking the cover glass is provided with a suction surface with a certain inclination angle, so that bubbles can be reduced in the cover glass placing process.
With the structure shown in fig. 5, the cell separation device of the present invention comprises a cell filtration assembly 6 and a cleaning assembly 7 adapted to enrich the CTCs tumor cells in the sample to be separated and separate the CTCs tumor cells for preparing a slide. Specifically, the cell filter 61 is fixed on the working support 10 of the whole system through the cell filter interface 62. Cell separation subassembly 6 includes that inside is provided with filtration membrane's cell filter 61, and the CTCs tumor cell of treating the separation is along with lotion subassembly 1 is sucked extremely in the cell filter 61 form the negative pressure under the effect of pipeline pinch valve 63, carry out the cell and subside and filter, and enrich in filtration membrane department, and the setting pipeline monitoring module 64 then is used for monitoring the pressure in the pipeline and the circulation condition.
In order to avoid that a lot of non-CTCs tumor cells are easily left on the back surface of the filtering membrane after cell separation in the cell enrichment process and bring great interference to the reading process after staining, the cleaning assembly 7 of the system comprises a membrane-below cleaning needle 71 arranged below the cell filter 61, so that the back surface of the enrichment surface of the filtering membrane can be cleaned, the water hammer effect can be reduced, the impact on the filtering membrane during sample filtration is avoided, and the cleaned liquid is temporarily stored in a waste liquid temporary storage cavity 72 communicated with the cell filter 61.
As shown in fig. 6-8, the specimen slide preparation device of the present invention comprises a heating assembly 4 disposed on a workbench and a cover glass extraction assembly 3 for controlling automatic extraction of a desired cover glass, and is used for staining, preparing and mounting the enriched target cells to obtain a specimen sample that can be directly used for observation.
In the structure shown in fig. 6-8, the cover glass extracting assembly 3 comprises a cover glass placing frame 31 arranged on a workbench, and the inner space of the cover glass placing frame 31 forms a cover glass stacking area 33 capable of accommodating the cover glass; coverslips are collectively stacked in the coverslip stack area 33, biased for on-demand output. The bottom position of the cover glass placing frame 31 and a gap which can only be pushed out by a single cover glass is arranged between the planes of the working tables, a cover glass push rod 35 is arranged on one side of the cover glass placing frame 31, the cover glass push rod 35 is arranged on the surface of the working table and corresponds to the gap position of the cover glass placing frame 31, and the cover glass push rod 35 can reciprocate on the surface of the working table along a preset direction under the control of a cover glass push rod motor 36 (an optional linear motor) arranged on the working table, so that the cover glass push rod 35 can penetrate through the gap to push the cover glass on the lowest layer in the cover glass stacking area 33 to move out and be pushed to the preset position, and is deviated from the picking of the cover glass picking sucker 21.
In order to control the number of the cover slips during operation, in the system of the present invention, the cover slip extracting assembly 3 further includes a cover slip pressing block 32 disposed in the cover slip placing frame 31 and capable of sliding along the inner wall of the cover slip placing frame 31, when the cover slip placing frame 31 is filled with the cover slips, the cover slip pressing block 32 is pushed to the top end position of the cover slips and slides down along with continuous pushing of the cover slips, and when the cover slip pressing block 32 descends to a preset height, a cover slip allowance alarm 34 disposed on the side surface of the cover slip placing frame 31 can be triggered to remind an operator to replenish the cover slips.
In the configuration shown in fig. 6-8, the heating assembly 4 includes a heating stage 42 disposed on the stage, and a slide placement area 41 is disposed on the heating stage 42. The heating table 42 may be a conventional heating table disposed on the upper surface of the working table, and is usually electrically controlled to heat the specimen after the staining solution is added and the cover glass is covered, so as to cure the staining solution and complete the sealing process. The heating temperature of the heating table 42 is preferably controlled so as not to affect the shape of the specimen, and can be controlled by itself. In order to facilitate the efficiency of the automatic operation, the slide placing areas 41 may be arranged in multiple groups or multiple groups, and more preferably, the arrangement of the slide placing areas 41 is arranged in a matrix structure as shown in fig. 6 to 8, and the arrangement of the slide placing areas 41 is preferably arranged in the same regular arrangement as the arrangement of the TIP head picking holes 11, so that multiple groups of samples can be conveniently added at the same time.
In order to collect and recover the dyeing solution waste liquid after the slide is produced, waste liquid sucker slide rails 43 are further arranged on two side faces of the heating table top 42, and can drive the waste liquid suckers 44 to the specified slide glass position to pump and remove the waste reagents produced during the dyeing.
As shown in the structure of figure 1, the integrated system for cell enrichment, separation, staining and slide production further comprises a consumable material placing assembly, so that various consumable materials involved in the whole operation process can be placed on corresponding supports for standardized management and corresponding automatic control. Specifically, the consumable holding assembly comprises a TIP head placing rack 81, a reagent placing rack 82 and/or a sample placing rack 83.
As shown in the structure of figure 1, the integrated system for cell enrichment, separation, staining and slide production further comprises a waste liquid storage component which is mainly used for collecting and storing consumables or reagents after use. In particular, the waste storage assembly includes a TIP waste chamber 91 and/or a waste reagent collection barrel 92.
Each consumable placing component and each waste liquid storage component can be placed at a designated position according to a specifically set automatic program, and more preferably, each consumable placing component and each waste liquid storage component are arranged at the positions shown in fig. 1 in consideration of the occupied space of the whole system, and writing and operation of a corresponding automatic control program are performed according to the arranged positions.
When the integrated system for cell enrichment, separation, staining and slide production is used, a user places a glass slide with a specimen on a glass slide placing area 41 of a heating table 42 and puts a cover glass into a cover glass placing frame 31 in a laminated manner; and reagents such as TIP heads, cell samples to be treated, staining solutions and the like involved in the whole operation process are respectively placed in a designated placing rack or a designated reagent area. After the device is started, the washing liquid assembly 1 is controlled to move to a TIP head placing frame 81 through a mechanical arm slide rail 5, the TIP head picking hole 11 sucks a TIP head 12 at a specified position, the TIP head 12 is used for sucking a cell sample to be separated at the sample placing frame 83 under the control of the washing liquid assembly 1, the cell sample to be separated is placed in the cell filter 61, the cell sample to be separated is subjected to sedimentation filtration of cells, the treated cells are enriched on a filtering membrane in the cell filter 61, and then the membrane under-membrane cleaning needle 71 is started to clean the back surface of the filtering membrane. The filter membrane in the cell filter 61 is then manually removed and placed on the slide at the slide placement area 41. At this time, the TIP pick-up hole 11 pushes out the used TIP and places the TIP into the TIP discard chamber 91, and the mechanical arm slide rail 5 picks up the TIP again and then sucks the staining solution from the reagent rack 82 to stain the filter membrane at a specified position. Meanwhile, a cover glass push rod motor 36 drives the cover glass push rod 35 to perform horizontal linear motion, so as to push the cover glass in the cover glass placing frame 31 to move out of a gap at the lower part of the cover glass placing frame and push the cover glass to a preset position; at this moment, the cover glass picks up the sucking disc driving motor 22 and drives the cover glass picks up the sucking disc 21 motion, and be the negative pressure in order to extract the pipeline through starting the diaphragm pump, realize picking up of cover glass, the cover glass that is absorb moves to slide glass department along with the arm device, through controlling diaphragm pump stop work and opening the release valve, make pipeline internal pressure reply, the cover glass that absorbs can break away from this moment the cover glass picks up sucking disc 21 and places in slide glass department, because the cover glass picks up sucking disc 21 and has certain inclination to get, put the cover glass, can reduce when the mounting with the bubble sealed in the sample, control afterwards heating mesa 42 heaies up in order to solidify the staining solution, accomplish automatic mounting process promptly, and the waste liquid suction head 44 that sets up on heating element 4 moves on waste liquid slide rail suction head 43 when absorbing the negative pressure and siphons away the staining solution after dyeing and to abandonment reagent collecting vessel 92 side by side, complete the whole process of cell enrichment, separation, staining and flaking.
It should be understood that the above examples are only for clarity of illustration and are not intended to limit the embodiments. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications therefrom are within the scope of the invention.
Claims (10)
1. An integrated system for cell enrichment, separation, staining and slide production is characterized by comprising an automatic control module, and a mechanical control device, a cell separation device and a specimen slide production device which can be controlled by the automatic control module in a linkage manner; wherein the content of the first and second substances,
the mechanical control device is used for picking up consumables and reagents required in the whole operation process and conveying the consumables and the reagents to a specified position for corresponding operation; the mechanical control device comprises a mechanical arm slide rail assembly (5), a liquid suction assembly (1) which can move to a designated position along the mechanical arm slide rail (5) and is used for picking up required consumables and reagents, and a cover glass capping assembly (2) used for picking up required cover glass;
the cell separation device comprises a cell filtering component (6) and a cleaning component (7) which is adapted to enrich target cells in a sample to be separated and separate the target cells to obtain target cells which can be used for preparing slices;
the specimen slide making device comprises a heating assembly (4) arranged on a workbench and a cover glass extracting assembly (3) for controlling automatic extraction of required cover glass, and is used for dyeing, making a slide and sealing the enriched target cells to obtain specimen samples which can be directly used for observation.
2. The integrated system for cell enrichment, separation, staining and flaking according to claim 1, wherein the robot arm slide rail assembly (5) comprises a set of fixed rails (51) disposed in parallel, and a slide rail (52) disposed vertically between the fixed rails (51) and movable in a vertical direction along the fixed rails (51);
the liquid suction assembly (1) and the cover glass capping assembly (2) are respectively arranged on two sides of the sliding rail (52) and are respectively conveyed to a specified working position along with the movement of the sliding rail (52).
3. The integrated system for cell enrichment, separation, staining and slide production according to claim 2, wherein the pipetting module (1) comprises a TIP pick-up hole (11), a TIP head (12) is required to pick up and push out under the control of a linear motor (13), and a reagent extraction injection pump (17) communicated with the TIP pick-up hole (11) is used to extract and fill a required reagent at a designated position at a fixed point.
4. The integrated system for cell enrichment, separation, staining and slide production according to claim 2 or 3, wherein the cover glass capping assembly (2) comprises a cover glass picking suction cup (21), and realizes the fixed-point picking of the required cover glass and caps the cover glass at the corresponding slide under the control of a cover glass picking suction cup driving motor (22);
the cover glass picking suction disc (21) is respectively communicated with the diaphragm pump (23) and the corresponding air release valve through a pipeline, and the suction and the placement of the required cover glass are realized by controlling the alternate work of the diaphragm pump (23) and the corresponding air release valve.
5. The integrated system for cell enrichment, separation, staining and tableting according to any one of claims 1 to 4, wherein the cell separation module (6) comprises a cell filter (61) with a filtering membrane arranged inside, and after target cells are injected into the cell filter (61), negative pressure is formed under the action of a pipeline pinch valve (63) to perform cell sedimentation filtration and enrich at the filtering membrane.
6. The integrated system for cell enrichment, separation, staining and flaking according to claim 5, characterized in that the cleaning assembly (7) comprises a membrane-under-membrane cleaning needle (71) disposed below the cell filter (61) and capable of cleaning the back surface of the enrichment surface of the filtration membrane, and the cleaned liquid is temporarily stored in a waste liquid temporary storage chamber (72) communicated with the cell filter (61).
7. Integrated system for cell enrichment, separation, staining, production according to any of claims 1-6, characterized in that the coverslip extraction module (3) comprises a coverslip rack (31) arranged on a work bench, inside which a coverslip stacking zone (33) is formed, which can accommodate the coverslips;
a gap which can only be pushed out by a single cover glass is arranged between the cover glass placing frame (31) and the plane of the workbench, and a cover glass push rod (35) which is arranged on the workbench and corresponds to the gap pushes the required cover glass to move out of the cover glass placing frame (31) to a fixed point position;
be provided with in cover glass rack (31) and follow the gliding cover glass briquetting (32) of cover glass rack (31) inner wall, cover glass briquetting (32) along with the continuous release of cover glass and the decline that slides work as cover glass briquetting (32) descend to when predetermineeing the height can trigger set up in cover glass surplus alarm (34) of cover glass rack (31) side reminds the replenishment cover glass.
8. The integrated system for cell enrichment, separation, staining and production according to claim 7, wherein the heating assembly (4) comprises a heating table top (42) disposed on the work stage, and a slide placement area (41) is disposed on the heating table top (42);
the heating table top (42) is also provided with a waste liquid sucker slide rail (43) which can drive a waste liquid sucker (44) to a designated glass slide position and pump and remove waste reagents generated during dyeing.
9. The integrated system for cell enrichment, separation, staining and pelletisation according to any of claims 1-8, wherein the system further comprises a consumable placement assembly;
the consumable placing assembly comprises a TIP head placing frame (81), a reagent placing frame (82) and/or a sample placing frame (83).
10. The integrated system for cell enrichment, separation, staining and sheet production according to any one of claims 1 to 9, wherein the system further comprises a waste storage assembly;
the waste storage assembly includes a TIP waste chamber (91) and/or a waste reagent collection barrel (92).
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