Flow cytometer, liquid taking method and system thereof and storage medium

Abstract

The invention relates to a flow cytometer, a liquid taking method and system thereof and a storage medium, and relates to the technical field of biological detection.The method comprises the steps that according to the detection volume, a constant delivery pump is controlled to work in a liquid suction mode, and a detection sample enters a liquid storage pipeline after passing through a valve; detecting a first sample flow at the valve; under the condition that the first sample flow is smaller than a first flow threshold value, the current sample volume in a liquid storage pipeline is obtained, and a constant displacement pump is controlled to discharge the detection sample in a liquid discharging mode; detecting a second sample flow at the valve; when the second sample flow is changed from a state smaller than a second flow threshold value to a state larger than the second flow threshold value, the constant displacement pump is controlled to discharge the detection sample with the preset volume in the liquid discharging mode; obtaining a sample volume difference value according to the detection volume, the current sample volume and a preset volume; and according to the sample volume difference value, controlling the metering pump to suck the detection sample in a liquid suction mode. The flow cytometer has the effect of improving the liquid taking accuracy of the flow cytometer.

Description

Technical Field

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

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

[0003] When the related technology uses a flow cytometer, it is necessary to load the test sample into a sample tube and ensure that the sample tube is not blocked, and then adjust the sample flow so that the individual samples in the single test sample are analyzed by the laser beam in turn to obtain the test results.

[0004] Regarding the above-mentioned related technologies, when the flow cytometer aspirates the test sample, in order to ensure the accuracy of the test result, the test sample needs to be quantitatively aspirated. However, in the process of aspirating the test sample, the flow cytometer cannot determine whether there is still test sample in the sample tube, resulting in the intake of some air, affecting the accuracy of the test result. Summary of the invention

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

[0006] In a first aspect, the present application provides a method for collecting liquid for a flow cytometer, which adopts the following technical solution: A method for collecting liquid for a flow cytometer, comprising: 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, the quantitative pump is controlled to aspirate the test sample in the liquid aspiration mode.

[0007] By adopting the above technical solution, after detecting that the flow rate of the first sample at the valve is too small, the quantitative pump will be controlled to discharge a preset volume of the test sample in the discharge mode, so that the bubbles in the test sample are also excluded, thereby improving the accuracy of the flow cytometer liquid collection and improving the accuracy of the flow cytometer detection results.

[0008] Optionally, the preset volume is obtained, the preset volume is greater than the volume of a liquid suction pipeline, and 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; The plunger in the metering pump is controlled to move the plunger advancement distance so that the detection sample of the preset volume passes through the valve.

[0009] 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, it is helpful to promote the bubbles to leave from the inside of the valve, facilitate the discharge of bubbles, and thus improve the accuracy of liquid collection by the flow cytometer, so as to improve the accuracy of the flow cytometer detection results.

[0010] Optionally, when the preset volume is greater than the current sample volume, a first timestamp is recorded; 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.

[0011] By adopting the above technical solution, the bubble length is obtained 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. The bubble moving distance is determined by the bubble length, so that the bubble can enter the flow chamber through the adapter and be discharged through the flow chamber. In this way, the accuracy of the flow cytometer liquid collection is improved, so as to improve the accuracy of the flow cytometer detection results.

[0012] Optionally, 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.

[0013] By adopting the above technical solution, when the bubbles are long, some bubbles will be discharged through the flow chamber first, and then the bubbles will be completely discharged for the second time to prevent the bubbles from remaining inside. The accuracy of the flow cytometer liquid collection can be improved, so as to improve the accuracy of the flow cytometer detection results.

[0014] Optionally, 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.

[0015] By adopting the above technical solution, when the bubbles in the test sample are too small, the test sample needs to be sucked according to the target sample flow rate to ensure that the bubbles are completely sucked into the flow chamber, and then the bubbles are removed by the flow chamber. This is conducive to improving the accuracy of the flow cytometer liquid collection, so as to improve the accuracy of the flow cytometer test results.

[0016] Optionally, 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.

[0017] By adopting the above technical solution, a method for calculating the target sample flow rate is provided, so that the target sample flow rate is large enough to completely absorb 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 guaranteed to be large enough.

[0018] Optionally, if it is not detected that the second sample flow rate is less than the second flow rate threshold within a preset time threshold, a preset sample flow rate and a preset liquid volume are obtained; 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.

[0019] 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 time threshold, the quantitative pump is controlled to work in the liquid discharge mode until the test sample in the liquid storage pipeline is completely discharged. It is ensured that the bubbles in the test sample can be completely removed, which is conducive to improving the accuracy of the flow cytometer liquid collection, so as to improve the accuracy of the flow cytometer test results.

[0020] In a second aspect, the present application provides a liquid collection system for a flow cytometer, which adopts the following technical solution: A liquid collection system for a flow cytometer, comprising: 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.

[0021] By adopting the above technical solution, after detecting that the flow rate of the first sample at the valve is too small, the quantitative pump will be controlled to discharge a preset volume of the test sample in the discharge mode, so that the bubbles in the test sample are also excluded, thereby improving the accuracy of the flow cytometer liquid collection and improving the accuracy of the flow cytometer detection results.

[0022] In a third aspect, the present application provides a flow cytometer, which adopts the following technical solution: A flow cytometer comprises a memory and a processor, wherein the memory stores a computer program which can be loaded by the processor and execute any one of the above-mentioned methods.

[0023] In a fourth aspect, the present application provides a computer storage medium capable of storing corresponding programs, which has the characteristic of facilitating the improvement of the accuracy of liquid collection by a flow cytometer, and adopts the following technical solution: A computer-readable storage medium stores a computer program that can be loaded by a processor and execute any of the above-mentioned liquid collection methods for a flow cytometer.

[0024] In summary, the present application includes at least one of the following beneficial technical effects: 1. After detecting that the flow rate of the first sample at the valve is too small, the quantitative pump will be controlled to discharge the preset volume of the test sample in the discharge mode, so that the bubbles in the test sample are also removed, thereby improving the accuracy of the flow cytometer liquid collection and improving the accuracy of the flow cytometer test results; 2. Turn the valve so that the first end of the valve is lower than the second end of the valve, which is conducive to promoting the bubbles to leave the inside of the valve, making it easier to discharge the bubbles, thereby improving the accuracy of the flow cytometer liquid collection and improving the accuracy of the flow cytometer test results; 3. The length of the bubble is obtained 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. The moving distance of the bubble is determined by the length of the bubble, so that the bubble can enter the flow chamber through the adapter and be discharged through the flow chamber. This improves the accuracy of the flow cytometer liquid collection and the accuracy of the flow cytometer detection results. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 It is a schematic diagram of a liquid collection device for a flow cytometer provided in an embodiment of the present application.

[0026] Figure 2 It is a flow chart of a method for collecting liquid for a flow cytometer provided in an embodiment of the present application.

[0027] Figure 3 It is a flow chart of a first air removal method based on liquid drainage provided in an embodiment of the present application.

[0028] Figure 4 It is a flow chart of a second de-airing method based on liquid drainage provided in an embodiment of the present application.

[0029] Figure 5 It is a flow chart of a third de-airing method based on liquid drainage provided in an embodiment of the present application.

[0030] Figure 6It is a flow chart of a fourth de-airing method based on liquid drainage provided in an embodiment of the present application.

[0031] Figure 7 It is a flow chart of a method for calculating a sample flow rate provided in an embodiment of the present application.

[0032] Figure 8 It is a flow chart of a fifth de-airing method based on liquid drainage provided in an embodiment of the present application.

[0033] Fig. 9 It is a structural schematic diagram of a liquid collection system of a flow cytometer provided in an embodiment of the present application. DETAILED DESCRIPTION

[0034] In order to make the purpose, technical solutions and advantages of this application more clear, the following Figure 1 To Attachment Fig. 9 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.

[0035] This application discloses a schematic diagram of a liquid collection device for a flow cytometer. Figure 1 The flow cytometer includes a quantitative pump 11, a liquid storage pipeline 12, a flow chamber 13 and a valve 14.

[0036] The quantitative 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 quantitative pump 11 can adjust the suction volume and suction flow rate of the test sample 161. For example, Figure 1 In the embodiment, the quantitative 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 quantitative pump 11 can also discharge the test sample 161 in the liquid storage pipeline 12. The liquid suction port of the quantitative 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 chamber 13. In some embodiments, a one-way valve is provided at the third end of the adapter 15 or the entrance of the flow chamber 13, so that the test sample 161 can only move toward the flow chamber 13.

[0037] The liquid storage pipeline 12 is used to temporarily store a certain amount of test sample 161. The volume of the test sample 161 in the liquid storage pipeline 12 can be controlled by the quantitative pump 11. Figure 1 When the quantitative pump 11 draws the test sample 161 , the test sample 161 passes through the valve 14 and the adapter 15 and enters the liquid storage pipeline 12 .

[0038] The flow chamber 13 is used to detect the test sample 161. Exemplarily, after the test sample 161 enters the flow chamber 13, the test sample 161 is irradiated by laser. After the laser passes through the test sample 161, a light spot is collected in the detection area. Various characteristics of cells in the test sample 161 are studied based on the light spot.

[0039] When the valve 14 is open, the quantitative pump 11 is controlled to work in the 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, the quantitative pump 11 is controlled to work in the discharge mode, so that the test sample 161 enters the flow chamber 13.

[0040] The present application embodiment discloses a method for collecting liquid for a flow cytometer. Figure 2 , the method comprising: Step S201: According to the detection volume, control the quantitative pump to operate 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.

[0041] The detection volume is the volume of sample that the flow cytometer needs to test the sample. The detection volume can be set by the relevant personnel.

[0042] In the suction mode, the metering pump will absorb the test sample, so that the test 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 suction mode, the plunger in the metering pump moves along the suction stroke, generating negative pressure in the metering pump, and at the same time opening the valve, so that the test sample enters the liquid storage pipeline along the valve.

[0043] On the other hand, the movement of the plunger in the quantitative pump can be adjusted to quantitatively absorb the test sample of the test volume. Exemplarily, the plunger movement distance is determined in a preset volume-distance mapping relationship according to the test volume, and the volume-distance mapping relationship is used to record the mapping relationship between the test volume and the plunger movement distance.

[0044] Step S202: Detect a first sample flow at a valve.

[0045] The first sample flow rate specifies the sample flow rate through the valve when the volumetric pump aspirates the test sample of the test volume in the aspiration mode.

[0046] 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.

[0047] Step S203: when the first sample flow rate is less than the first flow rate threshold, the current sample volume in the liquid storage pipeline is obtained, and the quantitative pump is controlled to discharge the test sample in the liquid discharge mode.

[0048] The first flow rate threshold is a preset empirical value, and 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 means that the flow rate of the test sample passing through the valve becomes smaller, and air is mixed in the test sample.

[0049] When the metering pump is a plunger pump, the current moving distance of the plunger can be obtained, and the current sample volume is determined from a volume-distance mapping relationship preset according to the current moving distance.

[0050] In the discharge mode, the metering pump will discharge the test sample, so that the test 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 discharge mode, the plunger in the metering pump moves along the extrusion stroke, generating positive pressure in the metering pump, and closing the valve at the same time, so that the test sample enters the flow chamber.

[0051] Step S204: Detect the second sample flow at the valve.

[0052] The second sample flow rate specifies the sample flow rate through the valve when the pump is discharging the test sample in the dispensing mode.

[0053] The second sample flow rate can also be obtained through the flow sensor on the valve. The second sample flow rate and the first sample flow rate are both the flow rates of the detection samples passing through the valve, and the two only represent the flow values ​​in different scenarios.

[0054] Step S205: When the second sample flow rate changes from less than the second flow rate threshold to greater than the second flow rate threshold, controlling the quantitative pump to discharge a preset volume of the test sample in the discharge mode, and the second flow rate threshold is greater than the first flow rate threshold.

[0055] The second flow threshold is a preset empirical value, and relevant personnel can adjust the specific value of the second flow threshold according to actual needs. When the second sample flow changes from a state less than the second flow threshold to a state greater than the second flow threshold, it means that the air in the test sample has completely entered the liquid aspiration line between the valve and the sample tube through the valve. Furthermore, the preset volume needs to be greater than the volume of the liquid aspiration line to ensure that the air is completely discharged.

[0056] Step S206: Obtaining a sample volume difference according to the detection volume, the current sample volume and the preset volume.

[0057] The sample volume difference refers to the volume of the test sample that needs to be drawn in order to make the volume of the drawn test sample reach the test volume after excluding the preset volume of the test sample.

[0058] Exemplarily, assuming that the detection volume is V0, the current sample volume is V1, and the preset volume is V2, the sample volume difference is ΔV=V0-(V1-V2).

[0059] Step S207: According to the sample volume difference, control the quantitative pump to aspirate the test sample in the liquid aspiration mode.

[0060] Exemplarily, a plunger movement distance difference is determined in a preset volume-distance mapping relationship according to the sample volume difference, and a plunger in a metering pump is controlled to move according to the plunger movement distance difference.

[0061] By adopting the above technical solution, after detecting that the flow rate of the first sample at the valve is too small, the quantitative pump will be controlled to discharge a preset volume of the test sample in the discharge mode, so that the bubbles in the test sample are also excluded, thereby improving the accuracy of the flow cytometer liquid collection and improving the accuracy of the flow cytometer detection results.

[0062] In the following embodiments, the air in the test sample can be discharged by discharging a certain volume of the test sample. Therefore, the present application embodiment discloses a de-airing method based on the liquid discharge method. Figure 3 , the method comprising: Step S301: obtaining a preset volume, the preset volume being greater than the volume of a liquid suction pipeline connected to the second end of the valve.

[0063] Exemplarily, the preset volume is pre-stored in a storage space of the flow cytometer.

[0064] Exemplarily, the preset volume may be input by relevant personnel.

[0065] Exemplarily, in response to the input operation of the pipette pipeline parameter, the specification parameters of the pipette pipeline are obtained, and the specification parameters include the inner diameter and length of the pipette pipeline. The volume of the pipette pipeline is calculated according to the specification parameters. The volume of the pipette pipeline and the sum of the preset volume error are calculated to obtain the preset volume.

[0066] 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.

[0067] For example, please refer to Figure 1 , the valve can be rotated around the center point of the valve on 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 lower than the density of the test sample, and the air will float into the liquid suction line, which is conducive to removing air.

[0068] Step S303: setting the plunger advancement distance of the metering pump according to the preset volume.

[0069] Exemplarily, the plunger advancement distance is determined in a preset volume-distance mapping relationship according to a preset volume. Since the bubbles need to completely leave the pipe and the bubbles themselves will occupy a certain volume, the volume of the test sample discharged by the metering pump needs to be at least greater than the preset volume after the plunger of the metering pump moves the above plunger advancement distance. Therefore, assuming that the preset volume is V1 and the plunger advancement distance is L, 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. V2 can be set by the technician, for example, V2 is 2mL or 3mL.

[0070] Step S304: Control 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.

[0071] Control the plunger movement in the dosing pump to advance the plunger distance, so that positive pressure is formed inside the dosing pump to discharge the test sample. 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, it is helpful to promote the bubbles to leave from the inside of the valve, facilitate the discharge of bubbles, and thus improve the accuracy of liquid collection by the flow cytometer, so as to improve the accuracy of the flow cytometer detection results.

[0072] In the following embodiments, when the preset volume is larger than the current sample volume, the air cannot be removed by removing the preset volume, so other methods are needed to remove the air. Therefore, the present application embodiment discloses a second air removal method based on the liquid discharge method. Figure 4 , the method comprising: Step S401: When the preset volume is greater than the current sample volume, a first timestamp is recorded.

[0073] The first timestamp refers to the time information when the second sample flow rate is detected to change 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 test sample has just completely left the valve and entered the liquid suction pipeline.

[0074] Step S402: Control the quantitative pump to absorb the test sample in the liquid aspiration mode, and detect the flow rate of the third sample at the valve.

[0075] The third sample flow rate refers to the sample flow rate through the valve when the second sample flow rate is detected to change from less than the second flow rate threshold to greater than the second flow rate threshold, and the metering pump is in the 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 test samples passing through the valve, and the three only represent the flow values ​​in different scenarios.

[0076] Step S403: when the third sample flow rate is greater than the second flow rate threshold, record a second timestamp.

[0077] 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.

[0078] Step S404: Obtain 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.

[0079] Exemplarily, according to the working power of the quantitative pump between the first timestamp and the second timestamp, the plunger moving speed of the quantitative pump is obtained. According to the product of the difference between the first timestamp and the second timestamp and the plunger moving speed, the bubble length is obtained. For example, the first real time corresponding to the first timestamp and the second real time corresponding to the second timestamp are obtained. The difference between the first real time and the second real time is calculated to obtain the timestamp difference. After determining the working power of the quantitative pump between the first timestamp and the second timestamp, the plunger moving speed is determined by the working power, wherein the working power here specifies the actual output power of the quantitative pump, and the corresponding relationship between the plunger moving speed and the working power can be obtained by technicians repeatedly measuring the plunger moving speed of the quantitative pump under different working powers to form a mapping table. Further, after obtaining the plunger moving speed and the timestamp difference, the product of the plunger moving speed and the timestamp difference is calculated to obtain the plunger moving distance. The detection sample volume discharged by the quantitative pump is obtained according to the plunger moving distance. On the other hand, because the movement of the bubble at this time depends on the volume of the test sample sucked in by 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 pipe corresponding to the first end of the valve, the bubble movement distance in the period between the first timestamp and the second timestamp is the same as the bubble length. Therefore, after obtaining the volume of the test sample discharged by the metering pump, the inner diameter of the pipe corresponding to the first end of the valve is obtained. The cross-sectional area of ​​the pipe is calculated based on the inner diameter of the aforementioned pipe. The ratio of the test sample volume to the pipe cross-sectional area is calculated to obtain the bubble length.

[0080] 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. According to the distance between the first plunger position and the second plunger position, the plunger movement distance is obtained. According to the plunger movement distance, the volume of the test sample discharged by the metering pump is obtained. Thus, the bubble length is obtained according to the test sample. The corresponding relationship between the coefficient and the opening in the opening-coefficient mapping relationship table can be obtained by technicians repeatedly measuring the values ​​of the conversion parameters under different valve openings to form a mapping table. In some other embodiments, the conversion parameter can also be a fixed value, which is independent of the valve opening.

[0081] Furthermore, the valve opening is obtained, a conversion parameter corresponding to the valve opening is determined in a preset opening-coefficient mapping relationship, and the product of the conversion parameter and the bubble length is calculated to achieve correction of the bubble length.

[0082] Step S405: Obtain the bubble movement distance according to the distance from the valve to the adapter and the bubble length.

[0083] Calculate the difference between the distance from the valve to the adapter and the length of the bubble to get the bubble travel distance.

[0084] In some other embodiments, the bubble movement distance includes a first bubble movement distance and a second bubble movement distance.

[0085] Step S406: 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.

[0086] Exemplarily, the pipe volume corresponding to the bubble movement distance is calculated, the plunger movement distance corresponding to the pipe volume is determined in the volume-distance mapping relationship, and the plunger movement in the metering pump is controlled according to the plunger movement distance.

[0087] Furthermore, in order to ensure that the bubbles can completely enter the flow chamber through the adapter later, it is necessary to control the power of the quantitative 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 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 quantitative pump needing to control the test sample flow rate again in the discharge mode to ensure that the bubbles can completely enter the flow chamber, thereby completely removing the bubbles. Therefore, in order to avoid subsequent power control and improve efficiency, it is necessary to control the power of the quantitative 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.

[0088] In some other embodiments, the first power and the first working time are obtained according to the first bubble moving distance. The second power and the second working time are obtained according to the second bubble moving distance, and the second power is less than the first power. According to the first preset power, the metering pump is controlled to work for the first working time. According to the second preset power, the metering pump is controlled to work for the second working time. After the metering pump works for the first working time according to the first preset power, one end of the bubble can just enter the adapter. At this time, the metering pump needs to be reduced to the second power to slow down the movement speed of the bubble to prevent the bubble from moving too fast and directly entering the liquid storage pipeline.

[0089] Step S407: close the valve, and control the metering pump to operate in the liquid discharge mode for a first preset time.

[0090] The first preset time is a preset empirical value, and relevant personnel can adjust the first preset time according to actual needs. Further, after the quantitative pump works in the liquid discharge mode for the first preset time, the test sample in the liquid storage pipeline can be completely entered into the flow chamber and discharged from the flow chamber to avoid air bubbles interfering with the operation of the flow cytometer.

[0091] By adopting the above technical solution, the bubble length is obtained 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. The bubble moving distance is determined by the bubble length, so that the bubble can enter the flow chamber through the adapter and be discharged through the flow chamber. In this way, the accuracy of the flow cytometer liquid collection is improved, so as to improve the accuracy of the flow cytometer detection results.

[0092] In actual scenarios, the bubbles may be too long, which may prevent them from entering the pipe corresponding to the third section of the adapter. To solve the above problem, the present application discloses a third air removal method based on liquid drainage. Figure 5 , the method comprising: Step S501: When the length of the bubble is greater than a first preset length, the metering pump is controlled to operate in a liquid suction mode according to a preset bubble movement distance.

[0093] 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.

[0094] 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.

[0095] Exemplarily, the preset pipeline volume is calculated based on the preset bubble moving distance. The quantitative pump is controlled to work in the liquid aspiration mode so that the volume of the test sample aspirated by the quantitative pump reaches the preset pipeline volume. At this time, a part of the bubbles is located in the pipeline corresponding to the third end of the adapter, and this part of the bubbles is defined as the third end bubbles, while the other part is located in the pipeline corresponding to the first end of the adapter, and this part of the bubbles is defined as the first end bubbles.

[0096] Step S502: close the valve, and control the metering pump to operate in the liquid discharge mode for a first preset time.

[0097] The first preset time is a preset empirical value. If the metering pump works in the discharge mode for the first preset time, the bubbles at the third end can be discharged through the flow chamber, but due to the structural design of the adapter, the bubbles at the first end cannot be completely discharged, so the remaining part of the bubbles at the first end needs to be discharged again through subsequent steps.

[0098] Step S503: Open the valve and obtain the remaining bubble length according to the preset bubble movement distance and bubble length.

[0099] Exemplarily, the difference between the bubble length and the preset bubble moving distance is calculated to obtain the remaining bubble length. The remaining bubble length refers to the length of the bubble remaining at the first end of the adapter in the pipeline.

[0100] Step S504: Controlling the metering pump to operate in the liquid suction mode according to the remaining bubble length.

[0101] Exemplarily, the remaining pipe volume is calculated based on the remaining bubble length, and the metering pump is controlled to work in the liquid aspiration mode so that the volume of the test sample aspirated by the metering pump reaches the remaining pipe volume.

[0102] Step S505: close the valve, and control the metering pump to operate in the liquid discharge mode for a second preset time period.

[0103] The second preset time is a preset empirical value. If the metering pump works in the liquid discharge mode for the second preset time, all the residual bubbles in the pipeline can be discharged through the flow chamber.

[0104] By adopting the above technical solution, when the bubbles are long, some bubbles will be discharged through the flow chamber first, and then the bubbles will be completely discharged for the second time to prevent the bubbles from remaining inside. The accuracy of the flow cytometer liquid collection can be improved, so as to improve the accuracy of the flow cytometer detection results.

[0105] In the following embodiments, when the volume of the bubbles in the test sample is too small, the fluidity of the bubbles in the test sample becomes worse, so that the quantitative pump needs to work at a higher power to discharge the bubbles. Therefore, the present application embodiment discloses a fourth air removal method based on the liquid discharge method. Figure 6 , the method comprising: 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, 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.

[0106] When the first sample flow rate is greater than the third flow rate threshold and less than the first flow rate threshold, it means that the change in the first sample flow rate can be used to determine that there are bubbles in the test sample, but the volume of the bubbles is 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 test sample is small.

[0107] Step S602: Close the valve.

[0108] In the present application, when the bubbles are too small, since the volume of the liquid storage pipeline is relatively limited, even if the test sample has filled the liquid storage pipeline, it is difficult to discharge the bubbles through the liquid aspiration pipeline only by the test sample in the liquid storage pipeline. Therefore, in the embodiment of the present application, it is selected to discharge the bubbles from the flow chamber to increase the probability of bubble discharge.

[0109] Step S603: Control the metering pump to operate in the liquid discharge mode until the test sample in the liquid storage pipeline is completely discharged.

[0110] Exemplarily, the metering pump is controlled to continue operating in the liquid discharge mode until all the test samples in the liquid storage pipeline are discharged.

[0111] By adopting the above technical solution, when the bubbles in the test sample are too small, the test sample needs to be sucked according to the target sample flow rate to ensure that the bubbles are completely sucked into the flow chamber, and then the bubbles are removed by the flow chamber. This is conducive to improving the accuracy of the flow cytometer liquid collection, so as to improve the accuracy of the flow cytometer test results.

[0112] In the following embodiments, the target sample flow rate needs to be limited so that the target sample flow rate is large enough to discharge small bubbles. Therefore, the present application embodiment discloses a method for calculating the sample flow rate. Figure 7 , the method comprising: Step S701: Obtain the liquid type of the test sample.

[0113] The liquid type can be input by relevant personnel. In some other embodiments, the liquid type can be uniformly set to a preset type.

[0114] Step S702: predicting the bubble radius according to the bubble length.

[0115] Exemplarily, the bubble length is regarded as the diameter of the bubble, thereby obtaining the bubble radius.

[0116] Step S703: Obtain the predicted sample flow rate according to the liquid type and bubble radius.

[0117] Exemplarily, the predicted sample flow rate is ; where r represents the bubble radius, is the density of the test sample, is the air density, g is the acceleration due to gravity, To detect the liquid viscosity of the sample. In some embodiments, due to Much greater than , so we can The value of is simplified to , in order to reduce the amount of calculation. Further, to simplify the calculation, the density of the test sample and the viscosity of the liquid can adopt preset values.

[0118] Step S704: Calculate the sum of the predicted sample flow rate and the flow rate error to obtain the target sample flow rate.

[0119] 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.

[0120] In some other embodiments, the target sample flow rate is required to be less than the maximum sample flow rate, where the maximum sample flow rate is ,in, is the liquid surface tension of the test sample, r is the bubble radius, is the density of the test sample. Further, to simplify the calculation, the liquid surface tension and density of the test sample can adopt preset values.

[0121] By adopting the above technical solution, a method for calculating the target sample flow rate is provided, so that the target sample flow rate is large enough to completely absorb 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 guaranteed to be large enough.

[0122] In the following embodiment, when the air is discharged, the bubbles may directly enter the pipe corresponding to the third end of the adapter, resulting in the metering pump still not being able to detect that the second sample flow rate is greater than the second flow threshold after working in the liquid discharge mode for a period of time. At this time, the bubbles in the test sample need to be discharged through the flow chamber. Therefore, the embodiment of the present application discloses a method for degassing based on the liquid discharge method. Figure 8 , the method comprising: Step S801: If it is not detected that the second sample flow rate is less than the second flow rate threshold within the preset time threshold, a preset sample flow rate and a preset liquid volume are obtained.

[0123] The preset time threshold may be a preset empirical value or may be related to the current sample volume in the liquid storage pipeline. For example, the product of the current sample volume and the preset ratio is calculated to obtain the detection discharge volume. According to the detection discharge volume and the working power of the metering pump, the time required for the metering pump to discharge the detection sample of the detection discharge volume is obtained, and the time is used as the preset time threshold.

[0124] 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 input by relevant personnel.

[0125] Step S802: Control 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.

[0126] Exemplarily, the plunger movement speed of the metering pump is determined according to the preset sample flow rate. The working power of the metering pump is determined according to the plunger movement speed. According to the aforementioned working power, the metering pump is controlled to work in the washing liquid mode.

[0127] Step S803: Close the valve.

[0128] Since the bubbles need to be discharged through the flow chamber in the subsequent steps, the valve needs to be closed in this step to prevent part of the test sample from returning to the sample tube from the valve, resulting in a poor bubble discharge effect.

[0129] Step S804: Control the metering pump to operate in the liquid discharge mode until the test sample in the liquid storage pipeline is completely discharged.

[0130] Exemplarily, the position information of the plunger when the test sample in the liquid storage pipeline is obtained, and the position of the plunger is adjusted to the position corresponding to the above position information to achieve the effect of draining the test sample in the liquid storage pipeline.

[0131] 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 time threshold, the quantitative pump is controlled to work in the liquid discharge mode until the test sample in the liquid storage pipeline is completely discharged. It is ensured that the bubbles in the test sample can be completely removed, which is conducive to improving the accuracy of the flow cytometer liquid collection, so as to improve the accuracy of the flow cytometer test results.

[0132] Based on the same inventive concept, the present application embodiment provides a liquid collection system for a flow cytometer, please refer to Fig. 9 The system comprises: An acquisition module 901 is 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; Memory 902, used to store the program of the liquid collection method of the flow cytometer; Processor 903, the program in the memory can be loaded and executed by the processor to implement the above-mentioned liquid collection method of the flow cytometer.

[0133] By adopting the above technical solution, after detecting that the flow rate of the first sample at the valve is too small, the quantitative pump will be controlled to discharge a preset volume of the test sample in the discharge mode, so that the bubbles in the test sample are also excluded, thereby improving the accuracy of the flow cytometer liquid collection and improving the accuracy of the flow cytometer detection results.

[0134] Those skilled in the art can clearly understand that for the convenience and simplicity 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 assigned 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 process of the system, device and unit described above can refer to the corresponding process in the aforementioned method embodiment, and will not be repeated here.

[0135] An embodiment of the present application provides a computer-readable storage medium storing a computer program that can be loaded by a processor and executed for a method of extracting liquid in a flow cytometer.

[0136] 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 disks.

[0137] Based on the same inventive concept, an embodiment of the present application provides an intelligent terminal, including a memory and a processor, wherein the memory stores a computer program that can be loaded by the processor and execute a liquid collection method of a flow cytometer.

[0138] Those skilled in the art can clearly understand that for the convenience and simplicity 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 assigned 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 process of the system, device and unit described above can refer to the corresponding process in the aforementioned method embodiment, and will not be repeated here.

[0139] The above are all preferred embodiments of the present application, and are not intended to limit the protection scope of the present application. Any feature disclosed in this specification (including the abstract and drawings), unless otherwise stated, can be replaced by other equivalent or alternative features with similar purposes. That is, unless otherwise stated, 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, the quantitative pump is controlled to aspirate the test sample in the liquid aspiration mode.

2. The method for collecting liquid for a flow cytometer according to claim 1, characterized in that: 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; The plunger in the metering pump is controlled to move the plunger advancement distance so that the detection sample of the preset volume passes through the valve.

3. The method for collecting liquid for a flow cytometer according to claim 2, characterized in that: 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.

4. The method for collecting liquid for a flow cytometer according to claim 3, 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.

5. The method for collecting liquid for a flow cytometer according to claim 3, 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.

6. The method for collecting liquid for a flow cytometer according to claim 5, 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.

7. The method for collecting liquid for a flow cytometer according to claim 2, characterized in that: After detecting the second sample flow at the valve, the method further includes: If it is not detected that the second sample flow rate is less than the second flow rate threshold within the preset time 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.

8. 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 7, 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.

9. 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 execute the method according to any one of claims 1 to 7.

10. 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 7.

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