Flow cytometry hemolysis degree detection method, device, equipment and storage medium

Abstract

The application discloses a flow cytometry hemolysis degree detection method, device, equipment and storage medium, and the method comprises the steps of placing a sample test tube after antigen-antibody reaction into an automatic incubation module for timed incubation; adding red blood cell lysate into the sample test tube after incubation through a liquid treatment module; transferring the sample test tube after lysate addition to a mixing module for mechanical oscillation treatment to obtain a treated sample; and performing hemolysis degree detection on the treated sample through an optical detection module to obtain a hemolysis degree detection result. Due to the pipeline connection of the automatic incubation module, the liquid treatment module and the mixing module, parallel and continuous treatment of multiple sample test tubes is realized, and online detection of the lysed sample is performed through the optical detection module, so that the efficiency and accuracy of hemolysis degree detection are improved compared with the prior art.

Description

Technical Field

[0001] This invention relates to the field of data processing technology, and in particular to a method, apparatus, equipment, and storage medium for detecting the degree of hemolysis in flow cytometry cells. Background Technology

[0002] Flow cytometry is a high-throughput, multi-parameter analysis technique for single cells or microparticles, widely used in the diagnosis, classification, treatment monitoring, prognostic assessment, and screening of therapeutic targets for hematological malignancies. In routine experiments, clinical samples such as bone marrow, peripheral blood, pleural effusion, and peritoneal effusion are prepared into single-cell suspensions, followed by fluorescent antibody labeling, erythrocyte lysis, fixation, and washing before final analysis. Erythrocyte lysis is a crucial step determining sample quality and detection accuracy.

[0003] Currently, laboratories generally use manual or semi-automatic methods to complete red blood cell lysis. Although there have been attempts to replace manual liquid addition, timing, and centrifugation with multi-channel pipetting workstations or simple automated platforms, there is still a lack of modules for objectively, quantitatively, and online interpreting the degree of hemolysis. This results in limited improvement in overall efficiency and fails to fundamentally solve the problem of insufficient detection accuracy.

[0004] Therefore, there is an urgent need for a flow cytometry method to detect the degree of hemolysis, which can improve the efficiency and accuracy of hemolysis detection. Summary of the Invention

[0005] The main objective of this invention is to provide a method, apparatus, device, and storage medium for detecting the degree of hemolysis by flow cytometry, aiming to solve the technical problems of low efficiency and insufficient detection accuracy of existing hemolysis degree detection methods.

[0006] To achieve the above objectives, the present invention provides a method for detecting the degree of hemolysis in flow cytometry cells. The method is applied to a flow cytometry system for detecting the degree of hemolysis in flow cytometry cells. The system includes an automated incubation module, a liquid handling module, a mixing module, and an optical detection module. The method includes the following steps:

[0007] The sample tubes that have completed the antigen-antibody reaction are placed in the automated incubation module for timed incubation;

[0008] The liquid processing module quantitatively adds red blood cell lysis buffer to the incubated sample tubes.

[0009] The sample tube containing the lysis buffer is transferred to the mixing module for mechanical agitation to obtain the processed sample.

[0010] The degree of hemolysis is detected by the optical detection module to obtain the hemolysis degree detection result.

[0011] Optionally, the step of detecting the degree of hemolysis of the processed sample using the optical detection module to obtain the hemolysis degree detection result includes:

[0012] The absorbance of the processed sample at different wavelengths is detected by the optical detection module using multi-wavelength detection technology within a preset wavelength range, thereby obtaining absorbance detection results.

[0013] The fluorescence signal intensity of each substance in the processed sample is detected using fluorescence detection technology to obtain fluorescence signal intensity detection results.

[0014] Based on the absorbance detection results and the fluorescence signal intensity detection results, the degree of hemolysis of the processed sample is detected to obtain the degree of hemolysis detection results.

[0015] Optionally, the step of detecting the degree of hemolysis of the processed sample based on the absorbance detection result and the fluorescence signal intensity detection result to obtain the hemolysis degree detection result includes:

[0016] The processed samples are classified using a pre-trained clustering model based on the absorbance detection results and the fluorescence signal intensity detection results to obtain classification results.

[0017] The degree of hemolysis of the processed sample is evaluated based on the classification results to obtain the hemolysis degree evaluation result, and the hemolysis degree evaluation result is used as the hemolysis degree detection result.

[0018] Optionally, the liquid processing module includes a test tube positioning module and a multi-channel pipette. The step of quantitatively adding red blood cell lysis buffer to the incubated sample test tube through the liquid processing module includes:

[0019] The test tube positioning module identifies the position information of the sample test tubes on the test tube rack.

[0020] Based on the sample tube position information, the multi-channel pipette is controlled to simultaneously dispense a preset volume of erythrocyte lysis buffer into multiple sample tubes;

[0021] Record the batch number and addition timestamp of the red blood cell lysis buffer, and update the sample processing progress database.

[0022] Optionally, after the step of detecting the degree of hemolysis of the processed sample using the optical detection module and obtaining the hemolysis degree detection result, the method further includes:

[0023] The preset threshold is compared with the hemolysis degree detection result, and a determination result of the hemolysis degree corresponding to the processed sample is generated based on the comparison result. The sample tubes are then classified and transferred according to the determination result.

[0024] If the judgment result is unqualified, the corresponding sample tube will be transferred to the secondary processing station;

[0025] After extending the lysis reaction time of the corresponding sample tube, the step of detecting the degree of hemolysis of the processed sample through the optical detection module and obtaining the hemolysis degree detection result is performed.

[0026] Optionally, the system further includes a data acquisition module;

[0027] The data acquisition module is used to detect the status and processing parameters of the sample tubes in each module of the system;

[0028] The status and processing parameters of each module in the system are stored in the sample processing progress database, and detection feedback data is generated based on the status and processing parameters of each module in the system.

[0029] Optionally, after the step of transferring the sample tube containing the lysis buffer to the mixing module for mechanical shaking to obtain the processed sample, the method further includes:

[0030] The processed sample is filtered using a flow cytometer to remove impurities.

[0031] Furthermore, to achieve the above objectives, the present invention also proposes a flow cytometry device for detecting the degree of hemolysis, the device comprising:

[0032] An automated incubation module is used to place sample tubes that have completed the antigen-antibody reaction and perform timed incubation;

[0033] The liquid processing module is used to quantitatively add red blood cell lysis buffer to the incubated sample tubes;

[0034] The mixing module is used to mechanically agitate the sample tubes containing the lysis buffer to obtain the processed samples.

[0035] An optical detection module is used to detect the degree of hemolysis in the processed sample and obtain the hemolysis degree detection result.

[0036] Furthermore, to achieve the above objectives, the present invention also proposes a flow cytometry hemolysis degree detection device, the device comprising: a memory, a processor, and a flow cytometry hemolysis degree detection program stored in the memory and executable on the processor, the flow cytometry hemolysis degree detection program being configured to implement the steps of the flow cytometry hemolysis degree detection method described above.

[0037] In addition, to achieve the above objectives, the present invention also proposes a storage medium storing a flow cytometry hemolysis degree detection program, wherein when the flow cytometry hemolysis degree detection program is executed by a processor, it implements the steps of the flow cytometry hemolysis degree detection method described above.

[0038] This invention discloses a method for incubating sample tubes that have completed antigen-antibody reactions in an automated incubation module for timed incubation; quantitatively adding red blood cell lysis buffer to the incubated sample tubes via a liquid processing module; transferring the sample tubes with added lysis buffer to a mixing module for mechanical agitation to obtain processed samples; and detecting the degree of hemolysis in the processed samples using an optical detection module to obtain hemolysis degree detection results. Because this invention achieves parallel and continuous processing of multiple sample tubes through the streamlined connection of the automated incubation module, liquid processing module, and mixing module, and performs online detection of the lysed samples using an optical detection module, compared to existing technologies, this invention improves the efficiency and accuracy of hemolysis degree detection. Attached Figure Description

[0039] Figure 1 This is a flowchart illustrating the first embodiment of the flow cytometry method for detecting the degree of hemolysis in cells according to the present invention.

[0040] Figure 2 This is a schematic flowchart of the second embodiment of the flow cytometry method for detecting the degree of hemolysis in cells according to the present invention;

[0041] Figure 3 This is a schematic flowchart of the third embodiment of the flow cytometry method for detecting the degree of hemolysis in cells according to the present invention;

[0042] Figure 4 This is a structural block diagram of the first embodiment of the flow cytometry hemolysis detection device of the present invention;

[0043] Figure 5 This is a schematic diagram of the structure of a flow cytometry device for detecting the degree of hemolysis in the hardware operating environment involved in the embodiments of the present invention.

[0044] The realization of the objective, functional features and advantages of the present invention will be further explained in conjunction with the embodiments and with reference to the accompanying drawings. Detailed Implementation

[0045] It should be understood that the specific embodiments described herein are for illustrative purposes only and are not intended to limit the scope of the invention.

[0046] This invention provides a flow cytometry method for detecting the degree of hemolysis in cells, referring to... Figure 1 , Figure 1 This is a schematic flowchart of the first embodiment of the flow cytometry method for detecting the degree of hemolysis in cells according to the present invention.

[0047] In this embodiment, the flow cytometry method for detecting the degree of hemolysis is applied to a flow cytometry system for detecting the degree of hemolysis. The system includes an automated incubation module, a liquid handling module, a mixing module, and an optical detection module. The method includes steps S10 to S40:

[0048] Step S10: Place the sample tube that has completed the antigen-antibody reaction into the automated incubation module for timed incubation.

[0049] It should be noted that the executing entity in this embodiment can be a computer server device with data processing, network communication, and program execution functions applied in the flow cytometry hemolysis degree detection scenario, such as a server, tablet computer, personal computer, etc., or an electronic device capable of realizing the above functions (such as a flow cytometry hemolysis degree detection device). The following uses a flow cytometry hemolysis degree detection system (hereinafter referred to as the system) including a flow cytometry hemolysis degree detection device as an example to illustrate this embodiment and the following embodiments.

[0050] It should be understood that the aforementioned sample tubes that have completed the antigen-antibody reaction can refer to test tubes in which the antibody has bound to its corresponding antigen to form an antigen-antibody complex, which can be directly used for subsequent red blood cell lysis and flow cytometry detection.

[0051] Understandably, timed incubation refers to the process of placing "sample tubes that have completed the antigen-antibody reaction" at a set temperature (usually room temperature or 37°C) and maintaining it for a preset time within an automated incubation module. This process is automatically timed by the system to ensure sufficient formation of the antigen-antibody complex and controllable reaction time.

[0052] Step S20: Add red blood cell lysis buffer quantitatively to the incubated sample tube through the liquid processing module.

[0053] Understandably, the incubated sample tubes can be those that have already undergone "timed incubation" in the automated incubation module, at which point the antigen-antibody complex reaction in the tubes has reached the preset duration.

[0054] It should be understood that quantitative addition of red blood cell lysis buffer can refer to the automatic and precise addition of red blood cell lysis buffer to each "incubated sample tube" according to a preset volume by a liquid processing module (such as a multi-channel pipette), ensuring that the amount added to each tube is consistent and avoiding volume deviations caused by manual operation.

[0055] It should be noted that the liquid processing module includes a tube positioning module and a multi-channel pipette. In specific implementation, the tube positioning module can identify the position information of sample tubes on the tube rack; based on the sample tube position information, the multi-channel pipette can be controlled to simultaneously dispense a preset volume of erythrocyte lysis buffer into multiple sample tubes; the batch number information and addition timestamp of the erythrocyte lysis buffer are recorded, and the sample processing progress database is updated.

[0056] Understandably, sample tube location information can refer to the physical coordinate data of each sample tube in the test tube rack and the entire system, which is obtained in real time by the test tube positioning module and is used to uniquely identify and locate each sample tube.

[0057] It should be understood that the sample processing progress database refers to the electronic data set within the system used to record and track the status of all sample tubes at each stage of the flow cytometry hemolysis detection process.

[0058] Step S30: Transfer the sample tube containing the lysis buffer to the mixing module for mechanical shaking to obtain the processed sample.

[0059] It should be noted that mechanical vibration treatment can refer to the controlled vibration of the sample tube after the addition of red blood cell lysis buffer by the mixing module through mechanical movement (such as horizontal, vertical or orbital oscillation), so as to promote the thorough mixing of lysis buffer and sample, accelerate the rupture of red blood cell membrane and uniform dispersion, thereby obtaining a homogeneous processed sample.

[0060] To avoid impurities forming precipitates during centrifugation and affecting cell recovery rate and final analysis results, in this specific implementation, after transferring the sample tube containing the lysis buffer to the mixing module for mechanical shaking to obtain the processed sample, the process further includes: filtering the processed sample using a flow cytometer to remove impurities from the processed sample.

[0061] Step S40: The hemolysis degree of the processed sample is detected by the optical detection module to obtain the hemolysis degree detection result.

[0062] It should be understood that the aforementioned optical detection module may refer to an optical unit integrated into a flow cytometry hemolysis degree detection system, which is capable of emitting light within a specific wavelength range and receiving transmitted light, scattered light, and / or fluorescence signals generated by the sample, and detecting the degree of hemolysis by analyzing changes in these optical signals.

[0063] It should be explained that hemolysis degree detection can refer to all measurement and analysis operations performed on a sample after red blood cell lysis using an optical detection module, including absorbance measurement, fluorescence signal acquisition and data processing, to obtain a process that reflects the degree of red blood cell damage.

[0064] Furthermore, the hemolysis degree detection result can refer to the quantitative or classification index output by the system after the above-mentioned "hemolysis degree detection" is completed, which is used to characterize whether the red blood cell lysis in the sample reaches the preset standard.

[0065] It should be noted that, in order to quickly locate the problem link based on the database records when abnormal test results or equipment alarms are detected, reducing troubleshooting time, the system also includes a data acquisition module. This module is used to detect the status and processing parameters of the sample tubes in each module of the system, store the status and processing parameters of each module in the sample processing progress database, and generate test feedback data based on the status and processing parameters of each module.

[0066] This embodiment discloses a method for incubating sample tubes that have completed antigen-antibody reactions in an automated incubation module for timed incubation; quantitatively adding red blood cell lysis buffer to the incubated sample tubes via a liquid processing module; transferring the sample tubes with added lysis buffer to a mixing module for mechanical agitation to obtain processed samples; and detecting the degree of hemolysis in the processed samples using an optical detection module to obtain hemolysis degree detection results. Because this embodiment achieves parallel and continuous processing of multiple sample tubes through the streamlined connection of the automated incubation module, liquid processing module, and mixing module, and performs online detection of the lysed samples using an optical detection module, this embodiment improves the efficiency and accuracy of hemolysis degree detection compared to existing technologies.

[0067] refer to Figure 2 , Figure 2 This is a flowchart illustrating the second embodiment of the flow cytometry method for detecting the degree of hemolysis in cells according to the present invention.

[0068] Based on the first embodiment described above, in this embodiment, step S40 includes steps S401 to S403:

[0069] Step S401: The absorbance of the processed sample at different wavelengths is detected by the optical detection module using multi-wavelength detection technology within a preset wavelength range to obtain absorbance detection results;

[0070] It should be explained that multi-wavelength detection technology refers to the optical detection module emitting and receiving light of at least two different wavelengths sequentially or simultaneously within the same measurement cycle, in order to obtain optical response data of the sample at different wavelengths, thereby enhancing the ability to distinguish specific components such as hemoglobin.

[0071] Accordingly, detecting the absorbance of the processed sample at different wavelengths refers to using the above-mentioned multi-wavelength detection technology to collect the transmittance or absorbance values ​​of the sample at each selected wavelength, forming an absorbance-wavelength dataset.

[0072] Furthermore, the absorbance detection result can refer to the set of absorbance values ​​of the sample at various selected wavelengths output by the above detection steps, which can be used as input parameters for subsequent hemolysis degree analysis and determination.

[0073] Step S402: Use fluorescence detection technology to detect the fluorescence signal intensity of each substance in the processed sample and obtain the fluorescence signal intensity detection results;

[0074] It should be explained that fluorescence detection technology can refer to the technique of using an optical detection module to irradiate a processed sample with light of a specific excitation wavelength, causing cells or components in the sample labeled with fluorescent antibodies to emit fluorescence signals of a longer wavelength, and then collecting and analyzing these fluorescence signals.

[0075] Correspondingly, fluorescence signal intensity detection refers to using the aforementioned fluorescence detection technology to measure the amplitude or integral value of the fluorescence signal of different cell populations or biomolecules labeled with fluorescent antibodies in the sample, so as to obtain fluorescence intensity data corresponding to each component.

[0076] Furthermore, the fluorescence signal intensity detection result can refer to the set of quantitative data output by the fluorescence detection step, which characterizes the intensity of fluorescence signals of each fluorescently labeled substance in the sample, and is used together with the absorbance detection result to assess the degree of hemolysis of the sample.

[0077] Step S403: Based on the absorbance detection result and the fluorescence signal intensity detection result, the hemolysis degree of the processed sample is detected to obtain the hemolysis degree detection result.

[0078] It should be noted that detecting the degree of hemolysis in processed samples based on absorbance and fluorescence signal intensity results can mean simultaneously inputting the absorbance and fluorescence signal intensity results into a trained clustering model (or an equivalent multi-parameter analysis model); the model classifies or scores the samples according to the preset hemolysis degree category boundaries, thereby outputting the corresponding hemolysis degree detection result for the sample.

[0079] Understandably, multi-wavelength absorbance detection can distinguish free hemoglobin from other interfering pigments, while fluorescence signal intensity detection can reflect the remaining intact cells and the binding status of labeled antibodies. The combination of the two significantly reduces misjudgments caused by a single indicator.

[0080] In a specific implementation, a pre-trained clustering model can be used to classify the processed sample based on the absorbance detection results and the fluorescence signal intensity detection results to obtain classification results; the degree of hemolysis of the processed sample can be evaluated based on the classification results to obtain a hemolysis degree evaluation result, and the hemolysis degree evaluation result can be used as the hemolysis degree detection result.

[0081] This embodiment discloses a method for detecting the absorbance of a processed sample at different wavelengths using a multi-wavelength detection technique within a preset wavelength range via an optical detection module, thereby obtaining absorbance detection results. Fluorescence detection technology is then used to detect the fluorescence signal intensity of each component in the processed sample, yielding fluorescence signal intensity detection results. Finally, based on the absorbance and fluorescence signal intensity detection results, the degree of hemolysis in the processed sample is detected, resulting in a hemolysis degree detection result. Compared to existing technologies, this embodiment improves the accuracy of hemolysis degree detection by using both absorbance and fluorescence signal intensity detection results to determine the degree of hemolysis.

[0082] refer to Figure 3 , Figure 3 This is a flowchart illustrating the third embodiment of the flow cytometry method for detecting the degree of hemolysis in cells according to the present invention.

[0083] Based on the above embodiments, in this embodiment, after step S40, steps S50 to S70 are further included:

[0084] Step S50: Compare the preset threshold with the hemolysis degree detection result, generate a determination result of the hemolysis degree corresponding to the processed sample based on the comparison result, and classify and transfer the sample test tubes according to the determination result.

[0085] Step S60: If the judgment result is unqualified, the corresponding sample tube is transferred to the secondary processing station.

[0086] Step S70: After extending the lysis reaction time of the corresponding sample tube, perform the step of detecting the degree of hemolysis of the processed sample through the optical detection module to obtain the hemolysis degree detection result.

[0087] It should be understood that classifying and transferring sample tubes according to the judgment results can refer to mechanical handling mechanisms (such as grippers or pipetting robots) transferring the tubes to different workstations according to the judgment results output by the system.

[0088] It should be noted that if the judgment result is qualified, the corresponding sample tube will be transferred to the centrifugation waiting station.

[0089] Understandably, a secondary processing station can refer to a physical location or functional module within the system specifically designed to further process sample tubes that were initially deemed "unqualified" in the hemolysis test. This typically includes operations such as extending the standing time, adding lysis buffer, remixing, or retesting to promote further lysis of red blood cells and reassess the degree of hemolysis.

[0090] Furthermore, extending the corresponding sample tube lysis reaction time can refer to keeping the sample tubes that were initially deemed unqualified by the hemolysis test in a static state or adding slight mixing at the secondary processing station, so that the contact time between the red blood cell lysis solution and the sample is increased compared to the initial setting, until the minimum lysis time required for the next test is met.

[0091] This embodiment discloses a method for comparing a preset threshold with the hemolysis degree detection result, generating a hemolysis degree determination result for the processed sample based on the comparison result, and classifying and transferring sample tubes according to the determination result. If the determination result is unqualified, the corresponding sample tube is transferred to a secondary processing station. After extending the lysis reaction time of the corresponding sample tube, the step of performing hemolysis degree detection on the processed sample through the optical detection module to obtain the hemolysis degree detection result is executed. Compared with the prior art, this embodiment not only allows samples determined to be unqualified to automatically enter the secondary processing station and extend the lysis time within the system, reducing manpower input and sample waste, but also allows some samples that are misjudged due to insufficient lysis to be corrected within the same batch, improving the overall first-pass yield.

[0092] Furthermore, this embodiment of the invention also proposes a storage medium storing a flow cytometry hemolysis degree detection program, which, when executed by a processor, implements the steps of the flow cytometry hemolysis degree detection method described above.

[0093] Reference Figure 4 , Figure 4 This is a structural block diagram of the first embodiment of the flow cytometry hemolysis detection device of the present invention.

[0094] like Figure 4 As shown, the flow cytometry hemolysis detection device proposed in this embodiment of the invention includes: an automated incubation module 501, a liquid processing module 502, a mixing module 503, and an optical detection module 504.

[0095] The automated incubation module 501 is used to place the sample tubes that have completed the antigen-antibody reaction and perform timed incubation.

[0096] The liquid processing module 502 is used to quantitatively add red blood cell lysis buffer to the incubated sample tube.

[0097] The mixing module 503 is used to mechanically agitate the sample tube containing the lysis buffer to obtain the processed sample.

[0098] The optical detection module 504 is used to detect the degree of hemolysis of the processed sample and obtain the hemolysis degree detection result.

[0099] The liquid processing module 502 is also used to identify the position information of sample tubes on the test tube rack through the test tube positioning module; control the multi-channel pipette to simultaneously distribute a preset volume of red blood cell lysis buffer to multiple sample tubes according to the sample tube position information; record the batch number information and addition timestamp of the red blood cell lysis buffer; and update the sample processing progress database.

[0100] The mixing module 503 is also used to filter the processed sample using a flow cytometer to remove impurities from the processed sample.

[0101] The system also includes a data acquisition module, which is used to detect the status and processing parameters of the sample tubes in each module of the system; store the status and processing parameters of each module of the system in a sample processing progress database; and generate detection feedback data based on the status and processing parameters of each module of the system.

[0102] This device embodiment discloses a process where sample tubes that have completed antigen-antibody reactions are placed in an automated incubation module for timed incubation; red blood cell lysis buffer is quantitatively added to the incubated sample tubes via a liquid processing module; the sample tubes with added lysis buffer are transferred to a mixing module for mechanical agitation to obtain processed samples; and the degree of hemolysis is detected by an optical detection module to obtain the hemolysis degree detection result. Because this device embodiment achieves parallel and continuous processing of multiple sample tubes through the streamlined connection of the automated incubation module, liquid processing module, and mixing module, and performs online detection of the lysed samples via the optical detection module, compared to existing technologies, this device embodiment improves the efficiency and accuracy of hemolysis degree detection.

[0103] Based on the first embodiment of the flow cytometry hemolysis degree detection device of the present invention, a second embodiment of the flow cytometry hemolysis degree detection device of the present invention is proposed.

[0104] In this embodiment, the optical detection module 504 is further configured to detect the absorbance of the processed sample at different wavelengths using multi-wavelength detection technology within a preset wavelength range, and obtain absorbance detection results; to detect the fluorescence signal intensity of each component in the processed sample using fluorescence detection technology, and obtain fluorescence signal intensity detection results; and to detect the degree of hemolysis of the processed sample based on the absorbance detection results and the fluorescence signal intensity detection results, and obtain hemolysis degree detection results.

[0105] The optical detection module 504 is further configured to classify the processed sample based on the absorbance detection result and the fluorescence signal intensity detection result using a pre-trained clustering model, and obtain a classification result; evaluate the degree of hemolysis of the processed sample based on the classification result, and obtain a hemolysis degree evaluation result, and use the hemolysis degree evaluation result as the hemolysis degree detection result.

[0106] Other embodiments or specific implementations of the flow cytometry hemolysis degree detection device of the present invention can be referred to the above-described method embodiments, and will not be repeated here.

[0107] This application provides a flow cytometry hemolysis degree detection device, which includes: at least one processor; and a memory communicatively connected to the at least one processor; wherein the memory stores instructions executable by the at least one processor, and the instructions are executed by the at least one processor to enable the at least one processor to perform the flow cytometry hemolysis degree detection method in the above embodiment 1.

[0108] The following is for reference. Figure 5 This document illustrates a structural schematic diagram of a flow cytometry hemolysis detection device suitable for implementing embodiments of this application. The flow cytometry hemolysis detection device in the embodiments of this application may include, but is not limited to, mobile terminals such as mobile phones, laptops, digital radio receivers, PDAs (Personal Digital Assistants), PADs (Portable Application Description), PMPs (Portable Media Players), in-vehicle terminals (e.g., in-vehicle navigation terminals), and fixed terminals such as digital TVs and desktop computers. Figure 5 The flow cytometry device for detecting the degree of hemolysis shown is merely an example and should not be construed as limiting the functionality and scope of the embodiments of this application.

[0109] like Figure 5As shown, the flow cytometry hemolysis level detection device may include a processing unit 1001 (e.g., a central processing unit, a graphics processing unit, etc.), which can perform various appropriate actions and processes according to a program stored in a read-only memory 1002 or a program loaded from a storage device 1003 into a random access memory 1004. The random access memory 1004 also stores various programs and data required for the operation of the flow cytometry hemolysis level detection device. The processing unit 1001, the read-only memory 1002, and the random access memory 1004 are interconnected via a bus 1005. An input / output interface 1006 is also connected to the bus. Typically, the following systems can be connected to the input / output interface 1006: input devices 1007 including, for example, a touch screen, touchpad, keyboard, mouse, image sensor, microphone, accelerometer, gyroscope, etc.; output devices 1008 including, for example, a liquid crystal display (LCD), speaker, vibrator, etc.; storage devices 1003 including, for example, magnetic tape, hard disk, etc.; and communication devices 1009. The communication device 1009 allows the flow cytometry hemolysis level detection device to communicate wirelessly or wiredly with other devices to exchange data. Although the figure shows flow cytometry hemolysis level detection devices with various systems, it should be understood that it is not required to implement or possess all of the systems shown. More or fewer systems may be implemented alternatively.

[0110] Specifically, according to the embodiments disclosed in this application, the processes described above with reference to the flowcharts can be implemented as computer software programs. For example, embodiments disclosed in this application include a computer program product comprising a computer program carried on a computer-readable medium, the computer program containing program code for performing the methods shown in the flowcharts. In such embodiments, the computer program can be downloaded and installed from a network via a communication device, or installed from storage device 1003, or installed from read-only memory 1002. When the computer program is executed by processing device 1001, it performs the functions defined in the methods of the embodiments disclosed in this application.

[0111] The flow cytometry hemolysis detection device provided in this application, employing the flow cytometry hemolysis detection method described in the above embodiments, can solve the technical problems of low efficiency and insufficient detection accuracy in existing hemolysis detection methods. Compared with the prior art, the beneficial effects of the flow cytometry hemolysis detection device provided in this application are the same as those of the flow cytometry hemolysis detection method provided in the above embodiments, and other technical features of this flow cytometry hemolysis detection device are the same as those disclosed in the previous embodiment method, and will not be repeated here.

[0112] It should be understood that the various parts disclosed in this application can be implemented using hardware, software, firmware, or a combination thereof. In the description of the above embodiments, specific features, structures, materials, or characteristics can be combined in any suitable manner in one or more embodiments or examples.

[0113] The above description is merely a specific embodiment of this application, but the scope of protection of this application is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the scope of the technology disclosed in this application should be included within the scope of protection of this application. Therefore, the scope of protection of this application should be determined by the scope of the claims.

[0114] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or system that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or system. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or system that includes that element.

[0115] The sequence numbers of the above embodiments of the present invention are for descriptive purposes only and do not represent the superiority or inferiority of the embodiments.

[0116] Through the above description of the embodiments, those skilled in the art can clearly understand that the methods of the above embodiments can be implemented by means of software plus necessary general-purpose hardware platforms. Of course, they can also be implemented by hardware, but in many cases the former is a better implementation method. Based on this understanding, the technical solution of the present invention, in essence, or the part that contributes to the prior art, can be embodied in the form of a software product. This computer software product is stored in a storage medium (such as read-only memory / random access memory, magnetic disk, optical disk) and includes several instructions to cause a terminal device (which may be a mobile phone, computer, server, air conditioner, or network device, etc.) to execute the methods described in the various embodiments of the present invention.

[0117] The above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Any equivalent structural or procedural transformations made based on the description and drawings of the present invention, or direct or indirect applications in other related technical fields, are similarly included within the protection scope of the present invention.

Claims

1. A flow cytometry method for detecting the degree of hemolysis, characterized in that, The method is applied to a flow cytometry system for detecting the degree of hemolysis, the system comprising an automated incubation module, a liquid handling module, a mixing module, and an optical detection module, and the method comprising: The sample tubes that have completed the antigen-antibody reaction are placed in the automated incubation module for timed incubation; The liquid processing module quantitatively adds red blood cell lysis buffer to the incubated sample tubes. The sample tube containing the lysis buffer is transferred to the mixing module for mechanical agitation to obtain the processed sample. The hemolysis degree of the processed sample is detected by the optical detection module to obtain the hemolysis degree detection result. The preset threshold is compared with the hemolysis degree detection result, and a judgment result of the hemolysis degree corresponding to the processed sample is generated based on the comparison result. The sample tubes are then classified and transferred according to the judgment result. If the judgment result is unqualified, the corresponding sample tube is transferred to the secondary processing station. After extending the lysis reaction time of the corresponding sample tube, the step of performing hemolysis degree detection on the processed sample through the optical detection module to obtain the hemolysis degree detection result is executed. The step of detecting the degree of hemolysis of the processed sample using the optical detection module to obtain the hemolysis degree detection result includes: using the optical detection module to detect the absorbance of the processed sample at different wavelengths using multi-wavelength detection technology within a preset wavelength range to obtain absorbance detection results; using fluorescence detection technology to detect the fluorescence signal intensity of each component in the processed sample to obtain fluorescence signal intensity detection results; and based on the absorbance detection results and the fluorescence signal intensity detection results, detecting the degree of hemolysis of the processed sample to obtain the hemolysis degree detection result.

2. The flow cytometry method for detecting the degree of hemolysis as described in claim 1, characterized in that, The step of detecting the degree of hemolysis of the processed sample based on the absorbance detection result and the fluorescence signal intensity detection result, and obtaining the hemolysis degree detection result, includes: The processed samples are classified using a pre-trained clustering model based on the absorbance detection results and the fluorescence signal intensity detection results to obtain classification results. The degree of hemolysis of the processed sample is evaluated based on the classification results to obtain the hemolysis degree evaluation result, and the hemolysis degree evaluation result is used as the hemolysis degree detection result.

3. The flow cytometry method for detecting the degree of hemolysis as described in claim 1, characterized in that, The liquid processing module includes a tube positioning module and a multi-channel pipette. The step of quantitatively adding red blood cell lysis buffer to the incubated sample tube through the liquid processing module includes: The test tube positioning module identifies the position information of the sample test tubes on the test tube rack. Based on the sample tube position information, the multi-channel pipette is controlled to simultaneously dispense a preset volume of erythrocyte lysis buffer into multiple sample tubes; Record the batch number and addition timestamp of the red blood cell lysis buffer, and update the sample processing progress database.

4. The flow cytometry method for detecting the degree of hemolysis as described in claim 1, characterized in that, The system also includes a data acquisition module; The data acquisition module is used to detect the status and processing parameters of the sample tubes in each module of the system; The status and processing parameters of each module in the system are stored in the sample processing progress database, and detection feedback data is generated based on the status and processing parameters of each module in the system.

5. The flow cytometry method for detecting the degree of hemolysis as described in any one of claims 1-4, characterized in that, After the step of transferring the sample tube containing the lysis buffer to the mixing module for mechanical agitation to obtain the processed sample, the method further includes: The processed sample is filtered using a flow cytometer to remove impurities.

6. A flow cytometry device for detecting the degree of hemolysis, characterized in that, The device includes: An automated incubation module is used to place sample tubes that have completed the antigen-antibody reaction and perform timed incubation; The liquid processing module is used to quantitatively add red blood cell lysis buffer to the incubated sample tubes; The mixing module is used to mechanically agitate the sample tubes containing the lysis buffer to obtain the processed samples. An optical detection module is used to detect the degree of hemolysis in the processed sample and obtain the hemolysis degree detection result. The optical detection module is further configured to compare a preset threshold with the hemolysis degree detection result, generate a hemolysis degree determination result corresponding to the processed sample based on the comparison result, and classify and transfer sample tubes according to the determination result; if the determination result is unqualified, the corresponding sample tube is transferred to a secondary processing station; after extending the lysis reaction time of the corresponding sample tube, the step of performing hemolysis degree detection on the processed sample through the optical detection module to obtain the hemolysis degree detection result is executed; The optical detection module is further configured to detect the absorbance of the processed sample at different wavelengths using multi-wavelength detection technology within a preset wavelength range, and obtain absorbance detection results; to detect the fluorescence signal intensity of each component in the processed sample using fluorescence detection technology, and obtain fluorescence signal intensity detection results; and to detect the degree of hemolysis of the processed sample based on the absorbance detection results and the fluorescence signal intensity detection results, and obtain hemolysis degree detection results.

7. A flow cytometry device for detecting the degree of hemolysis, characterized in that, The device includes: a memory, a processor, and a flow cytometry hemolysis degree detection program stored in the memory and executable on the processor, the flow cytometry hemolysis degree detection program being configured to implement the steps of the flow cytometry hemolysis degree detection method as described in any one of claims 1 to 5.

8. A storage medium, characterized in that, The storage medium stores a flow cytometry hemolysis degree detection program, which, when executed by a processor, implements the steps of the flow cytometry hemolysis degree detection method as described in any one of claims 1 to 5.

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