A fully automatic fluorescent labeled single cell suspension preparation device and matching sample tube

Abstract

The present invention discloses a fully automatic fluorescently labeled single cell suspension preparation device and its matching sample tube, the device is composed of a sample tube rack rotating device, a sample tube rack, a sample tube rack top cover, a sample adding device and an integrated controller, etc., and the matching sample tube is composed of a sample tube body, an incubation cup, a syringe rod, a syringe rod push plate, a syringe rod sealing ring, a syringe rod cavity, a filter membrane support frame, a filter membrane support frame sealing ring and a filter membrane, etc. The device and the matching sample tube of the present invention can automatically process human or animal peripheral blood specimens, including steps such as adding samples, staining, hemolysis, separation of white blood cells and suspension of white blood cells, reducing labor intensity and human errors. The matching sample tube of the present invention adopts the membrane filtration principle, and replaces the vacuum pump with a syringe to remove red blood cell fragments and waste liquid in batches, and can complete the separation of fluorescently labeled white blood cells in a very short time, reducing the volume of the corresponding equipment, eliminating the cost of the vacuum pump, and saving space and cost.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical equipment, and in particular relates to a fully automatic fluorescently labeled single cell suspension preparation device controlled by a PLC and a matching sample loading tube, which are used for automatically processing peripheral blood samples of humans or animals. Background Art

[0002] At present, for the detection of peripheral blood leukocyte surface antigens in humans or animals, the collected peripheral blood samples must be processed before the test. The existing processing methods are roughly divided into five steps: drawing blood samples into experimental vessels, staining with fluorescent labeled antibodies, hemolyzing with hemolysin (or hemolytic agent), separating labeled leukocytes and suspending labeled leukocytes in liquid, etc., and the experimental process of the processing has the characteristics of precious experimental consumables, cumbersome experimental process, and high requirements for the accuracy of sample volume and reaction time.

[0003] For the step of separating and labeling white blood cells, the commonly used separation methods include centrifugation, density gradient separation liquid method, magnetic bead adsorption method and membrane filtration method. Although centrifugation is economical and convenient, it requires a centrifuge and will cause damage and loss of white blood cells; density gradient separation liquid method and magnetic bead adsorption method require relatively expensive separation liquid and adsorption magnetic beads, which is time-consuming and cannot avoid the loss of white blood cells; membrane filtration method requires additional filter vessels and vacuum pumps. Centrifuges are standing equipment in laboratories, so centrifugation is currently the most common method for separating white blood cells, but this method requires manual transfer of experimental vessels to the centrifuge for centrifugation, and after centrifugation, manual sorting of the experimental vessels is still required in order to continue loading samples for testing, which is labor-intensive and prone to errors.

[0004] Currently, some sample processing devices that can replace manual work have been launched 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 the machine. This is called the no-wash mode. However, this method will have an adverse effect on subsequent test results due to excessive red blood cell fragments after hemolysis, and it is easy to cause malfunctions such as blockage of the detection device. Therefore, the promotion and application effect of this method is poor. Summary of the invention

[0005] In response to the problems existing in the above-mentioned prior art, the present invention provides a fully automatic fluorescently labeled single cell suspension preparation device and a matching sample loading tube, which can automatically process human or animal peripheral blood samples including steps such as sample addition, staining, hemolysis, separation of leukocytes and suspension of leukocytes in liquid, so that the specimens can be directly tested on the machine.

[0006] In order to solve the above technical problems and achieve the above technical effects, the present invention is implemented through the following technical solutions:

[0007] A fully automatic fluorescent labeled single cell suspension preparation device, which is composed of a sample tube rack rotating device, a sample tube rack, a sample tube rack top cover, a sample adding device and an integrated controller;

[0008] The sample tube rack rotating device comprises a sample tube rack support rod for mounting the sample tube rack, a sample tube rack rotating mechanism for driving the sample tube rack to rotate on the sample tube rack support rod is arranged at the lower part of the sample tube rack support rod, a sample tube rack chassis is movably sleeved on the middle part of the sample tube rack support rod, a sample tube rack chassis rotating mechanism for driving the sample tube rack chassis to rotate on the sample tube rack support rod is arranged on the lower surface of the sample tube rack chassis, and the sample tube rack chassis A loading tube rack chassis vertical movement mechanism is provided on the lower surface of the disk rotating mechanism for driving the loading tube rack chassis to move vertically on the loading tube rack support rod; a plurality of loading tube syringe rod push plate through holes that penetrate vertically are evenly provided on the circumferential edge of the loading tube rack chassis, and a loading tube syringe rod limiting hole that penetrates vertically is provided on the clockwise side of each loading tube syringe rod push plate through hole, and each loading tube syringe rod limiting hole is connected to the corresponding loading tube syringe rod push plate through hole;

[0009] The sample tube rack can be coaxially rotatably arranged on the upper part of the sample tube rack support rod, and a sample tube rack top cover positioning column for installing the sample tube rack top cover is arranged at the center of the circle on the upper surface of the sample tube rack, and a circle of bosses are arranged on the circumferential edge of the upper surface of the sample tube rack, and a plurality of specimen tube mounting holes are evenly arranged on the upper surface of the bosses, and a plurality of sample tube mounting holes are arranged on the upper surface of the sample tube rack along the inner side of the bosses, and a sample tube syringe rod through hole is arranged at the bottom of each of the sample tube mounting holes, and each of the sample tube syringe rod through holes is arranged The tube mounting holes are respectively coaxial with the corresponding sample tube syringe rod through holes; the number of the specimen tube mounting holes corresponds to the number of the sample tube syringe rod through holes, and when the sample tube rack is fixed on the sample tube rack support rod, each of the specimen tube mounting holes is respectively coaxial with the corresponding sample tube syringe rod through holes; the number of the sample tube mounting holes corresponds to the number of the specimen tube mounting holes, and the distribution position of each of the sample tube mounting holes corresponds to the distribution position of the corresponding specimen tube mounting holes;

[0010] The top cover of the sample tube rack can be coaxially rotatably arranged on the sample tube rack, a cover handle of the sample tube rack top cover is arranged at the center of the circle on the upper surface of the sample tube rack top cover, and a plurality of sample tube through holes that pass through the top cover are evenly arranged on the circumferential edge of the sample tube rack top cover, the number of the sample tube through holes corresponds to the number of the sample tube mounting holes, and when the top cover of the sample tube rack is fixed on the sample tube rack, each of the sample tube through holes is coaxial with the corresponding sample tube mounting hole in the upper and lower directions;

[0011] The sample loading device is arranged above the top cover of the sample loading tube rack, and the sample loading device includes an overall vertical moving mechanism of the sample loading device, and the lower part of the overall vertical moving mechanism of the sample loading device is provided with a hemolysin / buffer microinjection pump fixing rod, a specimen microinjection pump moving rod and an antibody microinjection pump fixing rod arranged side by side; a hemolysin / buffer microinjection pump is arranged at the front end of the hemolysin / buffer microinjection pump fixing rod through a hemolysin / buffer microinjection pump fixing block, a hemolysin / buffer microinjection pump is arranged at the lower end of the hemolysin / buffer microinjection pump with a hemolysin / buffer sample loading needle, and a hemolysin / buffer sample loading needle vertical movement for driving the hemolysin / buffer sample loading needle to extend and retract up and down is arranged at the upper end of the hemolysin / buffer microinjection pump. The hemolysin / buffer injection needle is provided with a vertically movable hemolysin / buffer injection needle cleaning mechanism; the front end of the antibody microinjection pump fixing rod is provided with a No. 1 antibody microinjection pump and a No. 2 antibody microinjection pump through an antibody microinjection pump fixing block, the No. 1 antibody microinjection pump and the No. 2 antibody microinjection pump are respectively provided with a No. 1 antibody injection needle and a No. 2 antibody injection needle at the lower ends, and the No. 1 antibody microinjection pump and the No. 2 antibody injection needle are respectively provided with a No. 1 antibody injection needle vertical moving mechanism and a No. 2 antibody injection needle vertical moving mechanism for driving the No. 1 antibody injection needle and the No. 2 antibody injection needle to extend and retract up and down at the upper ends; the specimen microinjection pump moving rod is provided with a movable A specimen microinjection pump horizontal moving mechanism, wherein a specimen microinjection pump is arranged on the specimen microinjection pump horizontal moving mechanism, a specimen injection needle is arranged at the lower end of the specimen microinjection pump, a specimen injection needle vertical moving mechanism for driving the specimen injection needle to extend and retract up and down is arranged at the upper end of the specimen microinjection pump, and a specimen injection needle cleaning mechanism that can move vertically is arranged on the specimen injection needle; when the specimen microinjection pump horizontal moving mechanism moves to the rear positioning point of the specimen microinjection pump moving rod, the hemolysin / buffer injection needle, the specimen injection needle, the No. 1 antibody injection needle and the No. 2 antibody injection needle are arranged in an arc shape, and are respectively aligned with the planes of any four adjacent sample tube through holes on the top cover of the sample tube rack The positions correspond one to one up and down. When the horizontal moving mechanism of the specimen microinjection pump moves to the front positioning point of the moving rod of the specimen microinjection pump, the specimen injection needle extends forward and corresponds up and down to one of the specimen tube mounting holes on the sample tube rack; a reagent bottle base is arranged in the middle of the overall vertical moving mechanism of the injection device, and a hemolysin reagent bottle, a buffer 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 reagent bottle are respectively connected to the hemolysin / buffer microinjection pump through pipelines, the No. 1 antibody reagent bottle is connected to the No. 1 antibody microinjection pump through a pipeline, and the No. 2 antibody reagent bottle is connected to the No. 2 antibody microinjection pump through a pipeline;

[0012] The integrated controller is a controller with PLC as the core, which is respectively electrically connected to the sample tube rack rotating mechanism, the sample tube rack chassis rotating mechanism, the sample tube rack chassis vertical movement mechanism, the overall vertical movement mechanism of the sample loading device, the hemolysin / buffer microinjection pump, the hemolysin / buffer injection needle vertical movement mechanism, the hemolysin / buffer injection needle cleaning mechanism, the No. 1 antibody microinjection pump, the No. 2 antibody microinjection pump, the No. 1 antibody injection needle vertical movement mechanism, the No. 2 antibody injection needle vertical movement mechanism, the specimen microinjection pump horizontal movement mechanism, the specimen microinjection pump, the specimen injection needle vertical movement mechanism and the specimen injection needle cleaning mechanism to coordinate the linkage work of the above-mentioned various motion mechanisms.

[0013] Furthermore, the number of the specimen tube mounting hole, the sample loading tube mounting hole, the syringe rod through hole, the syringe rod push plate through hole, the syringe rod push plate limiting hole and the sample loading tube through hole is 40 each.

[0014] Furthermore, the integrated controller is also electrically connected to a multimedia interactive device to facilitate operators to set up experiments and display experimental progress.

[0015] Furthermore, the side surface of the upper part of the sample tube rack support rod is relatively provided with two sample tube rack clamping grooves for fixing the sample tube rack, the center of the circle of the lower surface of the sample tube rack is provided with a sample tube rack support rod extending into a groove, and two retractable sample tube rack clamping columns are relatively provided on the groove wall where the sample tube rack support rod extends into the groove, and the side surface of the sample tube rack top cover positioning column is relatively provided with two sample tube rack clamping column buttons for controlling the extension and retraction of the two sample tube rack clamping columns, the sample tube rack is sleeved on the upper part of the sample tube rack support rod through the sample tube rack support rod extending into the groove, and is fixed to the sample tube rack support rod by the cooperation of the two sample tube rack clamping columns and the two sample tube rack clamping grooves.

[0016] The top cover of the loading tube rack is installed in the sample tube rack top cover fixing groove of the sample tube rack top cover positioning column through the sample tube rack top cover support rod, and the top cover of the sample tube rack is fixed with two sample tube rack top cover slots on the groove wall of the sample tube rack top cover fixing groove, and the top cover of the sample tube rack is fixed with two sample tube rack top cover slots on the groove wall of the sample tube rack top cover fixing groove. The top cover of the sample tube rack is fixed with two sample tube rack top cover slots on the groove wall of the sample tube rack top cover fixing groove, and the top cover of the sample tube rack is fixed with the top cover positioning column of the sample tube rack top cover through the cooperation of the two sample tube rack top cover slots and the two sample tube rack top cover slots.

[0017] A matching sample loading tube for a fully automatic fluorescently labeled single-cell suspension preparation device comprises a sample loading tube body, a puncturable incubation cup for cell fluorescent antibody staining is arranged in the middle of the sample loading tube body, a movable syringe rod is arranged in the lower part of the sample loading tube body, a syringe rod push plate is arranged at the lower part of the syringe rod, and the syringe rod push plate is exposed below the sample loading tube body, a syringe rod sealing ring is arranged on the circumferential surface of the upper part of the syringe rod and cooperates with the inner wall of the sample loading tube body, the interior of the syringe rod is a cavity for collecting waste liquid, the cavity extends upward to form an opening on the upper plane of the syringe rod, a filter membrane support frame with a hollow middle is arranged on the upper plane of the syringe rod, a filter membrane support frame sealing ring cooperating with the opening of the cavity is arranged on the lower surface of the filter membrane support frame, and a filter membrane for filtering out red blood cell fragments and waste liquid is arranged on the upper surface of the filter membrane support frame.

[0018] Furthermore, a plurality of legs are provided on the outer wall of the incubation cup, and the incubation cup is arranged in the middle of the body of the sample loading tube through the contact between the legs and the inner wall of the body of the sample loading tube.

[0019] Furthermore, a cross support structure is provided in the hollow portion of the filter membrane support frame, the cross support structure is connected to the main portion of the filter membrane support frame, and the filter membrane is jointly covered on the main portion of the filter membrane support frame and the cross support structure.

[0020] 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 the size of common cell loading tubes on the market.

[0021] Furthermore, the incubation cup is made of polyvinyl chloride.

[0022] Taking the 40-hole specimen tube mounting hole and the sample tube mounting hole of the present invention as an example, the working principle and operation method are as follows:

[0023] The operator first numbers the specimen tubes to be processed, and then places the numbered specimen tubes into the specimen tube installation holes in sequence, starting from position 1 of the specimen tube installation hole of the sample tube rack. A maximum of 40 specimen tubes can be placed.

[0024] Next, take the same number of matching sample tubes, and then, starting from position 1 of the sample tube mounting hole of the sample tube rack, sequentially place the matching sample tubes into the corresponding sample tube mounting holes on the sample tube rack, and ensure that the syringe rod of each matching sample tube passes through the corresponding syringe rod through hole on the sample tube rack so that its syringe rod push plate is exposed on the lower surface of the sample tube rack;

[0025] Place the sample tube rack containing the specimen tube and the matching sample tube on the sample tube rack support rod, press the sample tube rack clamping column buttons on both sides of the positioning column on the top cover of the sample tube rack, so that the sample tube rack support rod extends into the sample tube rack support rod at the bottom of the sample tube rack and extends into the groove, and then release the sample tube rack clamping column button to make the sample tube rack support rod extending into the groove clamp the sample tube rack clamping groove on the sample tube rack support rod, so as to fix the sample tube rack on the sample tube rack support rod, and at the same time, the syringe rod push plate at the bottom of each matching sample tube passes through the syringe rod push plate through hole in the corresponding sequence on the chassis of the sample tube rack;

[0026] Place the sample tube rack top cover on the sample tube rack, press the sample tube rack top cover clamping column buttons on both sides of the sample tube rack top cover handle, so that the sample tube rack top cover support rod on the lower surface of the sample tube rack top cover extends into the sample tube rack top cover fixing groove on the sample tube rack top cover positioning column, and then release the sample tube rack top cover clamping column button, so that the sample tube rack top cover clamping column on the sample tube rack top cover support rod is clamped with the sample tube rack top cover clamping slot in the sample tube rack top cover fixing groove, and fix the sample tube rack top cover on the sample tube rack;

[0027] Check whether the various reagents in the hemolysin reagent bottle, buffer reagent bottle, No. 1 antibody reagent bottle and No. 2 antibody reagent bottle are sufficient, confirm whether the pipelines are connected and normal, then select the corresponding experimental plan and quantity on the multimedia interactive device, and click Start to start the experiment;

[0028] The overall vertical moving mechanism of the sample loading device drives the overall sample loading device to move vertically downward into position. At this time, the No. 1 antibody loading needle is downwardly aligned with the matching loading tube at position 1, and then the No. 1 antibody loading needle injects antibody A into the matching loading tube at position 1 through the No. 1 antibody microinjection pump. Then the overall vertical moving mechanism of the sample loading device drives the overall sample loading device to move vertically upward back to position;

[0029] The sample tube rack rotating mechanism drives the sample tube rack to rotate 9 degrees counterclockwise, and the overall vertical movement mechanism of the sample loading device drives the sample loading device to move vertically downward into position. At this time, the No. 1 antibody sample loading needle is downwardly aligned with the matching sample loading tube at position 2, and the No. 2 antibody sample loading needle is downwardly aligned with the matching sample loading tube at position 1. Then, the No. 1 antibody sample loading needle injects antibody A into the matching sample loading tube at position 2 through the No. 1 antibody microinjection pump, and at the same time, the No. 2 antibody sample loading needle injects antibody B into the matching sample loading tube at position 1 through the No. 2 antibody microinjection pump, and then the overall vertical movement mechanism of the sample loading device drives the overall sample loading device to move vertically upward back to position;

[0030] The sample tube rack rotating mechanism drives the sample tube rack to rotate 9 degrees counterclockwise, and the specimen microinjection pump horizontal moving mechanism moves the specimen microinjection pump to the front positioning point of the specimen microinjection pump moving rod. At this time, the specimen injection needle is downwardly aligned with the specimen tube at position 1, and then the specimen injection needle vertical moving mechanism drives the specimen injection needle to move vertically downward and extend into the specimen tube at position 1. The specimen injection needle absorbs the specimen in the specimen tube at position 1 through the specimen microinjection pump, and then the specimen injection needle vertical moving mechanism drives the specimen injection needle to move vertically upward back to its position, and then the specimen microinjection pump horizontal moving mechanism moves the specimen microinjection pump back to the rear positioning point of the specimen microinjection pump moving rod;

[0031] The overall vertical moving mechanism of the sample loading device drives the overall vertical downward movement of the sample loading device into position, at which time the sample loading needle is downwardly aligned with the matching sample loading tube at position 1, the No. 2 antibody loading needle is downwardly aligned with the matching sample loading tube at position 2, and the No. 1 antibody loading needle is downwardly aligned with the matching sample loading tube at position 3, and then the sample loading needle vertical moving mechanism drives the sample loading needle to move vertically downward and extend into the matching sample loading tube at position 1, and the sample loading needle injects the absorbed sample into the matching sample loading tube at position 1 through the sample micro-injection pump, and then the sample loading needle is sucked once through the sample micro-injection pump to mix the antibody A, antibody B and sample in the matching sample loading tube at position 1, and at the same time, the No. 2 antibody loading needle injects antibody B into the matching sample loading tube at position 2, and the No. 1 antibody loading needle injects antibody A into the matching sample loading tube at position 3;

[0032] The specimen injection needle vertical moving mechanism drives the specimen injection needle to move vertically upward and back to the position, and then the overall vertical moving mechanism of the injection device drives the overall injection device to move vertically upward and back to the position, and the specimen injection needle cleaning mechanism cleans the specimen injection needle, and this cycle is repeated until all the matching injection tubes on the injection tube rack are added with antibody A, antibody B and specimen and mixed;

[0033] After all the matching sample tubes are added with antibody A, antibody B and specimens and mixed, the sample tube rack rotating mechanism drives the sample tube rack to rotate 9 degrees clockwise and then quickly rotate 9 degrees counterclockwise for 3 cycles to oscillate and mix the mixture of antibody A, antibody B and blood specimens in all the matching sample tubes, and then incubate the oscillated and mixed mixture of antibody A, antibody B and blood specimens for 15 minutes; the injection, mixing, oscillation and incubation of antibody A, antibody B and blood specimens in each matching sample tube are all carried out in the incubation cup inside it;

[0034] The sample tube rack rotating mechanism drives the sample tube rack to align the hemolysin / buffer sample injection needle downward with the matching sample tube at position 1, and then the hemolysin / buffer sample injection needle vertical movement mechanism drives the hemolysin / buffer sample injection needle to move vertically downward to a first height position, and the hemolysin / buffer sample injection needle injects hemolysin into the incubation cup of the matching sample tube at position 1 through the hemolysin / buffer microinjection pump;

[0035] The hemolysin / buffer injection needle vertical movement mechanism further drives the hemolysin / buffer injection needle to move vertically downward to a second height position. During the process of the hemolysin / buffer injection needle moving vertically downward to the second height position, the hemolysin / buffer injection needle pierces the incubation cup in the matching sample loading tube at position 1, and then the hemolysin / buffer injection needle is pumped once by the hemolysin / buffer microinjection pump to mix the hemolysin with the mixed solution of antibody A, antibody B and blood specimen;

[0036] The hemolysin / buffer injection needle vertical moving mechanism drives the hemolysin / buffer injection needle to move vertically upward, and returns to the first height position. The hemolysin / buffer injection needle cleaning mechanism cleans the hemolysin / buffer injection needle, and then the loading tube rack rotating mechanism drives the loading tube rack to rotate 9 degrees counterclockwise, so that the hemolysin / buffer injection needle is downwardly aligned with the matching loading tube at position 2, and this cycle is repeated until all matching loading tubes are injected with hemolysin and mixed, and the incubation cups in all matching loading tubes are punctured;

[0037] After all the matching sample tubes are injected with hemolysin and mixed, the sample tube rack rotating mechanism drives the sample tube rack to rotate clockwise 9 degrees and then quickly rotate counterclockwise 9 degrees for 3 cycles to oscillate and mix the mixed solution of hemolysin, antibody A, antibody B and blood specimen in all the matching sample tubes, and then incubate the oscillated and mixed mixed solution of hemolysin, antibody A, antibody B and blood specimen for 15 minutes;

[0038] The sample tube rack chassis rotating mechanism drives the sample tube rack chassis to rotate 9 degrees counterclockwise, so that the syringe rods of all matching sample tubes are inserted into the corresponding syringe rod limiting holes on the sample tube rack chassis, and then the sample tube rack chassis vertical moving mechanism drives the sample tube rack chassis and the sample tube rack chassis rotating mechanism to move vertically downward together, and the sample tube rack chassis pulls the syringe rods of all matching sample tubes to move vertically downward for a distance, causing negative pressure in the matching sample tubes;

[0039] Under the action of negative pressure, the mixed solution of the incubated hemolysin, antibody A, antibody B and blood specimen breaks through the perforations on the incubation cup and flows into the lower cavity of the matching sample loading tube between the incubation cup and the filter membrane, and under the action of gravity, the red blood cell fragments and waste liquid in the mixed solution of the hemolysin, antibody A, antibody B and blood specimen flow through the filter membrane into the cavity in the syringe rod, and the white blood cells in the mixed solution of the hemolysin, antibody A, antibody B and blood specimen are retained on the filter membrane;

[0040] The vertical moving mechanism of the sample tube rack chassis drives the sample tube rack chassis and the sample tube rack chassis rotating mechanism to move vertically upward and return to the original position. The sample tube rack chassis pushes the syringe rods of all matching sample tubes to move vertically upward and return to the original position. The sample tube rack chassis rotating mechanism drives the sample tube rack chassis to rotate 9 degrees clockwise, so that the syringe rods of all matching sample tubes are separated from the corresponding syringe rod limiting holes on the sample tube rack chassis and return to the original position.

[0041] The sample tube rack rotating mechanism drives the sample tube rack to align the hemolysin / buffer sample needle downward with the matching sample tube at position 1 again, and then the hemolysin / buffer sample needle vertical movement mechanism drives the hemolysin / buffer sample needle to move vertically downward to a first height position, and the hemolysin / buffer sample needle injects buffer into the matching sample tube at position 1 through the hemolysin / buffer microinjection pump, and then the hemolysin / buffer sample needle draws once through the hemolysin / buffer microinjection pump to mix the buffer with the leukocytes on the filter membrane;

[0042] The hemolysin / buffer injection needle vertical moving mechanism drives the hemolysin / buffer injection needle to move vertically upward back to its position, the hemolysin / buffer injection needle cleaning mechanism cleans the hemolysin / buffer injection needle, and then the loading tube rack rotating mechanism drives the loading tube rack to rotate 9 degrees counterclockwise, so that the hemolysin / buffer injection needle is downwardly aligned with the matching loading tube at position 2, and this cycle is repeated until the buffer is injected into all the matching loading tubes and mixed with the white blood cells on the corresponding filter membranes;

[0043] After all the matching sample tubes are injected with buffer and mixed, the sample tube rack rotating mechanism drives the sample tube rack to rotate 9 degrees clockwise and then quickly rotate 9 degrees counterclockwise, and continue for 3 cycles to oscillate and mix the buffer and white blood cells in all the matching sample tubes to form a fluorescently labeled white blood cell single cell suspension;

[0044] Finally, the operator removes the sample tube rack from the sample tube rack rotating device and moves it to the detection device for direct detection.

[0045] All the motion mechanisms on the fully automatic fluorescent labeled single cell suspension preparation device of the present invention are controlled by an integrated controller with PLC as the core. The integrated controller coordinates the work of all the motion mechanisms according to a pre-set experimental program, thereby ensuring the smooth completion of steps including sample addition, hemolysis, separation, separation of leukocytes and suspension of leukocytes, thereby realizing the automation of human or animal peripheral blood sample processing.

[0046] The beneficial effects of the present invention are:

[0047] 1. The fully automatic fluorescent labeled single cell suspension preparation device and its matching sample tube of the present invention can automatically process human or animal peripheral blood specimens, including the steps of adding samples, staining, hemolysis, separation of leukocytes and suspension of leukocytes, which greatly reduces the intensity of manual labor and can effectively reduce the errors caused by manual operation.

[0048] 2. The fully automatic fluorescent labeled single cell suspension preparation device of the present invention adopts a single-axis movement of the sample tube rack rotating device relative to the sample loading device, and the sample loading needles on the sample loading device are arranged in an arc shape, corresponding to the arc of the sample loading tube holes on the sample tube rack, which greatly reduces the positioning error of the two-axis two-dimensional movement during the sample suction and loading process of the traditional device, and reduces the configuration of the stepper motor, saving costs.

[0049] 3. The sample tube rack of the fully automatic fluorescent labeling single cell suspension preparation device of the present invention can be completely disassembled from the sample tube rack rotating device, so that after the sample is processed, the staff can directly place the sample tube rack together with the matching sample tube and specimen tube on the detection device for detection, which greatly reduces the work of manual transfer, numbering, etc. and reduces the intensity of manual labor.

[0050] 4. The matching sample tube of the present invention integrates the experimental steps of staining, hemolysis, separation of fluorescently labeled leukocytes, removal of waste liquid with red blood cell fragments, suspension of fluorescently labeled leukocytes, etc. into one sample tube. The matching sample tube is designed according to the size of common cell sample tubes currently on the market, thereby reducing the volume of the corresponding equipment and saving space.

[0051] 5. The matching sample tube of the present invention adopts the membrane filtration principle and uses a syringe instead of a vacuum pump to remove red blood cell fragments and waste liquid in batches, which can complete the separation of fluorescently labeled white blood cells in a very short time, which not only improves efficiency and reduces errors, but also reduces the volume of corresponding equipment, eliminates the cost of vacuum pumps, and saves space and cost.

[0052] 6. The syringe rod of the matching sample loading tube of the present invention is designed with a cavity, which can realize the functions of vacuuming and collecting waste liquid, thereby reducing the risk of biological hazards during the experiment.

[0053] The above description is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the present invention and implement it according to the contents of the specification, the following is a detailed description of the preferred embodiments of the present invention in conjunction with the accompanying drawings. The specific implementation of the present invention is given in detail by the following embodiments and their accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] The drawings described herein are used to provide a further understanding of the present invention and constitute a part of this application. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings:

[0055] Figure 1 It is a front stereoscopic view of the fully automatic fluorescent labeling single cell suspension preparation device of the present invention and the matching sample loading tube when used in combination;

[0056] Figure 2 It is a side stereoscopic view of the fully automatic fluorescent labeling single cell suspension preparation device of the present invention and the matching sample loading tube when used in combination;

[0057] Figure 3 It is a front view of the fully automatic fluorescent labeling single cell suspension preparation device of the present invention and the matching sample loading tube when used in combination;

[0058] Figure 4 It is a left view of the fully automatic fluorescent labeling single cell suspension preparation device of the present invention and the matching sample loading tube when used in combination;

[0059] Figure 5 It is a top view of the fully automatic fluorescent labeling single cell suspension preparation device of the present invention and the matching sample loading tube when used in combination;

[0060] Figure 6 A three-dimensional diagram of the sample tube rack of the present invention;

[0061] Figure 7 A top view of the sample tube rack of the present invention;

[0062] Figure 8 It is a front view of the sample tube rack of the present invention;

[0063] Fig. 9 A three-dimensional diagram of the top cover of the sample tube rack of the present invention;

[0064] Fig.10 It is a front view of the top cover of the sample tube rack of the present invention;

[0065] Fig.11An exploded view of the matching sample loading tube of the present invention;

[0066] Fig.12 It is a schematic diagram of the horizontal position relationship of various components of the matching sample loading tube of the present invention. DETAILED DESCRIPTION

[0067] The present invention will be described in detail below with reference to the accompanying drawings and in combination with embodiments. The descriptions herein are used to provide a further understanding of the present invention and constitute a part of this application. The illustrative embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute improper limitations on the present invention.

[0068] See also Figure 1-6 As shown, a fully automatic fluorescent labeled single cell suspension preparation device is composed of a sample tube rack rotating device, a sample tube rack 1, a sample tube rack top cover 2, a sample adding device and an integrated controller;

[0069] The sample tube rack rotating device includes a sample tube rack support rod 3 for mounting the sample tube rack 1, two sample tube rack clamping grooves 43 for fixing the sample tube rack 1 are relatively arranged on the side surface of the upper part of the sample tube rack support rod 3, a sample tube rack rotating mechanism 4 for driving the sample tube rack 1 to rotate on the sample tube rack support rod 3 is arranged at the lower part of the sample tube rack support rod 3, a sample tube rack chassis 5 is movably sleeved on the middle part of the sample tube rack support rod 3, and a sample tube rack chassis 5 is arranged on the lower surface of the sample tube rack chassis 5 for driving the sample tube rack chassis 5 to rotate on the sample tube rack support rod 3 A sample tube rack chassis rotating mechanism 6 is provided on the lower surface of the sample tube rack chassis rotating mechanism 6, and a sample tube rack chassis vertical moving mechanism 7 is provided for driving the sample tube rack chassis 5 to move vertically on the sample tube rack support rod 3; a plurality of sample tube syringe rod push plate through holes 8 are evenly provided on the circumferential edge of the sample tube rack chassis 5, and a sample tube syringe rod limiting hole 9 is provided on the clockwise side of each of the sample tube syringe rod push plate through holes 8, and each of the sample tube syringe rod limiting holes 9 is connected to the corresponding sample tube syringe rod push plate through holes 8;

[0070] The sample tube rack 1 is arranged on the sample tube rack rotating device, and a sample tube rack support rod extending into a groove 44 is arranged at the center of the circle on the lower surface of the sample tube rack 1. Two retractable sample tube rack clamping columns 45 are arranged on the groove wall of the sample tube rack support rod extending into the groove 44. The sample tube rack 1 is sleeved on the upper part of the sample tube rack support rod 3 through the sample tube rack support rod extending into the groove 44, and the two sample tube rack clamping columns 45 cooperate with the two sample tube rack clamping grooves 43 to realize the connection with the sample tube rack. The tube rack support rod 3 is fixed; a sample tube rack top cover positioning column 10 for mounting the sample tube rack top cover 2 is arranged at the center of the circle on the upper surface of the sample tube rack 1, and two sample tube rack clamping column buttons 46 for controlling the extension and retraction of the two sample tube rack clamping columns 45 are arranged on the side surface of the sample tube rack top cover positioning column 10; a sample tube rack top cover fixing groove 47 is arranged at the center of the circle on the upper surface of the sample tube rack top cover positioning column 10, and two sample tube rack top cover fixing grooves 47 are arranged on the groove walls of the sample tube rack top cover fixing grooves 47. The top cover of the sample tube rack has a slot 48; a circle of bosses 11 are arranged on the circumferential edge of the upper surface of the sample tube rack 1, and a plurality of specimen tube mounting holes 12 are evenly arranged on the upper surface of the bosses 11; a plurality of sample tube mounting holes 13 are arranged on the upper surface of the sample tube rack 1 along the inner side of the bosses 11, and a sample tube syringe rod through hole 14 is arranged at the bottom of each of the sample tube mounting holes 13, and each of the sample tube mounting holes 13 is coaxial with the corresponding sample tube syringe rod through hole 14; The number of the specimen tube mounting holes 12 corresponds to the number of the sample tube syringe rod through holes 14, and when the sample tube rack 1 is fixed on the sample tube rack support rod 3, each of the specimen tube mounting holes 12 is coaxial with the corresponding sample tube syringe rod through holes 14; the number of the sample tube mounting holes 13 corresponds to the number of the specimen tube mounting holes 12, and the distribution position of each of the sample tube mounting holes 13 corresponds to the distribution position of the corresponding specimen tube mounting holes 12;

[0071] The sample tube rack top cover 2 is arranged on the sample tube rack 1, and a sample tube rack top cover support rod 49 is arranged at the center of the circle of the lower surface of the sample tube rack top cover 2, and two retractable sample tube rack top cover clamping columns 50 are arranged on the side surfaces of the sample tube rack top cover support rod 49. The sample tube rack top cover 2 is installed in the sample tube rack top cover fixing groove 47 of the sample tube rack top cover positioning column 10 through the sample tube rack top cover support rod 49, and is fixed to the sample tube rack top cover positioning column 10 through the cooperation of the two sample tube rack top cover clamping columns 50 and the two sample tube rack top cover clamping grooves 48; the sample tube rack top cover A sample tube rack top cover handle 15 is arranged at the center of the circle on the upper surface of the tube rack top cover 2, and two sample tube rack top cover clamping column buttons 51 for controlling the extension and retraction of the two sample tube rack top cover clamping columns 50 are arranged oppositely on the side surface of the sample tube rack top cover handle 15; a plurality of sample tube through holes 16 penetrating vertically are evenly arranged on the circumferential edge of the sample tube rack top cover 2, and the number of the sample tube through holes 16 corresponds to the number of the sample tube mounting holes 13, and when the sample tube rack top cover 2 is fixed on the sample tube rack 1, each of the sample tube through holes 16 is coaxial with the corresponding sample tube mounting hole 13 in the vertical direction;

[0072] The sample loading device is arranged above the top cover 2 of the sample loading tube rack, and the sample loading device includes an overall vertical moving mechanism 17 of the sample loading device, and the lower part of the overall vertical moving mechanism 17 of the sample loading device is provided with a hemolysin / buffer microinjection pump fixing rod 18, a specimen microinjection pump moving rod 19 and an antibody microinjection pump fixing rod 20 arranged side by side; a hemolysin / buffer microinjection pump 22 is arranged at the front end of the hemolysin / buffer microinjection pump fixing rod 18 through a hemolysin / buffer microinjection pump fixing block 21, and a hemolysin / buffer microinjection pump 22 is provided with a hemolysin / buffer loading needle 23 at the lower end of the hemolysin / buffer microinjection pump 22. The upper end is provided with a hemolysin / buffer injection needle vertical moving mechanism 24 for driving the hemolysin / buffer injection needle 23 to extend and retract up and down, and the hemolysin / buffer injection needle 23 is provided with a vertically movable hemolysin / buffer injection needle cleaning mechanism 25; the front end of the antibody microinjection pump fixing rod 20 is provided with a No. 1 antibody microinjection pump 27 and a No. 2 antibody microinjection pump 28 through an antibody microinjection pump fixing block 26, and the lower ends of the No. 1 antibody microinjection pump 27 and the No. 2 antibody microinjection pump 28 are respectively provided with a No. 1 antibody injection needle 29 and a No. 2 antibody injection needle 30, and the upper ends of the No. 1 antibody microinjection pump 27 and the No. 2 antibody microinjection pump 28 are respectively provided with a No. 1 antibody injection needle 29 and a No. 2 antibody injection needle 30. The specimen microinjection pump moving rod 19 is provided with a movable specimen microinjection pump horizontal moving mechanism 33, and the specimen microinjection pump horizontal moving mechanism 33 is provided with a specimen microinjection pump 34, and the lower end of the specimen microinjection pump 34 is provided with a specimen injection needle 35, and the upper end of the specimen microinjection pump 34 is provided with a specimen injection needle vertical moving mechanism 36 for driving the specimen injection needle 35 to extend and retract up and down, and the specimen injection needle 35 is provided with a movable specimen injection needle horizontal moving mechanism 33. Specimen injection needle cleaning mechanism 37; when the specimen micro-injection pump horizontal moving mechanism 33 moves to the rear positioning point of the specimen micro-injection pump moving rod 19, the hemolysin / buffer injection needle 23, the specimen injection needle 35, the No. 1 antibody injection needle 29 and the No. 2 antibody injection needle 30 are arranged in an arc shape, and correspond to the plane positions of any four adjacent sample tube through holes 16 on the sample tube rack top cover 2 one by one, respectively. When the specimen micro-injection pump horizontal moving mechanism 33 moves to the front positioning point of the specimen micro-injection pump moving rod 19, the specimen injection needle 35 extends forward to correspond to one of the specimen tube mounting holes 12 on the sample tube rack 1;A reagent bottle base 38 is provided in the middle of the overall vertical moving mechanism 17 of the sample loading device, and a hemolysin reagent bottle 39, a buffer reagent bottle 40, a No. 1 antibody reagent bottle 41 and a No. 2 antibody reagent bottle 42 are respectively provided on the reagent bottle base 38, the hemolysin reagent bottle 39 and the buffer reagent bottle 40 are respectively connected to the hemolysin / buffer microinjection pump 22 through pipelines, the No. 1 antibody reagent bottle 41 is connected to the No. 1 antibody microinjection pump 27 through a pipeline, and the No. 2 antibody reagent bottle 42 is connected to the No. 2 antibody microinjection pump 28 through a pipeline;

[0073] The integrated controller is a controller with PLC as the core, which is respectively electrically connected to the sample tube rack rotating mechanism 4, the sample tube rack chassis rotating mechanism 6, the sample tube rack chassis vertical moving mechanism 7, the overall vertical moving mechanism 17 of the sample loading device, the hemolysin / buffer microinjection pump 22, the hemolysin / buffer injection needle vertical moving mechanism 24, the hemolysin / buffer injection needle cleaning mechanism 25, the No. 1 antibody microinjection pump 27, the No. 2 antibody microinjection pump 28, the No. 1 antibody injection needle vertical moving mechanism 31, the No. 2 antibody injection needle vertical moving mechanism 32, the specimen microinjection pump horizontal moving mechanism 33, the specimen microinjection pump 34, the specimen injection needle vertical moving mechanism 36 and the specimen injection needle cleaning mechanism 37 to coordinate the linkage work of the above-mentioned various motion mechanisms.

[0074] Furthermore, the number of the specimen tube mounting hole 12, the sample tube mounting hole 13, the sample tube syringe rod through hole 14, the sample tube syringe rod push plate through hole 8, the sample tube syringe rod push plate limiting hole 9 and the sample tube through hole 16 are all 40.

[0075] Furthermore, the integrated controller is also electrically connected to a multimedia interactive device to facilitate operators to set up experiments and display experimental progress.

[0076] A matching sample tube for a fully automatic fluorescently labeled single cell suspension preparation device comprises a sample tube body 52, wherein a puncturable incubation cup 53 for cell fluorescent antibody staining is arranged in the middle of the sample tube body 52, a movable syringe rod 54 is arranged in the lower part of the sample tube body 52, a syringe rod push plate 55 is arranged at the lower part of the syringe rod 54, and the syringe rod push plate 55 is exposed below the sample tube body 52, and a circumferential surface of the upper part of the syringe rod 54 is arranged with a syringe rod push plate 55 that is in contact with the sample tube body 52. The inner wall of the tube body 52 is matched with a syringe rod sealing ring 56, and the interior of the syringe rod 54 is a cavity 57 for collecting waste liquid. The cavity 57 extends upward to form an opening on the upper plane of the syringe rod 54, and the upper plane of the syringe rod 54 is provided with a filter membrane support frame 58 with a hollow middle. The lower surface of the filter membrane support frame 58 is provided with a filter membrane support frame sealing ring 59 that matches the opening 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 out red blood cell fragments and waste liquid.

[0077] Furthermore, a plurality of legs 61 are provided 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 legs 61 and the inner wall of the sample loading tube body 52 .

[0078] Furthermore, the hollow portion of the filter membrane support frame 58 is provided with a cross support structure 62 , and the cross support structure 62 is connected to the main portion of the filter membrane support frame 58 , and the filter membrane 60 is jointly covered on the main portion of the filter membrane support frame 58 and the cross support structure 62 .

[0079] Furthermore, the size of the sample loading tube body 52 is the same as the size of common cell loading tubes on the market.

[0080] Furthermore, the sample loading tube body 52 is made of polystyrene or polypropylene.

[0081] Furthermore, the incubation cup 53 is made of polyvinyl chloride.

[0082] Taking the 40-hole specimen tube mounting hole and the sample tube mounting hole of the present invention as an example, the working principle and operation method are as follows:

[0083] The operator first numbers the specimen tubes to be processed, and then sequentially places the numbered specimen tubes into the specimen tube installation holes 12 starting from position 1 of the specimen tube installation hole 12 of the sample tube rack 1, and a maximum of 40 specimen tubes can be placed;

[0084] Next, take the same number of matching sample tubes, and then, starting from position 1 of the sample tube mounting hole 13 of the sample tube rack 1, sequentially place the matching sample tubes into the corresponding sample tube mounting holes 13 on the sample tube rack 1, and ensure that the syringe rod 54 of each matching sample tube passes through the corresponding sample tube syringe rod through hole on the sample tube rack 1, so that its syringe rod push plate 55 is exposed on the lower surface of the sample tube rack 1;

[0085] Place the sample tube rack 1 with the specimen tube and the matching sample tube on the sample tube rack support rod 3, press the sample tube rack clamping column buttons 46 on both sides of the sample tube rack top cover positioning column 10, so that the sample tube rack support rod 3 extends into the sample tube rack support rod extending into the groove 44 at the bottom of the sample tube rack 1, and then release the sample tube rack clamping column button 46, so that the sample tube rack support rod extending into the groove 44 is clamped with the sample tube rack clamping groove 43 on the sample tube rack support rod 3, so that the sample tube rack 1 is fixed on the sample tube rack support rod 3, and at the same time, the syringe rod push plate 55 at the bottom of each matching sample tube passes through the corresponding sample tube syringe rod push plate through hole 8 on the sample tube rack chassis 5;

[0086] Place the sample tube rack top cover 2 on the sample tube rack 1, press the sample tube rack top cover clamping column buttons 51 on both sides of the sample tube rack top cover cover handle 15, so that the sample tube rack top cover support rod 49 on the lower surface of the sample tube rack top cover 2 extends into the sample tube rack top cover fixing groove 47 on the sample tube rack top cover positioning column 10, and then release the sample tube rack top cover clamping column button 51, so that the sample tube rack top cover clamping column 50 on the sample tube rack top cover support rod 49 is clamped with the sample tube rack top cover clamping groove 48 in the sample tube rack top cover fixing groove 47, and the sample tube rack top cover 2 is fixed on the sample tube rack 1;

[0087] Check whether the various reagents in the hemolysin reagent bottle 39, the buffer reagent bottle 40, the No. 1 antibody reagent bottle 41 and the No. 2 antibody reagent bottle 42 are sufficient, confirm whether the pipelines are connected and normal, then select the corresponding experimental plan and quantity on the multimedia interactive device, and click Start to start the experiment;

[0088] The whole vertical moving mechanism 17 of the sample loading device drives the whole sample loading device to move vertically downward to its position. At this time, the No. 1 antibody loading needle 29 is downwardly aligned with the matching loading tube at position 1. Then, the No. 1 antibody loading needle 29 injects antibody A into the matching loading tube at position 1 through the No. 1 antibody microinjection pump 27. Then, the whole vertical moving mechanism 17 of the sample loading device drives the whole sample loading device to move vertically upward back to its position.

[0089] The sample tube rack rotating mechanism 4 drives the sample tube rack 1 to rotate counterclockwise by 9 degrees, and the overall vertical moving mechanism 17 of the sample loading device drives the overall sample loading device to move vertically downward into position. At this time, the No. 1 antibody loading needle 29 is downwardly aligned with the matching sample loading tube at position 2, and the No. 2 antibody loading needle 30 is downwardly aligned with the matching sample loading tube at position 1, and then the No. 1 antibody loading needle 29 injects antibody A into the matching sample loading tube at position 2 through the No. 1 antibody micro-injection pump 27, and at the same time, the No. 2 antibody loading needle 30 injects antibody B into the matching sample loading tube at position 1 through the No. 2 antibody micro-injection pump 28, and then the overall vertical moving mechanism 17 of the sample loading device drives the overall sample loading device to move vertically upward back to position;

[0090] The sample tube rack rotating mechanism 4 drives the sample tube rack 1 to rotate counterclockwise by 9 degrees, and the specimen microinjection pump horizontal moving mechanism 33 moves the specimen microinjection pump 34 to the front positioning point of the specimen microinjection pump moving rod 19. At this time, the specimen injection needle 35 is downwardly aligned with the specimen tube at position 1, and then the specimen injection needle vertical moving mechanism 36 drives the specimen injection needle 35 to move vertically downward and extend into the specimen tube at position 1. The specimen injection needle 35 absorbs the specimen in the specimen tube at position 1 through the specimen microinjection pump 34, and then the specimen injection needle vertical moving mechanism 36 drives the specimen injection needle 35 to move vertically upward back to its position, and then the specimen microinjection pump horizontal moving mechanism 33 moves the specimen microinjection pump 34 back to the rear positioning point of the specimen microinjection pump moving rod 19;

[0091] The overall vertical moving mechanism 17 of the sample loading device drives the overall vertical downward movement of the sample loading device into position. At this time, the sample loading needle 35 is downwardly aligned with the matching sample loading tube at position 1, the No. 2 antibody loading needle 30 is downwardly aligned with the matching sample loading tube at position 2, and the No. 1 antibody loading needle 29 is downwardly aligned with the matching sample loading tube at position 3. Then, the sample loading needle vertical moving mechanism 36 drives the sample loading needle 35 to move vertically downward and extend into the matching sample loading tube at position 1. The sample loading needle 35 injects the absorbed sample into the matching sample loading tube at position 1 through the sample micro-injection pump 34. Then, the sample loading needle 35 is pumped once through the sample micro-injection pump 34 to mix the antibody A, antibody B and sample in the matching sample loading tube at position 1. At the same time, the No. 2 antibody loading needle 30 injects antibody B into the matching sample loading tube at position 2, and the No. 1 antibody loading needle 29 injects antibody A into the matching sample loading tube at position 3.

[0092] The specimen injection needle vertical moving mechanism 36 drives the specimen injection needle 35 to move vertically upward and back to the original position, and then the overall vertical moving mechanism 17 of the injection device drives the overall injection device to move vertically upward and back to the original position, and the specimen injection needle cleaning mechanism 37 cleans the specimen injection needle 35, and this cycle is repeated until all the matching injection tubes on the injection tube rack 1 are added with antibody A, antibody B and specimen and mixed;

[0093] After all the matching sample tubes are added with antibody A, antibody B and the specimen and mixed, the sample tube rack rotating mechanism 4 drives the sample tube rack 1 to rotate clockwise by 9 degrees and then quickly rotate counterclockwise by 9 degrees, and continues for 3 cycles, so as to oscillate and mix the mixture of antibody A, antibody B and blood specimen in all the matching sample tubes, and then incubate the oscillated and mixed mixture of antibody A, antibody B and blood specimen for 15 minutes; the injection, mixing, oscillation and incubation of antibody A, antibody B and blood specimen in each matching sample tube are all carried out in the incubation cup 53 inside it;

[0094] The sample tube rack rotating mechanism 4 drives the sample tube rack 1, so that the hemolysin / buffer injection needle 23 is downwardly aligned with the matching sample tube at position 1, and then the hemolysin / buffer injection needle vertical movement mechanism 24 drives the hemolysin / buffer injection needle 23 to move vertically downward to a first height position, and the hemolysin / buffer injection needle 23 injects hemolysin into the incubation cup 53 of the matching sample tube at position 1 through the hemolysin / buffer microinjection pump 22;

[0095] The hemolysin / buffer injection needle vertical moving mechanism 24 further drives the hemolysin / buffer injection needle 23 to move vertically downward to a second height position. During the process of the hemolysin / buffer injection needle 23 moving vertically downward to the second height position, the hemolysin / buffer injection needle 23 pierces the incubation cup 53 in the matching sample loading tube at position 1, and then the hemolysin / buffer injection needle 23 pumps once through the hemolysin / buffer microinjection pump 22 to mix the hemolysin with the mixed solution of antibody A, antibody B and blood sample;

[0096] The hemolysin / buffer injection needle vertical moving mechanism 24 drives the hemolysin / buffer injection needle 23 to move vertically upward, and returns to the first height position. The hemolysin / buffer injection needle cleaning mechanism 25 cleans the hemolysin / buffer injection needle 23, and then the loading tube rack rotating mechanism 4 drives the loading tube rack 1 to rotate 9 degrees counterclockwise, so that the hemolysin / buffer injection needle 23 is downwardly aligned with the matching loading tube at position 2, and this cycle is repeated until all the matching loading tubes are injected with hemolysin and mixed, and the incubation cups 53 in all the matching loading tubes are punctured;

[0097] After all the matching sample tubes are injected with hemolysin and mixed, the sample tube rack rotating mechanism 4 drives the sample tube rack 1 to rotate clockwise by 9 degrees and then quickly rotate counterclockwise by 9 degrees, and continue for 3 cycles to oscillate and mix the mixed solution of hemolysin, antibody A, antibody B and blood specimen in all the matching sample tubes, and then incubate the oscillated and mixed mixed solution of hemolysin, antibody A, antibody B and blood specimen for 15 minutes;

[0098] The sample tube rack chassis rotating mechanism 6 drives the sample tube rack chassis 5 to rotate 9 degrees counterclockwise, so that the syringe rods 54 of all matching sample tubes are inserted into the corresponding sample tube syringe rod limiting holes 9 on the sample tube rack chassis 5, and then the sample tube rack chassis vertical moving mechanism 7 drives the sample tube rack chassis 5 and the sample tube rack chassis rotating mechanism 6 to move vertically downward together, and the sample tube rack chassis 5 pulls the syringe rods 54 of all matching sample tubes to move vertically downward for a distance, causing negative pressure in the matching sample tubes;

[0099] Under the action of negative pressure, the mixed solution of the incubated hemolysin, antibody A, antibody B and blood specimen breaks through the perforations on the incubation cup 53 and flows into the lower cavity of the matching sample loading tube between the incubation cup 53 and the filter membrane 60. Under the action of gravity, the red blood cell fragments and waste liquid in the mixed solution of the hemolysin, antibody A, antibody B and blood specimen flow through the filter membrane 60 into the cavity 57 in the syringe rod 54, and the white blood cells in the mixed solution of the hemolysin, antibody A, antibody B and blood specimen are retained on the filter membrane 60.

[0100] The sample tube rack chassis vertical moving mechanism 7 drives the sample tube rack chassis 5 and the sample tube rack chassis rotating mechanism 6 to move vertically upward and return to their original positions. The sample tube rack chassis 5 pushes the syringe rods 54 of all matching sample tubes to move vertically upward and return to their original positions. The sample tube rack chassis rotating mechanism 6 drives the sample tube rack chassis 5 to rotate 9 degrees clockwise, so that the syringe rods 54 of all matching sample tubes are separated from the corresponding sample tube syringe rod limiting holes 9 on the sample tube rack chassis 5 and return to their original positions.

[0101] The sample tube rack rotating mechanism 4 drives the sample tube rack 1, so that the hemolysin / buffer injection needle 23 is again downwardly aligned with the matching sample tube at position 1, and then the hemolysin / buffer injection needle vertical movement mechanism 24 drives the hemolysin / buffer injection needle 23 to move vertically downward to a first height position, and the hemolysin / buffer injection needle 23 injects buffer into the matching sample tube at position 1 through the hemolysin / buffer microinjection pump 22, and then the hemolysin / buffer injection needle 23 is sucked once through the hemolysin / buffer microinjection pump 22 to mix the buffer with the white blood cells on the filter membrane 60;

[0102] The hemolysin / buffer injection needle vertical moving mechanism 24 drives the hemolysin / buffer injection needle 23 to move vertically upward back to its original position, and the hemolysin / buffer injection needle cleaning mechanism 25 cleans the hemolysin / buffer injection needle 23, and then the loading tube rack rotating mechanism 4 drives the loading tube rack 1 to rotate counterclockwise by 9 degrees, so that the hemolysin / buffer injection needle 23 is downwardly aligned with the matching loading tube at position 2, and the cycle is repeated until the buffer is injected into all the matching loading tubes and mixed with the white blood cells on the corresponding filter membranes 60;

[0103] After all the matching sample tubes are injected with buffer and mixed, the sample tube rack rotating mechanism 4 drives the sample tube rack 1 to rotate 9 degrees clockwise and then quickly rotate 9 degrees counterclockwise, and continue for 3 cycles to oscillate and mix the buffer and white blood cells in all the matching sample tubes to form a fluorescently labeled white blood cell single cell suspension;

[0104] Finally, the operator removes the sample tube rack 1 from the sample tube rack rotating device and moves it to the detection device for direct detection.

[0105] All the motion mechanisms on the fully automatic fluorescent labeled single cell suspension preparation device of the present invention are controlled by an integrated controller with PLC as the core. The integrated controller coordinates the work of all the motion mechanisms according to a pre-set experimental program, thereby ensuring the smooth completion of steps including sample addition, hemolysis, separation, separation of leukocytes and suspension of leukocytes, thereby realizing the automation of human or animal peripheral blood sample processing.

[0106] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A fully automatic fluorescent labeled single cell suspension preparation device, characterized in that: It is composed of a sample tube rack rotating device, a sample tube rack (1), a sample tube rack top cover (2), a sample adding device and an integrated controller; The sample tube rack rotating device comprises a sample tube rack support rod (3) for mounting the sample tube rack (1); a sample tube rack rotating mechanism (4) for driving the sample tube rack (1) to rotate on the sample tube rack support rod (3) is arranged at the lower part of the sample tube rack support rod (3); a sample tube rack chassis (5) is movably sleeved on the middle part of the sample tube rack support rod (3); a sample tube rack chassis rotating mechanism (6) for driving the sample tube rack chassis (5) to rotate on the sample tube rack support rod (3) is arranged on the lower surface of the sample tube rack chassis (5); A sample tube rack chassis vertical movement mechanism (7) is provided on the lower surface of the tube rack chassis rotation mechanism (6) for driving the sample tube rack chassis (5) to move vertically on the sample tube rack support rod (3); a plurality of sample tube syringe rod push plate through holes (8) extending vertically are evenly provided on the circumferential edge of the sample tube rack chassis (5); a sample tube syringe rod limiting hole (9) extending vertically is provided on the clockwise side of each sample tube syringe rod push plate through hole (8), and each sample tube syringe rod limiting hole (9) is connected to the corresponding sample tube syringe rod push plate through hole (8); The sample tube rack (1) is coaxially rotatably arranged on the upper part of the sample tube rack support rod (3); a sample tube rack top cover positioning column (10) for mounting the sample tube rack top cover (2) is arranged at the center of the circle on the upper surface of the sample tube rack (1); a circle of bosses (11) are arranged on the circumferential edge of the upper surface of the sample tube rack (1); a plurality of specimen tube mounting holes (12) are evenly arranged on the upper surface of the bosses (11); a plurality of sample tube mounting holes (13) are arranged on the upper surface of the sample tube rack (1) along the inner side of the bosses (11); a sample tube syringe rod through hole (14) is arranged at the bottom of each of the sample tube mounting holes (13) and is passed through from top to bottom; and each of the sample tube mounting holes (13) is provided with a sample tube syringe rod through hole (14) passing through from top to bottom. The sample tube mounting holes (13) are respectively coaxial with the corresponding sample tube syringe rod through holes (14); the number of the sample tube mounting holes (12) corresponds to the number of the sample tube syringe rod through holes (14), and when the sample tube rack (1) is fixed on the sample tube rack support rod (3), each of the sample tube mounting holes (12) is respectively coaxial with the corresponding sample tube syringe rod through holes (14); the number of the sample tube mounting holes (13) corresponds to the number of the sample tube mounting holes (12), and the distribution position of each of the sample tube mounting holes (13) corresponds to the distribution position of the corresponding sample tube mounting holes (12); The top cover (2) of the sample tube rack is coaxially rotatably arranged on the sample tube rack (1); a sample tube rack top cover handle (15) is arranged at the center of the circle on the upper surface of the sample tube rack top cover (2); a plurality of sample tube through holes (16) penetrating vertically are evenly arranged on the circumferential edge of the sample tube rack top cover (2); the number of the sample tube through holes (16) corresponds to the number of the sample tube mounting holes (13); and when the top cover (2) of the sample tube rack is fixed on the sample tube rack (1), each of the sample tube through holes (16) is vertically coaxial with the corresponding sample tube mounting hole (13); The sample loading device is arranged above the top cover (2) of the sample loading tube rack, and comprises an overall vertical moving mechanism (17) for the sample loading device, wherein a hemolysin / buffer microinjection pump fixing rod (18), a specimen microinjection pump moving rod (19) and an antibody microinjection pump fixing rod (20) are arranged side by side at the lower part of the overall vertical moving mechanism (17); a hemolysin / buffer microinjection pump fixing rod (18), a specimen microinjection pump moving rod (19) and an antibody microinjection pump fixing rod (20) are arranged side by side; a hemolysin / buffer microinjection pump (22) is arranged at the front end of the hemolysin / buffer microinjection pump fixing rod (18) via a hemolysin / buffer microinjection pump fixing block (21); The lower end of the hemolysin / buffer microinjection pump (22) is provided with a hemolysin / buffer injection needle (23), the upper end of the hemolysin / buffer injection pump (22) is provided with a hemolysin / buffer injection needle vertical movement mechanism (24) for driving the hemolysin / buffer injection needle (23) to extend and retract upward and downward, and the hemolysin / buffer injection needle (23) is provided with a hemolysin / buffer injection needle cleaning mechanism (25) that can be vertically moved; the front end of the antibody microinjection pump fixing rod (20) is provided with a No. 1 antibody via an antibody microinjection pump fixing block (26). A microinjection pump (27) and a No. 2 antibody microinjection pump (28), wherein the lower ends of the No. 1 antibody microinjection pump (27) and the No. 2 antibody microinjection pump (28) are respectively provided with a No. 1 antibody sample injection needle (29) and a No. 2 antibody sample injection needle (30), and the upper ends of the No. 1 antibody microinjection pump (27) and the No. 2 antibody microinjection pump (28) are respectively provided with a No. 1 antibody sample injection needle vertical movement mechanism (31) and a No. 2 antibody sample injection needle vertical movement mechanism (32) for driving the No. 1 antibody sample injection needle (29) and the No. 2 antibody sample injection needle (30) to extend and retract upward and downward; The specimen microinjection pump moving rod (19) is provided with a movable specimen microinjection pump horizontal moving mechanism (33), the specimen microinjection pump horizontal moving mechanism (33) is provided with a specimen microinjection pump (34), the lower end of the specimen microinjection pump (34) is provided with a specimen injection needle (35), the upper end of the specimen microinjection pump (34) is provided with a specimen injection needle vertical moving mechanism (36) for driving the specimen injection needle (35) to extend and retract upward and downward, and the specimen injection needle (35) is provided with a vertically movable specimen injection needle cleaning mechanism (37);When the specimen microinjection pump horizontal moving mechanism (33) moves to the rear positioning point of the specimen microinjection pump moving rod (19), the hemolysin / buffer injection needle (23), the specimen injection needle (35), the No. 1 antibody injection needle (29) and the No. 2 antibody injection needle (30) are arranged in an arc shape and correspond to the plane positions of any four adjacent sample tube through holes (16) on the top cover (2) of the sample tube rack, respectively. When the specimen microinjection pump horizontal moving mechanism (33) moves to the front positioning point of the specimen microinjection pump moving rod (19), the specimen injection needle (35) extends forward to contact with one of the sample tubes on the sample tube rack (1). The tube mounting holes (12) correspond to each other up and down; a reagent bottle base (38) is provided in the middle of the overall vertical moving mechanism (17) of the sample adding device, and a hemolysin reagent bottle (39), a buffer reagent bottle (40), a No. 1 antibody reagent bottle (41) and a No. 2 antibody reagent bottle (42) are respectively provided on the reagent bottle base (38); the hemolysin reagent bottle (39) and the buffer reagent bottle (40) are respectively connected to the hemolysin / buffer microinjection pump (22) through pipelines, the No. 1 antibody reagent bottle (41) is connected to the No. 1 antibody microinjection pump (27) through a pipeline, and the No. 2 antibody reagent bottle (42) is connected to the No. 2 antibody microinjection pump (28) through a pipeline; The integrated controller is electrically connected to the sample tube rack rotating mechanism (4), the sample tube rack chassis rotating mechanism (6), the sample tube rack chassis vertical moving mechanism (7), the sample loading device overall vertical moving mechanism (17), the hemolysin / buffer microinjection pump (22), the hemolysin / buffer injection needle vertical moving mechanism (24), the hemolysin / buffer injection needle cleaning mechanism (25), the No. 1 antibody microinjection pump (27), the No. 2 antibody microinjection pump (28), the No. 1 antibody injection needle vertical moving mechanism (31), the No. 2 antibody injection needle vertical moving mechanism (32), the specimen microinjection pump horizontal moving mechanism (33), the specimen microinjection pump (34), the specimen injection needle vertical moving mechanism (36) and the specimen injection needle cleaning mechanism (37); Two sample tube rack clamping grooves (43) for fixing the sample tube rack (1) are arranged on the side surface of the upper part of the sample tube rack support rod (3); a sample tube rack support rod extending groove (44) is arranged at the center of the circle of the lower surface of the sample tube rack (1); two retractable sample tube rack clamping columns (45) are arranged on the groove wall of the sample tube rack support rod extending groove (44); two sample tube rack clamping column buttons (46) for controlling the extension and retraction of the two sample tube rack clamping columns (45) are arranged on the side surface of the sample tube rack top cover positioning column (10); the sample tube rack (1) is sleeved on the upper part of the sample tube rack support rod (3) through the sample tube rack support rod extending groove (44), and is fixed to the sample tube rack support rod (3) by the cooperation of the two sample tube rack clamping columns (45) and the two sample tube rack clamping grooves (43); A sample tube rack top cover fixing groove (47) is arranged at the center of the upper surface of the sample tube rack top cover positioning column (10), two sample tube rack top cover clamping grooves (48) are arranged oppositely on the groove wall of the sample tube rack top cover fixing groove (47), a sample tube rack top cover support rod (49) is arranged at the center of the lower surface of the sample tube rack top cover (2), two retractable sample tube rack top cover clamping columns (50) are arranged oppositely on the side surface of the sample tube rack top cover support rod (49), and the sample tube rack top cover cover handle (15 ) are provided with two sample tube rack top cover clamping column buttons (51) for controlling the extension and retraction of the two sample tube rack top cover clamping columns (50) on the side surfaces thereof. The sample tube rack top cover (2) is installed in the sample tube rack top cover fixing groove (47) of the sample tube rack top cover positioning column (10) via the sample tube rack top cover supporting rod (49), and is fixed to the sample tube rack top cover positioning column (10) by the cooperation of the two sample tube rack top cover clamping columns (50) and the two sample tube rack top cover clamping grooves (48).

2. The fully automatic fluorescent labeled single cell suspension preparation device according to claim 1, characterized in that: The number of the specimen tube mounting hole (12), the sample tube mounting hole (13), the sample tube syringe rod through hole (14), the sample tube syringe rod push plate through hole (8), the sample tube syringe rod limiting hole (9) and the sample tube through hole (16) is 40.

3. The fully automatic fluorescent labeled single cell suspension preparation device according to claim 1, characterized in that: The integrated controller is also electrically connected to a multimedia interactive device, which is convenient for operators to set up experiments and display the progress of experiments.

4. A matching sample loading tube for the fully automatic fluorescent labeled single cell suspension preparation device according to any one of claims 1 to 3, characterized in that: The invention comprises a sample tube body (52), wherein a puncturable incubation cup (53) for cell fluorescent antibody staining is arranged in the middle of the sample tube body (52), a movable syringe rod (54) is arranged in the lower part of the sample tube body (52), a syringe rod push plate (55) is arranged in the lower part of the syringe rod (54), and the syringe rod push plate (55) is exposed below the sample tube body (52), and a syringe rod sealing ring (56) is arranged on the circumferential surface of the upper part of the syringe rod (54) and cooperates with the inner wall of the sample tube body (52), and the interior of the syringe rod (54) is a cavity (57) for collecting waste liquid, and the cavity (57) extends upward to form an opening on the upper plane of the syringe rod (54), and the upper plane of the syringe rod (54) is provided with a middle hollow A filter membrane support frame (58) is provided on the lower surface of the filter membrane support frame (58) with a filter membrane support frame sealing ring (59) that cooperates with the opening of the cavity (57), and a filter membrane (60) for filtering out red blood cell fragments and waste liquid is provided on the upper surface of the filter membrane support frame (58); a plurality of supporting feet (61) are provided on the outer wall of the incubation cup (53), and the incubation cup (53) is arranged in the middle part of the sample loading tube body (52) through the contact between the supporting feet (61) and the inner wall of the sample loading tube body (52); a cross supporting structure (62) is provided in the hollow part of the filter membrane support frame (58), and the cross supporting structure (62) is connected to the main part of the filter membrane support frame (58), and the filter membrane (60) is jointly covered on the main part of the filter membrane support frame (58) and the cross supporting structure (62).

5. The supporting sample loading tube of the fully automatic fluorescent labeled single cell suspension preparation device according to claim 4 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 the size of common cell loading tubes on the market.

6. The supporting sample loading tube of the fully automatic fluorescent labeled single cell suspension preparation device according to claim 4, characterized in that: The incubation cup (53) is made of polyvinyl chloride.

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