CN110988328A

CN110988328A - Full-automatic fluorescence labeling single cell suspension preparation device and matched sample loading tube

Info

Publication number: CN110988328A
Application number: CN201911413259.8A
Authority: CN
Inventors: 周汉宇; 李昕
Original assignee: Individual
Current assignee: Individual
Priority date: 2019-12-31
Filing date: 2019-12-31
Publication date: 2020-04-10

Abstract

The invention discloses a full-automatic fluorescence labeling single cell suspension preparation device and a matched upper sample tube thereof. The device and the matched sample loading tube can automatically process human or animal peripheral blood samples, and comprise the steps of sample loading, dyeing, hemolysis, separation of white blood cells and suspended white blood cells and the like, so that the labor intensity and human errors are reduced. The matched sample loading tube adopts the filter membrane filtration principle, and the injector replaces a vacuum pump to remove red blood cell fragments and waste liquid in batches, so that the separation of the fluorescence-labeled white blood cells can be completed in a short time, the volume of corresponding equipment is reduced, the cost of the vacuum pump is removed, and the space and the cost are saved.

Description

Full-automatic fluorescence labeling single cell suspension preparation device and matched sample loading tube

Technical Field

The invention belongs to the field of biomedical equipment, and particularly relates to a full-automatic fluorescence labeling single cell suspension preparation device controlled by a PLC (programmable logic controller) and a matched upper sampling tube, which are used for automatically treating peripheral blood samples of human beings or animals.

Background

At present, for detecting leukocyte surface antigens in peripheral blood of human or animals, collected peripheral blood samples are required to be processed before detection. The existing processing method is roughly divided into five steps of blood sample suction and adding into an experimental vessel, fluorescent labeling antibody staining, hemolysin (or hemolytic agent) adding, marked white blood cell separation and suspension marked white blood cell in liquid, and the like, and the experimental process of the processing has the characteristics of precious experimental consumables, complex experimental process, higher requirements on sample adding volume and reaction time accuracy, and the like.

For the step of separating the labeled leukocytes, several separation methods are commonly used at present, such as centrifugation, density gradient separation, magnetic bead adsorption, and filter membrane filtration. Although the centrifugation method is economical and convenient, a centrifuge is required to be equipped, and white blood cells are damaged and lost; the density gradient separation solution method and the magnetic bead adsorption method require expensive separation solution and adsorption magnetic beads, take longer time and cannot avoid the loss of white blood cells; the membrane filtration method requires an additional filtration vessel and a vacuum pump. The centrifuge is a stock device in a laboratory, so the separation of leukocytes by the centrifugation method is the most common at present, but the method needs to manually transfer a laboratory vessel into the centrifuge for centrifugation, and the laboratory vessel still needs to be manually sorted after centrifugation so as to continue to be loaded on a sample machine for detection, so that the labor consumption is high, and mistakes are easy to make.

At present, some sample processing devices replacing manual work are on the market, for example, after staining and hemolysis, the step of separating white blood cells is omitted, and the sample is directly tested on a computer, which is called as a washing-free mode.

Disclosure of Invention

Aiming at the problems in the prior art, the invention provides a full-automatic fluorescence labeling single cell suspension preparation device and a matched sample loading tube, which can automatically process human or animal peripheral blood samples including sample loading, dyeing, hemolysis, separation of white blood cells and suspension of white blood cells in liquid, so that a sample can be directly tested on a machine.

In order to solve the technical problems and achieve the technical effects, the invention is realized by the following technical scheme:

a full-automatic fluorescence labeling single cell suspension preparation device comprises a sample loading pipe frame rotating device, a sample loading pipe frame top cover, a sample adding device and a comprehensive controller;

the sample loading pipe rack rotating device comprises a sample loading pipe rack supporting rod used for installing the sample loading pipe rack, a sample loading pipe rack rotating mechanism used for driving the sample loading pipe rack to rotate on the sample loading pipe rack supporting rod is arranged at the lower part of the sample loading pipe rack supporting rod, a sample loading pipe rack chassis is movably sleeved at the middle part of the sample loading pipe rack supporting rod, a sample loading pipe rack chassis rotating mechanism used for driving the sample loading pipe rack chassis to rotate on the sample loading pipe rack supporting rod is arranged on the lower surface of the sample loading pipe rack chassis, and a sample loading pipe rack chassis vertical moving mechanism used for driving the sample loading pipe rack chassis to vertically move on the sample loading pipe rack supporting rod is arranged on the lower surface of the sample loading pipe rack chassis rotating mechanism; a plurality of sample loading tube injector rod push plate through holes which are communicated up and down are uniformly formed in the circumferential edge of the sample loading tube frame chassis, one side of each sample loading tube injector rod push plate through hole in the clockwise direction is provided with a sample loading tube injector rod limiting hole which is communicated up and down, and each sample loading tube injector rod limiting hole is communicated with the corresponding sample loading tube injector rod push plate through hole;

the sample loading pipe support is coaxially and rotatably arranged at the upper part of the sample loading pipe support supporting rod, a sample loading pipe support top cover positioning column used for installing a sample loading pipe support top cover is arranged on the circle center of the upper surface of the sample loading pipe support, a circle of boss is arranged on the circumferential edge of the upper surface of the sample loading pipe support, a plurality of sample pipe installation holes are uniformly formed in the upper surface of the boss, a plurality of sample loading pipe installation holes are formed in the upper surface of the sample loading pipe support along the inner side of the boss, a sample loading pipe injector rod through hole which penetrates up and down is formed in the bottom of each sample loading pipe installation hole, and each sample loading pipe installation hole is respectively coaxial with the corresponding sample loading pipe injector rod through hole up and down; the number of the sample tube mounting holes corresponds to that of the sample tube injector rod through holes, and when the sample tube rack is fixed on the sample tube rack supporting rod, each sample tube mounting hole is respectively coaxial with the sample tube injector rod through hole which corresponds to the sample tube rack mounting hole up and down; the number of the sample loading pipe mounting holes corresponds to the number of the sample pipe mounting holes, and the distribution position of each sample loading pipe mounting hole corresponds to the distribution position of the corresponding sample pipe mounting hole;

the top cover of the sample loading pipe rack is coaxially and rotatably arranged on the sample loading pipe rack, a top cover handle of the sample loading pipe rack is arranged on the circle center of the upper surface of the top cover of the sample loading pipe rack, a plurality of sample loading pipe through holes which are communicated up and down are uniformly arranged on the circumferential edge of the top cover of the sample loading pipe rack, the number of the sample loading pipe through holes corresponds to the number of the sample loading pipe mounting holes, and when the top cover of the sample loading pipe rack is fixed on the sample loading pipe rack, each sample loading pipe through hole is respectively coaxial with the corresponding sample loading pipe mounting hole up and down;

the sample adding device is arranged above the top cover of the sample adding pipe frame and comprises a sample adding device integral vertical moving mechanism, and a hemolysin/buffer solution micro-injection pump fixing rod, a sample micro-injection pump moving rod and an antibody micro-injection pump fixing rod are arranged side by side at the lower part of the sample adding device integral vertical moving mechanism; the front end of the hemolysin/buffer solution micro-injection pump fixing rod is provided with a hemolysin/buffer solution micro-injection pump through a hemolysin/buffer solution micro-injection pump fixing block, the lower end of the hemolysin/buffer solution micro-injection pump is provided with a hemolysin/buffer solution loading needle, the upper end of the hemolysin/buffer solution micro-injection pump is provided with a hemolysin/buffer solution loading needle vertical moving mechanism for driving the hemolysin/buffer solution loading needle to vertically extend and retract, and the hemolysin/buffer solution loading needle is provided with a hemolysin/buffer solution loading needle cleaning mechanism capable of vertically moving; the front end of the fixing rod of the antibody micro-injection pump is provided with a No. 1 antibody micro-injection pump and a No. 2 antibody micro-injection pump through an antibody micro-injection pump fixing block, the lower ends of the No. 1 antibody micro-injection pump and the No. 2 antibody micro-injection pump are respectively provided with a No. 1 antibody sampling needle and a No. 2 antibody sampling needle, and the upper ends of the No. 1 antibody micro-injection pump and the No. 2 antibody micro-injection pump are respectively provided with a No. 1 antibody sampling needle vertical moving mechanism and a No. 2 antibody sampling needle vertical moving mechanism which are used for driving the No. 1 antibody sampling needle and the No. 2 antibody sampling needle to vertically extend and retract; a movable horizontal sample micro-injection pump moving mechanism is arranged on the sample micro-injection pump moving rod, a sample micro-injection pump is arranged on the horizontal sample micro-injection pump moving mechanism, a sample loading needle is arranged at the lower end of the sample micro-injection pump, a vertical sample loading needle moving mechanism used for driving the sample loading needle to stretch up and down is arranged at the upper end of the sample micro-injection pump, and a vertically movable sample loading needle cleaning mechanism is arranged on the sample loading needle; when the horizontal moving mechanism of the sample micro-injection pump moves to a rear positioning point of the moving rod of the sample micro-injection pump, the hemolysin/buffer solution sample adding needle, the No. 1 antibody sample adding needle and the No. 2 antibody sample adding needle are arranged in an arc shape and respectively correspond to the plane positions of any four adjacent sample adding tube through holes on the top cover of the sample adding tube rack up and down one by one, and when the horizontal moving mechanism of the sample micro-injection pump moves to a front positioning point of the moving rod of the sample micro-injection pump, the sample adding needle extends forwards and vertically corresponds to one sample tube mounting hole on the sample adding tube rack; the sample adding device is characterized in that a reagent bottle base is arranged in the middle of the integral vertical moving mechanism, a hemolysin reagent bottle, a buffer solution reagent bottle, a No. 1 antibody reagent bottle and a No. 2 antibody reagent bottle are respectively arranged on the reagent bottle base, the hemolysin reagent bottle and the buffer solution reagent bottle are respectively connected with the hemolysin/buffer solution micro-injection pump through pipelines, the No. 1 antibody reagent bottle is connected with the No. 1 antibody micro-injection pump through a pipeline, and the No. 2 antibody reagent bottle is connected with the No. 2 antibody micro-injection pump through a pipeline;

the integrated controller takes a PLC as a core controller, and is respectively connected with the sample loading pipe frame rotating mechanism, the sample loading pipe frame chassis vertical moving mechanism, the sample adding device integral vertical moving mechanism, the hemolysin/buffer solution micro-injection pump, the hemolysin/buffer solution sample adding needle vertical moving mechanism and the hemolysin/buffer solution sample adding needle cleaning mechanism, the sample injection device comprises a No. 1 antibody micro-injection pump, a No. 2 antibody micro-injection pump, a No. 1 antibody sample injection needle vertical moving mechanism, a No. 2 antibody sample injection needle vertical moving mechanism, a sample micro-injection pump horizontal moving mechanism, a sample micro-injection pump, a sample injection needle vertical moving mechanism and a sample injection needle cleaning mechanism which are electrically connected so as to coordinate the linkage work of the various motion mechanisms.

Furthermore, the number of the sample tube mounting holes, the sample loading tube mounting holes, the injector rod via holes, the injector rod push plate limiting holes and the sample loading tube via holes is 40.

Furthermore, the integrated controller is electrically connected with one multimedia interaction device, so that an operator can set an experiment and display the experiment progress conveniently.

Further, the side surface on sample loading pipe support bracing piece upper portion is provided with two relatively and is used for fixing sample loading pipe support draw-in grooves of sample loading pipe support, it stretches into the recess to be provided with a sample loading pipe support bracing piece on the centre of a circle of sample loading pipe support lower surface, it is provided with two telescopic sample loading pipe support jamming columns relatively to stretch into on the cell wall of recess to go up sample loading pipe support bracing piece, the side surface of sample loading pipe support top cap reference column is provided with two relatively and is used for controlling two the flexible sample loading pipe support jamming column button of sample loading pipe support jamming column, the sample loading pipe support passes through sample loading pipe support bracing piece stretches into the recess cover and establishes the upper portion of sample loading pipe support bracing piece, and through two sample loading pipe support jamming columns and two the cooperation of sample loading pipe support draw-in grooves realize with sample loading pipe.

Further, be provided with a fixed recess of sample loading pipe support top cap on the centre of a circle of sample loading pipe support top cap reference column upper surface, be provided with two sample loading pipe support top cap draw-in grooves on the cell wall of sample loading pipe support top cap fixed recess relatively, be provided with a sample loading pipe support top cap bracing piece on the centre of a circle of sample loading pipe support top cap lower surface, the side surface of sample loading pipe support top cap bracing piece is provided with two telescopic sample loading pipe support top cap jamming posts relatively, the side surface of sample loading pipe support top cap handle is provided with two relatively and is used for controlling two the flexible sample loading pipe support top cap jamming post button of sample loading pipe support top cap jamming post, sample loading pipe support top cap passes through sample loading pipe support top cap bracing piece is installed in the sample loading pipe support top cap reference column in the fixed recess of sample loading pipe support top cap, and through two sample loading top cap jamming posts and two the cooperation of sample loading pipe support top cap draw-in.

The utility model provides a supporting appearance pipe of going up of full-automatic fluorescence labeling unicellular suspension preparation facilities, includes one goes up the appearance pipe body, the middle part in going up the appearance pipe body is provided with one and can be punctured be used for cell fluorescence antibody dyeing's incubation cup, the lower part in going up the appearance pipe body is provided with a mobilizable syringe pole, the lower part of syringe pole is provided with syringe pole push pedal, and syringe pole push pedal exposes the below of going up the appearance pipe body, the circumference surface on syringe pole upper portion be provided with one with go up appearance pipe body inner wall complex syringe pole sealing washer, the inside of syringe pole is a cavity that is used for collecting the waste liquid, the cavity upwards extends in the upper plane of syringe pole forms the trompil, the upper plane of syringe pole is provided with the filter membrane support frame of middle fretwork, the lower surface of filter membrane support frame be provided with one with the trompil complex filter membrane support frame sealing washer of cavity, the upper surface of the filter membrane support frame is provided with a layer of filter membrane for filtering erythrocyte fragments and waste liquid.

Furthermore, a plurality of support legs are arranged on the outer wall of the incubation cup, and the incubation cup is arranged in the middle of the sample loading tube body through the contact of the support legs and the inner wall of the sample loading tube body.

Furthermore, a cross supporting structure is arranged on the hollowed-out part of the filter membrane supporting frame, the cross supporting structure is connected with the main body part of the filter membrane supporting frame, and the filter membrane covers the main body part of the filter membrane supporting frame and the cross supporting structure together.

Furthermore, the sample loading tube body is made of polystyrene or polypropylene, and the size of the sample loading tube body is the same as that of a common cell sample loading tube in the market.

Furthermore, the incubation cup is made of polyvinyl chloride.

The 40-hole sample tube mounting hole and the sample loading tube mounting hole are taken as examples below, and the working principle and the operation method are as follows:

firstly numbering sample tubes to be processed by an operator, and then sequentially placing numbered sample tubes into sample tube mounting holes from the position 1 of the sample tube mounting hole of the sample tube rack, wherein 40 sample tubes can be placed at most;

then, taking the same number of matched sample loading tubes, sequentially placing the matched sample loading tubes into the corresponding sample loading tube mounting holes on the sample loading tube rack from the position No. 1 of the sample loading tube mounting holes of the sample loading tube rack, and ensuring that the injector rod push plate of each dominant sleeve of sample loading tubes is exposed out of the lower surface of the sample loading tube rack after the injector rod passes through the corresponding injector rod through hole on the sample loading tube rack;

placing a sample loading pipe rack with sample pipes and matched sample loading pipes on a sample loading pipe rack supporting rod, pressing sample loading pipe rack clamping column buttons on two sides of a sample loading pipe rack top cover positioning column to enable the sample loading pipe rack supporting rod to extend into a sample loading pipe rack supporting rod at the bottom of a sample loading pipe rack and extend into a groove, then loosening the sample loading pipe rack clamping column buttons to enable a sample loading pipe rack clamping column with the sample loading pipe rack supporting rod extending into the groove to be clamped with a sample loading pipe rack clamping groove on the sample loading pipe rack supporting rod, so that the sample loading pipe rack is fixed on the sample loading pipe rack supporting rod, and simultaneously, each injector rod push plate sleeved at the bottom of the sample loading pipe rack passes through an injector rod push plate through hole corresponding to the same position on a sample loading pipe rack;

placing a top cover of the sample loading pipe frame on the sample loading pipe frame, pressing sample loading pipe frame top cover clamping column buttons on two sides of a top cover handle of the sample loading pipe frame to enable a sample loading pipe frame top cover supporting rod on the lower surface of the top cover of the sample loading pipe frame to extend into a sample loading pipe frame top cover fixing groove on a sample loading pipe frame top cover positioning column, then loosening the sample loading pipe frame top cover clamping column buttons to enable a sample loading pipe frame top cover clamping column on the sample loading pipe frame top cover supporting rod to be clamped with a sample loading pipe frame top cover clamping groove in the sample loading pipe frame top cover fixing groove, and fixing the sample loading pipe frame top cover on the;

checking whether various reagents in the hemolysin reagent bottle, the buffer solution reagent bottle, the No. 1 antibody reagent bottle and the No. 2 antibody reagent bottle are sufficient or not, confirming whether pipelines are connected and normal or not, then selecting a corresponding experimental scheme and quantity on the multimedia interaction device, and starting an experiment at a point;

the integral vertical moving mechanism of the sample adding device drives the integral vertical downward movement of the sample adding device to be in place, the sample adding needle of the No. 1 antibody is downwards aligned with the sample adding tube on the No. 1 position, then the sample adding needle of the No. 1 antibody injects the antibody A into the sample adding tube on the No. 1 position through the micro-injection pump of the No. 1 antibody, and then the integral vertical moving mechanism of the sample adding device drives the integral vertical upward movement of the sample adding device to return;

the sample loading pipe frame rotating mechanism drives the sample loading pipe frame to rotate anticlockwise for 9 degrees, the integral vertical moving mechanism of the sample loading device drives the sample loading device to integrally vertically move downwards to be in place, at the moment, the No. 1 antibody sample loading needle is downwards aligned with the sample loading pipe matched with the No. 2 position, the No. 2 antibody sample loading needle is downwards aligned with the sample loading pipe matched with the No. 1 position, then the No. 1 antibody sample loading needle injects an antibody A into the sample loading pipe matched with the No. 2 position through the No. 1 antibody micro-injection pump, meanwhile, the No. 2 antibody sample loading needle injects an antibody B into the sample loading pipe matched with the No. 1 position through the No. 2 antibody micro-injection pump, and then the integral vertical moving mechanism of the sample loading;

the sample loading pipe frame rotating mechanism drives the sample loading pipe frame to rotate 9 degrees anticlockwise, the sample micro-injection pump horizontal moving mechanism moves the sample micro-injection pump to a front positioning point of a sample micro-injection pump moving rod, at the moment, a sample loading needle is downwards aligned to a sample pipe at the No. 1 position, then the sample loading needle vertical moving mechanism drives the sample loading needle to vertically move downwards and extend into the sample pipe at the No. 1 position, the sample loading needle sucks a sample in the sample pipe at the No. 1 position through the sample micro-injection pump, then the sample loading needle vertical moving mechanism drives the sample loading needle to vertically move upwards to return, and then the sample micro-injection pump horizontal moving mechanism moves the sample micro-injection pump back to a rear positioning point of the sample micro-injection pump moving rod;

the integral vertical moving mechanism of the sample adding device drives the sample adding device to integrally and vertically move downwards to be in place, at the moment, the sample adding needle is downwards aligned with the sample tube on the matched sleeve at the No. 1 position, the No. 2 antibody adding needle is downwards aligned with the sample tube on the matched sleeve at the No. 2 position, the No. 1 antibody adding needle is downwards aligned with the sample tube on the matched sleeve at the No. 3 position, then the sample adding needle vertical moving mechanism drives the sample adding needle to vertically move downwards and extend into the matched sample loading tube at the No. 1 position, the sample adding needle injects the sucked sample into the matched sample loading tube at the No. 1 position through a sample micro-injection pump, then the sample adding needle is pumped by a sample micro-injection pump once, the antibody A and the antibody B on the sample tube on the No. 1 position are uniformly mixed with the sample, simultaneously, the No. 2 antibody sampling needle injects the antibody B into the matched sample loading tube at the No. 2 position, and the No. 1 antibody sampling needle injects the antibody A into the matched sample loading tube at the No. 3 position;

the sample loading needle vertical moving mechanism drives the sample loading needle to vertically move upwards for returning, then the sample loading device integral vertical moving mechanism drives the sample loading device integral vertical moving upwards for returning, the sample loading needle cleaning mechanism cleans the sample loading needle, and the steps are repeated until all sample loading tubes on the sample loading tube rack are uniformly loaded with the antibody A, the antibody B and the sample and are uniformly mixed;

after the antibody A, the antibody B and the sample are added into all matched sample loading tubes and are uniformly mixed, the sample loading tube frame rotating mechanism drives the sample loading tube frame to rotate clockwise by 9 degrees and then rapidly rotate anticlockwise by 9 degrees for 3 cycles, the mixed solution of the antibody A, the antibody B and the blood sample in all matched sample loading tubes is uniformly oscillated, and then the mixed solution of the antibody A, the antibody B and the blood sample after uniform oscillation is incubated for 15 minutes; injecting, mixing, oscillating and incubating the antibody A and the antibody B and the blood sample in each matched sample loading tube in an incubation cup inside the sample loading tube;

the sample loading pipe frame rotating mechanism drives the sample loading pipe frame to enable the hemolysin/buffer solution sample adding needle to be downwards aligned to the sample loading pipe on the No. 1 position, then the hemolysin/buffer solution sample adding needle vertical moving mechanism drives the hemolysin/buffer solution sample adding needle to vertically move downwards to a first height position, and the hemolysin/buffer solution sample adding needle injects hemolysin into the incubation cup of the sample loading pipe on the No. 1 position through the hemolysin/buffer solution micro-injection pump;

the hemolysin/buffer solution sample adding needle vertical moving mechanism further drives the hemolysin/buffer solution sample adding needle to vertically move downwards to a second height position, in the process that the hemolysin/buffer solution sample adding needle vertically moves downwards to the second height position, the hemolysin/buffer solution sample adding needle punctures the incubation cup in the sample adding tube on the No. 1 position, then the hemolysin/buffer solution sample adding needle is pumped by the hemolysin/buffer solution micro-injection pump once, and the hemolysin is uniformly mixed with the mixed solution of the antibody A, the antibody B and the blood sample;

the hemolysin/buffer solution sample adding needle vertical moving mechanism drives the hemolysin/buffer solution sample adding needle to vertically move upwards, the hemolysin/buffer solution sample adding needle is reset after passing through a first height position, the hemolysin/buffer solution sample adding needle is cleaned by the hemolysin/buffer solution sample adding needle cleaning mechanism, then the sample loading tube rack rotating mechanism drives the sample loading tube rack to rotate anticlockwise for 9 degrees, so that the hemolysin/buffer solution sample adding needle is downwards aligned with the matched sample loading tube at the No. 2 position, the circulation is carried out until the hemolysin is injected into all the matched sample loading tubes and uniformly mixed, and all the incubation cups in the matched sample loading tubes are punctured;

injecting hemolysin into all matched sample loading tubes and uniformly mixing, driving the sample loading tube rack to rotate clockwise by 9 degrees and then rapidly rotate anticlockwise by 9 degrees by a rotating mechanism of the sample loading tube rack, continuing for 3 cycles, oscillating and uniformly mixing the mixed solution of the hemolysin, the antibody A, the antibody B and the blood sample in all matched sample loading tubes, and then incubating the mixed solution of the hemolysin, the antibody A, the antibody B and the blood sample after oscillating and uniformly mixing for 15 minutes;

the upper sample tube rack chassis rotating mechanism drives the upper sample tube rack chassis to rotate 9 degrees anticlockwise, so that the injector rods of all matched upper sample tubes are clamped into corresponding injector rod limiting holes on the upper sample tube rack chassis, then the upper sample tube rack chassis vertical moving mechanism drives the upper sample tube rack chassis and the upper sample tube rack chassis rotating mechanism to move together and vertically move downwards, and the upper sample tube rack chassis pulls all the injector rods of the matched upper sample tubes to vertically move downwards for a certain distance, so as to cause negative pressure in the matched upper sample tubes;

under the action of negative pressure, the mixed solution of the incubated hemolysin, the antibody A, the antibody B and the blood sample breaks through the perforation on the incubation cup and all flows into the cavity at the lower part of the matched sample loading tube between the incubation cup and the filter membrane, and under the action of gravity, the red cell fragments and the waste liquid in the mixed solution of the hemolysin, the antibody A, the antibody B and the blood sample flow into the cavity in the injector rod through the filter membrane, and the white cells in the mixed solution of the hemolysin, the antibody A, the antibody B and the blood sample are internally intercepted on the filter membrane;

the vertical moving mechanism of the upper sample pipe rack chassis drives the upper sample pipe rack chassis and the rotating mechanism of the upper sample pipe rack chassis to move together and vertically move upwards for returning, the upper sample pipe rack chassis pushes all the injector rods matched with the sample tubes to vertically move upwards for returning, and the rotating mechanism of the upper sample pipe rack chassis drives the upper sample pipe rack chassis to clockwise rotate 9 degrees, so that all the injector rods matched with the sample tubes and the corresponding injector rod limiting holes on the upper sample pipe rack chassis are separated for returning;

the sample loading tube frame rotating mechanism drives the sample loading tube frame to enable the hemolysin/buffer solution sample adding needle to be aligned to the sample loading tube on the No. 1 position downwards again, then the hemolysin/buffer solution sample adding needle vertical moving mechanism drives the hemolysin/buffer solution sample adding needle to vertically move downwards to a first height position, the hemolysin/buffer solution sample adding needle injects buffer solution into the sample loading tube on the No. 1 position through the hemolysin/buffer solution micro-injection pump, then the hemolysin/buffer solution sample adding needle pumps the buffer solution through the hemolysin/buffer solution micro-injection pump once, and the buffer solution and the white blood cells on the filter membrane are mixed uniformly;

the hemolysin/buffer solution sample adding needle vertical moving mechanism drives the hemolysin/buffer solution sample adding needle to vertically move upwards and return, the hemolysin/buffer solution sample adding needle cleaning mechanism cleans the hemolysin/buffer solution sample adding needle, then the sample loading pipe frame rotating mechanism drives the sample loading pipe frame to rotate anticlockwise for 9 degrees, so that the hemolysin/buffer solution sample adding needle is downwards aligned to the matched sample loading pipe at the No. 2 position, and the steps are repeated until buffer solution is injected into all the matched sample loading pipes and is uniformly mixed with the white blood cells on the corresponding filter membrane;

after all matched sample loading tubes are injected with buffer solution and uniformly mixed, the rotating mechanism of the sample loading tube rack drives the sample loading tube rack to rotate clockwise for 9 degrees and then rotate anticlockwise for 9 degrees rapidly, and the 3 cycles are continued, and the buffer solution and the white blood cells in all matched sample loading tubes are uniformly oscillated to form white blood cell single cell suspension marked by fluorescence;

and finally, the operator takes the sample loading pipe frame down from the sample loading pipe frame rotating device and moves the sample loading pipe frame to the detection device for direct detection.

All the movement mechanisms on the full-automatic fluorescence labeling single-cell suspension preparation device are controlled by a comprehensive controller taking a PLC as a core, and the comprehensive controller controls and coordinates the work of all the movement mechanisms according to a preset experiment program, so that the steps of sample adding, hemolysis, separation of white blood cells and suspended white blood cells and the like are smoothly finished, and the automation of the treatment of the peripheral blood sample of human or animals is realized.

The invention has the beneficial effects that:

1. the full-automatic fluorescence labeling single cell suspension preparation device and the matched set sample-sleeving tube thereof can automatically process human or animal peripheral blood samples, and comprise the steps of sample adding, dyeing, hemolysis, separation of white blood cells and suspended white blood cells and the like, thereby greatly reducing the labor intensity of workers and effectively reducing errors caused by manual operation.

2. The full-automatic fluorescence labeling single-cell suspension preparation device adopts the single-shaft movement of the sample loading pipe frame rotating device relative to the sample loading device, and the sample loading needles on the sample loading device are arranged in an arc shape and correspond to the arrangement radian of sample loading pipe holes on the sample loading pipe frame, so that the positioning error of two-dimensional movement of two shafts in the sample sucking and sample loading processes of the traditional device is greatly reduced, the configuration of a stepping motor is reduced, and the cost is saved.

3. The sample loading pipe frame of the full-automatic fluorescence labeling single-cell suspension preparation device can be integrally detached from the sample loading pipe frame rotating device, so that after a sample is processed, a worker can directly place the sample loading pipe frame, a matched sample loading pipe and a sample pipe on a detection device for detection, the work of manual transfer, numbering and the like is greatly reduced, and the manual labor intensity is reduced.

4. The matched upper sample tube integrates the experimental steps of dyeing, hemolysis, separating the white blood cells with fluorescent marks, removing the waste liquid with red blood cell fragments, suspending the white blood cells with fluorescent marks and the like into one sample tube for carrying out, and the matched upper sample tube is designed according to the size of the common cell sample tube on the market at present, thereby reducing the volume of corresponding equipment and saving space.

5. The matched sample loading tube adopts the filter membrane filtration principle, and the injector replaces a vacuum pump to remove red blood cell fragments and waste liquid in batches, so that the separation of the fluorescence-labeled white blood cells can be completed in a very short time, the efficiency can be improved, the error can be reduced, the volume of corresponding equipment can be reduced, the cost of the vacuum pump can be removed, and the space and the cost can be saved.

6. The injector rod matched with the sampling tube is provided with the cavity, so that the functions of vacuumizing and collecting waste liquid can be realized, and the risk of biohazard in the experimental process can be reduced.

The foregoing description is only an overview of the technical solutions of the present invention, and in order to make the technical solutions of the present invention more clearly understood and to implement them in accordance with the contents of the description, the following detailed description is given with reference to the preferred embodiments of the present invention and the accompanying drawings. The detailed description of the present invention is given in detail by the following examples and the accompanying drawings.

Drawings

The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this application, illustrate embodiment(s) of the invention and together with the description serve to explain the invention without limiting the invention. In the drawings:

FIG. 1 is a front perspective view of a fully automatic fluorescence-labeled single-cell suspension preparation device and a sample loading tube used in combination according to the present invention;

FIG. 2 is a side perspective view of the fully automatic fluorescence-labeled single-cell suspension preparation device of the present invention in combination with a sample loading tube;

FIG. 3 is a front view of the fully automatic fluorescence-labeled single-cell suspension preparation device and a sample loading tube used in combination according to the present invention;

FIG. 4 is a left side view of the fully automatic fluorescence-labeled single-cell suspension preparation device of the present invention in combination with a sample loading tube;

FIG. 5 is a top view of the fully automatic fluorescence-labeled single-cell suspension preparation device of the present invention in combination with a sample loading tube;

FIG. 6 is a perspective view of a loading tube rack of the present invention;

FIG. 7 is a top view of a sample loading tube rack of the present invention;

FIG. 8 is an elevation view of a loading tube rack of the present invention;

FIG. 9 is a perspective view of the top cover of the loading tube rack of the present invention;

FIG. 10 is a front view of a top cover of a loading tube rack of the present invention;

FIG. 11 is an exploded view of a mating sample tube of the present invention;

FIG. 12 is a horizontal position relationship of the components of a matching sleeve sample tube of the present invention.

Detailed Description

The present invention will be described in detail below with reference to the accompanying drawings in conjunction with embodiments. The description set forth herein is intended to provide a further understanding of the invention and forms a part of this application and is intended to be an exemplification of the invention and is not intended to limit the invention to the particular embodiments illustrated.

Referring to fig. 1-6, a full-automatic fluorescence labeling single cell suspension preparation device comprises a sample loading pipe frame rotating device, a sample loading pipe frame 1, a sample loading pipe frame top cover 2, a sample adding device and a comprehensive controller;

the sample loading pipe rack rotating device comprises a sample loading pipe rack supporting rod 3 used for installing the sample loading pipe rack 1, two sample loading pipe rack clamping grooves 43 for fixing the sample loading pipe rack 1 are oppositely arranged on the side surface of the upper part of the sample loading pipe rack supporting rod 3, the lower part of the sample loading pipe rack supporting rod 3 is provided with a sample loading pipe rack rotating mechanism 4 for driving the sample loading pipe rack 1 to rotate on the sample loading pipe rack supporting rod 3, the middle part of the upper sample tube rack supporting rod 3 is movably sleeved with an upper sample tube rack chassis 5, the lower surface of the upper sample tube rack chassis 5 is provided with an upper sample tube rack chassis rotating mechanism 6 for driving the upper sample tube rack chassis 5 to rotate on the upper sample tube rack supporting rod 3, the lower surface of the upper sample tube rack chassis rotating mechanism 6 is provided with an upper sample tube rack chassis vertical moving mechanism 7 for driving the upper sample tube rack chassis 5 to vertically move on the upper sample tube rack supporting rod 3; a plurality of upper sample tube injector rod push plate through holes 8 which are communicated up and down are uniformly formed in the circumferential edge of the upper sample tube rack chassis 5, an upper sample tube injector rod limiting hole 9 which is communicated up and down is formed in one side of each upper sample tube injector rod push plate through hole 8 in the clockwise direction, and each upper sample tube injector rod limiting hole 9 is communicated with the corresponding upper sample tube injector rod push plate through hole 8;

the sample loading pipe support 1 is arranged on the sample loading pipe support rotating device, a sample loading pipe support rod extending into the groove 44 is arranged in the circle center of the lower surface of the sample loading pipe support 1, two telescopic sample loading pipe support clamping columns 45 are oppositely arranged on the groove wall of the sample loading pipe support rod extending into the groove 44, the sample loading pipe support 1 is sleeved on the upper part of the sample loading pipe support rod 3 through the sample loading pipe support rod extending into the groove 44, and the sample loading pipe support 1 is fixed with the sample loading pipe support rod 3 through the matching of the two sample loading pipe support clamping columns 45 and the two sample loading pipe support clamping grooves 43; a sample loading pipe rack top cover positioning column 10 for mounting the sample loading pipe rack top cover 2 is arranged on the circle center of the upper surface of the sample loading pipe rack 1, two sample loading pipe rack clamping column buttons 46 for controlling the two sample loading pipe rack clamping columns 45 to stretch and retract are oppositely arranged on the side surface of the sample loading pipe rack top cover positioning column 10, a sample loading pipe rack top cover fixing groove 47 is arranged on the circle center of the upper surface of the sample loading pipe rack top cover positioning column 10, and two sample loading pipe rack top cover clamping grooves 48 are oppositely arranged on the groove wall of the sample loading pipe rack top cover fixing groove 47; a circle of bosses 11 are arranged on the circumferential edge of the upper surface of the upper sample tube rack 1, a plurality of sample tube mounting holes 12 are uniformly formed in the upper surface of each boss 11, a plurality of upper sample tube mounting holes 13 are formed in the upper surface of the upper sample tube rack 1 along the inner sides of the bosses 11, an upper sample tube injector rod through hole 14 which penetrates up and down is formed in the bottom of each upper sample tube mounting hole 13, and each upper sample tube mounting hole 13 is coaxial with the corresponding upper sample tube injector rod through hole 14 up and down; the number of the sample tube mounting holes 12 corresponds to the number of the sample tube injector rod through holes 14, and when the sample tube rack 1 is fixed on the sample tube rack supporting rod 3, each sample tube mounting hole 12 is respectively coaxial with the sample tube injector rod through hole 14 which corresponds to each sample tube mounting hole up and down; the number of the sample loading pipe mounting holes 13 corresponds to the number of the sample pipe mounting holes 12, and the distribution position of each sample loading pipe mounting hole 13 corresponds to the distribution position of the corresponding sample pipe mounting hole 12;

the sample loading pipe rack top cover 2 is arranged on the sample loading pipe rack 1, a sample loading pipe rack top cover supporting rod 49 is arranged on the circle center of the lower surface of the sample loading pipe rack top cover 2, two telescopic sample loading pipe rack top cover clamping columns 50 are oppositely arranged on the side surface of the sample loading pipe rack top cover supporting rod 49, the sample loading pipe rack top cover 2 is installed in the sample loading pipe rack top cover fixing groove 47 of the sample loading pipe rack top cover positioning column 10 through the sample loading pipe rack top cover supporting rod 49, and the sample loading pipe rack top cover positioning column 10 is fixed through the matching of the two sample loading pipe rack top cover clamping columns 50 and the two sample loading pipe rack top cover clamping grooves 48; a sample loading pipe rack top cover handle 15 is arranged at the center of the circle of the upper surface of the sample loading pipe rack top cover 2, and two sample loading pipe rack top cover clamping column buttons 51 for controlling the two sample loading pipe rack top cover clamping columns 50 to stretch and retract are oppositely arranged on the side surface of the sample loading pipe rack top cover handle 15; a plurality of sample loading pipe through holes 16 which are communicated up and down are uniformly formed in the circumferential edge of the sample loading pipe rack top cover 2, the number of the sample loading pipe through holes 16 corresponds to the number of the sample loading pipe mounting holes 13, and when the sample loading pipe rack top cover 2 is fixed on the sample loading pipe rack 1, each sample loading pipe through hole 16 is respectively coaxial with the corresponding sample loading pipe mounting hole 13 up and down;

the sample adding device is arranged above the top cover 2 of the sample adding pipe frame and comprises a sample adding device integral vertical moving mechanism 17, and a hemolysin/buffer solution micro-injection pump fixing rod 18, a sample micro-injection pump moving rod 19 and an antibody micro-injection pump fixing rod 20 are arranged side by side at the lower part of the sample adding device integral vertical moving mechanism 17; the front end of the hemolysin/buffer solution micro-injection pump fixing rod 18 is provided with a hemolysin/buffer solution micro-injection pump 22 through a hemolysin/buffer solution micro-injection pump fixing block 21, the lower end of the hemolysin/buffer solution micro-injection pump 22 is provided with a hemolysin/buffer solution loading needle 23, the upper end of the hemolysin/buffer solution micro-injection pump 22 is provided with a hemolysin/buffer solution loading needle vertical moving mechanism 24 for driving the hemolysin/buffer solution loading needle 23 to vertically extend and retract, and the hemolysin/buffer solution loading needle 23 is provided with a hemolysin/buffer solution loading needle cleaning mechanism 25 capable of vertically moving; the front end of the antibody micro-injection pump fixing rod 20 is provided with a No. 1 antibody micro-injection pump 27 and a No. 2 antibody micro-injection pump 28 through an antibody micro-injection pump fixing block 26, the lower ends of the No. 1 antibody micro-injection pump 27 and the No. 2 antibody micro-injection pump 28 are respectively provided with a No. 1 antibody sampling needle 29 and a No. 2 antibody sampling needle 30, and the upper ends of the No. 1 antibody micro-injection pump 27 and the No. 2 antibody micro-injection pump 28 are respectively provided with a No. 1 antibody sampling needle vertical moving mechanism 31 and a No. 2 antibody sampling needle vertical moving mechanism 32 for driving the No. 1 antibody sampling needle 29 and the No. 2 antibody sampling needle 30 to vertically extend and retract; a movable horizontal sample micro-injection pump moving mechanism 33 is arranged on the sample micro-injection pump moving rod 19, a sample micro-injection pump 34 is arranged on the horizontal sample micro-injection pump moving mechanism 33, a sample loading needle 35 is arranged at the lower end of the sample micro-injection pump 34, a vertical sample loading needle moving mechanism 36 for driving the sample loading needle 35 to stretch up and down is arranged at the upper end of the sample micro-injection pump 34, and a vertically movable sample loading needle cleaning mechanism 37 is arranged on the sample loading needle 35; when the horizontal movement mechanism 33 of the sample microinjection pump moves to a rear positioning point of the moving rod 19 of the sample microinjection pump, the hemolysin/buffer solution sample adding needle 23, the sample adding needle 35, the sample adding needle 29 of antibody No. 1 and the sample adding needle 30 of antibody No. 2 are arranged in an arc shape and respectively correspond to the plane positions of any four adjacent sample adding tube through holes 16 on the top cover 2 of the sample adding tube rack up and down one by one, and when the horizontal movement mechanism 33 of the sample microinjection pump moves to a front positioning point of the moving rod 19 of the sample microinjection pump, the sample adding needle 35 extends forwards and vertically corresponds to one sample tube mounting hole 12 on the sample adding tube rack 1; a reagent bottle base 38 is arranged in the middle of the integral vertical moving mechanism 17 of the sample adding device, a hemolysin reagent bottle 39, a buffer solution reagent bottle 40, a No. 1 antibody reagent bottle 41 and a No. 2 antibody reagent bottle 42 are respectively arranged on the reagent bottle base 38, the hemolysin reagent bottle 39 and the buffer solution reagent bottle 40 are respectively connected with the hemolysin/buffer solution micro-injection pump 22 through pipelines, the No. 1 antibody reagent bottle 41 is connected with the No. 1 antibody micro-injection pump 27 through a pipeline, and the No. 2 antibody reagent bottle 42 is connected with the No. 2 antibody micro-injection pump 28 through a pipeline;

the integrated controller takes a PLC as a core controller, and is electrically connected with the sample loading pipe frame rotating mechanism 4, the sample loading pipe frame chassis rotating mechanism 6, the sample loading pipe frame chassis vertical moving mechanism 7, the whole vertical moving mechanism 17 of the sample loading device, the hemolysin/buffer solution micro-injection pump 22, the hemolysin/buffer solution sample loading needle vertical moving mechanism 24, the hemolysin/buffer solution sample loading needle cleaning mechanism 25, the No. 1 antibody micro-injection pump 27, the No. 2 antibody micro-injection pump 28, the No. 1 antibody sample loading needle vertical moving mechanism 31, the No. 2 antibody sample loading needle vertical moving mechanism 32, the sample micro-injection pump horizontal moving mechanism 33, the sample micro-injection pump 34, the sample loading needle vertical moving mechanism 36 and the sample loading needle cleaning mechanism 37 respectively, so as to coordinate the linkage work of the various motion mechanisms.

Further, the number of the sample tube mounting hole 12, the sample tube mounting hole 13, the sample tube injector rod via hole 14, the sample tube injector rod push plate via hole 8, the sample tube injector rod push plate limiting hole 9 and the sample tube via hole 16 is 40.

A sample loading tube matched with a full-automatic fluorescence labeling single cell suspension preparation device comprises a sample loading tube body 52, wherein a punctured incubation cup 53 for staining cell fluorescent antibodies is arranged in the middle of the sample loading tube body 52, a movable injector rod 54 is arranged at the lower part of the sample loading tube body 52, an injector rod push plate 55 is arranged at the lower part of the injector rod 54, the injector rod push plate 55 is exposed below the sample loading tube body 52, an injector rod sealing ring 56 matched with the inner wall of the sample loading tube body 52 is arranged on the circumferential surface of the upper part of the injector rod 54, a cavity 57 for collecting waste liquid is arranged inside the injector rod 54, the cavity 57 extends upwards to form an open hole on the upper plane of the injector rod 54, and a filter membrane support frame 58 with a hollow middle is arranged on the upper plane of the injector rod 54, the lower surface of the filter membrane support frame 58 is provided with a filter membrane support frame sealing ring 59 matched with the open pore of the cavity 57, and the upper surface of the filter membrane support frame 58 is provided with a layer of filter membrane 60 for filtering erythrocyte fragments and waste liquid.

Further, a plurality of support legs 61 are arranged on the outer wall of the incubation cup 53, and the incubation cup 53 is arranged in the middle of the sample loading tube body 52 through the contact between the support legs 61 and the inner wall of the sample loading tube body 52.

Furthermore, a cross-shaped supporting structure 62 is arranged at the hollowed part of the filter membrane supporting frame 58, the cross-shaped supporting structure 62 is connected with the main part of the filter membrane supporting frame 58, and the filter membrane 60 covers the main part of the filter membrane supporting frame 58 and the cross-shaped supporting structure 62 together.

Further, the size of the loading tube body 52 is the same as the size of the cell loading tube commonly available in the market.

Further, the sample loading tube 52 is made of polystyrene or polystyrene.

Further, the incubation cup 53 is made of polyvinyl chloride.

firstly numbering sample tubes to be processed by an operator, and then sequentially placing numbered sample tubes into the sample tube mounting holes 12 from the position 1 of the sample tube mounting hole 12 of the sample tube rack 1, wherein 40 sample tubes can be placed at most;

then, taking the same number of matched sample tubes, sequentially placing the matched sample tubes into the corresponding sample tube mounting holes 13 on the sample tube rack 1 from the position No. 1 of the sample tube mounting holes 13 of the sample tube rack 1, and ensuring that the injector rod 54 of each domination sleeve sample tube passes through the corresponding sample tube injector rod through hole on the sample tube rack 1, so that the injector rod push plate 55 is exposed out of the lower surface of the sample tube rack 1;

placing a sample loading pipe rack 1 with sample pipes and sample loading pipes matched with the sample loading pipe rack on a sample loading pipe rack support rod 3, pressing sample loading pipe rack clamping column buttons 46 at two sides of a sample loading pipe rack top cover positioning column 10 to enable the sample loading pipe rack support rod 3 to extend into a sample loading pipe rack support rod at the bottom of the sample loading pipe rack 1 to extend into a groove 44, then loosening the sample loading pipe rack clamping column buttons 46 to enable a sample loading pipe rack clamping column 45 with the sample loading pipe rack clamping column extending into the groove 44 to be clamped with a sample loading pipe rack clamping groove 43 on the sample loading pipe rack support rod 3, so that the sample loading pipe rack 1 is fixed on the sample loading pipe rack support rod 3, and simultaneously, an injector rod push plate 55 at the bottom of each dominant sleeve sample loading pipe penetrates through a sample loading pipe injector rod push plate through hole 8 corresponding to the position on a sample loading;

placing a top cover 2 of the sample tube rack on a sample tube rack 1, pressing sample tube rack top cover clamping column buttons 51 on two sides of a top cover handle 15 of the sample tube rack to enable a sample tube rack top cover supporting rod 49 on the lower surface of the top cover 2 of the sample tube rack to extend into a sample tube rack top cover fixing groove 47 on a sample tube rack top cover positioning column 10, then loosening the sample tube rack top cover clamping column buttons 51 to enable a sample tube rack top cover clamping column 50 on the sample tube rack top cover supporting rod 49 to be clamped with a sample tube rack top cover clamping groove 48 in the sample tube rack top cover fixing groove 47, and fixing the sample tube rack top cover 2 on the sample tube rack 1;

checking whether various reagents in a hemolysin reagent bottle 39, a buffer solution reagent bottle 40, a No. 1 antibody reagent bottle 41 and a No. 2 antibody reagent bottle 42 are sufficient or not, confirming whether pipelines are connected and normal or not, then selecting a corresponding experimental scheme and quantity on a multimedia interaction device, and starting an experiment at a point;

the integral vertical moving mechanism 17 of the sample adding device drives the integral vertical moving mechanism 17 of the sample adding device to move downwards to be in place, at the moment, the No. 1 antibody sample adding needle 29 is aligned downwards to the No. 1 matched sample adding tube, then the No. 1 antibody sample adding needle 29 injects the antibody A into the No. 1 matched sample adding tube through the No. 1 antibody micro-injection pump 27, and then the integral vertical moving mechanism 17 of the sample adding device drives the integral vertical moving mechanism 17 of the sample adding device to move upwards to return;

the sample loading pipe frame rotating mechanism 4 drives the sample loading pipe frame 1 to rotate anticlockwise for 9 degrees, the sample loading device integral vertical moving mechanism 17 drives the sample loading device to integrally vertically move downwards to be in place, at the moment, the No. 1 antibody sample loading needle 29 is downwards aligned with the matched sample loading pipe at the No. 2 position, the No. 2 antibody sample loading needle 30 is downwards aligned with the matched sample loading pipe at the No. 1 position, then the No. 1 antibody sample loading needle 29 injects an antibody A into the matched sample loading pipe at the No. 2 position through the No. 1 antibody micro-injection pump 27, meanwhile, the No. 2 antibody sample loading needle 30 injects an antibody B into the matched sample loading pipe at the No. 1 position through the No. 2 antibody micro-injection pump 28, and then the sample loading device integral vertical moving mechanism 17 drives the sample;

the upper sample tube rack rotating mechanism 4 drives the upper sample tube rack 1 to rotate 9 degrees anticlockwise, the sample micro-injection pump horizontal moving mechanism 33 moves the sample micro-injection pump 34 to a front positioning point of the sample micro-injection pump moving rod 19, at the moment, the sample loading needle 35 is aligned downwards to the sample tube at the No. 1 position, then the sample loading needle vertical moving mechanism 36 drives the sample loading needle 35 to vertically move downwards and extend into the sample tube at the No. 1 position, the sample loading needle 35 sucks a sample in the sample tube at the No. 1 position through the sample micro-injection pump 34, then the sample loading needle vertical moving mechanism 36 drives the sample loading needle 35 to vertically move upwards for returning, and then the sample micro-injection pump 34 is moved backwards by the sample micro-injection pump horizontal moving mechanism 33 to a rear positioning point of the sample micro-injection pump moving rod 19;

the whole vertical moving mechanism 17 of the sample adding device drives the whole sample adding device to vertically move downwards to be in place, at this time, the sample adding needle 35 is downwards aligned with the sample tube on the matched sleeve on the No. 1 position, the No. 2 antibody adding needle 30 is downwards aligned with the sample tube on the matched sleeve on the No. 2 position, the No. 1 antibody adding needle 29 is downwards aligned with the sample tube on the matched sleeve on the No. 3 position, then the sample adding needle vertical moving mechanism 36 drives the sample adding needle 35 to vertically move downwards and extend into the matched sample adding tube at No. 1, the sample adding needle 35 injects the sucked sample into the matched sample adding tube at No. 1 through the sample micro-injection pump 34, then the sample adding needle 35 sucks the sample once through the sample micro-injection pump 34, the antibody A and the antibody B on the sample tube on the position 1 are uniformly mixed with the sample, simultaneously, the No. 2 antibody sample adding needle 30 injects the antibody B into the matched sample adding pipe at the No. 2 position, and the No. 1 antibody sample adding needle 29 injects the antibody A into the matched sample adding pipe at the No. 3 position;

the sample loading needle vertical moving mechanism 36 drives the sample loading needle 35 to vertically move upwards for returning, then the whole sample loading device vertical moving mechanism 17 drives the whole sample loading device to vertically move upwards for returning, the sample loading needle cleaning mechanism 37 cleans the sample loading needle 35, and the steps are repeated until all sample loading tubes on the sample loading tube rack 1 are uniformly loaded with the antibody A, the antibody B and the sample and are uniformly mixed;

after the antibody A, the antibody B and the sample are added into all matched sample loading tubes and are uniformly mixed, the sample loading tube frame rotating mechanism 4 drives the sample loading tube frame 1 to rotate clockwise by 9 degrees and then rapidly rotate anticlockwise by 9 degrees for 3 cycles, the mixed solution of the antibody A, the antibody B and the blood sample in all matched sample loading tubes is uniformly oscillated, and then the mixed solution of the antibody A, the antibody B and the blood sample after uniform oscillation is incubated for 15 minutes; the injection, mixing, oscillation and incubation of the antibody A and the antibody B and the blood sample in each matched sample loading tube are all carried out in an incubation cup 53 in the sample loading tube;

the sample loading pipe frame rotating mechanism 4 drives the sample loading pipe frame 1 to enable the hemolysin/buffer solution sample adding needle 23 to be downwards aligned to the sample adding pipe on the No. 1 position, then the hemolysin/buffer solution sample adding needle vertical moving mechanism 24 drives the hemolysin/buffer solution sample adding needle 23 to vertically move downwards to a first height position, and the hemolysin/buffer solution sample adding needle 23 injects hemolysin into the incubation cup 53 of the sample adding pipe on the No. 1 position through the hemolysin/buffer solution micro-injection pump 22;

the hemolysin/buffer solution sample adding needle vertical moving mechanism 24 further drives the hemolysin/buffer solution sample adding needle 23 to vertically move downwards to a second height position, in the process that the hemolysin/buffer solution sample adding needle 23 vertically moves downwards to the second height position, the hemolysin/buffer solution sample adding needle 23 punctures the incubation cup 53 in the sample adding tube on the position 1, then the hemolysin/buffer solution sample adding needle 23 sucks once through the hemolysin/buffer solution micro-injection pump 22, and the hemolysin, the mixed solution of the antibody A, the antibody B and the blood sample are uniformly mixed;

the hemolysin/buffer solution sample adding needle vertical moving mechanism 24 drives the hemolysin/buffer solution sample adding needle 23 to vertically move upwards, the hemolysin/buffer solution sample adding needle 23 returns after passing through a first height position, the hemolysin/buffer solution sample adding needle cleaning mechanism 25 cleans the hemolysin/buffer solution sample adding needle 23, then the sample adding pipe frame rotating mechanism 4 drives the sample adding pipe frame 1 to rotate anticlockwise for 9 degrees, so that the hemolysin/buffer solution sample adding needle 23 is downwards aligned with the matched sample adding pipe at the No. 2 position, the circulation is carried out until all the matched sample adding pipes are injected with hemolysin and mixed evenly, and all the incubation cups 53 in the matched sample adding pipes are punctured;

injecting hemolysin into all matched sample loading tubes and uniformly mixing, driving the sample loading tube rack 1 to rotate clockwise by 9 degrees and then rapidly rotate anticlockwise by 9 degrees by a sample loading tube rack rotating mechanism 4, continuing for 3 cycles, oscillating and uniformly mixing the mixed solution of the hemolysin, the antibody A, the antibody B and the blood sample in all matched sample loading tubes, and then incubating the mixed solution of the hemolysin, the antibody A, the antibody B and the blood sample after oscillating and uniformly mixing for 15 minutes;

the upper sample tube rack chassis rotating mechanism 6 drives the upper sample tube rack chassis 5 to rotate 9 degrees anticlockwise, so that the injector rods 54 of all matched upper sample tubes are clamped into corresponding upper sample tube injector rod limiting holes 9 on the upper sample tube rack chassis 5, then the upper sample tube rack chassis vertical moving mechanism 7 drives the upper sample tube rack chassis 5 and the upper sample tube rack chassis rotating mechanism 6 to move together vertically downwards, the upper sample tube rack chassis 5 pulls all the injector rods 54 of the matched upper sample tubes to move vertically downwards for a certain distance, and negative pressure is caused in the matched upper sample tubes;

under the action of negative pressure, the mixed liquid of the incubated hemolysin, the antibody A, the antibody B and the blood sample breaks through the perforation on the incubation cup 53 and all flows into the cavity at the lower part of the matched sample loading tube between the incubation cup 53 and the filter membrane 60, and under the action of gravity, the red cell fragments and the waste liquid in the mixed liquid of the hemolysin, the antibody A, the antibody B and the blood sample flow into the cavity 57 in the injector rod 54 through the filter membrane 60, and the white blood cells in the mixed liquid of the hemolysin, the antibody A, the antibody B and the blood sample are internally trapped on the filter membrane 60;

the vertical movement mechanism 7 of the upper sample tube rack chassis drives the upper sample tube rack chassis 5 and the rotating mechanism 6 of the upper sample tube rack chassis to move together and vertically move upwards for returning, the upper sample tube rack chassis 5 pushes all the injector rods 54 matched with the upper sample tubes to vertically move upwards for returning, and the rotating mechanism 6 of the upper sample tube rack chassis drives the upper sample tube rack chassis 5 to rotate clockwise for 9 degrees, so that all the injector rods 54 matched with the upper sample tubes are separated from the corresponding upper sample tube injector rod limiting holes 9 on the upper sample tube rack chassis 5 for returning;

the sample loading tube frame rotating mechanism 4 drives the sample loading tube frame 1 to enable the hemolysin/buffer solution sample adding needle 23 to be aligned with the sample loading tube on the No. 1 position downwards again, then the hemolysin/buffer solution sample adding needle vertical moving mechanism 24 drives the hemolysin/buffer solution sample adding needle 23 to vertically move downwards to a first height position, the hemolysin/buffer solution sample adding needle 23 injects a buffer solution into the sample loading tube on the No. 1 position through the hemolysin/buffer solution micro-injection pump 22, then the hemolysin/buffer solution sample adding needle 23 sucks once through the hemolysin/buffer solution micro-injection pump 22, and the buffer solution and the white blood cells on the filter membrane 60 are uniformly mixed;

the hemolysin/buffer solution sample adding needle vertical moving mechanism 24 drives the hemolysin/buffer solution sample adding needle 23 to vertically move upwards and return, the hemolysin/buffer solution sample adding needle cleaning mechanism 25 cleans the hemolysin/buffer solution sample adding needle 23, then the sample loading tube rack rotating mechanism 4 drives the sample loading tube rack 1 to rotate anticlockwise for 9 degrees, so that the hemolysin/buffer solution sample adding needle 23 is downwards aligned to the matched sample loading tube at the No. 2 position, and the circulation is carried out until buffer solution is injected into all the matched sample loading tubes and is uniformly mixed with the white blood cells on the corresponding filter membrane 60;

after all matched sample loading tubes are injected with buffer solution and mixed uniformly, the sample loading tube frame rotating mechanism 4 drives the sample loading tube frame 1 to rotate clockwise by 9 degrees and then rotate rapidly counterclockwise by 9 degrees, and the 3 cycles are continued, and the buffer solution and the white blood cells in all matched sample loading tubes are oscillated and mixed uniformly to form white blood cell single cell suspension marked by fluorescence;

and finally, the operator takes down the sample loading pipe frame 1 from the sample loading pipe frame rotating device and moves the sample loading pipe frame 1 to the detection device for direct detection.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A full-automatic fluorescence labeling single cell suspension preparation device is characterized in that: the device comprises a sample loading pipe frame rotating device, a sample loading pipe frame (1), a sample loading pipe frame top cover (2), a sample adding device and a comprehensive controller;

the sample loading pipe frame rotating device comprises a sample loading pipe frame supporting rod (3) used for installing the sample loading pipe frame (1), the lower part of the sample loading pipe rack supporting rod (3) is provided with a sample loading pipe rack rotating mechanism (4) for driving the sample loading pipe rack (1) to rotate on the sample loading pipe rack supporting rod (3), the middle part of the upper sample tube rack supporting rod (3) is movably sleeved with an upper sample tube rack chassis (5), the lower surface of the sample loading pipe frame chassis (5) is provided with a sample loading pipe frame chassis rotating mechanism (6) for driving the sample loading pipe frame chassis (5) to rotate on the sample loading pipe frame support rod (3), the lower surface of the sample loading pipe frame chassis rotating mechanism (6) is provided with a sample loading pipe frame chassis vertical moving mechanism (7) for driving the sample loading pipe frame chassis (5) to vertically move on the sample loading pipe frame supporting rod (3); a plurality of sample loading tube injector rod push plate through holes (8) which are communicated up and down are uniformly formed in the circumferential edge of the sample loading tube frame chassis (5), one side of each sample loading tube injector rod push plate through hole (8) in the clockwise direction is provided with a sample loading tube injector rod limiting hole (9) which is communicated up and down, and each sample loading tube injector rod limiting hole (9) is communicated with the corresponding sample loading tube injector rod push plate through hole (8);

the sample loading pipe support (1) is coaxially and rotatably arranged at the upper part of the sample loading pipe support rod (3), a sample loading pipe rack top cover positioning column (10) used for installing the sample loading pipe rack top cover (2) is arranged on the circle center of the upper surface of the sample loading pipe rack (1), a circle of lug boss (11) is arranged on the circumferential edge of the upper surface of the sample loading pipe frame (1), a plurality of specimen tube mounting holes (12) are uniformly arranged on the upper surface of the boss (11), a plurality of sample loading pipe mounting holes (13) are formed in the upper surface of the sample loading pipe frame (1) along the inner side of the boss (11), a sample loading pipe injector rod through hole (14) which is vertically communicated is formed in the bottom of each sample loading pipe mounting hole (13), each sampling tube mounting hole (13) is respectively coaxial with the corresponding sampling tube injector rod through hole (14) up and down; the number of the sample tube mounting holes (12) corresponds to that of the sample tube injector rod through holes (14), and when the sample tube rack (1) is fixed on the sample tube rack supporting rod (3), each sample tube mounting hole (12) is respectively coaxial with the sample tube injector rod through hole (14) which corresponds to the sample tube mounting hole up and down; the number of the sampling pipe mounting holes (13) corresponds to the number of the sample pipe mounting holes (12), and the distribution position of each sampling pipe mounting hole (13) corresponds to the distribution position of the corresponding sample pipe mounting hole (12);

the upper sample tube rack top cover (2) is coaxially and rotatably arranged on the upper sample tube rack (1), a top cover handle (15) of the upper sample tube rack top cover is arranged at the circle center of the upper surface of the upper sample tube rack top cover (2), a plurality of upper sample tube through holes (16) which are communicated up and down are uniformly formed in the circumferential edge of the upper sample tube rack top cover (2), the number of the upper sample tube through holes (16) corresponds to the number of the upper sample tube mounting holes (13), and when the upper sample tube rack top cover (2) is fixed on the upper sample tube rack (1), each upper sample tube through hole (16) is respectively coaxial with the corresponding upper sample tube mounting hole (13) up and down;

the sample adding device is arranged above the top cover (2) of the sample adding pipe frame and comprises a sample adding device integral vertical moving mechanism (17), and a hemolysin/buffer solution micro-injection pump fixing rod (18), a sample micro-injection pump moving rod (19) and an antibody micro-injection pump fixing rod (20) are arranged side by side on the lower part of the sample adding device integral vertical moving mechanism (17); the front end of the hemolysin/buffer solution micro-injection pump fixing rod (18) is provided with a hemolysin/buffer solution micro-injection pump (22) through a hemolysin/buffer solution micro-injection pump fixing block (21), the lower end of the hemolysin/buffer solution micro-injection pump (22) is provided with a hemolysin/buffer solution loading needle (23), the upper end of the hemolysin/buffer solution micro-injection pump (22) is provided with a hemolysin/buffer solution loading needle vertical moving mechanism (24) for driving the hemolysin/buffer solution loading needle (23) to stretch up and down, and the hemolysin/buffer solution loading needle (23) is provided with a hemolysin/buffer solution loading needle cleaning mechanism (25) capable of moving vertically; the front end of the antibody micro-injection pump fixing rod (20) is provided with a No. 1 antibody micro-injection pump (27) and a No. 2 antibody micro-injection pump (28) through an antibody micro-injection pump fixing block (26), the lower ends of the No. 1 antibody micro-injection pump (27) and the No. 2 antibody micro-injection pump (28) are respectively provided with a No. 1 antibody sampling needle (29) and a No. 2 antibody sampling needle (30), and the upper ends of the No. 1 antibody micro-injection pump (27) and the No. 2 antibody micro-injection pump (28) are respectively provided with a No. 1 antibody sampling needle vertical moving mechanism (31) and a No. 2 antibody sampling needle vertical moving mechanism (32) which are used for driving the No. 1 antibody sampling needle (29) and the No. 2 antibody sampling needle (30) to vertically extend and retract; a movable sample micro-injection pump horizontal moving mechanism (33) is arranged on the sample micro-injection pump moving rod (19), a sample micro-injection pump (34) is arranged on the sample micro-injection pump horizontal moving mechanism (33), a sample loading needle (35) is arranged at the lower end of the sample micro-injection pump (34), a sample loading needle vertical moving mechanism (36) used for driving the sample loading needle (35) to stretch up and down is arranged at the upper end of the sample micro-injection pump (34), and a sample loading needle cleaning mechanism (37) capable of moving vertically is arranged on the sample loading needle (35); when the sample micro-injection pump horizontal moving mechanism (33) moves to a rear positioning point of the sample micro-injection pump moving rod (19), the hemolysin/buffer solution sample adding needle (23), the sample adding needle (35), the No. 1 antibody sample adding needle (29) and the No. 2 antibody sample adding needle (30) are arranged in an arc shape and respectively correspond to the plane positions of any four adjacent sample adding tube through holes (16) on the sample adding tube rack top cover (2) up and down one by one, and when the sample micro-injection pump horizontal moving mechanism (33) moves to a front positioning point of the sample micro-injection pump moving rod (19), the sample adding needle (35) extends forwards to correspond to one sample adding tube mounting hole (12) on the sample adding tube rack (1) up and down; a reagent bottle base (38) is arranged in the middle of the integral vertical moving mechanism (17) of the sample adding device, a hemolysin reagent bottle (39), a buffer solution reagent bottle (40), a No. 1 antibody reagent bottle (41) and a No. 2 antibody reagent bottle (42) are respectively arranged on the reagent bottle base (38), the hemolysin reagent bottle (39) and the buffer solution reagent bottle (40) are respectively connected with the hemolysin/buffer solution micro-injection pump (22) through pipelines, the No. 1 antibody reagent bottle (41) is connected with the No. 1 antibody micro-injection pump (27) through a pipeline, and the No. 2 antibody reagent bottle (42) is connected with the No. 2 antibody micro-injection pump (28) through a pipeline;

the integrated controller is electrically connected with the sample loading pipe frame rotating mechanism (4), the sample loading pipe frame chassis rotating mechanism (6), the sample loading pipe frame chassis vertical moving mechanism (7), the whole vertical moving mechanism (17) of the sample loading device, the hemolysin/buffer solution micro-injection pump (22), the hemolysin/buffer solution sample loading needle vertical moving mechanism (24), the hemolysin/buffer solution sample loading needle cleaning mechanism (25), the No. 1 antibody micro-injection pump (27), the No. 2 antibody micro-injection pump (28), the No. 1 antibody sample loading needle vertical moving mechanism (31), the No. 2 antibody sample loading needle vertical moving mechanism (32), the sample micro-injection pump horizontal moving mechanism (33), the sample micro-injection pump (34), the sample loading needle vertical moving mechanism (36) and the sample loading needle cleaning mechanism (37) respectively, so as to coordinate the linkage work of the various motion mechanisms.

2. The apparatus for preparing a full-automatic fluorescence-labeled single-cell suspension according to claim 1, wherein: the number of the sample tube mounting holes (12), the sample tube mounting holes (13), the sample tube injector rod via holes (14), the sample tube injector rod push plate via holes (8), the sample tube injector rod limiting holes (9) and the sample tube via holes (16) is 40.

3. The apparatus for preparing a full-automatic fluorescence-labeled single-cell suspension according to claim 1, wherein: the integrated controller is also electrically connected with one multimedia interaction device, so that an operator can set an experiment and display the experiment progress conveniently.

4. The apparatus for preparing a full-automatic fluorescence-labeled single-cell suspension according to claim 1, wherein: two sample loading pipe rack clamping grooves (43) for fixing the sample loading pipe rack (1) are oppositely arranged on the side surface of the upper part of the sample loading pipe rack supporting rod (3), a circle center of the lower surface of the sample loading pipe frame (1) is provided with a sample loading pipe frame support rod extending into the groove (44), two telescopic sample loading pipe frame clamping columns (45) are oppositely arranged on the wall of the groove (44) where the sample loading pipe frame supporting rod extends into, two sample loading pipe rack clamping column buttons (46) for controlling the two sample loading pipe rack clamping columns (45) to stretch and retract are oppositely arranged on the side surface of the sample loading pipe rack top cover positioning column (10), the sample loading pipe frame (1) is sleeved on the upper part of the sample loading pipe frame supporting rod (3) through a sample loading pipe frame supporting rod extending into the groove (44), and the sample loading pipe frame support rods (3) are fixed through the matching of the two sample loading pipe frame clamping columns (45) and the two sample loading pipe frame clamping grooves (43).

5. The apparatus for preparing a full-automatic fluorescence-labeled single-cell suspension according to claim 1, wherein: a sample loading pipe rack top cover fixing groove (47) is formed in the circle center of the upper surface of the sample loading pipe rack top cover positioning column (10), two sample loading pipe rack top cover clamping grooves (48) are oppositely formed in the groove wall of the sample loading pipe rack top cover fixing groove (47), a sample loading pipe rack top cover supporting rod (49) is arranged in the circle center of the lower surface of the sample loading pipe rack top cover (2), two telescopic sample loading pipe rack top cover clamping columns (50) are oppositely arranged on the side surface of the sample loading pipe rack top cover supporting rod (49), two sample loading pipe rack top cover clamping column buttons (51) used for controlling the two sample loading pipe rack top cover clamping columns (50) to be telescopic are oppositely arranged on the side surface of the sample loading pipe rack top cover handle (15), the sample loading pipe rack top cover (2) is installed in the sample loading pipe rack top cover fixing groove (47) of the sample loading pipe rack top cover positioning column (10) through the sample loading top cover supporting rod, and the sample loading pipe frame top cover positioning column (10) is fixed by matching two sample loading pipe frame top cover clamping columns (50) with two sample loading pipe frame top cover clamping grooves (48).

6. A kit-on-sample tube for use in the fully automatic fluorescence-labeled single-cell suspension preparation apparatus according to any one of claims 1 to 5, wherein: comprises a sample feeding tube body (52), a punctured incubation cup (53) for cell fluorescent antibody staining is arranged in the middle of the interior of the sample feeding tube body (52), a movable injector rod (54) is arranged at the lower part of the interior of the sample feeding tube body (52), an injector rod push plate (55) is arranged at the lower part of the injector rod (54), the injector rod push plate (55) is exposed below the sample feeding tube body (52), an injector rod sealing ring (56) matched with the inner wall of the sample feeding tube body (52) is arranged on the circumferential surface of the upper part of the injector rod (54), a cavity (57) for collecting waste liquid is arranged in the injector rod (54), the cavity (57) upwards extends on the upper plane of the injector rod (54) to form an open pore, a filter membrane support frame (58) with a hollowed middle is arranged on the upper plane of the injector rod (54), the lower surface of the filter membrane support frame (58) is provided with a filter membrane support frame sealing ring (59) matched with the open hole of the cavity (57), and the upper surface of the filter membrane support frame (58) is provided with a layer of filter membrane (60) for filtering erythrocyte fragments and waste liquid.

7. The matching sleeve sampling tube of the full-automatic fluorescence labeling single-cell suspension preparation device according to claim 6, which is characterized in that: a plurality of support legs (61) are arranged on the outer wall of the incubation cup (53), and the incubation cup (53) is arranged in the middle of the sample loading tube body (52) through the contact of the support legs (61) and the inner wall of the sample loading tube body (52).

8. The matching sleeve sampling tube of the full-automatic fluorescence labeling single-cell suspension preparation device according to claim 6, which is characterized in that: the hollow part of the filter membrane support frame (58) is provided with a cross support structure (62), the cross support structure (62) is connected with the main body part of the filter membrane support frame (58), and the filter membrane (60) covers the main body part of the filter membrane support frame (58) and the cross support structure (62) together.

9. The matching sleeve sampling tube of the full-automatic fluorescence labeling single-cell suspension preparation device according to claim 6, which is characterized in that: the sample loading tube body (52) is made of polystyrene or polypropylene, and the size of the sample loading tube body (52) is the same as that of a common cell sample loading tube in the market.

10. The matching sleeve sampling tube of the full-automatic fluorescence labeling single-cell suspension preparation device according to claim 6, which is characterized in that: the incubation cup (53) is made of polyvinyl chloride.