Sterile sampling method, device and medium

Abstract

The invention relates to the technical field of biomedical treatment, and discloses a sterile sampling method, equipment and a medium, the method comprises the following steps: after receiving a sampling instruction, controlling a gas pumping and blowing device to be in a gas pumping state, a to-be-sampled liquid in the centrifugal container in a rotary centrifugal state is pumped into the buffer container through the first sterile one-way valve by the gas pumping and blowing device; acquiring the real-time sampling volume of the extracted to-be-sampled liquid; when it is confirmed that the real-time sampling volume reaches the preset sampling volume, the gas pumping and blowing device is controlled to be in a gas blowing state, so that gas filtered by the air filter is blown into the buffer container, and then the to-be-sampled liquid with the preset sampling volume in the buffer container is blown into the sampling container through the second sterile one-way valve. According to the invention, automatic extraction and sterile sampling are realized, the risk that cells are polluted is greatly reduced, accurate control of the sterile sampling volume is further realized, and the accuracy of the sterile sampling volume is improved.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technologies, and in particular, to a sterile sampling method, device, and medium. Background Art

[0002] Currently, cell therapy is a key area of focus in the international medical forefront. During the process of cell therapy, due to the characteristics of the production process of cell-based drugs, such as multiple preparation steps and complex processes, the complex process operations and the ex vivo environment of cells have a significant impact on the biological activity of cells. Therefore, during the production process of cell-based drugs, it is necessary to regularly perform sterile sampling of cells to monitor indicators such as cell viability, loss rate, and density.

[0003] In the prior art, in order to complete sterile sampling of cells, experimental personnel need to use a sterile sampling device for manual sterile sampling, that is, move the device into a sterile environment and open the lid for sterile sampling. Therefore, the existing sterile sampling process has problems such as the need for manual operation, cumbersome operation process, inability to precisely control the sterile sampling volume, and easy contamination caused by frequently opening the device lid. Therefore, how to achieve prevention of cell fluid sampling contamination and precise cell fluid sampling is an urgent problem for those skilled in the art in the current field. Summary of the Invention

[0004] Embodiments of the present invention provide a sterile sampling method, device, and medium to solve the problems of contamination risk and poor sampling accuracy in the sampling operation process in the prior art.

[0005] A sterile sampling method includes:

[0006] After receiving a sampling instruction, controlling a gas blowing device to be in a pumping state to extract a liquid to be sampled in a centrifugal container in a rotating and centrifuging state through the gas blowing device to a buffer container via a first sterile one-way valve;

[0007] Obtaining the real-time sampling volume of the extracted liquid to be sampled;

[0008] When it is confirmed that the real-time sampling volume reaches a preset sampling volume, controlling the gas blowing device to be in a blowing state to blow gas filtered by an air filter into the buffer container, and then blowing the preset sampling volume of the liquid to be sampled in the buffer container into a sampling container via a second sterile one-way valve.

[0009] A sterile sampling device includes a first sterile one-way valve, a second sterile one-way valve, a gas pumping and blowing device, an air filter, a buffer container, a centrifugal container, a sampling container, and a controller. The controller is used to execute the sampling method described above. The gas pumping and blowing device is communicatively connected to the controller. The first sterile one-way valve is connected between the centrifugal container and the buffer container. The second sterile one-way valve is connected between the buffer container and the sampling container. The gas pumping and blowing device is connected to a side of the buffer container away from the first sterile one-way valve and the second sterile one-way valve. The air filter is connected to a side of the gas pumping and blowing device away from the buffer container.

[0010] A computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, it implements the above-mentioned sterile sampling method.

[0011] For the above-mentioned sterile sampling method, device, and medium, the present invention realizes the automatic extraction of the liquid to be sampled in the centrifugal container under the rotating centrifugal state by controlling the gas pumping and blowing device to be in the air extraction state and the first sterile one-way valve, improving the sampling efficiency. By extracting the liquid to be sampled into the buffer container and obtaining the sampling volume, the accurate control of the sampling volume is realized, improving the sampling accuracy. By controlling the gas pumping and blowing device to be in the air blowing state, the liquid to be sampled is blown into the sampling container. Further, the entire sampling process is carried out in a sealed sterile environment (all devices are sealed and connected, and an air filter is provided to filter the input air), achieving the purpose of sterile sampling, thus ensuring that the sampling process will not be contaminated by other environmental factors, and further greatly reducing the risk of cell contamination. Description of the Drawings

[0012] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required to be used in the description of the embodiments of the present invention will be briefly introduced below. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can be obtained based on these drawings.

[0013] Figure 1 It is a flowchart of the sterile sampling method in an embodiment of the present invention;

[0014] Figure 2 It is a schematic diagram of the sterile sampling device in an embodiment of the present invention.

[0015] The reference numerals in the specification are as follows:

[0016] 1. Centrifugal container; 2. Buffer container; 3. Sampling container; 4. Gas pumping and blowing device; 5. Air filter; 6. First sterile one-way valve; 7. Second sterile one-way valve; 8. Flow sensor. Detailed implementation manner

[0017] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative efforts shall fall within the protection scope of the present invention.

[0018] In one embodiment, as Figure 1 shown, a sterile sampling method is provided, including the following steps:

[0019] S10: After receiving the sampling instruction, control the gas pumping and blowing device to be in the air extraction state, so as to extract the liquid to be sampled in the centrifugal container in the rotating centrifugal state through the first sterile one-way valve into the buffer container.

[0020] Understandably, the sampling instruction refers to an instruction to extract the liquid to be sampled. The gas pumping and blowing device 4 refers to a device that can extract gas and blow in gas, for example, a syringe tube, etc. The centrifugal container 1 is a container for containing the liquid to be sampled, including but not limited to a centrifuge cup, a centrifugal tank or a centrifugal bottle, etc. The first sterile one-way valve 6 is used to prevent the liquid to be sampled from flowing into the centrifugal container during subsequent air blowing, and a solenoid valve can also be used for control.

[0021] Specifically, when sampling is required, the controller sends a sampling instruction. After receiving the sampling instruction, first control the control valves at the inlet and outlet of the four-way joint on the centrifugal container 1 to close. Then, set the gas pumping and blowing device 4 according to the parameters in the sampling instruction, that is, control the gas pumping and blowing device 4 to operate with these parameters and be in the air extraction state, that is, extract the gas in the buffer container 2 and the centrifugal container 1 to extract the liquid to be sampled in the centrifugal container 1 in the rotating centrifugal state through the first sterile one-way valve 6 into the buffer container 2.

[0022] S20: Obtain the real-time sampling volume of the liquid to be sampled extracted.

[0023] Furthermore, the real-time sampling volume refers to the volume of the liquid to be sampled at each moment during the sampling process. When starting to draw the liquid to be sampled into the buffer container 2, by calculating the volume of the liquid in the buffer container 2, that is, when the buffer container 2 is transparent and provided with scale lines, taking a photo of the buffer container 2 through the photographing device on one side of the buffer container 2, so as to determine the scale line where the liquid is located, and thus obtain the real-time sampling volume of the liquid to be sampled being drawn. It is also possible to determine the liquid height in the buffer container 2 by setting a height sensor in the buffer container 2, and calculate the real-time sampling volume through the liquid height and the parameters of the buffer container 2.

[0024] In another embodiment, there will be some residual liquid to be sampled in the pipeline. This residual volume can be determined according to the pipeline length and pipeline diameter from the pipeline to the flow sensor 8, that is, by calculating the volume of the pipeline, the pipeline residual volume can be obtained. Furthermore, the sum of the pipeline residual volume and the liquid volume in the buffer container is the real-time sampling volume. That is to say, in this embodiment, the accuracy of the volume of the liquid to be sampled being drawn is ensured, and the precise control of the volume of the liquid to be sampled being drawn is realized.

[0025] S30: When it is confirmed that the real-time sampling volume reaches the preset sampling volume, control the gas blowing device to be in the blowing state, so as to blow the gas filtered by the air filter into the buffer container, and then blow the liquid to be sampled with the preset sampling volume in the buffer container into the sampling container through the second sterile one-way valve.

[0026] It can be understood that the preset sampling volume refers to the volume of the extracted solution set in advance. The sampling container refers to a container for storing the liquid to be sampled, such as a sampling bag, etc.

[0027] Specifically, when it is confirmed that the real-time sampling volume reaches the preset sampling volume, trigger a stop instruction to control the gas blowing device 4 to stop pumping air. And trigger a reverse instruction to control the gas blowing device 4 to be in the blowing state, and filter the blown gas through the air filter 5, so as to blow the gas filtered by the air filter 5 into the buffer container 2, and blow the liquid to be sampled with the preset sampling volume in the buffer container 2 into the sampling container 3 through the second sterile one-way valve 7. In this way, aseptic sampling can be completed.

[0028] In the present invention, by controlling the gas pumping and blowing device 4 to be in the air extraction state and the first sterile one-way valve 6, the automatic extraction of the liquid to be sampled in the centrifuge container 1 under the rotating and centrifuging state is realized, improving the sampling efficiency. By pumping the liquid to be sampled into the buffer container 2 and obtaining the sampling volume, the accurate control of the sampling volume is realized, improving the sampling accuracy. By controlling the gas pumping and blowing device 4 to be in the air blowing state, the liquid to be sampled is blown into the sampling container 3. Further, the whole sampling process is carried out in a sealed and sterile environment (all devices are hermetically connected, and an air filter is provided to filter the input air), realizing the purpose of sterile sampling, thus ensuring that the sampling process will not be contaminated by other environmental factors, and further greatly reducing the risk of cell contamination.

[0029] In one embodiment, after the step S30, that is, after blowing the preset sampling volume of the liquid to be sampled in the buffer container into the sampling container through the second sterile one-way valve, it includes:

[0030] S40, counting the cells in the sampling container to obtain a cell count result.

[0031] S50, determining the cell detection result of the liquid to be sampled according to the preset cell index and the cell count result, and the cell detection result is used to characterize whether the liquid to be sampled meets the preset cell index.

[0032] It can be understood that the cell count result refers to the number of cells in the liquid to be sampled. The cell detection result is used to characterize whether the liquid to be sampled meets the preset cell index. The preset cell index refers to a standard index preset for evaluating the cell state or function.

[0033] Specifically, after blowing 2 preset sampling volumes of the liquid to be sampled in the buffer container into the sampling container 3 through the second sterile one-way valve 7, the cells in the sampling container 3 are counted. That is, the flow cytometry method can be used. The liquid to be sampled in the sampling container 3 is injected into a flow cytometer, and the cells in the liquid to be sampled are counted by the flow cytometer to obtain the cell counting result. Further, the cell detection result of the liquid to be sampled is determined according to the preset cell index and the cell counting result. That is, the preset cell index is obtained. By comparing the cell counting result with the number of cells in the preset cell index, when the cell counting result is greater than or equal to the number of cells in the preset cell index, it is determined that the liquid to be sampled meets the preset cell index. When the cell counting result is less than the number of cells in the preset cell index, that is, when it is determined that the liquid to be sampled does not meet the preset cell index, the liquid to be sampled in the centrifugation container is sampled again, and the sampling result is detected again to obtain a new cell detection result. That is, in this embodiment, the cell counting result is obtained by counting the cells in the liquid to be sampled. By comparing with the preset cell index, the determination of the cell detection result is realized, and the evaluation of the cell quality in the liquid to be sampled is realized.

[0034] In one embodiment, in step S40, that is, counting the cells in the sampling container to obtain the cell counting result, further includes:

[0035] S401, disconnect the connection between the sampling container and the second sterile one-way valve, and transfer the liquid to be sampled in the sampling container to a counting container.

[0036] S402, photograph the cells in the counting container to obtain a cell photographed image.

[0037] S403, perform cell recognition on the cell photographed image to obtain the cell counting result.

[0038] Understandably, a counting container refers to a container for counting cells, for example, a counting dish, etc. A cell photographed image refers to a photographed cell image.

[0039] Specifically, after blowing the to-be-sampled liquid with a preset sampling volume in the buffer container 2 into the sampling container 3 through the second sterile one-way valve 7, the connection between the sampling container 3 and the second sterile one-way valve 7 is fused, and the to-be-sampled liquid in the sampling container 3 is transferred to the counting container. Then, the cells in the counting container are photographed by a photographing device installed above the counting container, that is, the photographing device can photograph the entire counting container, so as to ensure that all cells are within the photographing range. After obtaining the photographed image, boundary recognition is performed on the photographed image, that is, a pre-trained boundary extraction model is used to extract the circular boundary features of the photographed image, so as to obtain the circular boundary features. Then, feature recognition is performed according to the extracted circular boundary features, that is, the boundary recognition model can be used to perform feature recognition on the circular boundary features, so as to recognize the boundary area of the counting container. Based on the boundary area, the range of the counting container is determined, that is, the image outside the range of the counting container in the photographed image is cropped through the boundary area, so as to obtain the cell photographed image.

[0040] Furthermore, cell recognition is performed on the cell photographed image, that is, the cell photographed image can be input into a pre-trained neural network model to recognize and count the cells in the cell photographed image, that is, the neural network model first extracts the cell shape features in the cell photographed image, and then the neural network model recognizes the cell area through the cell shape features, and counts the cell area to obtain the cell counting result in the cell photographed image. Among them, in the counting process, the number of stacked or adjacent regions of cells is output as the corresponding number, so as to obtain the actual number of cells. For example, if the stacked region is within the region size range of one to two cells, it is considered that this region contains two cells after conversion. In this way, the number of cells can be recognized more accurately, and the reliability of cell number recognition is improved. That is, in this embodiment, by transferring the to-be-sampled liquid to the counting container for recognition and counting, photographing of the cells in the to-be-sampled liquid is realized, the clarity of the cell photographed image is ensured, and then the recognition of the number of cells is realized, and the accuracy of the cell counting result recognition is improved.

[0041] In one embodiment, in step S50, the preset cell index includes density index data and viability index data;

[0042] That is, determining the cell detection result of the to-be-sampled liquid according to the preset cell index and the cell counting result further includes:

[0043] S60, determining the density detection result corresponding to the to-be-sampled liquid in the sampling container according to the preset sampling volume and the cell counting result;

[0044] S70, performing a viability detection on the cells in the sampling container to obtain the viability detection result corresponding to the to-be-sampled liquid in the sampling container;

[0045] S80. When it is determined that the density detection result is greater than or equal to the density index data and the viability detection result is greater than or equal to the viability index data, confirm that the cell detection result is that the to-be-sampled liquid meets the preset cell index.

[0046] S90. When it is determined that the density detection result is less than the density index data, or / and the viability detection result is less than the viability index data, resample the to-be-sampled liquid in the centrifuge container and obtain a new cell detection result.

[0047] Understandably, the density detection result refers to the number of cells detected per unit volume. The viability detection result refers to the data obtained through the cell viability detection method, which reflects information such as the growth status, function, and metabolic level of cells under specific conditions. The preset cell index includes the density index data and the viability index data. The density index data refers to the set density requirement. The viability index data refers to the set viability requirement.

[0048] Specifically, after obtaining the cell counting result, according to the preset sampling volume and the cell counting result, determine the density detection result corresponding to the to-be-sampled liquid in the sampling container 3, that is, by calculating the division result of the cell counting result and the preset sampling volume, the density detection result corresponding to the extracted to-be-sampled liquid can be obtained. Then, perform a viability detection on the cells in the sampling container 3, that is, the staining counting method or the fluorescence staining method can be used. That is, by using the different effects of dead cells and live cells on dyes or fluorescent staining agents, detect the cell viability through a fluorescence microscope or a flow cytometer, and through cell counting, the viability detection result corresponding to the to-be-sampled liquid in the sampling container can be obtained. Further, compare the density index data with the density detection result, and compare the viability index data with the viability detection result. And when it is determined that the density detection result is greater than or equal to the density index data and the viability detection result is greater than or equal to the viability index data, confirm that the cell detection result is that the to-be-sampled liquid meets the preset cell index. When it is determined that the density detection result is less than the density index data, or / and the viability detection result is less than the viability index data, that is, when it is confirmed that the cell detection result is that the to-be-sampled liquid does not meet the preset cell index, resample the to-be-sampled liquid in the centrifuge container and re-detect the sampling result, so as to obtain a new cell detection result. That is, in this embodiment, by performing density detection and viability detection on the to-be-sampled liquid, the determination of the density detection result and the viability detection result is realized. By comparing the results with the preset cell index, the evaluation of the cell quality in the to-be-sampled liquid is realized, and further the conditions for subsequent cell culture are optimized.

[0049] In one embodiment, in step S10, the gas blowing device includes an air pump.

[0050] That is, the control gas pumping and blowing device is in the air extraction state, including:

[0051] S101, obtain the rotation speed and the number of rotation cycles of the air pump, determine the air extraction volume of the air pump according to the rotation speed and the number of rotation cycles, and control the air pump to start and enter the air extraction state with the air extraction volume, so as to extract the liquid to be sampled in the centrifuge container under the rotating centrifugal state.

[0052] Understandably, the number of rotation cycles refers to the number of cycles that the air pump rotates. The air extraction volume refers to the volume of gas extracted by the air pump. Among them, in a sealed environment, the air extraction volume is equal to the preset sampling volume.

[0053] Specifically, after receiving the sampling instruction, the rotation speed and the number of rotation cycles of the air pump can be obtained from the sampling instruction, or the rotation speed and the number of rotation cycles of the air pump can be obtained from the controller. Then, according to the rotation speed and the number of rotation cycles, the air extraction volume of the air pump is determined, that is, the air pump volume is obtained, and by calculating the product result of the rotation speed, the number of rotation cycles and the air pump volume, the air extraction volume of the air pump can be obtained, that is, the volume of the liquid to be sampled extracted. Next, control the air pump to start and enter the air extraction state with the air extraction volume, so as to extract the preset sampling volume of the liquid to be sampled in the centrifuge container 1 under the rotating centrifugal state. That is, in this embodiment, by controlling the air pump to start and enter the air extraction state with the air extraction volume, the extraction of the liquid to be sampled in the centrifuge container 1 under the rotating centrifugal state is realized, and thus the precise control of the volume of the liquid to be sampled is realized, and the sampling efficiency is improved.

[0054] The control of the gas pumping and blowing device to be in the blowing state to blow the gas filtered by the air filter into the buffer container includes:

[0055] S301, after controlling the air pump to shut down to stop air extraction, control the air pump to start in reverse and enter the blowing state, so as to blow the gas filtered by the air filter into the buffer container.

[0056] Understandably, the air filter 5 is a filtering device that can filter substances such as fine particles in the air.

[0057] Specifically, when it is confirmed that the real-time sampling volume reaches the preset sampling volume, control the air pump to shut down to stop air extraction. Then, trigger the reverse instruction, control the air pump to start in reverse and enter the blowing state, and filter the blown gas through the air filter 5, and then blow the gas filtered by the air filter 5 into the buffer container 2, so that the liquid to be sampled in the buffer container 2 is blown into the sampling container 3 through the second sterile one-way valve 7. That is, in this embodiment, the state of the automatic control gas pumping and blowing device 4 is realized, the gas filtered by the air filter 5 is blown into the buffer container 2, and thus the liquid to be sampled is blown into the sampling container 3.

[0058] In one embodiment, in step S20, that is, obtaining the real-time sampling volume of the to-be-sampled liquid to be extracted, includes:

[0059] S201, obtaining the rotation speed and rotation torque of the air pump, and determining the air extraction speed of the air pump according to the rotation speed and the rotation torque.

[0060] S202, obtaining the real-time extraction duration of the to-be-sampled liquid, and determining the real-time sampling volume according to the real-time extraction duration and the air extraction speed.

[0061] It can be understood that the rotation speed refers to the speed of the air pump rotation. The rotation torque refers to the torque for the air pump to rotate. The real-time sampling volume refers to the volume of the to-be-sampled liquid to be extracted monitored in real time during the sampling process.

[0062] Specifically, after receiving the sampling instruction and controlling the gas pumping and blowing device 4 to be in the air extraction state, obtain the rotation speed and rotation torque of the air pump, and obtain the air pump volume. Among them, the rotation speed, rotation torque, and air pump volume can be obtained from the sampling instruction. Then, according to the rotation speed, air pump volume, and rotation torque, determine the air extraction speed of the air pump, that is, divide the product of the rotation speed and rotation torque by the air pump volume, and the air extraction speed of the air pump can be calculated. Further, obtain the real-time extraction duration of the to-be-sampled liquid, and determine the real-time sampling volume according to the real-time extraction duration and the air extraction speed, that is, by calculating the product of the real-time extraction duration and the air extraction speed, the volume of the to-be-sampled liquid to be extracted can be calculated and determined as the real-time sampling volume. That is, in this embodiment, through the air extraction speed of the air pump and the real-time extraction duration, the calculation of the real-time sampling volume is realized, and further the precise control of the extraction volume of the to-be-sampled liquid is realized, improving the accuracy of sampling.

[0063] In another embodiment, when a stop instruction is triggered and the gas pumping and blowing device 4 is controlled to stop air extraction, at this time, part of the to-be-sampled liquid will remain in the pipeline. Therefore, when determining the real-time sampling volume, the pipeline residual volume needs to be considered. The pipeline length between the sampling port and the flow sensor can be determined in advance, the pipeline diameter can be obtained, and the pipeline residual volume can be determined by calculating the volume of the pipeline. When the extraction volume obtained by multiplying the air extraction speed and the real-time extraction duration plus the pipeline residual volume is equal to the preset sampling volume, trigger the stop instruction and control the gas pumping and blowing device 4 to stop air extraction. That is, in this embodiment, the accuracy of the volume of the to-be-sampled liquid to be extracted is ensured, and the precise control of the volume of the to-be-sampled liquid to be extracted is realized. Among them, the channel residual volume in the centrifuge container 1 can refer to the embodiment of step S203, and will not be elaborated here one by one.

[0064] In one embodiment, in step S20, that is, obtaining the real-time sampling volume of the to-be-sampled liquid to be extracted, further includes:

[0065] S203, determine the real-time sampling volume of the to-be-sampled liquid being extracted through a flow sensor disposed between the centrifugation container and the buffer container.

[0066] Understandably, the flow sensor 8 is a detection sensor that can detect the flow parameters of a liquid and convert them into signals in other forms for output.

[0067] Specifically, when the gas blowing device 4 is in the air extraction state, extract the gas in the entire pipeline so that the to-be-sampled liquid in the centrifugation container 1 flows through the connecting pipeline and passes through the first sterile one-way valve 6 into the buffer container 2. When it passes through the flow sensor 8 disposed between the centrifugation container 1 and the buffer container 2, record the flow rate of the to-be-sampled liquid to obtain the recorded flow rate. Then, obtain the real-time extraction duration of the to-be-sampled liquid, and calculate the volume of the to-be-sampled liquid in real time through the recorded flow rate and the real-time extraction duration, that is, calculate the product of the recorded flow rate and the real-time extraction duration, so as to determine the real-time sampling volume of the to-be-sampled liquid being extracted. That is, in this embodiment, through the flow sensor 8, the determination of the real-time sampling volume of the to-be-sampled liquid is realized, improving the accuracy of sampling.

[0068] Furthermore, the flow sensor 8 and the air pump can be used simultaneously to monitor the volume of the to-be-sampled liquid. That is, after the sampling volume corresponding to the real-time extraction duration reaches the preset sampling volume, determine the difference between the first sampling volume measured by the flow sensor 8 and the second sampling volume measured by the air extraction speed of the air pump, and when the difference does not exceed the preset error value, control the gas blowing device 4 to stop air extraction. When the difference exceeds the preset error value, send an alarm message.

[0069] In one embodiment, when a stop instruction is triggered to control the gas blowing device 4 to stop air extraction. At this time, a part of the to-be-sampled liquid will remain in the pipeline. Thus, the sampling volume exceeds the preset sampling volume. Therefore, the volume remaining in the pipeline needs to be considered. The length of the pipeline between the sampling port and the flow sensor 8 can be determined in advance, the pipeline diameter can be obtained, and the remaining volume can be determined by calculating the volume of the pipeline. When the volume measured by the flow sensor 8 plus the remaining volume is equal to the preset sampling volume, trigger the stop instruction to control the gas blowing device 4 to stop air extraction. That is, in this embodiment, the accuracy of the volume of the to-be-sampled liquid being extracted is ensured, and the precise control of the volume of the to-be-sampled liquid being extracted is realized.

[0070] In one embodiment, when there is a negative pressure in the centrifugal container 1, during sampling, the liquid to be sampled in the channel of the centrifugal container 1 cannot return to the centrifugal container under the action of gravity. At this time, the volume of the liquid to be sampled remaining in the channel needs to be considered. That is, when it is determined that there is a negative pressure in the centrifugal container 1, the average value is obtained through multiple tests to determine the channel volume of the liquid remaining in the channel. Then, the sum of the channel volume of the liquid remaining in the channel and the volume of the liquid remaining in the pipeline in the above embodiment is determined as the residual volume. When the volume measured by the flow sensor 8 plus the residual volume is equal to the preset sampling volume, a stop instruction is triggered to control the gas blowing device 4 to stop pumping air.

[0071] In another embodiment, at the end of sampling, a stop instruction is triggered to control the gas blowing device 4 to stop pumping air, and the control valve at the sampling port automatically closes. When there is residual liquid in the channel of the centrifugal container 1, a preset dose of solution is input into the centrifugal container 1 through the inlet so that the residual liquid in the channel is pushed into the centrifugal container 1. In this way, precise control of the sampling volume is achieved, and the accuracy of the sampling volume is improved.

[0072] It should be understood that the magnitudes of the sequence numbers of the steps in the above embodiments do not mean the order of execution. The order of execution of each process should be determined according to its function and internal logic, and should not constitute any limitation to the implementation process of the embodiments of the present invention.

[0073] A sterile sampling device includes a first sterile one-way valve, a second sterile one-way valve, a gas blowing device, an air filter, a buffer container, a centrifugal container, a sampling container, and a controller. The controller is used to execute the sampling method described above; the gas blowing device is communicatively connected to the controller; the first sterile one-way valve is connected between the centrifugal container and the buffer container; the second sterile one-way valve is connected between the buffer container and the sampling container; the gas blowing device is connected to one side of the buffer container away from the first sterile one-way valve and the second sterile one-way valve; the air filter is connected to one side of the gas blowing device away from the buffer container.

[0074] Specifically, a four-way joint is provided on the centrifuge container. One end is connected to the centrifuge container, one end is an inlet, one end is an outlet, and one end is a sampling port. A control valve for controlling the opening and closing of the interface is provided at each interface. The sampling port is connected to one end of the first sterile one-way valve through the first consumable pipeline. The other end of the first sterile one-way valve is connected to the first end of the three-way joint. The second end of the three-way joint is connected to one end of the buffer container through the second consumable pipeline, and a flow sensor for monitoring the real-time sampling volume is provided on the second consumable pipeline. The third end of the three-way joint is connected to one end of the second sterile one-way valve through the third consumable pipeline. The other end of the second sterile one-way valve is communicated with the sampling container through the fourth consumable pipeline. The other end of the buffer container is connected to the gas blowing device through the fifth consumable pipeline. An air filter for filtering the blown air is provided on the fifth consumable pipeline, and the air filter is connected to the side of the gas blowing device away from the buffer container. The gas blowing device is communicatively connected to the controller.

[0075] The sterile sampling device further includes: a flow sensor is provided between the second sterile one-way valve and the buffer container for monitoring the real-time sampling volume.

[0076] The centrifuge container further includes: a container body having a centrifugation space and a first channel; the first channel includes a first channel provided at the top of the container body and a second channel provided at the bottom of the container body, and both the first channel and the second channel are communicated with the centrifugation space. A filter membrane is provided at the top or bottom of the centrifugation space. The filter membrane can prevent external bacteria from entering the centrifugation space. At the same time, the filter membrane can realize gas exchange between the centrifugation space and the outside, ensuring the normal respiration of cells in the centrifugation space.

[0077] A controller includes a processor and a memory. The memory stores an executable program, and the processor is used to execute the executable program to implement the sterile sampling method.

[0078] Specific limitations on the controller and its various units and modules can be referred to the limitations on the aseptic sampling method in the above text, and will not be elaborated here. Each module in the above controller can be implemented in whole or / and in part by software, hardware, and their combination. Understandably, the controller includes a processor, a memory, a network interface, and a database connected through a device bus. Each module of the controller can be embedded in or / and independent of the processor in the form of hardware, or stored in the memory in the form of software, so as to facilitate the processor to call and execute the operations corresponding to each of the above modules. Among them, the processor is used to provide computing and control capabilities. The memory includes a non-volatile storage medium and an internal memory. The non-volatile storage medium stores an operating device, a computer program, and a database. The internal memory provides an environment for the operation of the operating device and the computer program in the non-volatile storage medium. The database is used to store the data used in the aseptic sampling method in the above embodiments. The network interface is used to communicate with an external terminal through a network connection. When the computer program is executed by the processor, it implements an aseptic sampling method.

[0079] In one embodiment, a computer-readable storage medium is provided, and the computer-readable storage medium stores a computer program, and when the computer program is executed by a processor, the above aseptic sampling method is implemented.

[0080] Those of ordinary skill in the art can understand that all or / and part of the processes in the methods of the above embodiments can be completed by instructing relevant hardware through a computer program. The computer program can be stored in a non-volatile computer-readable storage medium. When the computer program is executed, it can include the processes of the embodiments of the above methods. Among them, any reference to the memory, storage, database, or / and other media used in the various embodiments provided in the present application can include non-volatile and / or / and volatile memories. Non-volatile memory can include read-only memory (ROM), programmable ROM (PROM), electrically programmable ROM (EPROM), electrically erasable programmable ROM (EEPROM), or / and flash memory. Volatile memory can include random access memory (RAM) or / and an external cache memory. By way of illustration and not limitation, RAM is available in many forms, such as static RAM (SRAM), dynamic RAM (DRAM), synchronous DRAM (SDRAM), double data rate SDRAM (DDR SDRAM), enhanced SDRAM (ESDRAM), synchronous link (Synchlink) DRAM (SLDRAM), memory bus (Rambus) direct RAM (RDRAM), direct memory bus dynamic RAM (DRDRAM), and memory bus dynamic RAM (RDRAM), etc.

[0081] Those skilled in the art can clearly understand that, for the convenience and conciseness of description, only the above-mentioned division of each functional unit and module is used as an example. In actual applications, the above functions can be allocated to different functional units and modules according to needs, that is, the internal structure of the device can be divided into different functional units or / and modules to complete all or / and part of the functions described above.

[0082] The above-described embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions recorded in the foregoing embodiments, or / and perform equivalent replacements for some of the technical features; and these modifications or / and replacements do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention, and should all be included in the protection scope of the present invention.

Claims

1. A sterile sampling method, characterized in that, Including: After receiving a sampling instruction, controlling the gas blowing device to be in a gas extraction state, so as to extract the liquid to be sampled in the centrifuge container in a rotating and centrifugal state to the buffer container through the gas blowing device via a first sterile one-way valve; Obtaining the real-time sampling volume of the liquid to be sampled that has been extracted; When it is confirmed that the real-time sampling volume reaches a preset sampling volume, controlling the gas blowing device to be in a gas blowing state, so as to blow the gas filtered by the air filter into the buffer container, and then blowing the liquid to be sampled with the preset sampling volume in the buffer container into the sampling container through a second sterile one-way valve.

2. The aseptic sampling method according to claim 1, wherein After blowing the liquid to be sampled with the preset sampling volume in the buffer container into the sampling container through the second sterile one-way valve, it includes: Counting the cells in the sampling container to obtain a cell counting result; Determining the cell detection result of the liquid to be sampled according to a preset cell index and the cell counting result, and the cell detection result is used to characterize whether the liquid to be sampled reaches the preset cell index.

3. The aseptic sampling method according to claim 2, wherein The counting the cells in the sampling container to obtain a cell counting result further includes: Disconnecting the connection between the sampling container and the second sterile one-way valve, and transferring the liquid to be sampled in the sampling container to a counting container; Taking a picture of the cells in the counting container to obtain a cell picture; Performing cell recognition on the cell picture to obtain a cell counting result.

4. The aseptic sampling method according to claim 2, wherein The preset cell index includes density index data and viability index data; The determining the cell detection result of the liquid to be sampled according to a preset cell index and the cell counting result further includes: Determining the density detection result corresponding to the liquid to be sampled in the sampling container according to the preset sampling volume and the cell counting result; Performing a viability detection on the cells in the sampling container to obtain the viability detection result corresponding to the liquid to be sampled in the sampling container; When it is determined that the density detection result is greater than or equal to the density index data and the viability detection result is greater than or equal to the viability index data, confirming that the cell detection result is that the liquid to be sampled reaches the preset cell index; When it is determined that the density detection result is less than the density index data, or / and the viability detection result is less than the viability index data, resampling the liquid to be sampled in the centrifuge container and obtaining a new cell detection result.

5. The aseptic sampling method according to claim 1, wherein The gas blowing device includes an air pump; The controlling the gas blowing device to be in a gas extraction state includes: Obtaining the rotation speed and the number of rotation turns of the air pump, determining the air extraction volume of the air pump according to the rotation speed and the number of rotation turns, and controlling the air pump to start with the air extraction volume and enter the gas extraction state to extract the liquid to be sampled in the centrifuge container in a rotating and centrifugal state; The controlling the gas blowing device to be in a gas blowing state to blow the gas filtered by the air filter into the buffer container includes: After controlling the air pump to shut down to stop gas extraction, controlling the air pump to start in reverse and enter the gas blowing state to blow the gas filtered by the air filter into the buffer container.

6. The aseptic sampling method according to claim 5, characterized in that, Obtaining the real-time sampling volume of the to-be-sampled liquid to be extracted includes: Obtaining the rotation speed and rotation torque of the air pump, and determining the air extraction speed of the air pump according to the rotation speed and the rotation torque; Obtaining the real-time extraction duration of the to-be-sampled liquid, and determining the real-time sampling volume according to the real-time extraction duration and the air extraction speed.

7. The aseptic sampling method according to claim 1, wherein Obtaining the real-time sampling volume of the to-be-sampled liquid to be extracted includes: Determining the real-time sampling volume of the to-be-sampled liquid to be extracted through a flow sensor arranged between the centrifugal container and the buffer container.

8. A sterile sampling device, characterized in that, It includes a first sterile one-way valve, a second sterile one-way valve, a gas pumping and blowing device, an air filter, a buffer container, a centrifugal container, a sampling container and a controller. The controller is used to execute the sampling method according to any one of claims 1 to 7; the gas pumping and blowing device is communicatively connected to the controller; the first sterile one-way valve is connected between the centrifugal container and the buffer container; the second sterile one-way valve is connected between the buffer container and the sampling container; the gas pumping and blowing device is connected to a side of the buffer container away from the first sterile one-way valve and the second sterile one-way valve; the air filter is connected to a side of the gas pumping and blowing device away from the buffer container.

9. The aseptic sampling device according to claim 8, characterized in that, The sterile sampling device further includes: A flow sensor is arranged between the second sterile one-way valve and the buffer container for monitoring the real-time sampling volume.

10. A computer-readable storage medium storing a computer program, characterized in that, When the computer program is executed by a processor, it implements the sterile sampling method according to any one of claims 1 to 7.

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