A flow cytometer and its liquid sampling method, system, and storage medium

Abstract

The present application relates to a flow cytometer, a liquid extraction method, a system and a storage medium thereof, belonging to the technical field of biological detection. The method includes: controlling a metering pump to work in a liquid suction mode according to a detection volume, so that a detection sample enters a liquid storage pipeline after passing through a valve; detecting a first sample flow rate at the valve; when the first sample flow rate is less than a first flow rate threshold, obtaining a current sample volume in the liquid storage pipeline, and controlling the metering pump to discharge the detection sample in a liquid discharge mode; detecting a second sample flow rate at the valve; when the second sample flow rate changes from a state of being less than a second flow rate threshold to a state of being greater than the second flow rate threshold, controlling the metering pump to discharge a preset volume of the detection sample; obtaining a sample volume difference according to the detection volume, the current sample volume and the preset volume; and controlling the metering pump to suck the detection sample in a liquid suction mode according to the sample volume difference. The present application has the effect of improving the accuracy of liquid extraction of the flow cytometer.

Description

Technical Field

[0001] The present application relates to the technical field of biological detection, and in particular, to a flow cytometer and its liquid extraction method, system, and storage medium. Background Art

[0002] A flow cytometer is a device for automatically analyzing and sorting cells. It can quickly measure, store, and display a series of important biophysical and biochemical characteristic parameters of dispersed cells suspended in a liquid.

[0003] When using a flow cytometer in the related art, it is necessary to load a test sample into a sample tube and ensure that the sample tube is not blocked. Then, the sample flow is adjusted so that individual samples in a single test sample pass through a laser beam in sequence for analysis to obtain a test result.

[0004] In view of the above related art, when a flow cytometer aspirates a test sample, in order to ensure the accuracy of the test result, it is necessary to quantitatively aspirate the test sample. However, during the process of aspirating the test sample, the flow cytometer cannot determine whether there is still a test sample in the sample tube, resulting in the aspiration of some air and affecting the accuracy of the test result. Summary of the Invention

[0005] In order to improve the accuracy of liquid extraction of a flow cytometer, the present application provides a flow cytometer and its liquid extraction method, system, and storage medium.

[0006] In a first aspect, the present application provides a liquid extraction method for a flow cytometer, adopting the following technical solution:

[0007] A liquid extraction method for a flow cytometer includes:

[0008] According to the detection volume, control a metering pump to operate in a liquid suction mode, so that the test sample enters a liquid storage pipeline after passing through a valve. The liquid suction port of the metering pump is connected to the first end of the liquid storage pipeline, the second end of the liquid storage pipeline is connected to the first end of an adapter, the second end of the adapter is connected to the first end of the valve, and the third end of the adapter leads to a flow chamber;

[0009] Detect the first sample flow rate at the valve;

[0010] In the case where the first sample flow rate is less than a first flow rate threshold, obtain the current sample volume in the liquid storage pipeline, and control the metering pump to discharge the test sample in a liquid discharge mode;

[0011] Detect the second sample flow rate at the valve;

[0012] When the second sample flow rate changes from a state less than the second flow rate threshold to a state greater than the second flow rate threshold, control the metering pump to discharge a preset volume of the detection sample in the liquid discharge mode, where the second flow rate threshold is greater than the first flow rate threshold;

[0013] Obtain a sample volume difference based on the detected volume, the current sample volume, and the preset volume;

[0014] Control the metering pump to aspirate the detection sample in the liquid aspiration mode according to the sample volume difference.

[0015] By adopting the above technical solution, after detecting that the first sample flow rate at the valve is too small, the metering pump will be controlled to discharge a preset volume of the detection sample in the liquid discharge mode, so that the bubbles in the detection sample are also excluded, thereby improving the accuracy of liquid aspiration of the flow cytometer and improving the accuracy of the detection results of the flow cytometer.

[0016] Optionally, obtain the preset volume, where the preset volume is greater than the volume of the liquid aspiration pipeline, and the liquid aspiration pipeline is connected to the second end of the valve;

[0017] Rotate the valve so that the first end of the valve is lower than the second end of the valve, and the second end of the valve is the inlet end of the detection sample;

[0018] Set the plunger propulsion distance of the metering pump according to the preset volume;

[0019] Control the plunger in the metering pump to move the plunger propulsion distance so that the preset volume of the detection sample passes through the valve.

[0020] By adopting the above technical solution, rotating the valve so that the first end of the valve is lower than the second end of the valve is beneficial to promoting the bubbles to leave the inside of the valve, facilitating the discharge of the bubbles, and further improving the accuracy of liquid aspiration of the flow cytometer and improving the accuracy of the detection results of the flow cytometer.

[0021] Optionally, when the preset volume is greater than the current sample volume, record the first timestamp;

[0022] Control the metering pump to aspirate the detection sample in the liquid aspiration mode and detect the third sample flow rate at the valve;

[0023] When the third sample flow rate is greater than the second flow rate threshold, record the second timestamp;

[0024] Obtain the bubble length based on the working power of the metering pump between the first timestamp and the second timestamp and the difference between the first timestamp and the second timestamp;

[0025] Obtain the bubble movement distance based on the distance from the valve to the adapter and the bubble length.

[0026] Control the metering pump to operate in the liquid suction mode according to the bubble movement distance, so that the bubble enters the flow cell through the adapter.

[0027] Close the valve and control the metering pump to operate in the liquid discharge mode for a first preset duration.

[0028] By adopting the above technical solution, obtain the bubble length according to the working power of the metering pump between the first timestamp and the second timestamp and the difference between the first timestamp and the second timestamp. And determine the bubble movement distance based on the bubble length, so that the bubble can enter the flow cell through the adapter and be discharged through the flow cell. Furthermore, improve the accuracy of liquid extraction of the flow cytometer to improve the accuracy of the detection result of the flow cytometer.

[0029] Optionally, when the bubble length is greater than the first preset length, control the metering pump to operate in the liquid suction mode according to the preset bubble movement distance.

[0030] Close the valve and control the metering pump to operate in the liquid discharge mode for a first preset duration.

[0031] Open the valve and obtain the remaining bubble length according to the preset bubble movement distance and the bubble length.

[0032] Control the metering pump to operate in the liquid suction mode according to the remaining bubble length.

[0033] Close the valve and control the metering pump to operate in the liquid discharge mode for a second preset duration.

[0034] By adopting the above technical solution, when the bubble is long, part of the bubble will be discharged through the flow cell first, and then the bubble will be completely discharged for the second time to prevent the bubble from remaining inside. It can improve the accuracy of liquid extraction of the flow cytometer to improve the accuracy of the detection result of the flow cytometer.

[0035] Optionally, when the first sample flow rate is greater than the third flow threshold and less than the first flow threshold, or the bubble length is less than the second preset length, control the metering pump to operate in the liquid suction mode for a third preset duration according to the target sample flow rate, and the second preset length is less than the first preset length.

[0036] Close the valve.

[0037] Control the metering pump to operate in the liquid discharge mode until the detection sample in the liquid storage pipeline is drained.

[0038] By adopting the above technical solution, when the bubbles in the detection sample are too small, it is necessary to aspirate the detection sample at the target sample flow rate to ensure that the bubbles will be completely sucked into the flow cell, and then the bubbles will be removed through the flow cell. This is beneficial to improving the accuracy of liquid extraction of the flow cytometer, so as to improve the accuracy of the detection results of the flow cytometer.

[0039] Optionally, obtain the liquid type of the detection sample;

[0040] Predict the bubble radius according to the bubble length;

[0041] Obtain the predicted sample flow rate according to the liquid type and the bubble radius;

[0042] Calculate the sum of the predicted sample flow rate and the flow rate error to obtain the target sample flow rate.

[0043] By adopting the above technical solution, a calculation method for the target sample flow rate is provided, making the target sample flow rate large enough to completely suck in the bubbles. Moreover, since the target sample flow rate is obtained by calculating the sum of the predicted sample flow rate and the flow rate error, it can be ensured that the target sample flow rate is large enough.

[0044] Optionally, if the second sample flow rate is not detected to be less than the second flow rate threshold within the preset duration threshold, obtain the preset sample flow rate and the preset liquid volume;

[0045] Control the metering pump to work in the liquid suction mode according to the preset sample flow rate until the volume of the detection sample in the liquid storage pipeline reaches the preset liquid volume;

[0046] Close the valve;

[0047] Control the metering pump to work in the liquid discharge mode until the detection sample in the liquid storage pipeline is completely drained.

[0048] By adopting the above technical solution, if the second sample flow rate is not detected to be less than the second flow rate threshold within the preset duration threshold, control the metering pump to work in the liquid discharge mode until the detection sample in the liquid storage pipeline is completely drained. This ensures that the bubbles in the detection sample can be completely removed, which is beneficial to improving the accuracy of liquid extraction of the flow cytometer, so as to improve the accuracy of the detection results of the flow cytometer.

[0049] In a second aspect, the present application provides a liquid extraction system for a flow cytometer, adopting the following technical solution:

[0050] A liquid extraction system for a flow cytometer, comprising:

[0051] An acquisition module, configured to acquire a detection volume, a first sample flow rate, a second sample flow rate, a preset volume, a first timestamp, a second timestamp, a first preset length, a second preset length, a first preset duration, a second preset duration, a third preset duration, and a preset duration threshold;

[0052] A memory, configured to store a program for the liquid extraction method of the flow cytometer;

[0053] A processor, the program in the memory can be loaded and executed by the processor and implement the liquid extraction method of the flow cytometer.

[0054] By adopting the above technical solution, after detecting that the first sample flow rate at the valve is too small, the metering pump will be controlled to discharge a preset volume of the detection sample in the liquid discharge mode, so that the bubbles in the detection sample are also excluded, thereby improving the accuracy of liquid extraction of the flow cytometer and improving the accuracy of the detection result of the flow cytometer.

[0055] In a third aspect, the present application provides a flow cytometer, adopting the following technical solution:

[0056] A flow cytometer, including a memory and a processor, and a computer program capable of being loaded and executed by the processor is stored on the memory and implements any one of the above methods.

[0057] In a fourth aspect, the present application provides a computer storage medium, which can store a corresponding program and has the characteristic of facilitating the improvement of the accuracy of liquid extraction of the flow cytometer, adopting the following technical solution:

[0058] A computer-readable storage medium stores a computer program capable of being loaded and executed by the processor and implementing any one of the liquid extraction methods of the flow cytometer.

[0059] In summary, the present application includes at least one of the following beneficial technical effects:

[0060] 1. After detecting that the first sample flow rate at the valve is too small, the metering pump will be controlled to discharge a preset volume of the detection sample in the liquid discharge mode, so that the bubbles in the detection sample are also excluded, thereby improving the accuracy of liquid extraction of the flow cytometer and improving the accuracy of the detection result of the flow cytometer;

[0061] 2. Rotating the valve so that the first end of the valve is lower than the second end of the valve is beneficial to promoting the bubbles to leave the inside of the valve, facilitating the discharge of the bubbles, and further improving the accuracy of liquid extraction of the flow cytometer and improving the accuracy of the detection result of the flow cytometer;

[0062] 3. Obtain the bubble length based on the working power of the metering pump between the first timestamp and the second timestamp and the difference between the first timestamp and the second timestamp. Determine the bubble movement distance based on the bubble length, so that the bubble can enter the flow cell through the adapter, and the bubble is discharged through the flow cell. Furthermore, improve the accuracy of liquid extraction of the flow cytometer to improve the accuracy of the detection result of the flow cytometer. Description of the Drawings

[0063] Figure 1 It is a schematic diagram of a liquid extraction device of a flow cytometer provided by an embodiment of the present application.

[0064] Figure 2 It is a schematic flowchart of a liquid extraction method of a flow cytometer provided by an embodiment of the present application.

[0065] Figure 3 It is a schematic flowchart of a method for removing air based on the liquid discharge method 1 provided by an embodiment of the present application.

[0066] Figure 4 It is a schematic flowchart of a method for removing air based on the liquid discharge method 2 provided by an embodiment of the present application.

[0067] Figure 5 It is a schematic flowchart of a method for removing air based on the liquid discharge method 3 provided by an embodiment of the present application.

[0068] Figure 6 It is a schematic flowchart of a method for removing air based on the liquid discharge method 4 provided by an embodiment of the present application.

[0069] Figure 7 It is a schematic flowchart of a method for calculating the sample flow rate provided by an embodiment of the present application.

[0070] Figure 8 It is a schematic flowchart of a method for removing air based on the liquid discharge method 5 provided by an embodiment of the present application.

[0071] Figure 9 It is a schematic structural diagram of a liquid extraction system of a flow cytometer provided by an embodiment of the present application. Detailed Embodiments

[0072] In order to make the objectives, technical solutions and advantages of the present application clearer, the following further describes the present application in detail with reference to the attached Figure 1 to the attached Figure 9 and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present application and are not used to limit the present application.

[0073] The present application discloses a schematic diagram of a liquid extraction device of a flow cytometer. Please refer to Figure 1, the flow cytometer includes a metering pump 11, a liquid storage pipeline 12, a flow cell 13 and a valve 14.

[0074] The metering pump 11 is used to collect a fixed volume of the test sample 161, and the test sample 161 is stored in the sample tube 16. Optionally, the test object of the test sample 161 is a cell suspension. Exemplarily, the metering pump 11 can adjust the aspiration volume and aspiration flow rate of the test sample 161. For example, in Figure 1 , the metering pump 11 can collect the test sample 161, and the test sample 161 in the sample tube 16 enters the liquid storage pipeline 12 through the valve 14. In addition, the metering pump 11 can also discharge the test sample 161 in the liquid storage pipeline 12. The liquid suction port of the metering pump 11 is connected to the first end of the liquid storage pipeline 12, the second end of the liquid storage pipeline 12 is connected to the first end of the adapter 15, the second end of the adapter 15 is connected to the first end of the valve 14, and the third end of the adapter 15 leads to the flow cell 13. In some embodiments, a one-way valve is provided at the third end of the adapter 15 or at the inlet of the flow cell 13, so that the test sample 161 can only move towards the flow cell 13.

[0075] The liquid storage pipeline 12 is used to temporarily store a certain amount of the test sample 161, and the volume of the test sample 161 in the liquid storage pipeline 12 can be controlled by the metering pump 11. For example, in Figure 1 , when the metering pump 11 aspirates the test sample 161, the test sample 161 enters the liquid storage pipeline 12 through the valve 14 and the adapter 15.

[0076] The flow cell 13 is used to detect the test sample 161. Exemplarily, after the test sample 161 enters the flow cell 13, the test sample 161 will be irradiated by a laser. After collecting the laser passing through the test sample 161, a light spot is collected in the detection area. Various characteristics of the cells in the test sample 161 are studied according to the light spot.

[0077] When the valve 14 is opened, control the metering pump 11 to work in the liquid suction mode, so that the test sample 161 can enter the liquid storage pipeline 12 through the valve 14. When the valve 14 is closed, control the metering pump 11 to work in the liquid discharge mode, so that the test sample 161 enters the flow cell 13.

[0078] The embodiment of the present application discloses a liquid extraction method for a flow cytometer. Referring to Figure 2 , the method includes:

[0079] Step S201: According to the detection volume, control the metering pump to work in the liquid suction mode, so that the test sample enters the liquid storage pipeline after passing through the valve. The liquid suction port of the metering pump is connected to the first end of the liquid storage pipeline, the second end of the liquid storage pipeline is connected to the first end of the adapter, the second end of the adapter is connected to the first end of the valve, and the third end of the adapter leads to the flow cell.

[0080] The detection volume is the sample volume required for the flow cytometer to test the sample to be detected. The detection volume can be set by the relevant personnel by entering it themselves.

[0081] In the liquid suction mode, the metering pump sucks the detection sample, and the detection sample enters the liquid storage pipeline after passing through the valve. Further, taking the metering pump as a plunger pump as an example, in the liquid suction mode, the plunger in the metering pump moves along the suction stroke, generating a negative pressure in the metering pump, and at the same time opening the valve, so that the detection sample enters the liquid storage pipeline along the valve.

[0082] On the other hand, the movement of the plunger in the metering pump can be adjusted to quantitatively suck the detection sample with the detection volume. Exemplarily, according to the detection volume, the plunger movement distance is determined in the preset volume-distance mapping relationship, and the volume-distance mapping relationship is used to record the mapping relationship between the detection volume and the plunger movement distance.

[0083] Step S202: Detect the first sample flow rate at the valve.

[0084] The first sample flow rate refers to the sample flow rate passing through the valve when the metering pump sucks the detection sample with the detection volume in the liquid suction mode.

[0085] A flow sensor is provided on the valve, and the first sample flow rate can be obtained through the flow sensor. For example, the flow sensor is a mechanical flow sensor.

[0086] Step S203: When the first sample flow rate is less than the first flow rate threshold, obtain the current sample volume in the liquid storage pipeline, and control the metering pump to discharge the detection sample in the liquid discharge mode.

[0087] The first flow rate threshold is a preset empirical value, and the relevant personnel can adjust the specific value of the first flow rate threshold according to actual needs. When the first sample flow rate is less than the first flow rate threshold, it indicates that the flow rate of the detection sample passing through the valve becomes smaller, and there is air mixed in the detection sample.

[0088] When the metering pump is a plunger pump, the current movement distance of the plunger can be obtained. The current sample volume is determined according to the current movement distance in the preset volume-distance mapping relationship.

[0089] In the liquid discharge mode, the metering pump discharges the detection sample, and the detection sample returns to the sample tube after passing through the valve. Further, taking the metering pump as a plunger pump as an example, in the liquid discharge mode, the plunger in the metering pump moves along the discharge stroke, generating a positive pressure in the metering pump, and at the same time closing the valve, so that the detection sample enters the flow cell.

[0090] Step S204: Detect the second sample flow rate at the valve.

[0091] When the second sample flow rate metering pump discharges the test sample in the liquid discharge mode, the sample flow rate through the valve.

[0092] The second sample flow rate can also be obtained through the flow sensor on the valve. Both the second sample flow rate and the first sample flow rate are the flow rates of the test sample passing through the valve, and the two only represent the flow rate values in different scenarios.

[0093] Step S205: When the second sample flow rate changes from a state less than the second flow rate threshold to a state greater than the second flow rate threshold, control the metering pump to discharge a preset volume of the test sample in the liquid discharge mode, and the second flow rate threshold is greater than the first flow rate threshold.

[0094] The second flow rate threshold is a preset empirical value, and relevant personnel can adjust the specific value of the second flow rate threshold according to actual needs. When the second sample flow rate changes from a state less than the second flow rate threshold to a state greater than the second flow rate threshold, it means that the air in the test sample completely enters the liquid suction pipeline between the valve and the sample tube through the valve. Further, the preset volume needs to be greater than the volume of the liquid suction pipeline to ensure that the air is completely discharged.

[0095] Step S206: Obtain the sample volume difference according to the test volume, the current sample volume, and the preset volume.

[0096] The sample volume difference refers to the volume of the test sample that still needs to be aspirated after discharging a preset volume of the test sample to make the volume of the aspirated test sample reach the test volume.

[0097] Exemplarily, let the test volume be V0, the current sample volume be V1, and the preset volume be V2, then the sample volume difference is ΔV = V0 - (V1 - V2).

[0098] Step S207: Control the metering pump to aspirate the test sample in the liquid suction mode according to the sample volume difference.

[0099] Exemplarily, determine the difference in the plunger movement distance according to the sample volume difference in the preset volume - distance mapping relationship, and control the plunger in the metering pump to move according to the difference in the plunger movement distance.

[0100] By adopting the above technical solution, after detecting that the first sample flow rate at the valve is too small, the metering pump will be controlled to discharge a preset volume of the test sample in the liquid discharge mode, so that the bubbles in the test sample are also excluded, thereby improving the accuracy of liquid aspiration of the flow cytometer and improving the accuracy of the detection results of the flow cytometer.

[0101] In the following embodiments, the air in the test sample can be discharged by discharging a certain volume of the test sample. Therefore, the embodiment of the present application discloses a de - air method based on the liquid discharge method one. Refer to Figure 3, the method includes:

[0102] Step S301: Obtain a preset volume, where the preset volume is greater than the volume of the liquid suction pipeline, and the liquid suction pipeline is connected to the second end of the valve.

[0103] Exemplarily, the preset volume is pre-stored in the storage space of the flow cytometer.

[0104] Exemplarily, the preset volume can be input by relevant personnel themselves.

[0105] Exemplarily, in response to an input operation of the liquid suction pipeline parameters, obtain the specification parameters of the liquid suction pipeline, where the specification parameters include the inner diameter and length of the liquid suction pipeline. Calculate the volume of the liquid suction pipeline according to the specification parameters. Calculate the sum of the volume of the liquid suction pipeline and the preset volume error to obtain the preset volume.

[0106] Step S302: Rotate the valve so that the first end of the valve is lower than the second end of the valve, and the second end of the valve is the inlet end of the test sample.

[0107] Exemplarily, please refer to Figure 1 , the valve can rotate around the center point of the valve in a vertical plane so that the first end of the valve is lower than the second end of the valve. At this time, the density of air is less than the density of the test sample, and the air will float into the liquid suction pipeline, which is beneficial to removing air.

[0108] Step S303: Set the plunger propulsion distance of the metering pump according to the preset volume.

[0109] Exemplarily, determine the plunger propulsion distance according to the preset volume in the preset volume-distance mapping relationship. Since it is necessary to make the bubbles completely leave the pipeline and the bubbles themselves will occupy a certain volume, after the plunger of the metering pump moves the above plunger propulsion distance, the volume of the test sample discharged by the metering pump needs to be at least greater than the preset volume. Therefore, let the preset volume be V1 and the plunger propulsion distance be L, then it is necessary to satisfy L*S > V1 + V2, where S represents the cross-sectional area inside the metering pump, and V2 is a preset fixed value, and V2 can be set by technicians themselves. For example, V2 takes 2 mL or 3 mL.

[0110] Step S304: Control the plunger in the metering pump to move the plunger propulsion distance so that the preset volume of the test sample passes through the valve.

[0111] Control the plunger in the metering pump to move the plunger propulsion distance to form a positive pressure inside the metering pump and discharge the test sample.

[0112] By adopting the above technical solution, rotating the valve so that the first end of the valve is lower than the second end of the valve is beneficial to promoting the bubbles to leave the inside of the valve, facilitating the discharge of the bubbles, thereby improving the accuracy of liquid extraction of the flow cytometer and improving the accuracy of the detection results of the flow cytometer.

[0113] In the following embodiments, when the preset volume is greater than the current sample volume, air cannot be removed by excluding the preset volume, so other methods need to be adopted to remove air. Therefore, the embodiments of the present application disclose a second air removal method based on the liquid drainage method. Refer to Figure 4 , the method includes:

[0114] Step S401: When the preset volume is greater than the current sample volume, record the first timestamp.

[0115] The first timestamp refers to the time information when it is detected that the second sample flow rate changes from a state less than the second flow rate threshold to a state greater than the second flow rate threshold. At this time, it is considered that the air in the detection sample just completely leaves the valve and enters the liquid suction pipeline.

[0116] Step S402: Control the metering pump to suck the detection sample in the liquid suction mode and detect the third sample flow rate at the valve.

[0117] The third sample flow rate refers to the sample flow rate passing through the valve when it is detected that the second sample flow rate changes from a state less than the second flow rate threshold to a state greater than the second flow rate threshold and the metering pump is in the liquid suction mode. The first sample flow rate, the second sample flow rate, and the third sample flow rate are all the flow rates of the detection sample passing through the valve, and the three only represent the flow rate values in different scenarios.

[0118] Step S403: When the third sample flow rate is greater than the second flow rate threshold, record the second timestamp.

[0119] When it is detected that the third sample flow rate is greater than the second flow rate threshold, it indicates that the air has passed through the valve and completely entered the pipeline corresponding to the first end of the valve.

[0120] Step S404: Obtain the bubble length according to the working power of the metering pump between the first timestamp and the second timestamp and the difference between the first timestamp and the second timestamp.

[0121] Exemplarily, the plunger movement speed of the metering pump is obtained based on the working power of the metering pump between the first timestamp and the second timestamp. The bubble length is obtained based on the product of the difference between the first timestamp and the second timestamp and the plunger movement speed. For example, the first actual time corresponding to the first timestamp and the second actual time corresponding to the second timestamp are obtained. The difference between the first actual time and the second actual time is calculated to obtain the timestamp difference. After determining the working power of the metering pump between the first timestamp and the second timestamp, the plunger movement speed is determined by the working power, where the working power here refers to the actual output power of the metering pump, and the corresponding relationship between the plunger movement speed and the working power can be obtained by technicians repeatedly measuring the plunger movement speed of the metering pump at different working powers to form a mapping table. Further, after obtaining the plunger movement speed and the timestamp difference, the product of the plunger movement speed and the timestamp difference is calculated to obtain the plunger movement distance. The volume of the detection sample discharged by the metering pump is obtained based on the plunger movement distance. On the other hand, since the movement of the bubble at this time depends on the volume of the detection sample inhaled by the working of the metering pump, and the first timestamp corresponds to the bubble completely leaving the valve, and the second timestamp corresponds to the bubble completely entering the pipeline corresponding to the first end of the valve, the bubble movement distance is the same as the bubble length during the period between the first timestamp and the second timestamp. Therefore, after obtaining the volume of the detection sample discharged by the metering pump, the inner diameter of the pipeline corresponding to the first end of the valve is obtained. The cross-sectional area of the pipeline is calculated based on the inner diameter of the aforementioned pipeline. The ratio of the detection sample volume to the cross-sectional area of the pipeline is calculated to obtain the bubble length.

[0122] In one embodiment, if a position sensor is installed on the metering pump, it is also possible to determine the first plunger position corresponding to the first timestamp and the second plunger position corresponding to the second timestamp. The plunger movement distance is obtained based on the distance between the first plunger position and the second plunger position. The volume of the detection sample discharged by the metering pump is obtained based on the plunger movement distance. Thus, the bubble length is obtained based on the detection sample. The corresponding relationship between the coefficient and the opening degree in the opening degree - coefficient mapping relationship table can be obtained by technicians repeatedly measuring the values of the conversion parameters at different valve opening degrees to form a mapping table. In some other embodiments, the conversion parameter can also be a fixed value and has nothing to do with the valve opening degree.

[0123] Further, the valve opening degree is obtained. In the preset opening degree - coefficient mapping relationship, the conversion parameter corresponding to the valve opening degree is determined. The product of the conversion parameter and the bubble length is calculated to correct the bubble length.

[0124] Step S405: Obtain the bubble movement distance based on the distance from the valve to the adapter and the bubble length.

[0125] Calculate the difference between the distance from the valve to the adapter and the bubble length to obtain the bubble movement distance.

[0126] In some other embodiments, the bubble moving distance includes a first bubble moving distance and a second bubble moving distance.

[0127] Step S406: According to the bubble moving distance, control the metering pump to work in the liquid suction mode so that the bubbles enter the flow cell through the adapter.

[0128] Exemplarily, calculate the pipe volume corresponding to the bubble moving distance. Determine the plunger moving distance corresponding to the pipe volume in the volume-distance mapping relationship. Control the movement of the plunger in the metering pump according to the plunger moving distance.

[0129] Further, to ensure that the bubbles can completely enter the flow cell through the adapter subsequently, it is necessary to control the power of the metering pump to be less than the preset power, so that the flow rate of the test sample is less than the preset flow rate threshold. If the flow rate of the test sample is greater than the flow rate threshold, the bubbles in the test sample may quickly pass through the adapter and directly enter the liquid storage pipeline, resulting in the need to control the flow rate of the test sample again in the liquid discharge mode of the metering pump to ensure that the bubbles can completely enter the flow cell, thereby completely removing the bubbles. Therefore, to avoid subsequent power control and improve efficiency, it is necessary to control the power of the metering pump to be less than the preset power to ensure that the bubbles can enter the pipeline corresponding to the third section of the adapter in this step.

[0130] In some other embodiments, obtain a first power and a first working duration according to the first bubble moving distance. Obtain a second power and a second working duration according to the second bubble moving distance, and the second power is less than the first power. Control the metering pump to work for the first working duration according to the first preset power. Control the metering pump to work for the second working duration according to the second preset power. After the metering pump works for the first working duration according to the first preset power, one end of the bubble can just enter the adapter. At this time, it is necessary to reduce the power of the metering pump to the second power to slow down the bubble moving speed and prevent the situation that the bubble moving speed is too fast and directly enters the liquid storage pipeline.

[0131] Step S407: Close the valve and control the metering pump to work for a first preset duration in the liquid discharge mode.

[0132] The first preset duration is a preset empirical value, and relevant personnel can adjust the first preset duration according to actual needs. Further, after the metering pump works for the first preset duration in the liquid discharge mode, the test sample in the liquid storage pipeline can be completely introduced into the flow cell and discharged from the flow cell to avoid interference of the bubbles with the operation of the flow cytometer.

[0133] By adopting the above technical solution, the bubble length is obtained according to the working power of the metering pump between the first timestamp and the second timestamp and the difference between the first timestamp and the second timestamp. And the bubble moving distance is determined by the bubble length, so that the bubble can enter the flow cell through the adapter, and the bubble is discharged through the flow cell. Furthermore, the accuracy of liquid extraction of the flow cytometer is improved, so as to improve the accuracy of the detection result of the flow cytometer.

[0134] In an actual scenario, the bubble length may be too long, resulting in the bubble being unable to smoothly enter the pipeline corresponding to the third section of the adapter. To solve the foregoing problem, an embodiment of the present application discloses a third air removal method based on the liquid discharge method. Refer to Figure 5 , the method includes:

[0135] Step S501: When the bubble length is greater than the first preset length, control the metering pump to work in the liquid suction mode according to the preset bubble moving distance.

[0136] The first preset length is a preset empirical value, and relevant personnel can adjust the specific value of the first preset length according to actual needs.

[0137] The preset bubble moving distance is a preset empirical value, and the preset bubble moving distance is not greater than the first preset distance. Relevant personnel can adjust the specific value of the preset bubble moving distance according to actual needs.

[0138] Exemplarily, the preset pipeline volume is calculated according to the preset bubble moving distance. Control the metering pump to work in the liquid suction mode so that the volume of the detection sample sucked by the metering pump reaches the preset pipeline volume. At this time, a part of the bubble is located in the pipeline corresponding to the third end of the adapter, and this part of the bubble is defined as the third-end bubble, while the other part is located in the pipeline corresponding to the first end of the adapter, and this part of the bubble is defined as the first-end bubble.

[0139] Step S502: Close the valve and control the metering pump to work in the liquid discharge mode for a first preset duration.

[0140] The first preset duration is a preset empirical value. After the metering pump works in the liquid discharge mode for the first preset duration, the third-end bubble can be discharged through the flow cell. Due to the structural design of the adapter, it cannot be guaranteed that the first-end bubble is completely discharged. Therefore, the remaining part of the first-end bubble needs to be discharged again through subsequent steps.

[0141] Step S503: Open the valve and obtain the remaining bubble length according to the preset bubble moving distance and the bubble length.

[0142] Exemplarily, calculate the difference between the bubble length and the preset bubble moving distance to obtain the remaining bubble length. The remaining bubble length refers to the length of the first-end bubble remaining in the pipeline of the adapter.

[0143] Step S504: Control the metering pump to work in the liquid suction mode according to the remaining bubble length.

[0144] Exemplarily, calculate the remaining pipeline volume according to the remaining bubble length. Control the metering pump to work in the liquid suction mode so that the volume of the detection sample sucked by the metering pump reaches the remaining pipeline volume.

[0145] Step S505: Close the valve and control the metering pump to work in the liquid discharge mode for a second preset duration.

[0146] The second preset duration is a preset empirical value. If the metering pump works in the liquid discharge mode for the second preset duration, the residual bubbles in the pipeline can be completely discharged through the flow cell.

[0147] By adopting the above technical solution, when the bubble is long, part of the bubble will be discharged through the flow cell first, and then the bubble will be completely discharged for the second time, preventing the bubble from remaining inside. The accuracy of liquid extraction by the flow cytometer can be improved, so as to improve the accuracy of the detection result of the flow cytometer.

[0148] In the following embodiments, when the volume of the bubble in the detection sample is too small, the fluidity of the bubble in the detection sample is worse, so that the metering pump needs to work with a greater power to discharge the bubble. Therefore, the embodiment of the present application discloses a method for removing air based on the liquid discharge method four. Refer to Figure 6 , this method includes:

[0149] Step S601: When the first sample flow rate is greater than the third flow rate threshold and less than the first flow rate threshold, or the bubble length is less than the second preset length, control the metering pump to work in the liquid suction mode for a third preset duration according to the target sample flow rate, and the second preset length is less than the first preset length.

[0150] When the first sample flow rate is greater than the third flow rate threshold and less than the first flow rate threshold, it indicates that the presence of bubbles in the detection sample can be determined by the change of the first sample flow rate, but the volume of the bubbles is relatively small. In addition, when the bubble length is less than the second preset length, it can also be clearly determined that the volume of the bubbles in the detection sample is small.

[0151] Step S602: Close the valve.

[0152] In the present application, when the bubble is too small, since the volume of the liquid storage pipeline is relatively limited, even if the detection sample has filled the liquid storage pipeline, it is difficult for the bubble to be discharged through the liquid suction pipeline only by the detection sample in the liquid storage pipeline. Therefore, in the embodiment of the present application, it is selected to let the bubble be discharged from the flow cell to increase the probability of bubble discharge.

[0153] Step S603: Control the metering pump to work in the liquid discharge mode until the detection sample in the liquid storage pipeline is completely discharged.

[0154] Exemplarily, control the metering pump to continuously work in the liquid discharge mode until all the detection samples in the liquid storage pipeline are discharged.

[0155] By adopting the above technical solution, when the bubbles in the detection sample are too small, it is necessary to aspirate the detection sample at the target sample flow rate to ensure that the bubbles will be completely sucked into the flow cell, and then the bubbles are removed through the flow cell. This is beneficial to improving the accuracy of liquid sampling of the flow cytometer, so as to improve the accuracy of the detection results of the flow cytometer.

[0156] In the following embodiments, it is necessary to limit the target sample flow rate so that the target sample flow rate is large enough to discharge the bubbles with a small volume. Therefore, the embodiments of the present application disclose a calculation method for the sample flow rate. Refer to Figure 7 , and the method includes:

[0157] Step S701: Obtain the liquid type of the detection sample.

[0158] The liquid type can be obtained by input from relevant personnel. In some other embodiments, the liquid type can be uniformly set to a preset type.

[0159] Step S702: Predict the bubble radius according to the bubble length.

[0160] Exemplarily, regard the bubble length as the diameter of the bubble, so as to obtain the bubble radius.

[0161] Step S703: Obtain the predicted sample flow rate according to the liquid type and the bubble radius.

[0162] Exemplarily, the predicted sample flow rate is ; where r represents the bubble radius, is the density of the detection sample, is the air density, g is the acceleration due to gravity, is the liquid viscosity of the detection sample. In some embodiments, since is much larger than , therefore, the value of can be simplified to to reduce the calculation amount. Further, for simplifying the operation, the density and liquid viscosity of the detection sample can adopt preset values.

[0163] Step S704: Calculate the sum of the predicted sample flow rate and the flow rate error to obtain the target sample flow rate.

[0164] The flow rate error is a preset empirical value, and relevant personnel adjust the specific value of the flow rate error according to actual needs.

[0165] In some other embodiments, it is also required that the target sample flow rate is less than the maximum sample flow rate, where the maximum sample flow rate is , where is the liquid surface tension of the detection sample, r is the bubble radius, is the density of the detection sample. Further, for simplifying the calculation, the liquid surface tension and density of the detection sample can adopt preset values.

[0166] By adopting the above technical solution, a calculation method for the target sample flow rate is provided, making the target sample flow rate large enough to completely suck in the bubbles. Moreover, the target sample flow rate is obtained by calculating the sum of the predicted sample flow rate and the flow rate error, so the target sample flow rate can be ensured to be large enough.

[0167] In the following embodiments, when discharging air, the bubbles may directly enter the pipeline corresponding to the third end of the adapter, resulting in that after the metering pump works in the liquid discharge mode for a period of time, the second sample flow rate still cannot be detected to be greater than the second flow rate threshold. At this time, it is necessary to discharge the bubbles in the detection sample through the flow cell. Therefore, the embodiments of the present application disclose a method for removing air based on the liquid discharge method five. Refer to Figure 8 , the method includes:

[0168] Step S801: If the second sample flow rate is not detected to be less than the second flow rate threshold within the preset duration threshold, obtain the preset sample flow rate and the preset liquid volume.

[0169] The preset duration threshold can be a preset empirical value or related to the current sample volume in the liquid storage pipeline. Exemplarily, calculate the product of the current sample volume and the preset ratio to obtain the detected discharge volume. According to the detected discharge volume and the working power of the metering pump, obtain the time required for the metering pump to discharge the detection sample of the detected discharge volume, and use this time as the preset time threshold.

[0170] The preset sample flow rate and the preset liquid volume are preset empirical values, and relevant personnel can adjust the specific values of the preset sample flow rate and the preset liquid volume according to actual needs. For example, the preset liquid volume is the maximum volume of the liquid storage pipeline. In some other embodiments, the preset sample flow rate and the preset liquid volume can also be obtained by relevant personnel input.

[0171] Step S802: Control the metering pump to work in the liquid suction mode according to the preset sample flow rate until the volume of the detection sample in the liquid storage pipeline reaches the preset liquid volume.

[0172] Exemplarily, determine the plunger movement speed of the metering pump according to the preset sample flow rate. Determine the working power of the metering pump according to the plunger movement speed. Control the metering pump to work in the washing liquid mode according to the foregoing working power.

[0173] Step S803: Close the valve.

[0174] Since air bubbles need to pass through the flow cell and be discharged in subsequent steps, the valve needs to be closed first in this step to prevent some of the test sample from returning to the sample tube through the valve, resulting in a poor effect of exhausting air bubbles.

[0175] Step S804: Control the metering pump to work in the liquid discharge mode until the test sample in the liquid storage pipeline is completely discharged.

[0176] Exemplarily, obtain the position information of the plunger when acquiring the test sample in the liquid storage pipeline. Adjust the position of the plunger to the position corresponding to the aforementioned position information to achieve the effect of completely discharging the test sample in the liquid storage pipeline.

[0177] By adopting the above technical solution, if the second sample flow rate is not detected to be less than the second flow rate threshold within the preset duration threshold, control the metering pump to work in the liquid discharge mode until the test sample in the liquid storage pipeline is completely discharged. Ensure that the air bubbles in the test sample can be completely removed, which is beneficial to improving the accuracy of liquid extraction by the flow cytometer, so as to improve the accuracy of the detection results of the flow cytometer.

[0178] Based on the same inventive concept, an embodiment of the present application provides a liquid extraction system for a flow cytometer. Please refer to Figure 9 , and this system includes:

[0179] An acquisition module 901, configured to acquire a detection volume, a first sample flow rate, a second sample flow rate, a preset volume, a first timestamp, a second timestamp, a first preset length, a second preset length, a first preset duration, a second preset duration, a third preset duration, and a preset duration threshold;

[0180] A memory 902, configured to store the program of the above-mentioned liquid extraction method of the flow cytometer;

[0181] A processor 903, the program in the memory can be loaded and executed by the processor and implement the above-mentioned liquid extraction method of the flow cytometer.

[0182] By adopting the above technical solution, after detecting that the first sample flow rate at the valve is too small, the metering pump will be controlled to discharge a preset volume of the test sample in the liquid discharge mode, so that the air bubbles in the test sample are also removed, thereby improving the accuracy of liquid extraction by the flow cytometer, so as to improve the accuracy of the detection results of the flow cytometer.

[0183] Those skilled in the art can clearly understand that, for the convenience and brevity of description, only the division of the above-mentioned functional modules is used as an example. In actual applications, the above-mentioned functions can be allocated to different functional modules as needed, that is, the internal structure of the device is divided into different functional modules to complete all or part of the functions described above. The specific working processes of the above-described systems, devices, and units can refer to the corresponding processes in the foregoing method embodiments and will not be elaborated herein.

[0184] The embodiment of the present application provides a computer-readable storage medium storing a computer program that can be loaded and executed by a processor to perform the liquid extraction method of a flow cytometer.

[0185] Computer storage media include, for example, various media that can store program codes such as USB flash drives, mobile hard disks, read-only memories (ROM), random access memories (RAM), magnetic disks, or optical discs.

[0186] Based on the same inventive concept, the embodiment of the present application provides an intelligent terminal, including a memory and a processor, and a computer program that can be loaded and executed by the processor to perform the liquid extraction method of a flow cytometer is stored on the memory.

[0187] Those skilled in the art can clearly understand that, for the convenience and brevity of description, only the division of the above-mentioned functional modules is used as an example. In actual applications, the above-mentioned functions can be allocated to different functional modules as needed, that is, the internal structure of the device is divided into different functional modules to complete all or part of the functions described above. The specific working processes of the above-described systems, devices, and units can refer to the corresponding processes in the foregoing method embodiments and will not be elaborated herein.

[0188] The above are all the preferred embodiments of the present application. Without limiting the protection scope of the present application accordingly, any feature disclosed in this specification (including the abstract and drawings), unless specifically described, can be replaced by other equivalent or features with similar purposes. That is, unless specifically described, each feature is only an example of a series of equivalent or similar features.

Claims

1. A method for collecting liquid for a flow cytometer, characterized in that: The method comprises: According to the detection volume, the quantitative pump is controlled to work in the liquid aspiration mode, so that the detection sample enters the liquid storage pipeline after passing through the valve, the liquid aspiration port of the quantitative pump is connected to the first end of the liquid storage pipeline, the second end of the liquid storage pipeline is connected to the first end of the adapter, the second end of the adapter is connected to the first end of the valve, and the third end of the adapter leads to the flow chamber; detecting a first sample flow at the valve; When the first sample flow rate is less than a first flow rate threshold, obtaining a current sample volume in the liquid storage pipeline, and controlling the quantitative pump to discharge the test sample in a liquid discharge mode; detecting a second sample flow at the valve; When the second sample flow rate changes from a state less than a second flow rate threshold to a state greater than the second flow rate threshold, controlling the metering pump to discharge a preset volume of the test sample in the liquid discharge mode, and the second flow rate threshold is greater than the first flow rate threshold; Obtaining a sample volume difference according to the detection volume, the current sample volume and the preset volume; According to the sample volume difference, controlling the quantitative pump to absorb the test sample in the liquid aspiration mode; The controlling the metering pump to discharge a preset volume of the test sample in the liquid discharge mode comprises: Obtaining the preset volume, wherein the preset volume is greater than the volume of a liquid suction pipeline, wherein the liquid suction pipeline is connected to the second end of the valve; Rotate the valve so that the first end of the valve is lower than the second end of the valve, and the second end of the valve is the inlet end of the test sample; Setting the plunger advancement distance of the metering pump according to the preset volume; Controlling the plunger in the metering pump to move the plunger advancement distance so that the detection sample of the preset volume passes through the valve; After obtaining the preset volume, the method further includes: When the preset volume is greater than the current sample volume, recording a first timestamp; Controlling the quantitative pump to absorb the test sample in the liquid aspiration mode, and detecting a third sample flow at the valve; When the third sample flow rate is greater than the second flow rate threshold, recording a second timestamp; Obtaining the bubble length according to the working power of the quantitative pump between the first timestamp and the second timestamp and the difference between the first timestamp and the second timestamp; Obtaining a bubble movement distance according to the distance from the valve to the adapter and the bubble length; According to the moving distance of the bubble, the quantitative pump is controlled to operate in the liquid aspiration mode, so that the bubble enters the flow chamber through the adapter; The valve is closed, and the metering pump is controlled to operate in the liquid discharge mode for a first preset time period.

2. The method for collecting liquid for a flow cytometer according to claim 1, characterized in that: The method further comprises: When the bubble length is greater than a first preset length, the metering pump is controlled to operate in the liquid suction mode according to a preset bubble movement distance; Close the valve, and control the metering pump to operate in the liquid discharge mode for a first preset time period; Opening the valve, and obtaining the remaining bubble length according to the preset bubble movement distance and the bubble length; According to the remaining bubble length, controlling the metering pump to operate in the liquid suction mode; The valve is closed, and the metering pump is controlled to operate in the liquid discharge mode for a second preset time period.

3. The method for collecting liquid for a flow cytometer according to claim 1, characterized in that: Said include: When the first sample flow rate is greater than a third flow rate threshold and less than the first flow rate threshold, or the bubble length is less than a second preset length, the quantitative pump is controlled to operate in the aspiration mode for a third preset time according to the target sample flow rate, and the second preset length is less than the first preset length; closing the valve; The metering pump is controlled to operate in the liquid discharge mode until the test sample in the liquid storage pipeline is completely discharged.

4. The method for collecting liquid for a flow cytometer according to claim 3, characterized in that: The method further comprises: Obtaining the liquid type of the test sample; predicting a bubble radius based on the bubble length; According to the liquid type and the bubble radius, a predicted sample flow rate is obtained; The sum of the predicted sample flow rate and the flow rate error is calculated to obtain the target sample flow rate.

5. The method for collecting liquid for a flow cytometer according to claim 1, characterized in that: After detecting the second sample flow at the valve, the method further includes: If it is not detected within the preset time threshold that the second sample flow rate is greater than the second flow threshold, obtaining a preset sample flow rate and a preset liquid volume; Controlling the quantitative pump to operate in the liquid aspiration mode according to the preset sample flow rate until the volume of the test sample in the liquid storage pipeline reaches the preset liquid volume; closing the valve; The metering pump is controlled to operate in the liquid discharge mode until the test sample in the liquid storage pipeline is completely discharged.

6. A liquid collection system for a flow cytometer, characterized in that: The system is used to perform the liquid collection method of the flow cytometer according to any one of claims 1 to 5, and the system comprises: An acquisition module, used to acquire a detection volume, a first sample flow rate, a second sample flow rate, a preset volume, a first timestamp, a second timestamp, a first preset length, a second preset length, a first preset duration, a second preset duration, a third preset duration, and a preset duration threshold; A memory, used to store a program of a liquid collection method of the flow cytometer; The program in the memory can be loaded and executed by the processor to implement the liquid collection method of the flow cytometer.

7. A flow cytometer, characterized in that: The method comprises a memory and a processor, wherein the memory stores a computer program that can be loaded by the processor and executes the method according to any one of claims 1 to 5.

8. A computer-readable storage medium, characterized in that: A computer program is stored which can be loaded by a processor and execute the method according to any one of claims 1 to 5.

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