CN115369021A - Multipurpose flow cytometry washing instrument, control method and application - Google Patents
Multipurpose flow cytometry washing instrument, control method and application Download PDFInfo
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Abstract
The invention belongs to the technical field of medical experimental equipment, and discloses a multipurpose flow cytometry washing instrument, a control method and application. A compressor is arranged on the left side of the bottom of the case body; four heat preservation water tanks are arranged in the middle of the case body and respectively provide common temperatures. The outer side of each heat-preservation water tank is provided with a water tank water injection port and a water tank overflow port, and is also provided with an evaporator and a temperature controller, and an electric heating tube is arranged inside the heat-preservation water tank. And a heat exchange coil is arranged in each heat preservation water tank, and the washing liquor in the heat preservation water tank and the water body of the heat preservation water tank perform non-contact heat exchange. The inlet of the heat exchange coil is connected with a washing liquid bag through a washing liquid connecting pipe; the outlet of the heat exchange coil is sequentially connected with a lotion distributing pipe, a washing kettle, a liquid discharge pipe, a waste liquid pipe and a waste liquid box through a channel switching pipe. The washing kettle is provided with a cell filter membrane, a sample inlet and a sample outlet. The invention can avoid centrifugal damage in traditional washing, can efficiently carry out open or closed washing on cells, and can meet the requirement of temperature control.
Description
Technical Field
The invention belongs to the technical field of medical experimental equipment, and particularly relates to a multipurpose flow cytometry washing instrument, a control method and application.
Background
Currently, transfusion departments and other medical/life science related laboratories in various medical institutions often require washing of cells, such as: (1) Carrying out an anti-human globulin test or an absorption and diffusion test by a test tube method; (2) Some frozen cells require a process of reviving and removing the protective agent; (3) A process is needed for preparing washed red blood cells to patients for clinical transfusion. For the case (1), open washing of the cells is generally required, and the common procedures are: adding appropriate amount of cell sap to be washed into 1 clean test tube or centrifuge tube, adding sufficient physiological saline or special washing solution, mixing, centrifuging, separating supernatant, and discarding supernatant. Physiological saline or a special washing solution is added again, the resuspended cells are centrifuged again and the supernatant is discarded. Repeating the steps for 3 to 6 times until the washing task is finished. For the case (2), if the cells are recovered and washed to remove the protective agent for the related detection, an open-type washing is usually adopted, and the operation process is the same as that of the case (1); if the frozen cells are used for culture, human therapy or animal experiments after washing, closed washing is required, and the operation principle is basically the same as that of the case (1), but a sterile cell bag is used, and sterile operation is required for both addition and removal of the washing solution. And (3) washing in the case of (3) also needs to adopt aseptic operation, adding physiological saline into the erythrocyte suspension bag, uniformly mixing and centrifuging, separating and removing supernatant, circulating for 3 times, adding proper amount of physiological saline or erythrocyte preservation solution, and resuspending to obtain a washed erythrocyte product.
At present, most laboratories finish washing in a manual operation mode, washing effects of different operators are obviously different, batch-to-batch differences of operation also exist in the same operator, and especially when the sample amount is large. In the case of aseptic washing, the process of frequently adding washing solution and removing supernatant after centrifugation has a great risk of contamination. Furthermore, manual operations are labor intensive, inefficient, time consuming, error prone, and difficult to perform in large batches, such as: when the antibody titer of the patient is detected by a test tube anti-human globulin method. It is more difficult to wash samples at different temperatures for absorption and diffusion tests, to eliminate self-agglutination or non-specific agglutination, etc., and even to wash the same sample cyclically with different temperature washes, which are essentially impossible to perform in manual mode of operation. Even if the method of placing the washing liquid in a refrigerator or a water bath box in advance is adopted, the temperature of the centrifugal link cannot be accurately controlled. Finally, the important disadvantage is that not only is the cleanliness of the wash limited, but the cells often suffer from various degrees of centrifugal damage, even rupture or hemolysis after centrifugal washing. Some laboratories adopt automatic centrifugal equipment, can realize totally enclosed washing, still have centrifugal damage, can't realize the washing of multiple temperature, can't wash a plurality of samples simultaneously, it is long consuming time, complete consumptive material can only disposable use lead to cost high grade to be not enough. In addition, some open automatic centrifugal washing devices cannot be used in an application environment with aseptic requirements for washing, and still have the defects of centrifugal damage, incapability of controlling temperature and the like. Some laboratories adopt needle filters or test tubes/centrifuge tubes with filter membranes to carry out manual filtration, some are still used with a suction filtration device, so that centrifugal damage is avoided, the time consumption is short, and the laboratory has the advantage of being more thorough than centrifugal washing. However, this method is only suitable for washing a small amount of cell samples, and cannot control the washing temperature precisely, and the risk of contamination is high (even in a fully open washing).
Through the above analysis, the main problems and defects of the prior art are as follows: the device is only suitable for washing a small amount of cell samples, the washing temperature cannot be controlled, the pollution risk is high, and centrifugal damage exists.
The difficulty in solving the above problems and defects is: a multi-channel washing device is needed which can wash multiple cell samples simultaneously; equipment capable of switching the temperature of the washing liquid at any time is required; instruments that can perform both open and closed washes as desired; it is necessary to abandon the centrifugal washing mode.
The significance of solving the problems and the defects is as follows: the invention designs a full-automatic washing instrument, which washes a cell sample by adopting a flowing temperature-control washing liquid and collects the washed cells by adopting a filter membrane. Thus, not only can centrifugal damage be avoided, but also open-type or closed-type washing can be efficiently carried out on cells, and the temperature control requirement is met. Through the multichannel setting, still can realize the quick processing of big sample size. The invention can meet the cell washing requirements of all relevant mechanisms and has very wide application prospect.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a multipurpose flow cytometry washing instrument, a control method and application.
The invention is realized in this way, a multi-purpose flow cytometer is provided with a case body;
a compressor is installed on the left side of the bottom of the case body, four heat-preservation water tanks are arranged in the middle of the case body and respectively provide a common temperature, an evaporator and a temperature controller are arranged on the outer sides of the heat-preservation water tanks, and an electric heating pipe is arranged in each heat-preservation water tank (or a temperature-changing part such as a semiconductor refrigerating sheet is used for replacing the compressor, the electric heating pipe and the like);
a water tank water filling port and a water tank overflow port are arranged on the outer side of each heat-preservation water tank, the water tank water filling port is also a water outlet when water is changed, a heat exchange coil is arranged in each heat-preservation water tank, and cleaning liquid in each heat-preservation water tank is in contactless heat exchange with a water body of the heat-preservation water tank;
the outlet of the heat exchange coil is connected with a washing liquid distribution pipe through a channel switching pipe, and the washing liquid distribution pipe is connected with a washing kettle; the liquid discharge pipe is connected with the outlet of the washing kettle, the waste liquid box is connected with the liquid discharge pipe through a waste liquid pipe, and the waste liquid box is arranged on the right side of the bottom of the case body and can be connected with a sewer interface.
Furthermore, an inlet of a heat exchange coil arranged in the heat-preservation water tank is connected with a washing liquid bag through a washing liquid connecting pipe, and the washing liquid bag is arranged at the upper part of the case body;
except that the inlet and the outlet of the heat exchange coil are exposed on the water surface of the heat-preservation water tank, the tube body is completely immersed under the water surface of the water tank, and the inside and the outside of the tube body are completely isolated.
Furthermore, the channel switching tube, the washing liquid distribution tube and the washing kettle are all arranged on a support inside the heat insulation structure, and the heat insulation structure is arranged on the upper part of the case body, so that the washing liquid flowing out of the heat exchange coil passes through the channel switching tube, the washing liquid distribution tube and the washing kettle at a temperature close to a set temperature to wash a cell sample in the kettle; in the heat insulation structure, each washing channel is sealed in the whole process, and no exchange occurs between the washing liquid in the pipeline and the heat insulation cavity.
Further, a first case top cover and a second case top cover are installed on the upper side of the case body, and a case front cabinet door is installed on the front side of the case body.
Furthermore, a touch display screen is embedded on the upper side of the case body.
Furthermore, a washing liquid distribution pipe is installed at the upper end of the washing kettle, an air inlet pipe with an air filter membrane, a peristaltic pump valve and an electromagnetic valve are installed at the upper end of the washing liquid distribution pipe, and a washing liquid main outlet and a washing liquid auxiliary outlet are formed in the right end of the washing liquid distribution pipe; the main outlet of the washing liquid is connected with the main inlet of the washing kettle, the auxiliary outlet of the washing liquid is connected with the inlet of the scouring pipe network in the washing kettle body, and the right side of the washing kettle is provided with a sample inlet and a sample outlet.
Furthermore, a first washing kettle bracket and a second washing kettle bracket are arranged on the outer side of the washing kettle, a liquid discharge pipe is arranged at the lower end of the washing kettle, and a peristaltic pump valve is arranged on the liquid discharge pipe;
the upper end of the washing kettle is provided with an upper side cell filter membrane for intercepting larger impurity cells, and the aperture of the upper side cell filter membrane can pass through the cells to be washed and simultaneously intercept the mixed cells;
the lower end of the washing kettle is provided with a lower cell filter membrane for intercepting cells to be washed, the types of the cells to be washed are different, and the pore diameters of the lower cell filter membrane are different;
an integrally formed scouring pipe network is arranged in the kettle wall of the washing kettle, and the scouring pipe network is provided with 1 inlet, namely a washing liquid auxiliary inlet, which is connected with an auxiliary outlet of a washing liquid distribution pipe; the scouring pipe network is provided with a plurality of outlets and is positioned on the upper pot wall of the lower side cell filter membrane to form a plurality of scouring holes; according to different application requirements, the washing kettle is divided into a plurality of specifications, and the shape size, the number of filter membranes and the aperture of the filter membranes are different.
Another object of the present invention is to provide a method for controlling the multi-purpose flow cytometer, comprising the steps of:
before washing, setting the temperature of each heat-preservation water tank at one time to keep the instrument in a standby state, and starting washing work according to work requirements at any time; simultaneously installing all parts of the washing instrument;
secondly, under the linkage control of each peristaltic pump valve and each electromagnetic valve, gas in each tube cavity of the washing liquid emptying passage is added through a sample inlet of a washing kettle, and cells to be washed are added through a pipettor or a dropper or are directly connected with a sterile cell storage bag for sample addition; washing is started, washing liquor with a specific temperature is selectively added into a main inlet and an auxiliary inlet of a washing kettle synchronously or alternatively according to program control, the washing kettle synchronously or alternatively rotates back and forth at a certain angle along with a support, and the speed is set to be 5-10 times/min; the washing liquid flows to a liquid discharge pipe after passing through the filter membrane, and flows to a waste liquid tank or a sewer through a waste liquid pipe;
step three, stopping the washing kettle from rotating back and forth and keeping the washing kettle in a vertical state, and stopping the washing liquor from entering the washing liquor auxiliary inlet; the main inlet continues to feed the washing liquid until all the cells to be washed reach the lower side cell filter membrane in the washing kettle cavity; the main liquid inlet stops feeding washing liquid, and the electromagnetic valve of the air inlet pipe is opened to enable filtered clean air to enter so as to empty the washing liquid in the washing kettle cavity; the washing machine can be set to single or multiple times of washing according to application requirements, and the same temperature or different temperatures can be set in each washing process;
step four, after the last washing 1 time is finished and the washing liquid is emptied, closing a drain pipe peristaltic pump valve; adding a proper amount of washing liquid through the auxiliary inlet of the washing kettle to resuspend the cells on the lower side filter membrane; or directly adding physiological saline or special cell preservation solution from the sample outlet to resuspend and wash the washed cells; collecting the washed cells from the outlet for subsequent detection or treatment; according to the experiment or treatment requirements, the cell collection mode can be an open suction mode such as a pipette, a suction pipe and the like; or directly connecting with a sterile cell preservation bag pre-filled with preservation solution or empty for collection;
and fifthly, disassembling and assembling a washing liquid distribution pipe, a washing kettle and a liquid discharge pipe, and discarding.
Further, the specific process of installing each component in the step one is as follows:
hoisting or placing a washing liquid bag, a washing liquid bottle or a washing liquid box, selecting and installing a plurality of heat exchange coil pipes with proper specifications according to needs, connecting an outlet of the washing liquid bag and an inlet of the heat exchange coil pipe through 1 washing liquid connecting pipe, and installing the heat exchange coil pipe on an instrument; installing a washing liquid channel adapter tube, and connecting an inlet of the washing liquid channel adapter tube with an outlet of the heat exchange coil; installing a washing liquid distribution pipe, and connecting an inlet of the washing liquid distribution pipe with an outlet of the channel adapter pipe; installing a washing kettle with proper specification, respectively connecting a main inlet and an auxiliary inlet of the washing kettle with a main outlet and an auxiliary outlet of a washing liquid distribution pipe, and adjusting a support of the washing kettle to an initial position to enable the washing kettle to be in a vertical state; installing a liquid discharge pipe, connecting the inlet of the liquid discharge pipe with the outlet of the washing kettle, and connecting the outlet of the liquid discharge pipe with the inlet of the waste liquid pipe; and a waste liquid pipe is arranged, and the outlet of the waste liquid pipe is connected with a waste liquid barrel, a waste liquid box, a waste liquid bag or a sewer.
It is another object of the present invention to provide a medical experimental apparatus employing the multi-purpose flow cytometer.
By combining all the technical schemes, the invention has the advantages and positive effects that:
the invention provides a full-automatic flow type cell washing device, through continuous washing and elutriation of washing liquid flow, the capacity of removing to-be-discarded components can reach about 100%, and the washing effect is excellent. The traditional centrifugal washing mode is essentially to repeatedly dilute and separate the discarded components, so that the cleanliness of the washed cells is limited, for example, the residual amount of plasma protein of the washed red blood cell products for clinical blood transfusion prepared by a centrifugal method can still reach about 3 percent.
The flow washing speed adopted by the invention is obviously higher than that of the traditional centrifugal washing mode, for example, 1 unit of red blood cell is prepared, the traditional centrifugal washing mode needs 3 times of physiological saline injection, 3 times of centrifugation and 3 times of supernatant separation, and the whole process generally takes more than 1 hour; the time consumed by washing for 3 times by adopting the method can be controlled within 15 minutes.
The invention can not cause cell damage basically, while the traditional centrifugal washing mode can cause obvious cell damage. For example, in the preparation of washed erythrocytes, since centrifugation is required at about 4000rpm for 6 minutes each time, the whole preparation process requires 3 times of centrifugation, and thus damage to erythrocytes is significant, and the hemolysis rate often reaches about 0.8%. The traditional centrifugal washing method needs repeated separation of cells and supernatant, which results in a large loss of washed cells. For example, the amount of red blood cells lost during the preparation of 1 bag of washed red blood cells is generally greater than 10% or even up to 20% of the total amount of red blood cells before washing. After the flow washing mode is adopted, a small amount of red blood cells are lost once only in the cell stage after the final collection and washing, and the loss rate can be controlled within 2 percent.
The invention can adopt multi-temperature washing, and washing liquids with different temperatures can be switched randomly according to requirements, thereby bringing great convenience to the completion of an anti-human globulin test, an absorption and diffusion test, the removal of interference of condensed agglutinin or autoantibodies, the elimination of self-agglutination in blood transfusion compatibility detection and the like, also obviously improving the standard operation level of the test and enhancing the comparability of the detection result.
The invention has flexible pipeline configuration, and designs combined and integrated pipeline consumables aiming at open type and closed type washing respectively, thereby reducing the use of disposable consumables to the maximum extent. Compared with the disposable complete pipeline system, the use cost of consumables can be reduced by about 60 percent. The pipeline specification and the use mode of the invention are diversified: the specification, the size and the pore diameter of the filter membrane of the washing kettle can be designed according to different washing requirements; the sample introduction and sample discharge modes are flexible and selectable, and open type or closed type washing can be carried out. Therefore, the application range of the instrument is very wide, and the general centrifugal cell washing equipment can only be applied to a certain single use situation.
The invention can be designed into a multi-channel structure, can synchronously wash a plurality of cell samples, and the washing mode (open or closed), the washing temperature, the times of repeated washing and the specification of the used consumable pipeline of each channel can be independently selected and independently set by a main control computer without mutual interference. The apparatus is thus particularly suitable for large-scale, multi-purpose washing, which conventional centrifugal washing apparatuses are unable to meet.
The above advantages of the present invention allow the scope of use of the instrument to cover almost all applications, such as transfusion department laboratories, blood station component laboratories and laboratories, other medical or life science laboratories, and the like. The domestic market demand can reach more than 2 thousands of devices, and the application prospect is very wide.
Drawings
FIG. 1 is a schematic diagram of a multi-purpose flow cytometer provided in accordance with an embodiment of the present invention;
FIG. 2 is a simplified diagram of a case and a thermal insulation water tank of the multi-purpose flow cytometer;
FIG. 3 is a schematic structural diagram of a heat preservation water tank and a temperature control system of the multi-purpose flow cytometer according to the embodiment of the present invention;
FIG. 4 is a schematic diagram showing the connection relationship between the components of the multi-purpose flow cytometer in the four-temperature single-channel washing process;
FIG. 5 is a schematic diagram of the connection method of the pipes and the flow direction of the washing solution in the four-temperature four-channel washing process of the multi-purpose flow cytometer provided in the embodiment of the present invention;
FIG. 6 is a schematic view of a structure of a washing kettle and a washing liquid distribution pipe according to an embodiment of the present invention;
FIG. 7 is a schematic flow chart of a multi-purpose flow cytometer control method provided by an embodiment of the present invention;
in the figure: 1. a washing liquid bag; 2. a washing liquid connecting pipe; 3. a first chassis top cover; 4. a channel switching tube; 5. an overflow port of the water tank; 6. a heat exchange coil; 7. a heat preservation water tank; 8. an electric heating tube; 9. a water filling port of the water tank; 10. an air return pipe; 11. a compressor; 12. a heat dissipation hole of the chassis; 13. an exhaust pipe; 14. a condenser; 15. drying the filter; 16. a capillary tube; 17. a waste liquid tank; 18. a temperature controller; 19. an evaporator; 20. a waste liquid pipe; 21. a cabinet door in front of the cabinet; 22. a liquid discharge pipe; 23. a heat preservation structure; 24. a washing kettle; 25. a washing liquid distribution pipe; 26. a second chassis top cover; 27. a touch-sensitive display screen; 28. an electromagnetic valve; 29. a peristaltic pump valve; 30. an air inlet pipe; 31. a heat exchange coil inlet; 32. a heat exchange coil outlet; 33. a secondary washing liquid inlet; 34. an upper cell filtration membrane; 35. scouring the pipe network; 36. a first washpot stand; 37. flushing the hole; 38. a lower cell filtration membrane; 39. a second washpot stand; 40. a main washing liquid inlet; 41. a sample inlet; 42. and a sample outlet.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
In view of the problems of the prior art, the present invention provides a multi-purpose flow cytometer, a control method and an application thereof, which will be described in detail with reference to the accompanying drawings.
As shown in fig. 1, the multipurpose flow cytometer provided by the embodiment of the present invention includes a case body, and 4 heat preservation water tanks 7 with different temperatures are disposed in parallel at the middle position of the case body. An electric heating pipe 8 is arranged in the heat preservation water tank 7, a compressor 11 is arranged on the left side of the bottom of the case body, the compressor 11 is sequentially connected with a condenser 14, a drying filter 15 and a capillary tube 16 through an exhaust pipe 13, the rear branches are respectively connected with evaporators 19 arranged on the outer sides of the heat preservation water tanks 7, and air return pipes of the evaporators 19 are converged into an air return pipe 10 and then return to the compressor 11. The outside of each heat preservation water tank 7 is provided with a temperature controller 18 for detecting the temperature in each water tank and controlling the operation of the electric heating tube 8 and the compressor 11 according to the program setting. The outside of the heat preservation water tank 7 is provided with a water tank water filling port 9 (which is a discharge port when water is replaced) and a water tank overflow port 5, and the heat exchange coil 6 is arranged inside the heat preservation water tank 7. The outlet of the heat exchange coil 6 is connected with a washing liquid distribution pipe 25 through a channel adapter pipe 4, and the washing liquid distribution pipe 25 is connected with a washing kettle 24. The channel adapter tube 4, the washing liquid distribution tube 25 and the washing kettle 24 are all installed on a support inside the heat insulation structure 23, and the heat insulation structure 23 is installed on the upper portion of the case body. An inlet of the heat exchange coil 6 is connected with a washing liquid bag 1 through a washing liquid connecting pipe 2, and the washing liquid bag 1 is arranged at the upper part of the case body; a waste liquid box 17 is arranged on the right side of the bottom of the case body, the waste liquid box 17 is connected with a liquid discharge pipe 22 through a waste liquid pipe 20, and the liquid discharge pipe 22 is connected with an outlet of a washing kettle 24. The first case top cover 3 and the second case top cover 26 are installed on the upper side of the case body, the touch display screen 27 is embedded on the upper side of the case body, the case front cabinet door 21 is installed on the front side of the case body, and the case heat dissipation holes 12 are formed in the left side of the case body.
As shown in fig. 2, the multipurpose flow cytometer provided by the embodiment of the present invention includes a case body, and four thermal water tanks 7 are disposed in parallel in the middle of the case body. The first case top cover 3 and the second case top cover 26 are installed on the upper side of the case body, the touch display screen 27 is embedded on the upper side of the case body, the front case door 21 is installed on the front side of the case body, and the case heat dissipation holes 12 are formed in the left side of the case body.
As shown in fig. 3, 4 heat preservation water tanks 7 with different temperatures are arranged in parallel at the middle position of the case body of the multipurpose flow cytometer provided by the embodiment of the present invention. An electric heating pipe 8 is arranged in the heat preservation water tank 7, a compressor 11 is arranged on the left side of the bottom of the case body, the compressor 11 is sequentially connected with a condenser 14, a drying filter 15 and a capillary tube 16 through an exhaust pipe 13, the pipeline branches are respectively connected with evaporators 19 arranged on the outer sides of the heat preservation water tanks 7, and air return pipes of the evaporators 19 are converged into an air return pipe 10 and then return to the compressor 11. The outside of each heat preservation water tank 7 is provided with a temperature controller 18 for detecting the temperature in each water tank and controlling the operation of the electric heating pipe 8 and the compressor 11 according to the program setting. The outside of the heat preservation water tank 7 is provided with a water tank overflow port 5 and a water tank water filling port 9 (which is a discharge port when water is changed at the same time), the water tank water filling port 9 branches and then enters the lower side of each heat preservation water tank 7, and each branch pipe is provided with an electromagnetic valve 28 for controlling the opening and closing of the branch pipe.
As shown in fig. 4, 4 heat preservation water tanks 7 with different temperatures are arranged in parallel at the middle position of the case body of the multipurpose flow cytometer provided by the embodiment of the present invention, and a heat exchange coil 6 is installed inside each heat preservation water tank 7. The inlet of the heat exchange coil 6 is connected with the washing liquid bag 1 through the washing liquid connecting pipe 2, and the washing liquid connecting pipe 2 is provided with a peristaltic pump valve 29 for controlling the flowing of the washing liquid. The outlet of the heat exchange coil 6 is connected with a washing liquid distribution pipe 25 through a channel adapter tube 4, and the washing liquid distribution pipe 25 is connected with a washing kettle 24. The outlet of the washing kettle 24 is connected with the waste liquid pipe 20 through the liquid discharge pipe 22, and the lower end of the waste liquid pipe 20 is connected with the waste liquid tank 17 (see figure 1) or a sewer. Each branch of the channel adapter tube 4 is provided with an electromagnetic valve 28 for selectively using washing liquid with different temperatures to wash cell samples; peristaltic pump valves 29 are arranged on the upstream of the main outlet and the auxiliary outlet of the washing liquid distribution pipe 25 and on the liquid discharge pipe 22 and are used for controlling liquid flow during washing, emptying and cell sample collection after washing; a solenoid valve 28 is also provided upstream of the main outlet of the wash liquor distribution pipe 25 and in the air inlet pipe 30 for selecting the washing liquor to enter the wash pot 24 for washing the sample or for selecting clean air to empty the wash pot 24.
As shown in fig. 5, 4 heat-preservation water tanks 7 with different temperatures are arranged in parallel at the middle position of the case body of the multipurpose flow-type cell washing instrument provided by the embodiment of the invention, a heat exchange coil 6 is installed in each heat-preservation water tank 7, and the main body of the heat exchange coil 6 is completely immersed below the water surface of each heat-preservation water tank 7 except for the heat exchange coil inlet 31 and the heat exchange coil outlet 32, but the internal washing liquid thereof is completely isolated from the water outside the pipes. The inlet of the heat exchange coil 6 is connected with a washing liquid connecting pipe 2, and a peristaltic pump valve 29 is arranged on the washing liquid connecting pipe 2 and used for controlling the flowing of the washing liquid. The outlet 32 of the heat exchange coil is connected with different channel switching tubes 4, and each branch of the channel switching tube 4 is provided with an electromagnetic valve 28 for selectively using washing liquid with different temperatures to wash cell samples. The bulk flow direction of the wash liquor was: the washing liquid connecting pipe 2 → the heat exchange coil inlet 31 → the heat exchange coil outlet 32 → the passage adapter pipe 4 → the washing liquid distributing pipe 25 (see fig. 4), wherein the washing liquid connecting pipe 2 is divided into a plurality of branches downstream. Through the linkage control of the electromagnetic valve 28, the temperature of the washing liquid flowing from any channel adapter tube 4 to the downstream can be independently and freely selected and can be switched at any time.
As shown in fig. 6, the washing liquid distributing pipe 25 is installed at the upper end of the washing kettle 24 provided by the embodiment of the present invention, the air inlet pipe 30, the peristaltic pump valve 29 and the electromagnetic valve 28 are installed at the upper end of the washing liquid distributing pipe 25, and the main washing liquid outlet at the right end of the washing liquid distributing pipe 25 is connected with the main washing liquid inlet 40 in the washing kettle 24; a washing liquid auxiliary outlet at the right end of the washing liquid distribution pipe 25 is connected with a washing liquid auxiliary inlet 33 in the washing kettle 24, and 2 peristaltic pump valves 29 and 2 electromagnetic valves 28 are arranged on the washing liquid distribution pipe 25; a sample inlet 41 and a sample outlet 42 are arranged on the right side of the washing kettle 24, a first washing kettle bracket 36 and a second washing kettle bracket 39 are arranged on the outer side of the washing kettle 24, a liquid discharge pipe 22 is arranged at the lower end of the washing kettle 24, and a peristaltic pump valve 29 is arranged on the liquid discharge pipe 22; the upper end of the washing kettle 24 is provided with an upper side cell filter membrane 34 for intercepting larger impurity cells, and the aperture of the upper side cell filter membrane 34 can pass through the cells to be washed and simultaneously intercept the impurity cells; the lower end of the washing kettle 24 is provided with a lower cell filter membrane 38 for intercepting cells to be washed, the types of the cells to be washed are different, and the pore diameters of the lower cell filter membrane 38 are different; an integrally formed scouring pipe network 35 is arranged in the wall of the washing kettle 24, and the scouring pipe network 35 is provided with 1 inlet, namely a washing liquid auxiliary inlet 33 which is connected with an auxiliary outlet of the washing liquid distribution pipe 25; the scouring pipe network 35 is provided with a plurality of outlets, which are positioned on the upper pot wall of the lower side cell filter membrane 38 to form a plurality of scouring holes 37. According to different application requirements, the washing kettle is divided into a plurality of specifications, and the shape size, the number of filter membranes and the aperture of the filter membranes are different.
As shown in fig. 7, a method for controlling a multi-purpose flow cytometer according to an embodiment of the present invention includes:
s101: before washing, the temperature of each water tank is set at one time, so that the instrument is continuously in a standby state, and the washing work is started at any time according to the work requirement; and simultaneously, installing all parts of the washing instrument.
S102: under the linkage control of each peristaltic pump valve, electromagnetic valve and the like, gas in each lumen of the washing liquid emptying passage is added through a sample inlet of a washing kettle, cells to be washed are added through a pipettor or, a dropper and the like, or are directly connected with a sterile cell preservation bag for sample addition; washing is started, washing liquor with a specific temperature is selectively added into a main inlet and an auxiliary inlet of a washing kettle synchronously or alternatively according to program control, the washing kettle synchronously or alternatively rotates back and forth at a certain angle along with a support, and the speed is set to be 5-10 times/min; the washing liquid flows to a liquid discharge pipe after passing through the filter membrane, and flows to a waste liquid tank or a sewer through a waste liquid pipe.
S103: the washing kettle stops rotating back and forth and keeps a vertical state, and the washing liquid is stopped entering the washing liquid auxiliary inlet; the main inlet continues to feed the washing liquid until all the cells reach the cell filter membrane in the washing kettle cavity; the main washing liquid inlet stops feeding washing liquid, the electromagnetic valve on the upper air inlet pipe upstream of the washing liquid distribution pipe is closed, and meanwhile, the electromagnetic valve of the air inlet pipe is opened, so that filtered clean air enters to empty the washing liquid in the washing kettle cavity; the washing device can be set to be single or multiple times of washing, and each washing process can be set to be the same temperature or different temperatures.
S104: after the last 1 time of washing is finished and the washing liquid is emptied, closing the drain pipe peristaltic pump valve; adding a proper amount of washing liquid to resuspend the cells on the filter membrane through the auxiliary inlet of the washing kettle; collecting the washed cells from the sample outlet for subsequent detection or treatment, or directly adding physiological saline or special cell preservation solution from the sample outlet for resuspension and collecting the cells; according to the experiment or treatment requirements, the cell collection mode is an open suction mode such as a pipettor, a suction pipe and the like; or directly connecting with a sterile cell preservation bag pre-filled with preservation solution or empty for collection.
S105: the washing liquid distribution pipe, the washing kettle and the liquid discharge pipe are disassembled and discarded.
In S101 provided by the embodiment of the present invention, a specific process of installing each component is as follows:
hoisting or placing a washing liquid bag, a washing liquid bottle or a washing liquid box, selecting and installing a plurality of heat exchange coil pipes with proper specifications according to needs, connecting an outlet of the washing liquid bag and an inlet of the heat exchange coil pipes through 1 washing liquid connecting pipe, and installing the heat exchange coil pipes on an instrument; installing a washing liquid channel adapter tube, and connecting an inlet of the washing liquid channel adapter tube with an outlet of the heat exchange coil; installing a washing liquid distribution pipe, and connecting an inlet of the washing liquid distribution pipe with an outlet of the channel adapter pipe; installing a washing kettle with proper specification, respectively connecting a main inlet and an auxiliary inlet of the washing kettle with a main outlet and an auxiliary outlet of a washing liquid distribution pipe, and adjusting a support of the washing kettle to an initial position to enable the washing kettle to be in a vertical state; installing a liquid discharge pipe, connecting an inlet of the liquid discharge pipe with an outlet of the washing kettle, and connecting an outlet of the liquid discharge pipe with an inlet of the waste liquid pipe; and a waste liquid pipe is arranged, and the outlet of the waste liquid pipe is connected with a waste liquid barrel, a waste liquid box, a waste liquid bag or a sewer.
As shown in fig. 5, the outlets of the first to fourth channel adapter tubes 4 are connected to the inlets of the first to fourth channel washing liquid distribution tubes, respectively.
The technical solution of the present invention will be described in detail with reference to the following specific examples.
The multipurpose flow cytometry washing instrument provided by the embodiment of the invention is divided into two main structures, wherein the first part is a temperature changing structure and is used for heating or cooling washing liquor as required; the second part is a washing kettle with a filter membrane and an accessory structure. The two parts of structures are integrated in the same case, and are uniformly controlled and operated by the setting program of the operation panel.
1. Temperature changing structure
According to the experimental requirement, the temperature changing structure is divided into 4 temperature zones, each temperature zone is composed of an independent heat preservation water tank with a cover, the interior of the temperature zone is filled with pure water, and the water temperature can be controlled by a semiconductor refrigeration sheet (Peltier group) or a combination of a compressor and an electric heating tube. The temperatures of the 4 water tanks are respectively set to be 4 ℃, 22 ℃, 37 ℃ and 42 ℃; other temperatures may also be set by the main operating panel as required by the particular experiment. The bottom or the lower part of each water tank is provided with a water inlet channel for injecting pure water into the water tank; or periodically to empty the tank with this passage in order to clean the tank. The inner wall of the water tank is made of stainless steel or other heat-resistant and water-resistant materials, and the outer wall of the water tank is made of heat-insulating and flame-retardant materials. Every water tank upper portion all is equipped with 1 overflow hole, and every water tank top and upper portion all are provided with a plurality of pore, can supply the lotion pipeline to pass, and its figure is the same with the passageway figure of instrument. Each tank is provided with 1 openable upper cover to allow an operator to install or remove the heat exchange coil. The upper part of the instrument is provided with a plurality of hooks or a platform for hanging or placing a bag/bottle/box type sterile or open washing liquid container (filled with normal saline or various special washing liquids). The outlet of the washing liquid container is connected with the inlet of a washing liquid connecting pipe, the washing liquid connecting pipe is a single pipeline or a branch pipe with one division, each outlet of the washing liquid connecting pipe is respectively connected with the inlets of 1 heat exchange coil, and each heat exchange coil is respectively arranged in water tanks with different temperatures according to experimental requirements. Except for the inlet and the outlet, the main body of the heat exchange coil pipe needs to be completely immersed under water so as to achieve the effect of fully changing the temperature. The outlets of the heat exchange coil pipes are respectively connected with the adapter pipes of different channels, and the washing liquid is conveyed into the washing kettle of the appointed channel according to program control. All tubes and heat exchange coils in direct contact with the wash solution are made of biocompatible materials, such as but not limited to those used in medical blood transfusion/infusion. The washing liquid can be subjected to temperature change through the temperature change structure and is conveyed to different washing channels according to requirements. Because the washing liquid is always in the closed pipeline system and is not in direct contact with the water body and the heat insulation structure of each heat insulation water tank, the problems that the washing liquid is polluted by microorganisms, the washing liquid leaks out to corrode the water tanks and other parts are avoided.
2 washing kettle and accessory structure
According to the different volume of a single washing sample, the washing kettle is designed into different specifications, and the volume of the inner cavity of the washing kettle is different from 5 ml to 1000 ml. The washing pots of different specifications are made of biocompatible materials, such as but not limited to materials used for medical blood transfusion/fluid bags. The shape of the washing kettle is similar to that of a drip kettle at the middle section of the medical infusion apparatus. When the washing kettle is vertically arranged, a main washing liquid inlet is arranged above the washing kettle, the washing liquid inlet is connected with an outlet of the heat exchange coil in the temperature changing structure through a washing liquid distribution pipe and a channel switching pipe, and the temperature of washing liquid entering the washing kettle can be switched at any time and the flow rate of the washing liquid can be adjusted through the control of a group of program control electromagnetic valves and peristaltic pump valves. The outlet of the washing kettle is positioned at the bottom of the kettle body, 1 cell filter membrane is arranged at the lower side of the kettle body, and the cell filter membrane is also made of biocompatible materials, and can be selected from but not limited to the same materials as a medical leukocyte filter. The pore size of the filter membrane meets the requirement that the filter membrane can pass through blood plasma, culture solution, washing solution, cell debris and the like but cannot pass through cells to be washed. The washing kettle for washing human erythrocytes has a cell filter membrane (erythrocyte filter membrane) with a pore size of 1.0-5.0 um, and the specific numerical value can be optimized according to cell washing data. The washing kettle is used for washing other cells, and the aperture of the cell filter membrane at the lower side of the kettle body is set according to the size of the cells to be washed. The washing kettle for preparing the special washing erythrocyte product for clinical transfusion has the advantages that a layer of leukocyte filter membrane is additionally arranged on the upper side of the kettle body to intercept and remove leukocytes, and the leukocyte filter membrane is positioned below the sample inlet and positioned above the cell filter membrane and the sample outlet on the lower side of the kettle body. The aperture of the leukocyte filter membrane is smaller than the diameter of leukocytes, but larger than the diameter of erythrocytes, and can be set to be 8.5-9.0 um, and the specific numerical value can be optimized according to cell washing data.
The upper part of the washing kettle is provided with 1 cell sample inlet, and a cell sample to be washed can be added into the kettle cavity through the inlet in the modes of micropipette sample adding, dropper sample adding or test tube dumping and the like (open type washing), and can also be connected with a cell storage bag in the mode of aseptic connection (closed type aseptic washing). The pot wall position above the cell filter membrane at the lower side of the washing pot is provided with 1 cell sample outlet, and the washed cells can be directly sucked by a suction pipe or the like through the outlet or flow to the sterile collection bag connected with the outlet.
In order to enhance the washing effect and prevent the filter membrane from being blocked in the washing process, an integrally formed scouring pipe network is additionally arranged in the wall of the washing kettle, the inlet of the scouring pipe network is the auxiliary inlet of the washing kettle, the scouring pipe network is connected with the auxiliary outlet of the washing liquid distribution pipe and enters the wall of the washing kettle from the upper part of the washing kettle, then the scouring pipe network is divided into a plurality of washing holes to form a net-shaped flow distribution system, and finally the scouring holes are communicated with the inner cavity of the washing kettle above the lower side cell filter membrane. The scouring holes are annularly distributed on the kettle wall, and the orifices are close to the upper side of the cell filter membrane and are approximately parallel to the plane of the filter membrane. The liquid inlet quantity of the flushing pipe network system is controlled by an independent peristaltic pump valve. When washing, it and main inlet of washing liquor are fed simultaneously or alternatively, and when the cell is contacted with lower side cell filter membrane under the action of main liquid flow, it is raised by high-speed secondary liquid flow jetted from flushing hole. Thus the cells are more thoroughly washed and the chance of clogging the pores of the lower cell filter is reduced.
In order to further enhance the washing effect, prevent the filter membrane from being blocked in the washing process and facilitate the sample to flow to the sterile collection bag through the sample outlet after washing, the whole washing kettle, a part of upstream washing liquid distribution pipe and a part of downstream liquid discharge pipe can rotate back and forth at a certain angle (such as-150 degrees) along with the program-controlled washing kettle bracket in the washing process and after washing, and the rotation can be synchronous with or alternate with the washing liquid injection process. In the washing process, washing liquor enters the cavity of the washing kettle through the main inlet and the auxiliary inlet, penetrates through the filtering membrane under the action of hydraulic pressure and gravity, finally flows to the outlet at the bottom of the washing kettle, then is converged into the waste liquid pipe through the liquid discharge pipe connected with the washing kettle, and then flows to a waste liquid box or a sewer.
In order to facilitate the thorough evacuation of residual washing liquid in a washing kettle cavity after washing, 1 air inlet pipe is arranged on a washing liquid distribution pipe at the near end of a main inlet of the washing liquid, the opening and the closing of the air inlet pipe are controlled by an electromagnetic valve, and an air filter is required to be arranged at an air inlet of the air inlet pipe so as to ensure that the cleanliness of the air inlet reaches the relevant quality requirements of cell products. In addition, a peristaltic pump valve is arranged at the position of the liquid discharge pipe, and can timely separate the upstream part of the liquid discharge pipe from the downstream waste liquid pipe under the control of a program so as to ensure that the downstream sewage cannot flow backwards to cause pollution in the washing kettle cavity under any condition; meanwhile, the peristaltic pump valve can be used for appropriately pumping the washing liquid in the washing kettle cavity according to program control in the washing process so as to accelerate liquid flow emptying.
3 case and control panel
The case provides components such as a power supply, a semiconductor refrigeration sheet or a compressor and an electric heating tube, a circuit board, a control computer host, a touch display screen and the like. An openable case top cover is arranged above the washing machine, and an openable cabinet door is arranged on the front side of the washing machine, so that washing consumables can be conveniently loaded and unloaded. In order to guarantee the temperature control precision, a heat insulation structure is arranged in the case, and the passage switching pipe, the washing liquid distribution pipe and the washing kettle are wrapped in the heat insulation structure so as to guarantee that the washing liquid completes the washing work under the condition of approaching the set temperature. The heat insulation structure is composed of a plurality of pieces of heat insulation materials such as foam plastics, sponge and the like, and is a cavity-shaped combination body. The heat-insulating combination body can be opened when the pipeline and the washing kettle are assembled and disassembled, and the combination body is closed to form a relatively closed heat-insulating cavity when the pipeline and the washing kettle are washed. Different washing channels are arranged at different positions in the heat preservation structure and are insulated from each other. In the heat insulation structure, the whole process of the pipeline of each washing channel is closed, and no matter exchange occurs between the washing liquid in the pipeline and the heat insulation cavity. According to the actual needs of users, the instrument can be set to a single washing channel (namely only 1 washing kettle is installed, and only 1 sample can be washed at the same time), and also can be set to a plurality of channels (namely a plurality of washing kettles are installed, and a plurality of samples can be washed at the same time). Each channel can be washed at a single temperature, and washing liquids at different temperatures can be alternately washed according to actual needs. When a plurality of channels work simultaneously, the washing temperature of each channel can be independently selected and switched at any time without mutual influence.
The working principle of the invention is as follows: before washing, the power supply of the flow cytometry washing instrument is firstly turned on, the temperature of each heat preservation water tank 7 is set, the instrument can be continuously in a standby state after the temperature of each heat preservation water tank 7 is set at one time, and the washing work can be started at any time according to the work requirement. Hoisting or placing the washing liquid bag 1/the washing liquid bottle/the washing liquid box, selecting and installing a plurality of heat exchange coil pipes 6 with proper quantity and specification as required, connecting the outlet of the washing liquid bag 1 and the inlets of the heat exchange coil pipes 6 by using washing liquid connecting pipes 2 (single pipes or branch pipes) with proper specification, and installing the washing liquid connecting pipes on the instrument. And a washing liquid channel adapter tube 4 is arranged, and the inlet of the washing liquid channel adapter tube is connected with the outlet of the heat exchange coil 6. A washing liquid distribution pipe 25 is arranged, and the inlet of the washing liquid distribution pipe is connected with the outlet of the channel adapter pipe 4; a washing kettle 24 is arranged, and the main inlet and the auxiliary inlet of the washing kettle are respectively connected with the main outlet and the auxiliary outlet of a washing liquid distribution pipe 25; the washpot stand is adjusted to an initial position so that the washpot 24 is in an upright position. A drain pipe 22 is installed, an inlet of which is connected with an outlet of the washing pot 24, and an outlet of which is connected with an inlet of the waste liquid pipe 20; a waste pipe 20 is installed with its outlet connected to a waste drum/waste tank/waste bag/sewer. Gas in each lumen of the washing liquid emptying passage is added through a sample inlet 41 (according to application conditions, cells to be washed can be added by a pipette, a dropper and the like, and can also be directly connected with a sterile cell preservation bag for adding samples); the washing is started, the main inlet and the auxiliary inlet are synchronously or alternatively controlled to be filled with washing liquor with specific temperature according to a program, the washing kettle 24 synchronously/alternatively rotates back and forth at a certain angle (such as-150 degrees) along with the support, and the speed can be set to be 5-10 times/minute. The washing liquid passes through the filter membrane and flows to the drain 22 and via the waste pipe 20 to the waste tank 17 or sewer.
The wash pot 24 stops rotating back and forth and maintains the upright state, and the washing liquid sub-inlet 33 stops the washing liquid. The main wash inlet 40 continues into the wash until all cells to be washed have reached the lower cell filter 38 in the wash pot chamber. The main wash liquor inlet 40 stops feeding wash liquor, the solenoid 28 controlling the inlet pipe 30 opens and closes the solenoid 28 on its upstream wash liquor dispensing pipe 25, allowing clean air to enter to empty the wash liquor in the chamber of the wash kettle 24 (the peristaltic pump valve 29 at the drain pipe can be actuated, speeding the emptying process). According to different washing requirements, the washing process can be set to be single washing or multiple washing, and each washing process can be set to be the same temperature or different temperatures.
After the last 1 washing is finished and the washing liquid is emptied, the peristaltic pump valve 29 at the drain pipe is closed. Adding a proper amount of washing solution through the flushing holes 37 to resuspend the cells on the lower cell filter membrane 38, and collecting the washed cells from the sample outlet 42 for subsequent detection or treatment; or adding physiological saline or special cell preservation solution directly from the sample outlet 42 to resuspend and collect cells. According to the requirements of experiments or treatment, the cell collection mode can adopt an open suction mode such as a pipette, a suction pipe and the like, or can be directly connected with a sterile cell preservation bag pre-filled with preservation solution for collection.
The wash liquor distribution pipe 25, wash kettle 24 and drain pipe 22 are removed and discarded. For open washing without asepsis or closed washing without asepsis requirement, they are disposable consumables, and the rest pipelines can be replaced after repeated use. For washing with higher requirement on sterility grade, the whole set of pipelines is suitable for disposable use, and the whole set of washing liquid pipelines is suitable for being designed into a whole without being connected step by step in the using process.
The invention can be designed into single channel or multiple channels, and when a multi-channel washing instrument is used for washing multiple samples at the same time, the operation process of each channel is the same. Under all conditions, except the processes of consumable material loading and unloading, sample adding, cell collection and washing, other processes are controlled by a preset program, and the washing process is fully automatically operated without human intervention.
In the description of the present invention, "a plurality" means two or more unless otherwise specified; the terms "upper", "lower", "left", "right", "inner", "outer", "front", "rear", "head", "tail", and the like, indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, are only for convenience in describing and simplifying the description, and do not indicate or imply that the device or element referred to must have a particular orientation, be constructed in a particular orientation, and be operated, and thus, should not be construed as limiting the invention. Furthermore, the terms "first," "second," "third," and the like are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
The above description is only for the purpose of illustrating the present invention and the appended claims are not to be construed as limiting the scope of the invention, which is intended to cover all modifications, equivalents and improvements that are within the spirit and scope of the invention as defined by the appended claims.
Claims (10)
1. A multipurpose flow cytometry washing instrument is characterized in that the multipurpose flow cytometry washing instrument is provided with a case body;
a compressor is installed on the left side of the bottom of the case body, four heat-preservation water tanks are arranged in the middle of the case body and respectively provide a common temperature, an evaporator and a temperature controller are arranged on the outer sides of the heat-preservation water tanks, and an electric heating pipe is arranged in each heat-preservation water tank (or a temperature-variable part such as a semiconductor refrigerating sheet is used for replacing the compressor, the electric heating pipe and the like);
a water tank water filling port and a water tank overflow port are arranged on the outer side of each heat-preservation water tank, and the water tank water filling port is also a water outlet when water is changed; a heat exchange coil is arranged in each heat preservation water tank, and the washing liquid in each heat preservation water tank and the water body of the heat preservation water tank perform non-contact heat exchange;
the outlet of the heat exchange coil is connected with a washing liquid distribution pipe through a channel switching pipe, and the washing liquid distribution pipe is connected with a washing kettle; the liquid discharge pipe is connected with the outlet of the washing kettle, the waste liquid box is connected with the liquid discharge pipe through a waste liquid pipe, and the waste liquid box is arranged on the right side of the bottom of the case body and can be connected with a sewer interface.
2. The multi-purpose flow cytometer as described in claim 1, wherein the inlet of the heat exchange coil installed in the heat-insulating water tank is connected to the washing liquid bag through the washing liquid connection tube, and the washing liquid bag is installed on the upper portion of the cabinet body;
except that the inlet and the outlet of the heat exchange coil are exposed above the water surface of the heat-preservation water tank, the tube body is completely immersed below the water surface of the water tank, and the inside and the outside of the tube body are completely isolated.
3. The multi-purpose flow cytometer of claim 1 wherein the channel adapter tube, the wash solution distribution tube and the wash pot are all mounted on a support inside a thermal insulation structure, the thermal insulation structure is mounted on the upper portion of the cabinet body, so that the wash solution flowing out of the heat exchange coil passes through the channel adapter tube, the wash solution distribution tube and the wash pot at a temperature close to a set temperature to wash the cell sample in the pot; in the heat insulation structure, each washing channel is sealed in the whole process, and no exchange occurs between the washing liquid in the pipeline and the heat insulation cavity.
4. The multi-purpose flow cytometer of claim 1 wherein the first housing top cap and the second housing top cap are mounted to the top side of the housing body and the front housing door is mounted to the front side of the housing body.
5. The multi-purpose flow cytometer of claim 1 wherein the top side of the housing body is embedded with a touch screen.
6. The multipurpose flow cytometer as described in claim 1, wherein the upper end of said wash pot is equipped with a wash liquor distribution tube, the upper end of said wash liquor distribution tube is equipped with an air inlet tube with an air filter membrane, a peristaltic pump valve and an electromagnetic valve, and the right end of said wash liquor distribution tube is equipped with a main wash liquor outlet and an auxiliary wash liquor outlet; the main outlet of the washing liquid is connected with the main inlet of the washing kettle, the auxiliary outlet of the washing liquid is connected with the inlet of the scouring pipe network in the washing kettle body, and the right side of the washing kettle is provided with a sample inlet and a sample outlet.
7. The multipurpose flow cytometer as described in claim 1, wherein a first wash pot holder and a second wash pot holder are installed outside the wash pot, a drain pipe is installed at the lower end of the wash pot, and a peristaltic pump valve is installed on the drain pipe;
the upper end of the washing kettle is provided with an upper side cell filter membrane for intercepting larger impurity cells, and the aperture of the upper side cell filter membrane can pass through the cells to be washed and simultaneously intercept the mixed cells;
the lower end of the washing kettle is provided with a lower side cell filter membrane for intercepting cells to be washed, the types of the cells to be washed are different, and the pore diameters of the lower side cell filter membrane are also different;
an integrally formed scouring pipe network is arranged in the kettle wall of the washing kettle, and the scouring pipe network is provided with 1 inlet, namely a washing liquid auxiliary inlet, which is connected with an auxiliary outlet of a washing liquid distribution pipe; the scouring pipe network is provided with a plurality of outlets and is positioned on the upper pot wall of the lower side cell filter membrane to form a plurality of scouring holes; according to different application requirements, the washing kettle is divided into a plurality of specifications, and the shape size, the number of filter membranes and the aperture of the filter membranes are different.
8. A method of controlling a multi-purpose flow cytometer as described in any one of claims 1-7, wherein the method of controlling the multi-purpose flow cytometer comprises the steps of:
before washing, setting the temperature of each heat-preservation water tank at one time to keep the instrument in a standby state, and starting washing work according to work requirements at any time; meanwhile, all parts of the washing instrument are installed;
secondly, under the linkage control of each peristaltic pump valve and each electromagnetic valve, gas in each tube cavity of the washing liquid emptying passage is added through a sample inlet of a washing kettle, and cells to be washed are added through a pipettor or a dropper or are directly connected with a sterile cell storage bag for sample addition; washing is started, washing liquor with a specific temperature is selectively added into a main inlet and an auxiliary inlet of a washing kettle synchronously or alternatively according to program control, the washing kettle synchronously or alternatively rotates back and forth at a certain angle along with a support, and the speed is set to be 5-10 times/min; the washing liquid flows to a liquid discharge pipe after passing through the filter membrane, and flows to a waste liquid tank or a sewer through a waste liquid pipe;
step three, stopping the washing kettle from rotating back and forth and keeping the washing kettle in a vertical state, and stopping the washing liquor from entering the washing liquor auxiliary inlet; the main inlet continues to feed the washing liquid until all the cells to be washed reach the lower side cell filter membrane in the washing kettle cavity; the main liquid inlet stops feeding washing liquid, and the electromagnetic valve of the air inlet pipe is opened to enable filtered clean air to enter so as to empty the washing liquid in the washing kettle cavity; the washing machine can be set to single or multiple times of washing according to application requirements, and the same temperature or different temperatures can be set in each washing process;
step four, after the last washing 1 time is finished and the washing liquid is emptied, closing a drain pipe peristaltic pump valve; adding a proper amount of washing liquid through the auxiliary inlet of the washing kettle to resuspend the cells on the lower side filter membrane; or directly adding physiological saline or special cell preservation solution from the sample outlet to resuspend and wash the washed cells; collecting the washed cells from the outlet for subsequent detection or treatment; according to the experiment or treatment requirements, the cell collection mode can be an open suction mode such as a pipette, a suction pipe and the like; or directly connecting with a sterile cell preservation bag pre-filled with preservation solution or empty for collection;
and fifthly, disassembling and assembling a washing liquid distribution pipe, a washing kettle and a liquid discharge pipe, and discarding.
9. The multi-purpose flow cytometer control method of claim 8, wherein in the first step, the steps of installing the components are as follows:
hoisting or placing a washing liquid bag, a washing liquid bottle or a washing liquid box, selecting and installing a plurality of heat exchange coil pipes with proper specifications according to needs, connecting an outlet of the washing liquid bag and an inlet of the heat exchange coil pipe through 1 washing liquid connecting pipe, and installing the heat exchange coil pipe on an instrument; installing a washing liquid channel adapter tube, and connecting an inlet of the washing liquid channel adapter tube with an outlet of the heat exchange coil; installing a washing liquid distribution pipe, and connecting an inlet of the washing liquid distribution pipe with an outlet of the channel adapter pipe; installing a washing kettle with proper specification, respectively connecting a main inlet and an auxiliary inlet of the washing kettle with a main outlet and an auxiliary outlet of a washing liquid distribution pipe, and adjusting a support of the washing kettle to an initial position to enable the washing kettle to be in a vertical state; installing a liquid discharge pipe, connecting an inlet of the liquid discharge pipe with an outlet of the washing kettle, and connecting an outlet of the liquid discharge pipe with an inlet of the waste liquid pipe; and a waste liquid pipe is arranged, and the outlet of the waste liquid pipe is connected with a waste liquid barrel, a waste liquid box, a waste liquid bag or a sewer.
10. A medical laboratory apparatus, wherein the multipurpose flow cytometer of any one of claims 1 to 7 is used in the medical laboratory apparatus.
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