Cell elution method, device and medium

Abstract

The invention provides a cell elution method, equipment and a medium, and the cell elution method comprises the following steps: after a magnetic field is confirmed to be generated in a magnetic separation column, controlling a sample liquid to flow through the magnetic separation column so as to adsorb magnetic labeled cells in the sample liquid into the magnetic separation column through the magnetic field; performing gas-liquid cleaning operation on the magnetic separation column for a first preset number of times; the gas-liquid cleaning operation comprises the following steps: controlling a first preset dosage of gas to clean the magnetic separation column at a first preset flow rate, and then controlling a second preset dosage of buffer solution to clean the magnetic separation column at a second preset flow rate, so as to remove impurities except the magnetically labeled cells in the magnetic separation column through gas-liquid alternate cleaning. According to the magnetic separation column, the magnetic labeled cells can be successfully separated, the purity and the separation precision of the magnetic labeled cells adsorbed by the magnetic separation column are also improved, and the cell separation efficiency is further improved.

Description

Technical Field

[0001] The invention belongs to the technical field of cell processing, and in particular relates to a cell elution method, equipment and medium. Background Art

[0002] At present, in cytological research, functional studies on certain cells require the cells to be purified into high-purity target cells. It is inevitable that other impurities will remain in the purification process. For example, in single-collected blood, peripheral blood and other blood samples, the sorting and purification of target cells is easily interfered by red blood cells. Therefore, reducing the residual impurities of miscellaneous cells in the purification process plays an important role in subsequent cell culture and cell function research.

[0003] In the prior art, cells can be magnetically sorted by cell sorting technology to obtain target cells. However, after magnetic sorting, there are often still many impurity cells in the selected target cells, which will contaminate the target cells and interfere with subsequent culture and cell research, ultimately resulting in low purity and inaccurate sorting accuracy. Summary of the invention

[0004] The present invention aims to solve the technical problems in the prior art that when magnetically sorting cells, the selected target cells often still contain a large number of impurity cells that contaminate the target cells, resulting in low purity and sorting accuracy, and provides a cell elution method, equipment and medium.

[0005] In view of the above technical problems, an embodiment of the present invention provides a cell elution method, comprising:

[0006] After confirming that a magnetic field is generated in the magnetic separation column, controlling the sample liquid to flow through the magnetic separation column, so that the magnetically labeled cells in the sample liquid are adsorbed in the magnetic separation column through the magnetic field;

[0007] A first preset number of gas-liquid cleaning operations are performed on the magnetic separation column; the gas-liquid cleaning operation includes: controlling a first preset dose of gas to clean the magnetic separation column at a first preset flow rate, and then controlling a second preset dose of buffer to clean the magnetic separation column at a second preset flow rate, so as to remove impurities other than magnetically labeled cells in the magnetic separation column through alternating gas-liquid cleaning.

[0008] A cell elution device comprises a controller, wherein the controller comprises a processor and a memory, wherein the memory stores an executable program, and the processor is used to execute the cell elution method.

[0009] A computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, the cell elution method is implemented.

[0010] In the present invention, the cell elution method includes: after confirming that a magnetic field is generated in a magnetic separation column, controlling the sample liquid to flow through the magnetic separation column so that the magnetically labeled cells in the sample liquid are adsorbed in the magnetic separation column through the magnetic field; performing a first preset number of gas-liquid cleaning operations on the magnetic separation column; the gas-liquid cleaning operation includes: after controlling a first preset dose of gas to clean the magnetic separation column at a first preset flow rate, controlling a second preset dose of buffer to clean the magnetic separation column at a second preset flow rate, so as to remove impurities other than the magnetically labeled cells in the magnetic separation column through alternating gas-liquid cleaning. The cell elution method provided by the present invention, after adsorbing the magnetically labeled cells in the sample liquid by a magnetic separation column, removes impurities (other cells except the magnetically labeled cells) remaining in the magnetic separation column by gas-liquid alternating cleaning in a gas-liquid cleaning operation. The present invention can not only successfully sort out the magnetically labeled cells, but also improve the purity and sorting accuracy of the magnetically labeled cells adsorbed by the magnetic separation column, reduce the contamination of impurities to the target cells, and thus ensure the smooth progress of subsequent culture and cell research; and avoid the need to improve the purity through multiple sorting, thereby improving the efficiency of cell sorting. BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The present invention is further described below in conjunction with the accompanying drawings and embodiments.

[0012] Figure 1 4 is a flow chart of the cell elution method provided in the first embodiment of the present invention.

[0013] Figure 2 4 is a flow chart of the cell elution method provided in the second embodiment of the present invention.

[0014] Figure 3 4 is a flow chart of a cell elution method provided in the third embodiment of the present invention.

[0015] Figure 4 It is a schematic diagram of the structure of a cell elution device provided in one embodiment of the present invention.

[0016] The reference numerals in the specification are as follows:

[0017] 100, magnetic separation column; 210, sample pipeline; 220, output pipeline; 230, gas pipeline; 240, cleaning pipeline; 250, auxiliary pipeline; 310, sample container; 320, negative selection container; 330, cleaning container; 340, positive selection container; 400, first driving pump; 510, sample switch valve; 520, negative selection switch valve; 530, gas switch valve; 540, cleaning switch valve; 550, circulation switch valve; 560, positive selection switch valve. DETAILED DESCRIPTION

[0018] In order to make the technical problems, technical solutions and beneficial effects solved by the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0019] It should be understood that the directions or positional relationships indicated by terms such as "upper", "lower", "left", "right", "front", "back", and "middle" are based on the directions or positional relationships shown in the accompanying drawings and are only for the convenience of describing the present invention and simplifying the description. They do not indicate or imply that the device or element referred to must have a specific direction, be constructed and operated in a specific direction, and therefore should not be understood as a limitation of the present invention.

[0020] like Figure 1 and Figure 4 As shown, an embodiment of the present invention provides a cell elution method, comprising steps S100-S200:

[0021] S100 , after confirming that a magnetic field is generated in the magnetic separation column 100 , controlling the sample liquid to flow through the magnetic separation column 100 , so that the magnetically labeled cells in the sample liquid are adsorbed in the magnetic separation column 100 through the magnetic field.

[0022] The cell elution method provided in this embodiment can be applied in Figure 4 On the cell elution device shown. For example, one end of the magnetic separation column 100 can be connected to the sample container 310 through the sample pipeline 210, and the other end of the magnetic separation column 100 can be connected to the negative separation container 320 through the output pipeline 220, and a first driving pump 400 (such as a peristaltic pump, etc.) for driving the flow of liquid in the output pipeline 220 is provided on the output pipeline 220, so that the sample liquid can be driven from the sample container 310 through the sample pipeline 210 to the magnetic separation column 100 through the control of the operation of the first driving pump 400, and from the magnetic separation column 100 to the negative separation container 320 through the output pipeline 220, so that the sample liquid flows through the magnetic separation column 100. A magnetic field generating mechanism can be arranged near the magnetic separation column 100, so that the magnetic field can be generated or eliminated in the magnetic separation column 100 by the magnetic field generating mechanism. Through the magnetic field generated in the magnetic separation column 100, the magnetically labeled cells in the sample liquid can be adsorbed in the magnetic separation column 100. The magnetic separation column 100 can also be connected to structural elements and pipelines, so that the sample liquid can flow through the magnetic separation column 100 through other elements or pipelines, so that the magnetically labeled cells in the sample liquid can be adsorbed in the magnetic separation column 100 through the magnetic field, which will not be repeated here.

[0023] It can be understood that the target cells in the sample liquid are magnetically labeled, thereby combining with the magnetic substance and thus having magnetism; that is, the target cells are magnetized, thereby generating immunomagnetic bead cells, that is, magnetically labeled cells (generally including immunomagnetic bead cells and bacteria labeled with magnetic beads, etc.). The magnetic field generating mechanism can generate a magnetic field in the magnetic separation column 100 by controlling the electromagnetic coil arranged near the magnetic separation column 100 to be energized, or controlling the magnet to be close to the magnetic separation column 100. The magnetic field generating mechanism can be arranged at a preset position outside the magnetic separation column 100 as needed, as long as the magnetic field generating mechanism can generate a magnetic field in the magnetic separation column 100 that can adsorb the magnetically labeled cells in the sample liquid flowing through into the magnetic separation column 100. After the magnetic field generating mechanism generates a magnetic field in the magnetic separation column 100, the magnetically labeled cells in the sample liquid flowing through can be adsorbed into the magnetic separation column 100 through the magnetic field. The flow rate of the sample liquid flowing through the magnetic separation column 100 can be set according to actual conditions, and the flow rate can be set according to factors such as the survival rate of the target cells (to avoid the destruction of the target cells due to excessive flow rate) and the adsorption rate of the target cells by the magnetic field of the magnetic separation column 100 (to avoid the flow rate being too fast and causing the target cells to flow out before being adsorbed in the magnetic separation column 100). In one embodiment, the sample liquid is controlled to flow through the magnetic separation column 100 at a flow rate of 5-100 mL / min, so that the sorting efficiency of the magnetic separation column 100 (including cell survival rate and cell adsorption rate, etc.) can be improved.

[0024] S200, performing a first preset number of gas-liquid cleaning operations on the magnetic separation column 100; the gas-liquid cleaning operation includes: controlling a first preset dose of gas to clean the magnetic separation column 100 at a first preset flow rate, and then controlling a second preset dose of buffer to clean the magnetic separation column 100 at a second preset flow rate, so as to remove impurities other than magnetically labeled cells in the magnetic separation column 100 through alternating gas-liquid cleaning. The first preset number of times can be set according to demand.

[0025] In this embodiment, if Figure 4As shown, one end of the magnetic separation column 100 connected to the sample pipeline 210 can be connected to the atmosphere through the gas pipeline 230, and a sample switch valve 510 for controlling the switch of the sample pipeline 210 is provided on the sample pipeline 210, so that after closing the sample switch valve 510, by controlling the operation of the first driving pump 400, the driving gas (which can be air after passing through a sterile filter) is driven from the outside through the gas pipeline 230 to clean the magnetic separation column 100; the end of the magnetic separation column 100 connected to the sample pipeline 210 can also be connected to the cleaning container 330 through the cleaning pipeline 240. , and a gas switch valve 530 for controlling the switch of the gas pipeline 230 is provided on the gas pipeline 230, so that after closing the sample switch valve 510 and the gas switch valve 530, by controlling the operation of the first driving pump 400, the buffer solution (such as nanomagnetic separation buffer solution, etc.) can be driven from the cleaning container 330 through the cleaning pipeline 240 to enter the magnetic separation column 100, and output from the magnetic separation column 100 through the output pipeline 220 to the negative separation container 320, thereby removing impurities other than magnetically labeled cells in the magnetic separation column 100 through gas-liquid alternating cleaning. In addition, a cleaning switch valve 540 for controlling the switch of the cleaning pipeline 240 can be provided on the cleaning pipeline 240, so that after closing the sample switch valve 510 and the cleaning switch valve 540, by controlling the operation of the first driving pump 400, the gas can be driven from the outside through the gas pipeline 230 to clean the magnetic separation column 100, thereby realizing the next round of gas-liquid cleaning operation. The gas-liquid alternating cleaning of the gas-liquid cleaning operation can better remove impurities other than magnetically labeled cells in the magnetic separation column 100 .

[0026] It can be understood that in the process of adsorbing the magnetically labeled cells in the sample liquid in the magnetic separation column 100 through the magnetic field, impurities other than the magnetically labeled cells in the sample liquid may get stuck in the magnetic separation column 100 or stay in the magnetic separation column 100 together with the magnetically labeled cells (for example, if the magnetically labeled cells are white blood cells, impurities such as red blood cells that are smaller in volume than white blood cells in the sample liquid may get stuck in the gaps between multiple white blood cells in the magnetic separation column 100 or adhere to white blood cells and stay in the magnetic separation column 100 together), thereby affecting the purity of the magnetically labeled cells adsorbed by the magnetic separation column 100. Therefore, in this embodiment, the impurities other than the magnetically labeled cells in the magnetic separation column 100 can be better removed through the gas-liquid cleaning operation, thereby improving the purity of the magnetically labeled cells adsorbed by the magnetic separation column 100. The first preset dose can be set according to actual conditions, so that impurities other than the magnetically labeled cells in the magnetic separation column 100 can be blown away. The larger the first preset dose, the more impurities are blown away, but the lower the efficiency. In one embodiment, the first preset dose is 5-50mL, thereby improving the efficiency under the premise that the purity of the adsorbed magnetically labeled cells after blowing away the impurities meets the purity requirements. The second preset dose can be set according to the actual situation, so that impurities other than magnetically labeled cells in the magnetic separation column 100 can be removed. In one embodiment, the second preset dose is 5-50mL, so that impurities can be discharged from the magnetic separation column 100, and impurities can be discharged from the output pipeline 220 to avoid the impurities in the output pipeline 220 in the subsequent steps affecting the purity of the magnetically labeled cells adsorbed by the magnetic separation column 100. The second preset flow rate can be equal to or unequal to the first preset flow rate. In one embodiment, the second preset flow rate is equal to the first preset flow rate, so that the first drive pump 400 can control the gas and buffer respectively without changing the drive speed to remove impurities other than magnetically labeled cells in the magnetic separation column 100 through gas-liquid alternating cleaning. The first preset flow rate can be set according to the actual situation. In one embodiment, the first preset flow rate is 2-50 mL / min, thereby avoiding the flow rate being too high to blow the magnetically labeled cells away from the magnetic separation column 100, and avoiding the flow rate being too low to blow the impurities away from the magnetic separation column 100. This embodiment can reduce the interference of impurities such as red blood cells by about 20%, and significantly improve the purity index of the magnetically labeled cells adsorbed by the magnetic separation column 100. The first preset number of times can be set according to actual conditions, so that the impurities other than the magnetically labeled cells in the magnetic separation column 100 are removed by the first preset number of gas-liquid alternating cleanings to achieve the preset target (the preset target can be the purity requirement of the magnetically labeled cells adsorbed by the magnetic separation column 100). In one embodiment, the first preset number of times is once, so as to achieve efficient removal of impurities other than the magnetically labeled cells in the magnetic separation column 100.In another embodiment, in order to avoid the destruction of the activity of magnetically labeled cells due to a single long-term exposure to gas, when a gas-liquid cleaning operation of a preset duration is required and the preset duration exceeds a preset duration threshold, the preset duration can be divided into multiple shorter cleaning durations, and then a gas-liquid cleaning operation is performed in each cleaning duration. In this way, multiple gas-liquid cleaning operations will be performed within the preset duration. Furthermore, through multiple gas-liquid cleaning operations, the exposure time of the magnetically labeled cells to the gas in a single gas-liquid cleaning operation can be reduced, thereby improving the cell survival rate.

[0027] The cell elution method provided by the present invention adsorbs the magnetically labeled cells in the sample liquid through the magnetic separation column 100, and removes impurities (cells other than the magnetically labeled cells) remaining in the magnetic separation column 100 through gas-liquid alternating cleaning in a gas-liquid cleaning operation. The present invention can not only successfully sort out the magnetically labeled cells, but also improve the purity and sorting accuracy of the magnetically labeled cells adsorbed by the magnetic separation column, reduce the contamination of impurities to the target cells, and thus ensure the smooth progress of subsequent culture and cell research; and avoid the need to improve the purity through multiple sorting, thereby improving the efficiency of cell sorting.

[0028] In one embodiment, in the step S200, before performing the gas-liquid cleaning operation on the magnetic separation column 100 for a first preset number of times, the step further includes:

[0029] S300, controlling the buffer solution to flow through the pipeline before the magnetic separation column 100 and enter the magnetic separation column 100, so as to drive the sample liquid remaining in the pipeline before the magnetic separation column 100 to flow through the magnetic separation column 100, and then adsorbing the magnetically labeled cells in the residual sample liquid into the magnetic separation column 100 through the magnetic field.

[0030] It is understandable that after the sample liquid is controlled to flow through the magnetic separation column 100, a certain amount of sample liquid may remain in the pipeline before the magnetic separation column 100. Figure 4 In the cell elution device shown in FIG. 1 , after the sample liquid is driven from the sample container 310 to the magnetic separation column 100 through the sample pipeline 210, a certain amount of sample liquid will still remain in the sample pipeline 210. Figure 4As shown, the cleaning pipeline 240 can be connected to the sample pipeline 210, so as to be connected to the magnetic separation column 100 through the sample pipeline 210, and the buffer solution can flow through the sample pipeline 210 between the connection point between the cleaning pipeline 240 and the sample pipeline 210 and the magnetic separation column 100, thereby driving the sample liquid remaining in the sample pipeline 210 to flow through the magnetic separation column 100. In this embodiment, the buffer solution is controlled to flow through the pipeline before the magnetic separation column 100 and enter the magnetic separation column 100, so as to drive the sample liquid remaining in the pipeline before the magnetic separation column 100 to flow through the magnetic separation column 100, and then the magnetically labeled cells in the residual sample liquid are adsorbed in the magnetic separation column 100 through the magnetic field, thereby improving the utilization rate of the sample liquid and avoiding the influence of the sample liquid in the pipeline on the subsequent steps. The flow rate at which the buffer solution drives the sample liquid remaining in the pipeline before the magnetic separation column 100 to flow through the magnetic separation column 100, and the dosage of the buffer solution can be set according to actual conditions. The flow rate generally takes into account factors such as the survival rate of the target cells (to avoid the target cells being destroyed due to excessive flow rate) and the adsorption rate of the magnetic field of the magnetic separation column 100 on the target cells (to avoid the target cells flowing out of the magnetic separation column 100 before being adsorbed in the magnetic separation column 100 due to excessive flow rate). The above dosage is generally greater than the volume of the pipeline before the magnetic separation column 100, so that all the sample liquid remaining in the pipeline before the magnetic separation column 100 can flow through the magnetic separation column 100. In one embodiment, the buffer solution is controlled to flow through the pipeline before the magnetic separation column 100 at a flow rate of 5-100 mL / min and enter the magnetic separation column 100, thereby improving the sorting efficiency of the magnetic separation column 100 (including cell survival rate and cell adsorption rate, etc.); the dosage of the buffer solution is 100 mL, thereby ensuring that all the sample liquid remaining in the pipeline before the magnetic separation column 100 flows through the magnetic separation column 100.

[0031] like Figure 2 As shown, in one embodiment, the step S200, after performing the gas-liquid cleaning operation on the magnetic separation column 100 for a first preset number of times, further includes:

[0032] S400, controlling the magnetic field in the magnetic separation column 100 to be eliminated, and controlling the magnetic separation column 100 to be connected to the side attached pipeline 250 connected to the opposite ends of the magnetic separation column 100, so that the magnetically labeled cells are no longer adsorbed by the magnetic field in the magnetic separation column 100, and a ring-shaped internal circulation channel is formed between the magnetic separation column 100 and the side attached pipeline 250. It can be understood that the side attached pipeline 250 is connected to the opposite ends of the magnetic separation column 100, so as to form a ring-shaped internal circulation channel between the magnetic separation column 100 and the side attached pipeline 250.

[0033] S500, controlling the circulating flow of the buffer solution in the annular inner circulation channel, so as to flush the magnetically labeled cells that are no longer adsorbed by the magnetic field in the magnetic separation column 100 into the annular inner circulation channel and flow with the buffer solution. It can be understood that a second driving pump (such as a peristaltic pump, etc.) for driving the flow of fluid in the bypass pipeline 250 can be provided on the bypass pipeline 250, and the circulating flow of the buffer solution in the annular inner circulation channel can be controlled by controlling the second driving pump; Figure 4 As shown, one end of the bypass line 250 can be connected to the output line 220, the first driving pump 400 can be set on the output line 220 and located between the connecting point of the bypass line 250 and the output line 220 and the magnetic separation column 100, and a negative separation switch valve 520 for controlling the connection between the output line 220 and the negative separation container 320 can be set at the entrance of the negative separation container 320. After closing the negative separation switch valve 520, the magnetic separation column 100, the bypass line 250, and the output line 220 between the connecting point of the bypass line 250 and the output line 220 and the magnetic separation column 100 together form an annular internal circulation channel, and then the circulation of the buffer solution in the annular internal circulation channel is controlled by controlling the first driving pump 400. After the magnetic field in the magnetic separation column 100 is eliminated, the magnetically labeled cells that are no longer adsorbed by the magnetic field in the magnetic separation column 100 can be mixed with the buffer solution and flow along with the buffer solution under the circulating flow of the buffer solution in the annular inner circulation channel.

[0034] S600, control the magnetic field to be regenerated in the magnetic separation column 100, and control the circulation of the buffer solution in the annular inner circulation channel, so that when the buffer solution flows through the magnetic separation column 100, the magnetically labeled cells in the buffer solution are re-adsorbed in the magnetic separation column 100 through the magnetic field. It can be understood that after the magnetic field is regenerated in the magnetic separation column 100, the magnetically labeled cells in the buffer solution can be re-adsorbed in the magnetic separation column 100 through the magnetic field. The flow rate of the circulating flow of the buffer solution in the annular inner circulation channel can be set according to actual conditions. In one embodiment, the flow rate of the circulating flow of the buffer solution in the annular inner circulation channel is controlled to be 5-100mL / min, thereby improving the sorting efficiency (including cell viability and cell adsorption rate, etc.) of the magnetic separation column 100. In this embodiment, the magnetically labeled cells are detached and then re-adsorbed, so that the impurities stuck in the magnetic separation column 100 or staying in the magnetic separation column 100 together with the magnetically labeled cells can also be detached from the magnetic separation column 100. In the process of re-adsorbing the magnetically labeled cells on the magnetic separation column 100, the above impurities may no longer be stuck in the magnetic separation column 100 and detach from the magnetically labeled cells, so as not to be re-adsorbed in the magnetic separation column 100, thereby further improving the purity of the magnetically labeled cells adsorbed by the magnetic separation column 100. In this embodiment, the purity of the magnetically labeled cells is improved by 5%-15%, and finally the purity of the magnetically labeled cells can reach 97%-99%.

[0035] In one embodiment, in step 500, controlling the circulating flow of the buffer solution in the annular inner circulation channel includes:

[0036] S510, perform an internal circulation flushing operation of a preset number of cycles, the internal circulation flushing operation comprising: controlling the buffer to flow in the annular inner circulation channel along the first circulation direction and continue for a first preset time, and then controlling the buffer to flow in the annular inner circulation channel along the second circulation direction and continue for a second preset time; the second circulation direction is opposite to the first circulation direction. It can be understood that the first preset time and the second preset time can be set according to actual conditions, so that the magnetically labeled cells that are no longer adsorbed by the magnetic field in the magnetic separation column 100 are flushed into the annular inner circulation channel. The flow rate of the control buffer flowing in the annular inner circulation channel can be set according to actual conditions. In one embodiment, the above flow rate is 5-500mL / min, so that the separation of magnetically labeled cells and impurities can be promoted. The preset number of cycles can be set according to actual conditions. In one embodiment, the preset number of cycles is once, so as to achieve efficient separation of magnetically labeled cells and impurities. In another embodiment, the preset number of cycles can also be multiple times, so as to further promote the separation of magnetically labeled cells and impurities.

[0037] like Figure 3 As shown, in one embodiment, the step S200, after performing the gas-liquid cleaning operation on the magnetic separation column 100 for a first preset number of times, further includes:

[0038] S700, controlling the magnetic field in the magnetic separation column 100 to be eliminated so that the magnetically labeled cells are no longer adsorbed by the magnetic field in the magnetic separation column 100. It can be understood that after the magnetic field in the magnetic separation column 100 is eliminated, the magnetically labeled cells are no longer adsorbed by the magnetic field in the magnetic separation column 100.

[0039] S800 , performing a second preset number of gas-liquid flushing operations to flush the magnetically labeled cells in the magnetic separation column 100 that are no longer adsorbed by the magnetic field into the positive selection container 340 .

[0040] It can be understood that the gas-liquid flushing operation is used to flush the magnetically labeled cells in the magnetic separation column 100 that are no longer adsorbed by the magnetic field into the positive selection container 340, thereby completing the sorting of the magnetically labeled cells in the sample liquid. The second preset number of times can be set according to actual conditions. In one embodiment, the second preset number of times is once, so as to achieve efficient flushing of the magnetically labeled cells. In another embodiment, the second preset number of times can also be multiple times, so as to avoid the residue of the magnetically labeled cells.

[0041] In one embodiment, in step S800, the gas-liquid flushing operation includes:

[0042] S810, after controlling the third preset dose of buffer to flush the magnetic separation column 100 at the third preset flow rate, controlling the fourth preset dose of gas to flush the magnetic separation column 100 at the fourth preset flow rate, so that the magnetically labeled cells in the magnetic separation column 100 that are no longer adsorbed by the magnetic field are discharged along with the buffer through alternating gas-liquid flushing, and then collected in the positive selection container 340. The third preset flow rate is greater than the second preset flow rate; the fourth preset flow rate is greater than the first preset flow rate. It can be understood that the magnetic separation column 100 generally contains multiple flow channels inside and has a structure with a narrow entrance and a wide middle. By controlling the gas to flush the magnetic separation column 100, gravity can be used to make the gas-liquid interface corresponding to the gas-liquid alternating flushing parallel to the ground, thereby making the flow rate at each position under the gas-liquid interface relatively uniform, thereby ensuring that each flushed position will be re-eluted by a liquid at a certain speed (because the magnetic separation column 100 has a structure with a narrow entrance and a wide middle, if the gas-liquid alternating flushing is not performed, the magnetic separation column 100 may only have a certain impact force at the middle position relative to the entrance, while the position far from the entrance and close to the outer wall will not be impacted by the liquid with a certain speed. Since the magnetically labeled cells that need to be flushed are often attached to the side wall, this will result in a poor flushing effect). Therefore, the gas-liquid alternating flushing of this embodiment obviously improves the flushing effect.

[0043] That is, in this embodiment, after the magnetic field in the magnetic separation column 100 is eliminated, a gas-liquid flushing operation can be performed by buffer and gas, and the magnetically labeled cells no longer adsorbed by the magnetic field are transported to the positive selection container 340 after alternately flushing with buffer and gas, thereby completing the sorting of the magnetically labeled cells in the sample liquid. The third preset dose and the fourth preset dose can be set according to actual conditions, so that the magnetically labeled cells in the magnetic separation column 100 that are no longer adsorbed by the magnetic field can be all transported to the positive selection container 340 without being left in the pipeline. The second preset flow rate is the flow rate of the buffer solution to remove impurities other than the magnetically labeled cells in the magnetic separation column 100, and the third preset flow rate is the flow rate of the buffer solution to flush the magnetically labeled cells. Therefore, the second preset flow rate is less than the third preset flow rate, so that when the impurities in the magnetic separation column 100 are removed by the buffer solution, the magnetically labeled cells in the magnetic separation column 100 will not be removed, thereby avoiding the loss of magnetically labeled cells. The first preset flow rate is the flow rate of gas washing to remove impurities other than magnetically labeled cells in the magnetic separation column 100, and the fourth preset flow rate is the flow rate of gas washing the magnetically labeled cells. Therefore, the first preset flow rate is less than the fourth preset flow rate, and when the impurities in the magnetic separation column 100 are removed by gas washing, the magnetically labeled cells in the magnetic separation column 100 will not be removed, thereby avoiding the loss of magnetically labeled cells.

[0044] In one embodiment, the third preset flow rate is 50-500 mL / min, and the third preset dose is 0-500 mL. The fourth preset flow rate is 10-300 mL / min, and the fourth preset dose is 0-300 mL. In this embodiment, the magnetically labeled cells that have not been mixed with the buffer in the magnetic separation column 100 can be completely washed into the positive selection container 340 through alternating gas-liquid washing.

[0045] In this embodiment, if Figure 4 As shown, the magnetic separation column 100 can be connected to the cation selection container 340 through the output pipeline 220, the inlet of the cation selection container 340 can be provided with a cation selection switch valve 560 for controlling the connection between the output pipeline 220 and the cation selection container 340, and the bypass pipeline 250 can be provided with a circulation switch valve 550 for controlling the connection of the bypass pipeline 250. First, after the positive selection switch valve 560 and the cleaning switch valve 540 are opened and other switch valves are closed, the operation of the first driving pump 400 can be controlled to drive the buffer solution from the cleaning container 330 to the magnetic separation column 100 through the cleaning pipeline 240; secondly, after the positive selection switch valve 560 and the gas switch valve 530 are opened and other switch valves are closed, the operation of the first driving pump 400 can be controlled to drive the gas to be input into the magnetic separation column 100 through the gas pipeline 230, and then the magnetically labeled cells in the magnetic separation column 100 that are no longer adsorbed by the magnetic field are discharged along with the buffer solution through gas-liquid alternating flushing, and then collected in the positive selection container 340. The magnetic separation column 100 can also be connected to the positive selection container 340 through structural elements and pipelines, so that the magnetically labeled cells in the magnetic separation column 100 that are no longer adsorbed by the magnetic field are discharged along with the buffer solution through gas-liquid alternating flushing, and then collected in the positive selection container 340, which will not be repeated here.

[0046] In one embodiment, the step S100, after confirming that a magnetic field is generated in the magnetic separation column 100, controlling the sample liquid to flow through the magnetic separation column 100 so as to adsorb the magnetically labeled cells in the sample liquid into the magnetic separation column 100 through the magnetic field, comprises:

[0047] S900, control the buffer solution to be perfused into the magnetic separation column 100 and all the pipelines to discharge the gas in the magnetic separation column 100 and all the pipelines. It can be understood that discharging the gas in the magnetic separation column 100 and all the pipelines can avoid the presence of gas in the magnetic separation column 100 and the pipelines to affect the separation effect of the magnetic separation column 100.

[0048] In one embodiment, the step S900, before the control buffer is perfused into the magnetic separation column 100 and all pipelines to discharge the gas in the magnetic separation column 100 and all pipelines, includes:

[0049] S1000: Inspect the magnetic separation column 100 and all pipelines according to preset inspection rules and generate inspection results. It can be understood that inspecting the magnetic separation column 100 and all pipelines can avoid waste of sample liquid due to equipment failure.

[0050] It should be understood that the order of execution of the steps in the above embodiment does not necessarily mean the order of execution. The execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiment of the present invention.

[0051] The present invention also provides a cell elution device, comprising a controller, the controller comprising a processor and a memory, the memory storing an executable program, and the processor being used to execute the steps of the cell elution method. Figure 4 As shown, the cell elution device also includes a magnetic separation column 100, one end of which can be connected to the sample container 310 through the sample pipeline 210, and the other end of which can be connected to the negative separation container 320 through the output pipeline 220, and a first driving pump 400 (such as a peristaltic pump, etc., the first driving pump 400 is connected to the controller) for driving the flow of liquid in the output pipeline 220 is arranged on the output pipeline 220, so that the sample liquid can be driven from the sample container 310 through the sample pipeline 210 to the magnetic separation column 100 by controlling the operation of the first driving pump 400, and can be output from the magnetic separation column 100 to the negative separation container 320 through the output pipeline 220, so that the sample liquid flows through the magnetic separation column 100. A magnetic field generating mechanism connected to the controller can be arranged near the magnetic separation column 100, so that the controller can control the magnetic field generating mechanism to generate or eliminate a magnetic field in the magnetic separation column 100.

[0052] Furthermore, one end of the magnetic separation column 100 connected to the sample pipeline 210 can be connected to the atmospheric environment through the gas pipeline 230, and a sample switch valve 510 for controlling the switch of the sample pipeline 210 is provided on the sample pipeline 210, so that after closing the sample switch valve 510, by controlling the operation of the first driving pump 400, driving gas from the outside through the gas pipeline 230 to clean the magnetic separation column 100; one end of the magnetic separation column 100 connected to the sample pipeline 210 can also be connected to the cleaning container 330 through the cleaning pipeline 240, and in the gas The gas switch valve 530 for controlling the switch of the gas pipeline 230 is provided on the body pipeline 230, so that after closing the sample switch valve 510 and the gas switch valve 530, by controlling the operation of the first driving pump 400, the buffer solution (such as nanomagnetic separation buffer solution, etc.) can be driven from the cleaning container 330 through the cleaning pipeline 240 to enter the magnetic separation column 100, and output from the magnetic separation column 100 through the output pipeline 220 to the negative separation container 320, thereby removing impurities other than magnetically labeled cells in the magnetic separation column 100 through gas-liquid alternating cleaning. In addition, the cleaning pipeline 240 can be provided with a cleaning switch valve 540 for controlling the switch of the cleaning pipeline 240, so that after closing the sample switch valve 510 and the cleaning switch valve 540, by controlling the operation of the first driving pump 400, the gas can be driven from the outside through the gas pipeline 230 to clean the magnetic separation column 100, thereby realizing the next round of gas-liquid cleaning operation. The gas-liquid alternating cleaning in the gas-liquid cleaning operation can better remove impurities other than magnetically labeled cells in the magnetic separation column 100 .

[0053] It can be understood that the cell elution device further includes the side pipe 250 connected to the opposite ends of the magnetic separation column 100 , so that an annular internal circulation channel can be formed between the magnetic separation column 100 and the side pipe 250 .

[0054] Understandably, the execution function of the controller corresponds to the cell elution method in the above embodiment. For the specific definition of the controller, please refer to the definition of the cell elution method above, which will not be repeated here. Each submodule in the above controller can be implemented in whole or in part by software, hardware and a combination thereof. Each of the above submodules can be embedded in or independent of the processor in the computer device in hardware form, or can be stored in the memory in the computer device in software form, so that the processor can call and execute the operations corresponding to each of the above submodules.

[0055] The present invention also provides a computer-readable storage medium, wherein the computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, the steps of the cell elution method are implemented.

[0056] Those of ordinary skill in the art can understand that all or part of the processes in the above-mentioned embodiment methods can be implemented by instructing related hardware through computer-readable instructions, and the computer-readable instructions can be stored in a non-volatile readable storage medium or a volatile readable storage medium. When the computer-readable instructions are executed, they may include the processes of the embodiments of the above-mentioned methods. Among them, any reference to memory, storage, database or other media used in the embodiments provided in this application may include non-volatile and / or volatile memory. Non-volatile memory may include read-only memory (ROM), programmable ROM (PROM), electrically programmable ROM (EPROM), electrically erasable programmable ROM (EEPROM) or flash memory. Volatile memory may include random access memory (RAM) or external cache memory. By way of illustration and not limitation, RAM is available in many forms, such as static RAM (SRAM), dynamic RAM (DRAM), synchronous DRAM (SDRAM), double data rate SDRAM (DDRSDRAM), enhanced SDRAM (ESDRAM), synchronous link (Synchlink) DRAM (SLDRAM), memory bus (Rambus) direct RAM (RDRAM), direct memory bus dynamic RAM (DRDRAM), and memory bus dynamic RAM (RDRAM), etc.

[0057] Those skilled in the art will clearly understand that for the sake of convenience and brevity of description, only the division of the above-mentioned functional units and modules is used as an example. In actual applications, the above-mentioned functions can be distributed and completed by different functional units and modules as needed, that is, the internal structure of the system can be divided into different functional units or modules to complete all or part of the functions described above.

[0058] The embodiments described above are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. Such modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention, and should all be included in the protection scope of the present invention.

Claims

1. A cell elution method, characterized in that: include: After confirming that a magnetic field is generated in the magnetic separation column, controlling the sample liquid to flow through the magnetic separation column, so that the magnetically labeled cells in the sample liquid are adsorbed in the magnetic separation column through the magnetic field; A first preset number of gas-liquid cleaning operations are performed on the magnetic separation column; the gas-liquid cleaning operation includes: controlling a first preset dose of gas to clean the magnetic separation column at a first preset flow rate, and then controlling a second preset dose of buffer to clean the magnetic separation column at a second preset flow rate, so as to remove impurities other than magnetically labeled cells in the magnetic separation column through alternating gas-liquid cleaning.

2. The cell elution method according to claim 1, characterized in that: Before performing the gas-liquid cleaning operation on the magnetic separation column for a first preset number of times, the method further includes: The buffer solution is controlled to flow through the pipeline before the magnetic separation column and enter the magnetic separation column to drive the sample liquid remaining in the pipeline before the magnetic separation column to flow through the magnetic separation column, and then the magnetically labeled cells in the residual sample liquid are adsorbed in the magnetic separation column through the magnetic field.

3. The cell elution method according to claim 1, characterized in that: After the magnetic separation column is subjected to a first preset number of gas-liquid cleaning operations, the method further comprises: Controlling the elimination of the magnetic field in the magnetic separation column, and controlling the magnetic separation column to communicate with the side attached pipelines connected to the two opposite ends of the magnetic separation column, so that the magnetically labeled cells are no longer adsorbed by the magnetic field in the magnetic separation column, and a ring-shaped internal circulation channel is formed between the magnetic separation column and the side attached pipeline; Controlling the circulation of the buffer solution in the annular inner circulation channel to flush the magnetically labeled cells that are no longer adsorbed by the magnetic field in the magnetic separation column into the annular inner circulation channel and flow with the buffer solution; The magnetic field in the magnetic separation column is controlled to be regenerated, and the circulation of the buffer solution in the annular inner circulation channel is controlled so that when the buffer solution flows through the magnetic separation column, the magnetically labeled cells in the buffer solution are re-adsorbed into the magnetic separation column through the magnetic field.

4. The cell elution method according to claim 3, characterized in that: The controlling the circulating flow of the buffer solution in the annular inner circulation channel comprises: An internal circulation flushing operation is performed for a preset number of cycles, wherein the internal circulation flushing operation includes: controlling the buffer solution to flow in the annular internal circulation channel along a first circulation direction and continuing for a first preset time, and then controlling the buffer solution to flow in the annular internal circulation channel along a second circulation direction and continuing for a second preset time; the second circulation direction is opposite to the first circulation direction.

5. The cell elution method according to claim 1, characterized in that: After the magnetic separation column is subjected to a first preset number of gas-liquid cleaning operations, the method further comprises: Controlling the elimination of the magnetic field in the magnetic separation column so that the magnetically labeled cells are no longer adsorbed by the magnetic field in the magnetic separation column; A second preset number of gas-liquid flushing operations are performed to flush the magnetically labeled cells in the magnetic separation column that are no longer adsorbed by the magnetic field into the positive selection container.

6. The cell elution method according to claim 5, characterized in that: The gas-liquid flushing operation comprises: After controlling the third preset dose of buffer to flush the magnetic separation column at the third preset flow rate, controlling the fourth preset dose of gas to flush the magnetic separation column at the fourth preset flow rate, so that the magnetically labeled cells in the magnetic separation column that are no longer adsorbed by the magnetic field are discharged along with the buffer through alternating gas-liquid flushing, and then collected in the positive selection container; wherein the third preset flow rate is greater than the second preset flow rate; and the fourth preset flow rate is greater than the first preset flow rate.

7. The cell elution method according to claim 1, characterized in that: After confirming that a magnetic field is generated in the magnetic separation column, before controlling the sample liquid to flow through the magnetic separation column so as to adsorb the magnetically labeled cells in the sample liquid into the magnetic separation column through the magnetic field, the method comprises: The control buffer is perfused into the magnetic separation column and all pipelines to discharge the gas in the magnetic separation column and all pipelines.

8. The cell elution method according to claim 7, characterized in that: Before the control buffer is perfused into the magnetic separation column and all pipelines to discharge the gas in the magnetic separation column and all pipelines, the method comprises: The magnetic separation column and all pipelines are inspected according to preset inspection rules, and inspection results are generated.

9. A cell elution device, characterized in that: It comprises a controller, the controller comprises a processor and a memory, the memory stores an executable program, and the processor is used to execute the cell elution method according to any one of claims 1 to 8.

10. A computer-readable storage medium storing a computer program, characterized in that: When the computer program is executed by a processor, the cell elution method according to any one of claims 1 to 8 is implemented.

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