Cell concentration cyclic measurement method and tubing

Abstract

This application relates to the field of cell biology, providing a method and tubing for cyclic cell concentration measurement. The method includes: using a peristaltic pump to draw the cell solution to be tested from the outlet of a cell counting chip to the inlet of the cell counting chip; obtaining the cell concentration of the cell solution to be tested based on the number of cells in the shunt channel of the cell counting chip and the volume of the cell solution to be tested in the shunt channel; updating the cell concentration obtained in the Nth iteration using the cell concentration obtained in the Nth iteration, and determining the updated cell concentration obtained in the Nth iteration as the final cell concentration of the cell solution to be tested. The cyclic cell concentration measurement method and tubing provided in this application can repeatedly use the cell solution to be tested for multiple cell counting and cell concentration calculations, and determine the final cell concentration of the cell solution to be tested based on the multiple cell concentrations obtained, avoiding uncorrectable errors caused by a single cell concentration measurement, and improving the accuracy of cell concentration measurement.

Description

Technical Field

[0001] This application relates to the field of cell technology, specifically to a method and pipeline for circulating cell concentration measurement. Background Technology

[0002] Currently, when measuring the cell concentration of cell sap, a cell counter is usually used for cell counting. First, cell sap is manually extracted and dropped into the counting area of ​​the counting chamber of the counting chamber. Then, the counting chamber is placed under a microscope for manual cell counting. Finally, the cell concentration of the cell sap is calculated based on the number of cells and the volume of extracted cell sap.

[0003] However, in this method, the cell concentration is measured only once. If there is human error or cell contamination, the cell concentration measurement will be inaccurate. Summary of the Invention

[0004] This application provides a cell concentration cyclic measurement method and pipeline to solve the technical problem that traditional methods of cell concentration measurement are one-time, and that inaccurate cell concentration measurement can occur due to human error or cell contamination.

[0005] In a first aspect, embodiments of this application provide a method for cyclic measurement of cell concentration, comprising:

[0006] A peristaltic pump is used to extract the cell solution to be tested from the container into the cell counting chip;

[0007] The peristaltic pump is used to draw the cell solution to be tested from the outlet of the cell counting chip to the inlet of the cell counting chip;

[0008] Based on the number of cells in the shunt channel of the cell counting chip and the volume of the cell solution to be tested in the shunt channel, the cell concentration of the cell solution to be tested is obtained. Then, the process returns to the step of using the peristaltic pump to draw the cell solution to be tested from the outlet of the cell counting chip to the inlet of the cell counting chip, until the cell concentration of the cell solution to be tested is obtained again.

[0009] The cell concentration obtained in the Nth iteration is updated using the cell concentration obtained in the Nth iteration, and the updated cell concentration obtained in the Nth iteration is determined as the final cell concentration of the cell solution to be tested; where N is an integer greater than or equal to 2.

[0010] In one embodiment, updating the cell concentration obtained in the Nth iteration using the cell concentrations obtained in the Nth iteration includes:

[0011] Update the cell concentration obtained in the Nth iteration using the average of the cell concentrations obtained in the Nth iteration.

[0012] In one embodiment, updating the cell concentration obtained in the Nth iteration using the cell concentrations obtained in the Nth iteration includes:

[0013] The average of the cell concentrations obtained in the first and second tests was determined as the corrected cell concentration obtained in the second test.

[0014] The cell concentration obtained in the Nth iteration is updated using the average of the cell concentration obtained in the N-th iteration and the corrected cell concentration obtained in the (N-1)th iteration; where N is an integer greater than or equal to 3.

[0015] In one embodiment, before determining the cell concentration of the test cell solution based on the number of cells in the shunt channel of the cell counting chip and the volume of the test cell solution in the shunt channel, the following steps are included:

[0016] After the cell fluid to be tested is diverted into multiple diversion channels of different depths on the cell counting chip, microscope images of the multiple diversion channels of different depths are acquired to obtain the image to be processed.

[0017] The number of cells in the test fluid in each shunt channel is identified based on the image to be processed.

[0018] In one embodiment, obtaining the cell concentration of the test cell solution based on the number of cells in the shunt channel of the cell counting chip and the volume of the test cell solution in the shunt channel includes:

[0019] The cell concentration of the test cell solution is obtained based on the total number of cells in the test cell solution in all shunt channels and the total volume of the test cell solution in all shunt channels.

[0020] In one embodiment, the step of using a peristaltic pump to draw the cell solution to be tested from the container into the cell counting chip includes:

[0021] A peristaltic pump is used to draw the cell solution to be tested from the container into the first inlet of the mixing channel in the cell counting chip.

[0022] In one embodiment, before determining the cell concentration of the test cell solution based on the number of cells in the shunt channel of the cell counting chip and the volume of the test cell solution in the shunt channel, the following steps are included:

[0023] The staining agent is drawn into the second inlet of the mixing channel in the cell counting chip, so that the cell solution to be tested and the staining agent are mixed in the mixing channel; the outlet of the mixing channel is connected to the inlet of the diversion channel.

[0024] In one embodiment, the step of using a peristaltic pump to draw the cell solution to be tested from the container into the cell counting chip includes:

[0025] When the container is in a static state, a peristaltic pump is used to extract the cell fluid to be tested from the bottom of the container wall into the cell counting chip.

[0026] When the container is in a centrifugal state, a peristaltic pump is used to extract the test cell fluid near the top or bottom of the container wall into the cell counting chip.

[0027] In one embodiment, the step of using the peristaltic pump to draw the cell solution to be tested from the outlet of the cell counting chip to the inlet of the cell counting chip includes:

[0028] The peristaltic pump is used to draw the cell solution to be tested from the outlet of the diversion channel to the first inlet of the mixing channel.

[0029] Secondly, embodiments of this application provide a cell concentration circulating metering pipeline for implementing the cell concentration circulating metering method described in the first aspect, comprising: a container, a peristaltic pump, and a cell counting chip;

[0030] The cell counting chip includes a mixing channel and multiple diversion channels of different depths, and the outlet of the mixing channel is connected to the inlet of all diversion channels.

[0031] The outlet of the container is connected to the inlet of the peristaltic pump;

[0032] The outlet of the peristaltic pump is connected to the first inlet of the mixing channel;

[0033] The outlets of all the diversion channels are connected to the inlet of the peristaltic pump.

[0034] The cell concentration cyclic measurement method and tubing provided in this application utilizes a peristaltic pump to draw the test cell solution from a container into a cell counting chip. The peristaltic pump then draws the test cell solution from the outlet of the cell counting chip to the inlet. Based on the number of cells in the shunt channel of the cell counting chip and the volume of the test cell solution in the shunt channel, the cell concentration of the test cell solution is obtained. The process returns to the step of drawing the test cell solution from the outlet of the cell counting chip to the inlet, repeating until the cell concentration of the test cell solution is obtained again. The cell concentration obtained in the Nth iteration is updated using the cell concentration obtained in the Nth iteration. If N equals a threshold number of iterations, the updated cell concentration obtained in the Nth iteration is determined as the final cell concentration of the test cell solution. This application involves drawing the test cell solution from the container into a cell counting chip, and then drawing the test cell solution from the outlet of the cell counting chip to the inlet of the cell counting chip. This allows the test cell solution to circulate on the cell counting chip, enabling repeated cell counting and cell concentration calculations. The final cell concentration of the test cell solution is determined based on the cell concentrations obtained from multiple measurements, avoiding uncorrectable errors caused by a single cell concentration measurement and improving the accuracy of cell concentration measurement. Attached Figure Description

[0035] To more clearly illustrate the technical solutions in this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0036] Figure 1 This is one of the schematic flowcharts of the cell concentration cyclic measurement method provided in the embodiments of this application;

[0037] Figure 2 This is the second schematic flowchart of the cell concentration cyclic measurement method provided in the embodiments of this application;

[0038] Figure 3 This is the third schematic flowchart of the cell concentration cyclic measurement method provided in the embodiments of this application;

[0039] Figure 4 This is the fourth schematic flowchart of the cell concentration cyclic measurement method provided in the embodiments of this application;

[0040] Figure 5 This is a schematic diagram of the cell concentration circulation metering pipeline structure provided in the embodiments of this application.

[0041] Figure label:

[0042] 1-Container; 11-First opening; 12-Second opening; 2-Peristaltic pump; 3-Cell counting chip; 4-First control valve; 5-Second control valve. Detailed Implementation

[0043] To make the objectives, technical solutions, and advantages of this application clearer, the technical solutions of this application will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0044] Figure 1 This is one of the schematic flowcharts of the cell concentration cyclic metering method provided in the embodiments of this application. (Refer to...) Figure 1 This application provides a method for cyclic measurement of cell concentration, which may include:

[0045] 101. Use a peristaltic pump to draw the cell solution to be tested from the container into the cell counting chip;

[0046] 102. Use a peristaltic pump to draw the cell solution to be tested from the outlet of the cell counting chip to the inlet of the cell counting chip;

[0047] 103. Based on the number of cells in the shunt channel of the cell counting chip and the volume of the cell solution to be tested in the shunt channel, obtain the cell concentration of the cell solution to be tested, return to step 102, and repeat until the cell concentration of the cell solution to be tested is obtained again.

[0048] 104. Update the cell concentration obtained in the Nth test using the cell concentration obtained in the Nth test, and determine the updated cell concentration obtained in the Nth test as the final cell concentration of the cell solution to be tested.

[0049] Where N is an integer greater than or equal to 2;

[0050] In step 103, the cell concentration of the test cell solution can be obtained by dividing the number of cells in the shunt channel by the volume of the test cell solution in the shunt channel.

[0051] In step 104, a number of times threshold can be set. When N equals the number of times threshold, the cell concentration obtained in the updated Nth time is determined as the final cell concentration of the cell solution to be tested.

[0052] The cell concentration cyclic measurement method provided in this embodiment uses a peristaltic pump to draw the test cell solution from the container into the cell counting chip. The peristaltic pump also draws the test cell solution from the outlet of the cell counting chip to the inlet of the cell counting chip. Based on the number of cells in the shunt channel of the cell counting chip and the volume of the test cell solution in the shunt channel, the cell concentration of the test cell solution is obtained. The process returns to the step of drawing the test cell solution from the outlet of the cell counting chip to the inlet of the cell counting chip, until the cell concentration of the test cell solution is obtained again. The cell concentration obtained in the Nth iteration is updated using the cell concentration obtained in the Nth iteration. If N is equal to the number of iterations threshold, the updated cell concentration obtained in the Nth iteration is determined as the final cell concentration of the test cell solution. This embodiment involves drawing the test cell solution from the container into the cell counting chip, and then drawing the test cell solution from the outlet of the cell counting chip to the inlet of the cell counting chip. This allows the test cell solution to circulate on the cell counting chip, enabling repeated cell counting and cell concentration calculations. The final cell concentration of the test cell solution is determined based on the multiple cell concentrations obtained, avoiding uncorrectable errors caused by a single cell concentration measurement and improving the accuracy of cell concentration measurement.

[0053] In addition, in this embodiment, the extraction of the cell solution to be tested is carried out under sterile machine control without manual operation, which can avoid operational errors and contamination of the cell solution caused by manual extraction, and further improve the accuracy and efficiency of cell concentration measurement.

[0054] In one embodiment, updating the cell concentration obtained in the Nth iteration using the cell concentrations obtained in the Nth iteration includes:

[0055] Update the cell concentration obtained in the Nth iteration using the average of the cell concentrations obtained in the Nth iteration.

[0056] This embodiment uses the average value of the cell concentrations obtained N times to update the cell concentration obtained N times, which can balance the possible errors in the cell concentration obtained N times and make the final cell concentration measurement more accurate.

[0057] Figure 2 This is the second schematic flowchart of the cell concentration cyclic metering method provided in the embodiments of this application. (Refer to...) Figure 2 In one embodiment, updating the cell concentration obtained in the Nth iteration using the cell concentrations obtained in the Nth iteration may include:

[0058] 201. The average of the cell concentrations obtained in the first and second trials is taken as the corrected cell concentration obtained in the second trial.

[0059] 202. Update the cell concentration obtained in the Nth iteration using the average of the cell concentration obtained in the N-th iteration and the corrected cell concentration obtained in the N-1th iteration.

[0060] Where N is an integer greater than or equal to 3.

[0061] Assuming N equals 4, the cell concentration obtained in the first test is A, in the second test is B, in the third test is C, and in the fourth test is D, then the corrected cell concentration obtained in the second test is: The third corrected cell concentration was Then utilize Update the cell concentration obtained in the fourth iteration.

[0062] It should be noted that when N equals 2, the corrected cell concentration obtained the second time is used, that is, the average of the cell concentration obtained the first time and the cell concentration obtained the second time is used to update the cell concentration obtained the second time.

[0063] This embodiment uses the average of the cell concentration obtained in the Nth time and the corrected cell concentration obtained in the (N-1)th time to update the cell concentration obtained in the Nth time. It can continuously use the average value to correct the cell concentration until the correction of the cell concentration obtained in the Nth time is completed, thereby improving the accuracy of the final cell concentration measurement.

[0064] Figure 3 This is the third schematic flowchart of the cell concentration cyclic metering method provided in the embodiments of this application. (Refer to...) Figure 3 In one embodiment, before obtaining the cell concentration of the test cell solution based on the number of cells in the shunt channel of the cell counting chip and the volume of the test cell solution in the shunt channel, and before obtaining the cell concentration of the test cell solution based on the number of cells in the shunt channel of the cell counting chip and the volume of the test cell solution in the shunt channel, the process may include:

[0065] 301. After the cell fluid to be tested is diverted into multiple diversion channels of different depths on the cell counting chip, microscope images of the multiple diversion channels of different depths are acquired to obtain the image to be processed.

[0066] 302. Identify the number of cells in the cell solution to be tested in each shunt channel based on the image to be processed;

[0067] 303. Based on the total number of cells in the test cell solution in all shunt channels and the total volume of the test cell solution in all shunt channels, the cell concentration of the test cell solution is obtained.

[0068] In step 301, an image can be acquired below the cell counting chip using a microscope camera.

[0069] The number and depth of the shunt channels of the cell counting chip are not limited here and can be set according to the needs of cell morphology, size and other factors. In this embodiment, the number of shunt channels can be 5, and the depths of these 5 shunt channels can be 50 micrometers, 150 micrometers, 250 micrometers, 350 micrometers and 450 micrometers respectively.

[0070] In step 302, a deep learning model can be used to identify the number of cells in the image to be processed, so as to improve the accuracy of the identification.

[0071] In step 303, the cell concentration of the test cell solution can be obtained by dividing the total number of cells in the test cell solution in all shunt channels by the total volume of the test cell solution in all shunt channels.

[0072] In traditional cell concentration measurement methods, all grooves in the counting area of ​​a counting chamber are of the same depth. When the cell concentration in the cell solution is low, if the groove depth is low, it means there is less cell solution in the groove, i.e., the number of cells is correspondingly lower. Under a microscope, the number of cells may be less than the true value or even absent, leading to inaccurate cell counting and consequently inaccurate cell concentration measurement. Conversely, when the cell concentration in the cell solution is high, if the groove depth is high, it means there is more cell solution in the groove, i.e., the number of cells is correspondingly higher. Under a microscope, cell stacking and covering may cause missed cell counts, leading to inaccurate cell counting and consequently inaccurate cell concentration measurement.

[0073] The cell concentration measurement method provided in this embodiment utilizes multiple shunt channels of different depths to divert the extracted cell solution. This allows for the use of different shunt channel depths to meet the cell concentration requirements of different cell solutions. The cell concentration of the cell solution is calculated by using the total number of cells in multiple shunt channels of different depths and the total volume of the cell solution to be measured. This method can balance the adverse effects of excessively low or high depths in individual channels on cell counting and improve the accuracy of cell concentration measurement.

[0074] Figure 4 This is the fourth schematic flowchart of the cell concentration cyclic metering method provided in the embodiments of this application. (Refer to...) Figure 4 In one embodiment, a peristaltic pump is used to draw the test cell solution from the container into a cell counting chip. Before determining the cell concentration of the test cell solution based on the number of cells in the shunt channel of the cell counting chip and the volume of the test cell solution in the shunt channel, staining of the test cell solution may include:

[0075] 401. Use a peristaltic pump to draw the cell solution to be tested from the container into the first inlet of the mixing channel in the cell counting chip.

[0076] 402. Draw the staining agent into the second inlet of the mixing channel in the cell counting chip, so that the cell solution to be tested and the staining agent are mixed in the mixing channel.

[0077] The outlet of the mixing channel is connected to the inlet of the diversion channel.

[0078] By controlling the test cell solution and staining agent to flow synchronously into the mixing channel from different inlets, the liquid flow rate in the mixing channel is slowed down due to its bend compared to the straight channel. This allows the test cell solution and staining agent to come into full contact and mix in the mixing channel, resulting in uniform staining of cells in the test cell solution, highlighting cell morphology, and facilitating subsequent identification and counting.

[0079] It should be noted that the specific shape of the mixing channel is not limited here. Its bends can be right angles, arcs, or other shapes. In this embodiment, the mixing channel is a back-and-forth bend type, and its bends are right angles. In addition, bubble sensors can be set at the first and second inlets to sense the entry time of the cell solution and staining agent, and to control the two to enter the mixing channel at the same time to achieve a better mixing effect.

[0080] Since the outlet of the mixing channel is connected to the inlet of all the shunt channels, the test cell solution in the mixing channel flows into all the shunt channels after being mixed and stained.

[0081] In this embodiment, by drawing the cell solution to be tested and the staining agent into the mixing channel of the cell counting chip, the two can be mixed using the mixing channel to ensure the staining effect of the cells in the cell solution to be tested. By identifying the stained cells, viable cells can be detected. Subsequently, the cell solution to be tested can be reused to count the viable cells multiple times and calculate the cell concentration, as well as calculate the cell viability rate, and finally determine the true cell concentration of viable cells in the cell solution to be tested.

[0082] In one embodiment, a peristaltic pump is used to draw the cell solution to be tested from a container into a cell counting chip, including:

[0083] When the container is in a static state, a peristaltic pump is used to extract the cell fluid to be tested from the bottom of the container wall into the cell counting chip.

[0084] When the container is in a centrifugal state, a peristaltic pump is used to extract the test cell fluid near the top or bottom of the container wall into the cell counting chip.

[0085] When the container is in a static state, the cells in the test cell solution are more evenly distributed at the bottom of the container. The test cell solution at the bottom of the container is closer to the true indicator of the test cell solution than the test cell solution in other areas. In order to facilitate sampling, a peristaltic pump can be used to extract the test cell solution near the bottom of the container wall into the cell counting chip.

[0086] When the container is centrifuged, the cells in the test cell solution move towards the container wall during centrifugation and are distributed relatively evenly on the container wall. The test cell solution on the container wall is closer to the true test cell solution than the test cell solution in other areas. To facilitate sampling, a peristaltic pump can be used to extract the test cell solution near the top or bottom of the container wall into the cell counting chip.

[0087] It should be noted that by setting control valves and peristaltic pumps, the test cell fluid near the top or bottom of the container wall can be drawn into the cell counting chip.

[0088] This embodiment uses different methods to sample the test cell fluid according to the different states of the container, which can ensure that the sampled test cell fluid is as close as possible to its true index level.

[0089] The cell concentration circulating metering pipeline provided in the embodiments of this application is described below. The cell concentration circulating metering pipeline described below can be referred to in correspondence with the cell concentration circulating metering method described above.

[0090] Figure 5 This is a schematic diagram of the cell concentration circulating metering pipeline structure provided in an embodiment of this application. (Refer to...) Figure 5 This application provides a cell concentration circulating metering pipeline for implementing the aforementioned cell concentration circulating metering method, which may include: a container 1, a peristaltic pump 2, and a cell counting chip 3;

[0091] The cell counting chip 3 includes a mixing channel and multiple diversion channels of different depths, with the outlet of the mixing channel connected to the inlet of all diversion channels.

[0092] The outlet of container 1 is connected to the inlet of peristaltic pump 2;

[0093] The outlet of peristaltic pump 2 is connected to the first inlet of the mixing channel;

[0094] The outlets of all the diversion channels are connected to the inlet of the peristaltic pump 2.

[0095] See Figure 5 Assuming that container 1 is in a centrifugal state, the second opening 12 of container 1 can be determined as the outlet, and a peristaltic pump 2 can be set between the second opening 12 and the cell counting chip 3 to draw the test cell fluid near the top of the container wall of container 1 into the cell counting chip 3.

[0096] If it is necessary to extract the test cell fluid near the bottom of the container wall of container 1 into cell counting chip 3, the peristaltic pump 2 can be set between the first opening 11 and cell counting chip 3, and the first opening 11 of container 1 is determined as the outlet.

[0097] The peristaltic pump 2 draws the test cell solution from container 1 into cell counting chip 3. After the cell counting chip 3 is filled with the test cell solution, or after the cell concentration is obtained for the first time, a circulation pipeline is formed between cell counting chip 3 and peristaltic pump 2 by connecting the pipeline between the first control valve 4 and the second control valve 5. Then, the peristaltic pump 2 draws the test cell solution from the outlet of the diversion channel of cell counting chip 3 to the first inlet of the mixing channel, so that the test cell solution circulates in the circulation pipeline.

[0098] Among them, the first control valve 4 and the second control valve 5 are three-way valves. By switching the closure of the three channels, a continuously flowing circulation pipeline is formed between the cell counting chip 3 and the peristaltic pump 2. A sterile air filter can be installed in the circulation pipeline. For example, the sterile air filter is installed at the top of the circulation pipeline so that the pressure in the circulation pipeline is equal to the atmospheric pressure of the air, which can better allow the test cell fluid in the circulation pipeline to flow continuously.

[0099] By drawing the test cell solution from the outlet of the diversion channel to the first inlet of the mixing channel, it is ensured that the test cell solution on the cell counting chip 3 is drawn out and recirculated only after passing through the mixing channel and the diversion channel, thereby improving the utilization rate of the test cell solution on the cell counting chip 3.

[0100] The cell concentration circulating metering pipeline provided in this embodiment draws the test cell solution from the container into the cell counting chip, and then draws the test cell solution from the outlet of the cell counting chip to the inlet of the cell counting chip, so that the test cell solution circulates on the cell counting chip. This allows the test cell solution to be reused for multiple cell counting and cell concentration calculations, and the final cell concentration of the test cell solution is determined based on the cell concentrations obtained from multiple measurements. This avoids the uncorrectable error caused by a single cell concentration measurement and improves the accuracy of cell concentration measurement.

[0101] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this application, and are not intended to limit them. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this application.

Claims

1. A method for cyclic measurement of cell concentration, characterized in that, include: A peristaltic pump is used to extract the cell solution to be tested from the container into the cell counting chip; The peristaltic pump is used to draw the cell solution to be tested from the outlet of the cell counting chip to the inlet of the cell counting chip; After the cell fluid to be tested is diverted into multiple diversion channels of different depths on the cell counting chip, microscope images of the multiple diversion channels of different depths are acquired to obtain the image to be processed. The number of cells in the cell solution to be tested in each shunt channel is identified based on the image to be processed. The cell concentration of the test cell solution is obtained based on the number of cells in the shunt channel of the cell counting chip and the volume of the test cell solution in the shunt channel, including: The cell concentration of the test cell solution is obtained based on the total number of cells in the test cell solution in all shunt channels and the total volume of the test cell solution in all shunt channels. Return to the step of using the peristaltic pump to draw the test cell solution from the outlet of the cell counting chip to the inlet of the cell counting chip, until the cell concentration of the test cell solution is obtained again; The cell concentration obtained in the Nth iteration is updated using the cell concentration obtained in the Nth iteration, and the updated cell concentration obtained in the Nth iteration is determined as the final cell concentration of the cell solution to be tested; where N is an integer greater than or equal to 2.

2. The cell concentration cyclic measurement method according to claim 1, characterized in that, The process of updating the cell concentration obtained in the Nth iteration using the cell concentration obtained in the Nth iteration includes: Update the cell concentration obtained in the Nth iteration using the average of the cell concentrations obtained in the Nth iteration.

3. The cell concentration cyclic measurement method according to claim 1, characterized in that, The process of updating the cell concentration obtained in the Nth iteration using the cell concentration obtained in the Nth iteration includes: The average of the cell concentrations obtained in the first and second tests was determined as the corrected cell concentration obtained in the second test. The cell concentration obtained in the Nth iteration is updated using the average of the cell concentration obtained in the N-th iteration and the corrected cell concentration obtained in the (N-1)th iteration; where N is an integer greater than or equal to 3.

4. The cell concentration cyclic measurement method according to claim 1, characterized in that, The process of using a peristaltic pump to draw the cell solution to be tested from the container into the cell counting chip includes: A peristaltic pump is used to draw the cell solution to be tested from the container into the first inlet of the mixing channel in the cell counting chip.

5. The cell concentration cyclic measurement method according to claim 4, characterized in that, Before determining the cell concentration of the test cell solution based on the number of cells in the shunt channel of the cell counting chip and the volume of the test cell solution in the shunt channel, the following steps are included: The staining agent is drawn into the second inlet of the mixing channel in the cell counting chip, so that the cell solution to be tested is mixed with the staining agent in the mixing channel; the outlet of the mixing channel is connected to the inlet of the diversion channel.

6. The cell concentration cyclic measurement method according to claim 1, characterized in that, The process of using a peristaltic pump to draw the cell solution to be tested from the container into the cell counting chip includes: When the container is in a static state, a peristaltic pump is used to extract the cell fluid to be tested from the bottom of the container wall into the cell counting chip. When the container is in a centrifugal state, a peristaltic pump is used to extract the test cell fluid near the top or bottom of the container wall into the cell counting chip.

7. The cell concentration cyclic measurement method according to claim 4, characterized in that, The step of using the peristaltic pump to draw the cell solution to be tested from the outlet of the cell counting chip to the inlet of the cell counting chip includes: The peristaltic pump is used to draw the cell solution to be tested from the outlet of the diversion channel to the first inlet of the mixing channel.

8. A cell concentration circulating metering pipeline for implementing the cell concentration circulating metering method according to any one of claims 1 to 7, characterized in that, include: Containers, peristaltic pumps, and cell counting chips; The cell counting chip includes a mixing channel and multiple diversion channels of different depths, and the outlet of the mixing channel is connected to the inlet of all diversion channels. The outlet of the container is connected to the inlet of the peristaltic pump; The outlet of the peristaltic pump is connected to the first inlet of the mixing channel; The outlets of all the diversion channels are connected to the inlet of the peristaltic pump.

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