Cell analysis method, cell analyzer, and computer-readable storage medium

Abstract

A cell analysis method, cell analyzer, and computer-readable storage medium are disclosed. The method comprises: obtaining first and second original optical information of each cell to be tested in a sample solution to be tested; using the first original optical information as reference optical information to obtain a first set of valid detection data for each cell to be tested; using the second original optical information as reference optical information to obtain a second set of valid detection data for each cell to be tested; and determining, according to preset rules, final valid detection data for each cell to be tested from the first and second sets of valid detection data, for use in identifying target cells in the sample solution to be tested. The cell analysis method, cell analyzer, and computer-readable storage medium fully utilize cell characteristic information reflected by different optical signals, thereby improving the accuracy of cell classification, scatter plot morphology, and cell counting.

Description

Technical Field

[0001] The embodiments of the present invention relate to medical equipment technology, and in particular to a cell analysis method, a cell analyzer, and a computer-readable storage medium. Background Art

[0002] Currently, hematology analyzers typically use the laser scattering principle for cell detection and analysis, primarily employing low-angle scattered light signals (forward scattered light signal FS), side scattered light signals (SS), and side fluorescence signals (FL). FS characterizes cell volume, FS characterizes cell surface complexity, and FL characterizes the intracellular nuclear content. Light scattering cell detection technology combines the optical and biological properties of cells. Its rapidity, simplicity, accuracy, non-destructiveness, and reproducibility have led to significant development and widespread application in cell identification. When using the light scattering principle for detection, a light beam illuminates the cells to be detected, passing through the optical detection zone. The interaction between the light and the cells generates a scattered light signal, which contains information related to cell size and its distribution. If the cells to be detected are fluorescently stained, the irradiated cells also generate a fluorescent signal representing the intracellular nucleic acid content. When the cells to be tested pass rapidly through the optical detection zone, the intensities of the three optical signals (i.e., FS, SS, and FL) generated change instantaneously, generating light pulses. These pulses enter the photoelectric sensor and processing circuitry, which convert them into electrical pulses. The low-angle scattering signal is then used as a reference signal to identify the pulses generated by the cells to be tested, obtain the pulse amplitudes of the cells to be tested in the three optical signals, and count the number of pulses. By analyzing the pulse amplitude information, a scatter plot can be generated and cells can be classified, and the number of pulses can indicate the number of cells. However, currently, only low-angle scattering signals acquired at low angles are used for pulse identification. Since low-angle scattering signals represent cell volume information, they can better identify larger cells based on low-angle scattering signals. However, some very small cells may be missed or unable to be identified, leading to problems such as inaccurate cell classification, abnormal scatter plots, and inaccurate cell counts. Summary of the Invention

[0003] Embodiments of the present invention provide a cell analysis method, a cell analyzer, and a computer-readable storage medium to solve at least one of the above-mentioned problems.

[0004] According to a first aspect of the present invention, there is provided a cell analysis method comprising:

[0005] The step of acquiring original optical information: acquiring first original optical information detected by the first optical detector and second original optical information detected by the second optical detector when each cell to be tested in the sample liquid to be tested passes through the optical detection area of ​​the cell analyzer within a preset time period;

[0006] A first set of valid detection data acquisition step: using the first original optical information as reference optical information for identifying valid light pulses generated by each cell to be detected passing through the optical detection area, and determining first valid light pulse information and second valid light pulse information of each cell to be detected from the first original optical information and the second original optical information, respectively, to acquire a first set of valid detection data for each cell to be detected, including the first valid light pulse information and the second valid light pulse information;

[0007] A second set of valid detection data acquisition step: using the second original optical information as the reference optical information, determining third valid light pulse information and fourth valid light pulse information of each cell to be detected from the first original optical information and the second original optical information, respectively, to acquire a second set of valid detection data for each cell to be detected, including the third valid light pulse information and the fourth valid light pulse information;

[0008] Cell identification step: According to preset rules, the final valid detection data of each cell to be tested is determined from the first set of valid detection data and the second set of valid detection data of each cell to be tested, so as to identify the target cells in the sample liquid to be tested.

[0009] According to a second aspect of the present invention, there is provided a cell analysis method comprising:

[0010] Selecting the type of reference optical information according to the type of target cells, wherein the reference optical information is used to identify effective light pulses generated by each cell to be tested in the sample liquid to be tested passing through the optical detection area of ​​the cell analyzer;

[0011] Acquiring at least two types of optical information of each cell to be tested in the sample liquid to be tested when the cell passes through the optical detection area, wherein the at least two types of optical information include the reference optical information and at least one type of non-reference optical information;

[0012] determining, based on the reference optical information, valid detection data corresponding to the valid light pulse from the at least two types of optical information;

[0013] The target cells in the sample liquid to be tested are identified according to the effective detection data.

[0014] According to a third aspect of the present invention, there is provided a method for detecting platelets and / or reticulocytes, comprising:

[0015] Acquiring at least two types of optical information generated when each cell to be tested in the sample liquid to be tested passes through the optical detection area of ​​the cell analyzer, wherein the at least two types of optical information include side scattered light information or fluorescence information;

[0016] Using the side scattered light information or the fluorescence information as reference optical information for identifying effective light pulses generated by each of the cells to be detected passing through the optical detection area, and determining first effective detection data corresponding to the effective light pulse from the at least two types of optical information;

[0017] Platelets in the sample liquid to be tested are identified according to the first valid detection data.

[0018] According to a fourth aspect of the present invention, there is provided a cell analyzer comprising:

[0019] a sampling device having a pipette with a pipette nozzle and a driving device for driving the pipette to quantitatively draw a blood sample through the pipette nozzle;

[0020] A sample preparation device comprising a reaction pool and a reagent supply unit, wherein the reaction pool is used to receive a blood sample drawn by a sampling device, and the reagent supply unit provides a reagent to the reaction pool, so that the blood sample drawn by the sampling device and the reagent provided by the reagent supply unit are mixed in the reaction pool to prepare a sample liquid to be tested;

[0021] An optical detection device, comprising a light source, a flow chamber, and a light detector, wherein each cell to be tested in the sample liquid to be tested can flow in the flow chamber, the light emitted by the light source illuminates the cells in the flow chamber to generate optical information, and the light detector is used to collect the optical information; and

[0022] A data processing device is electrically connected to the optical detection device and includes a processor and a computer-readable storage medium storing a computer program, wherein the data processing device is configured to perform the various steps of the method described in the first to third aspects when the computer program is executed by the processor.

[0023] According to a fifth aspect of an embodiment of the present invention, there is provided a computer-readable storage medium, comprising: a program, wherein the program can be executed by a processor to implement the method as described in the first to third aspects.

[0024] The cell analysis method, cell analyzer, and computer-readable storage medium of the embodiments of the present invention use multiple different types of optical signals generated by cells as reference optical information for pulse identification, fully utilizing the cell characteristic information reflected by the different optical signals to avoid missed or unidentified target cells, thereby improving the accuracy of cell classification, scatter plot morphology, and cell counting, and thereby improving the accuracy of the blood cell analyzer. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0026] Figure 1 is a schematic structural diagram of a cell analyzer according to an embodiment of the present invention;

[0027] Figure 2 is a schematic diagram of a framework of an optical detection device according to an embodiment of the present invention;

[0028] Figure 3 is a schematic diagram of a framework of a control device according to an embodiment of the present invention;

[0029] Figure 4 It is a logical diagram of a cell analysis method in the prior art;

[0030] Figure 5 is a schematic flow chart of a cell analysis method according to an embodiment of the present invention;

[0031] Figures 6 to 8 is a logic diagram of a cell analysis method according to different embodiments of the present invention;

[0032] Figure 9 is a schematic flow chart of another cell analysis method according to an embodiment of the present invention;

[0033] Figure 10 FIG. 4 is a logic diagram of another cell analysis method according to an embodiment of the present invention. DETAILED DESCRIPTION

[0034] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0035] According to an embodiment of the present invention, a cell analyzer is provided. The cell analyzer includes a sampling device, a sample preparation device, an optical detection device, and a data processing device.

[0036] The sampling device comprises a pipette with a pipette nozzle and a driving device for driving the pipette to quantitatively draw a liquid sample to be measured through the pipette nozzle.

[0037] The sample preparation device comprises a reaction pool and a reagent supply unit, wherein the reaction pool is used to receive the liquid sample to be tested sucked by the sampling device, and the reagent supply unit provides at least one reagent to the reaction pool, so that the liquid sample to be tested sucked by the sampling device and the at least one reagent provided by the reagent supply unit are mixed in the reaction pool to prepare the sample liquid to be tested.

[0038] The optical detection device includes a light source, a flow chamber and at least two detectors. The cells in the sample liquid to be tested can flow in the flow chamber. The light emitted by the light source illuminates the cells in the flow chamber to generate original optical information. The detector is used to collect the original optical information.

[0039] The data processing device is electrically connected to the optical detection device and includes a processor and a computer-readable storage medium storing a computer program. The data processing device is configured to, when the computer program is executed by the processor, perform some, all, or any combination of the steps of the cell analysis method according to an embodiment of the present invention. The cell analysis method according to the present invention is described in detail below.

[0040] Alternatively, the processor may be a CPU, a GPU or other chip with computing power. In practical applications, the processor may be implemented by software, hardware, firmware or a combination thereof, and may be implemented by circuits, single or multiple application-specific integrated circuits (ASICs), digital signal processors (DSPs), digital signal processing devices (DSPDs), programmable logic devices (PLDs), field programmable gate arrays (FPGAs), central processing units (CPUs), controllers, microcontrollers, or at least one of microprocessors, so that the processor can execute some or all of the steps or any combination of the steps in the cell analysis methods of the various embodiments of the present application.

[0041] Optionally, the computer-readable storage medium may be a volatile memory or a non-volatile memory, or may include both volatile and non-volatile memories. The non-volatile memory may be a read-only memory, a programmable read-only memory, an erasable programmable read-only memory, an electrically erasable programmable read-only memory, a magnetic random access memory, a flash memory, a magnetic surface memory, an optical disc, or a read-only optical disc; and the magnetic surface memory may be a magnetic disk or a tape. The volatile memory may be a random access memory used as an external cache. By way of example and not limitation, many forms of RAM are available, such as static random access memory, synchronous static random access memory, dynamic random access memory, synchronous dynamic random access memory, double data rate synchronous dynamic random access memory, enhanced synchronous dynamic random access memory, synchronously linked dynamic random access memory, and direct memory bus random access memory. The memory described in the embodiments of the present invention is intended to include these and any other suitable types of memory.

[0042] In one embodiment, Figure 1 FIG2 shows a schematic diagram of the structure of a cell analyzer according to an embodiment of the present invention. Figure 1 The cell analyzer includes a first housing 100, a second housing 200, a sampling device 10, a sample preparation device 30, an optical detection device 50, a data processing device 70, and an output unit 90. In practical applications, the output unit 90 may serve as a user interface. The optical detection device 50 and the data processing device 70 are disposed within the second housing 200, on either side of the second housing 200. The sample preparation device 30 is disposed within the first housing 100, while the output unit 90 and the sampling device 10 are disposed on the exterior of the first housing 100.

[0043] It should be understood that the liquid sample to be tested can be any type of liquid sample containing cells, such as blood, body fluid, etc., and is not limited here. In this embodiment, the liquid sample to be tested is a blood sample as an example for description.

[0044] Optionally, the sampling device 10 has a sampling needle for collecting a blood sample and transferring the collected blood sample to the sample preparation device 30. According to different embodiments, the sampling device can collect multiple blood samples, provide them to different chambers of the sample preparation device for different processing, and then perform different tests.

[0045] Optionally, the sample preparation device 30 includes a reaction pool and a reagent supply unit. The reagent supply unit stores reagents for reacting with the blood sample, for example, at least a first reagent and / or a second reagent, and supplies the corresponding reagents to the reaction pool as needed. The first reagent may be a diluent or a hemolytic agent, wherein the hemolytic agent is used to break down red blood cells in the liquid sample to be tested into fragments while leaving the morphology of white blood cells in the liquid sample to be tested substantially intact. Furthermore, the second reagent may be a fluorescent dye used to stain cells in the liquid sample to be tested.

[0046] Optionally, the optical detection device 50 includes at least two of a forward scattered light detector, a side scattered light detector, and a side fluorescence detector. The forward scattered light detector is usually arranged on the straight line where the light source and the flow chamber are located, and is arranged on both sides of the flow chamber with the light source to detect the forward scattered light intensity or low-angle scattered light intensity of the cells flowing in the flow chamber. The side scattered light detector is usually arranged at a certain angle to the straight line where the light source and the flow chamber are located to detect the side scattered light intensity of the cells flowing in the flow chamber. The fluorescence detector is usually arranged at a certain angle to the straight line where the light source and the flow chamber are located to detect the side fluorescence intensity of the cells flowing in the flow chamber.

[0047] In one embodiment, see Figure 2 , Figure 2 An example of an optical detection device according to an embodiment of the present invention is shown. The optical detection device comprises a light source 101, a beam shaping assembly 102, a flow chamber 103, and a forward scattered light detector 104, arranged sequentially in a straight line. A dichroic mirror 106 is arranged on one side of the flow chamber 103 at a 45° angle to the straight line. A portion of the side light emitted by cells in the flow chamber 103 passes through the dichroic mirror 106 and is captured by a fluorescence detector 105, which is arranged at a 45° angle behind the dichroic mirror 106. Another portion of the side light is reflected by the dichroic mirror 106 and captured by a side scattered light detector 107, which is arranged at a 45° angle in front of the dichroic mirror 106.

[0048] It should be understood that the straight line where the light source and the flow chamber are located can be in a straight line with the forward scattered light detector, or can form a certain angle. The angle can be set as needed and is not limited here. Similarly, the angle between the straight line where the light source and the flow chamber are located and the above-mentioned dichroic mirror, as well as the angles between the dichroic mirror and the side scattered light detector and the fluorescence detector respectively, can all be set as needed and are not limited here.

[0049] The data processing device 70 is configured to detect the target cells flowing through the flow chamber based on the at least two light intensity signals of the scattered light, and obtain corresponding detection results.

[0050] The output unit 90 is configured to output the test results. The output unit 90 may be a display device (not shown) for displaying the test results, which may be provided on the cell analyzer or external to the cell analyzer and electrically connected to the data processing device 70. Furthermore, the display device may be a touch screen display, a liquid crystal display, or a display screen on an electronic device such as a mobile phone or tablet computer.

[0051] According to an embodiment of the present invention, the cell analyzer may also provide a corresponding operation interface for the operator to operate. The operation interface may include various corresponding controls, such as a selection box or a menu bar, so that the operator can enter operating instructions on the operation interface according to actual usage to perform cell analysis using the cell analyzer. Furthermore, the output unit 90 provides the operation interface.

[0052] In one embodiment, Figure 3 As shown, the data processing device 70 includes at least a processor 71, RAM 72, ROM 73, a communication interface 74, a memory 76, and an I / O interface 75. The processor 71, RAM 72, ROM 73, communication interface 74, memory 76, and I / O interface 75 communicate via a bus 77. The processor can be a CPU, GPU, or other chip with computing power. The memory 76 contains various computer programs, such as an operating system and application programs, for execution by the processor 71, as well as the data required to execute these computer programs. In addition, during the cell detection process, any data that needs to be stored locally can be stored in the memory 76. The I / O interface 75 is composed of serial interfaces such as USB, IEEE1394, or RS-232C, parallel interfaces such as SCSI, IDE, or IEEE1284, and analog signal interfaces composed of D / A converters and A / D converters. An input device, such as a keyboard, mouse, touch screen, or other control buttons, is connected to the I / O interface 75, allowing the user to directly input data into the data processing device 70 using the input device. Additionally, the I / O interface 75 can be connected to a display with a display function, such as an LCD screen, a touch screen, or an LED display. The data processing device 70 can output processed data, such as analytical data or instrument operating parameters, to the display as graphical display data for display. The communication interface 74 can be an interface that uses any currently known communication protocol. The communication interface 74 communicates with the outside world via a network. The data processing device 70 can transmit data to any device connected to the network using a specific communication protocol via the communication interface 74.

[0053] In current practical applications, such as Figure 4As shown, a conventional cell analysis method is illustrated. In the prior art, during the analysis of cells in a liquid sample to be tested, pulse identification is performed using forward scattered light information (FS signal) collected by a forward scattered light detector as reference optical information, obtaining pulse information of the forward scattered light information, including the number of pulses, FS pulse amplitude H1, and FS pulse position t (i.e., the time corresponding to the identified pulse). Then, based on the pulse position t corresponding to each pulse identified based on the forward scattered light information, the pulse amplitudes H2 and H3 corresponding to the side scattered light information (SS signal) collected by the side scattered light detector and the side fluorescence information (FL signal) collected by the side fluorescence detector at pulse position t are respectively obtained. For all identified pulses, three FS-based amplitude data (H1, H2, H3) corresponding to the forward scattered light information, side scattered light information, and side fluorescence information can be obtained. Cell classification is then performed based on the three FS-based amplitude data (H1, H2, H3) and a corresponding scatter plot is obtained. The number of target cells is also determined based on the number of pulses. However, as mentioned above, FS represents cell volume information. For some cells (such as platelets) that are very small but have a complex cell surface (i.e., high SS signal intensity) or a high nuclear count content (i.e., high FL signal intensity), using only the FS signal as a reference signal for pulse recognition will result in missed recognition or failure to recognize, leading to inaccurate cell classification, abnormal scatter plots, and inaccurate cell counts.

[0054] Based on the above considerations, a cell analysis method is provided according to an embodiment of the present invention. Figure 5 , which shows a schematic flow chart of a cell analysis method according to an embodiment of the present invention. Figure 5 As shown, the method 500 includes:

[0055] Original optical information acquisition step S510: acquiring first original optical information detected by the first optical detector and second original optical information detected by the second optical detector when each cell to be tested in the sample liquid to be tested passes through the optical detection area of ​​the cell analyzer within a preset time period;

[0056] A first set of valid detection data acquisition step S520: using the first original optical information as reference optical information for identifying valid light pulses generated by each cell to be detected passing through the optical detection area, determining first valid light pulse information and second valid light pulse information of each cell to be detected from the first original optical information and the second original optical information, respectively, to acquire a first set of valid detection data for each cell to be detected, including the first valid light pulse information and the second valid light pulse information;

[0057] A second set of valid detection data acquisition step S530: using the second original optical information as the reference optical information, determining third valid light pulse information and fourth valid light pulse information of each cell to be detected from the first original optical information and the second original optical information, to acquire a second set of valid detection data for each cell to be detected, including the third valid light pulse information and the fourth valid light pulse information;

[0058] Cell identification step S540: According to preset rules, final valid detection data of each cell to be tested is determined from the first set of valid detection data and the second set of valid detection data of each cell to be tested, so as to identify target cells in the sample liquid to be tested.

[0059] In an embodiment of the present invention, the original optical information may be a continuous signal, such as an electrical signal, detected and output by an optical detector within a predetermined time period. Reference optical information refers to original optical information whose identified pulse positions are used as a reference to obtain pulse information of other original optical information. Pulse information obtained from the reference optical information and pulse information obtained from other original optical information based on the reference optical information may be recorded as effective optical pulse information.

[0060] For example, when the first original optical information is used as the reference optical information, n pulses are identified from the first original optical information and n pulse information corresponding to these n pulses is obtained, which is recorded as the first valid optical pulse information. Then, based on the n pulse positions of these n pulses, pulse information corresponding to these n pulse positions is obtained from the second original optical information and recorded as the second valid optical pulse information. The first valid optical pulse information and the second valid optical pulse information are recorded as the first set of valid detection data. When the second original optical information is used as the reference optical information, m pulses are identified from the second original optical information and m pulse information corresponding to these m pulses is obtained, which is recorded as the fourth valid optical pulse information. Then, based on the m pulse positions of these m pulses, pulse information corresponding to these m pulse positions is obtained from the first original optical information and recorded as the third valid optical pulse information. The third valid optical pulse information and the fourth valid optical pulse information are recorded as the second set of valid detection data. Finally, based on the intelligent screening algorithm (i.e., pre-set rules), final valid detection data is obtained from the first and second sets of valid data to identify target cells.

[0061] The cell analysis method according to an embodiment of the present invention uses different optical information collected by at least two optical detectors as reference optical information for pulse recognition. This method leverages the principle that different optical information reflects different cellular characteristics, complementing each other in pulse recognition and preventing missed or unrecognized target cells. Compared to traditional cell analysis methods that use only forward scattered light as reference optical information, this method overcomes the limitations of pulse recognition based on a single optical signal and significantly improves the accuracy of cell analysis.

[0062] Optionally, the first original optical information and the second original optical information are different optical information, and are respectively selected from one of forward scattered light information, side scattered light information and fluorescence information.

[0063] In some embodiments, the first original optical information or the second original optical information is fluorescence information. When fluorescence information is used as reference optical information, it is particularly suitable for identifying small cells in the sample liquid to be tested, such as platelets, reticulocytes, etc.

[0064] In some embodiments, the first set of valid detection data acquisition step S520 includes:

[0065] Based on the first amplitude threshold, the pulses generated when each cell to be tested passes through the optical detection area are identified from the first original optical information and first effective light pulse information of each cell to be tested is obtained, wherein the first effective light pulse information includes first pulse amplitude information and first time information.

[0066] acquiring second effective optical pulse information of each cell to be tested from the second original optical information according to the first time information, wherein the second effective optical pulse information includes second pulse amplitude information;

[0067] Accordingly, the second set of valid detection data acquisition step S530 includes:

[0068] Based on the second amplitude threshold, the pulses generated when each cell to be tested passes through the optical detection area are identified from the second original optical information and fourth effective light pulse information of each cell to be tested is obtained, wherein the fourth effective light pulse information includes fourth pulse amplitude information and second time information.

[0069] According to the second time information, third effective light pulse information of each cell to be measured is acquired from the first original optical information, where the third effective light pulse information includes third pulse amplitude information.

[0070] Among them, for different original optical information, different amplitude thresholds can be set for identifying pulses. If the peak value of the pulse identified from the original optical information is greater than the amplitude threshold, the pulse is considered to be a pulse generated by the cell passing through the optical detection area; if the peak value of the pulse identified from the original optical information is less than the amplitude threshold, the pulse is considered not to be a pulse generated by the cell, but may be a pulse generated by signal interference. In other words, when no cells pass through the optical detection area, the first original optical information and the second original optical information are usually maintained below their respective DC baseline voltages. When a cell passes through the optical detection area and generates optical information, the amplitude of the first original optical information and / or the second original optical information changes. Whether it is a valid pulse can be identified by judging the trend of the rising and falling edges of the pulse, and the difference between the pulse peak and the baseline is recorded as the pulse amplitude and the corresponding peak position (i.e., the time when the pulse is generated) is recorded. The number of cells can be calculated by accumulating the number of valid pulses.

[0071] like Figure 6 As shown, in some embodiments, the first original optical information is forward scattered light information or side scattered light information, and the second original optical information is fluorescence information. Figure 1 and Figure 2 , the sampling device 10 in the cell analyzer collects a blood sample through a sampling needle and transports the collected blood sample to the sample preparation device 30. The reagent supply part in the sample preparation device 30 supplies the diluent and the fluorescent dye to the reaction pool as needed, and the blood sample reacts with the diluent and the fluorescent dye in the reaction pool in turn to obtain a liquid sample to be tested containing multiple cells, and makes the cells in the liquid sample to be tested line up one by one and flow through the flow chamber of the optical detection device 50. The light source of the optical detection device 50 emits a light beam to the optical detection area of ​​the flow chamber, and the cells passing through the optical detection area generate different optical information after being excited by light. The forward scattered light detector or the side scattered light detector detects the forward scattered light intensity or the side scattered light intensity of the cells flowing in the flow chamber to obtain the first original optical information, and the fluorescence detector detects the side fluorescence intensity of the cells flowing in the flow chamber to obtain the second original optical information. After obtaining the first original optical information and the second original optical information, the data processing device 70 performs cell analysis. When the forward scattered light information or the side scattered light information (for example Figure 6 When the forward scattered light information in the image is used as the reference optical information, n pulses (for example, Figure 6 The number of pulses in the image is 1) and n first effective optical pulse information corresponding to the n pulses is obtained, each first effective optical pulse information including first pulse amplitude information H11 (for example, Figure 6 FS pulse amplitude in ) and the first time information t1 (e.g., Figure 6FS pulse position in the fluorescence information); then, according to the first time information t1, the second pulse amplitude information H12 corresponding to the time t1 in the fluorescence information is obtained, thereby obtaining n second effective light pulse information including the second pulse amplitude information H12, and then obtaining n first groups of effective detection data, that is, n (t1, H11, H12) (for example, Figure 6 When the fluorescence information is used as the reference optical information, m pulses are identified from the fluorescence information (e.g., Figure 6 The number of pulses in 3) and obtain m fourth effective optical pulse information corresponding to the m pulses, each fourth effective optical pulse information including fourth pulse amplitude information H24 (for example, Figure 6 FL pulse amplitude in) and second time information t2 (e.g., Figure 6 FL pulse position in the forward scattered light information or the side scattered light information); then, according to the second time information t2, the third pulse amplitude information H23 corresponding to the time near t2 in the forward scattered light information or the side scattered light information is obtained, thereby obtaining m third effective light pulse information including the third pulse amplitude information H23, and further obtaining m second groups of effective detection data, i.e., m (t2, H23, H24) (for example, Figure 6 In other words, after receiving the two original optical information, the first original optical information is used to perform pulse identification and obtain pulse amplitude information and position information, and the number of pulses is counted, and the pulses at the same position on the second original optical information are found and the amplitude information is obtained; at the same time, the second original optical information is used to perform pulse identification and obtain pulse amplitude information and position information, and the number of pulses is counted, and the pulses at the same position on the first original optical information are found and the amplitude information is obtained. Finally, according to the preset rules (for example, Figure 6 The method uses an intelligent screening algorithm (e.g., an algorithm comprising a plurality of first and second valid detection data) to determine final valid detection data from the n first and m second valid detection data sets, for use in identifying and counting target cells and optionally generating a scatter plot. The pulse amplitude information in the final valid detection data is used to identify target cells, and the pulse amplitude information and time information (pulse position information) in the final valid detection data are used to determine the number of target cells.

[0072] In some embodiments, the first original optical information and the second original optical information may also be forward scattered light information and side scattered light information, respectively. Figure 1 and Figure 2, the sampling device 10 in the cell analyzer collects a blood sample through a sampling needle and transports the collected blood sample to the sample preparation device 30. The reagent supply part in the sample preparation device 30 supplies the diluent to the reaction pool as needed, and the blood sample reacts with the diluent to obtain a liquid sample to be tested containing multiple cells, and makes the cells in the liquid sample to be tested line up one by one and flow through the flow chamber of the optical detection device 50. The light source of the optical detection device 50 emits a light beam to the optical detection area of ​​the flow chamber, and the cells passing through the optical detection area generate different optical information after being excited by light. The forward scattered light detector and the side scattered light detector respectively detect the forward scattered light intensity and the side scattered light intensity of the cells flowing in the flow chamber to obtain the first original optical information and the second original optical information. After obtaining the first original optical information and the second original optical information, the data processing device 70 performs cell analysis. The specific cell analysis process can be referred to in combination with the above. Figure 6 The described embodiments will not be described in detail here.

[0073] In some embodiments, the original optical information acquisition step S510 further includes: acquiring third original optical information detected in the third optical detector when each cell to be tested in the sample liquid to be tested passes through the optical detection area within the preset time period; accordingly, the first set of valid detection data acquisition step S520 further includes: using the first original optical information as the reference optical information, determining the fifth valid light pulse information of each cell to be tested from the third original optical information, and the first set of valid detection data further includes the fifth valid light pulse information; accordingly, the second set of valid detection data acquisition step S530 further includes: using the second original optical information as the reference optical information, determining the sixth valid light pulse information of each cell to be tested from the third original optical information, and the second set of valid detection data further includes the sixth valid light pulse information.

[0074] Optionally, the first original optical information, the second original optical information and the third original optical information are different from each other and are respectively selected from one of forward scattered light information, side scattered light information and fluorescence information.

[0075] like Figure 7 As shown, in some embodiments, the first raw optical information is forward scattered light information, the second raw optical information is fluorescence information, and the third raw optical information is side scattered light information. When the forward scattered light information is used as the reference optical information, n pulses (i.e., Figure 7 The number of pulses in the image is 1) and n first effective optical pulse information corresponding to the n pulses is obtained, each first effective optical pulse information includes first pulse amplitude information H11 (ie, Figure 7 FS pulse amplitude in) and the first time information t1 (ie, Figure 7 FS pulse position in the fluorescence information); then, according to the first time information t1, the second pulse amplitude information H12 corresponding to the time t1 in the fluorescence information and the fifth pulse amplitude information H15 corresponding to the time t1 in the side scattered light information are obtained, thereby respectively obtaining n second effective light pulse information including the second pulse amplitude information H12 and n fifth effective light pulse information including the fifth pulse amplitude information H15, thereby obtaining n first groups of effective detection data, i.e., n (t1, H11, H12, H15) (i.e., Figure 7 When the fluorescence information is used as the reference optical information, m pulses are identified from the fluorescence information (i.e., Figure 7 The number of pulses in 3) and obtain m fourth effective light pulse information corresponding to the m pulses, each fourth effective light pulse information includes fourth pulse amplitude information H24 (ie, Figure 7 FL pulse amplitude in) and the second time information t2 (ie, Figure 7 FL pulse position in the forward scattered light information); then, according to the second time information t2, the third pulse amplitude information H23 corresponding to the time near t2 in the forward scattered light information and the sixth pulse amplitude information H26 corresponding to the time near t2 in the side scattered light information are obtained, thereby respectively obtaining m third valid light pulse information including the third pulse amplitude information H23 and m sixth valid light pulse information including the sixth pulse amplitude information H26, thereby obtaining m second groups of valid detection data, i.e., m (t2, H23, H24, H26) (i.e., Figure 7 (the three amplitude data based on FL in ).

[0076] It should be understood that the first, second, and third raw optical information are merely used to distinguish different types of raw optical information, and there is no ordering relationship between them. Therefore, the first raw optical signal could also be side scattered light information or fluorescence information, the second raw optical information could also be forward scattered light information or fluorescence information, and the third raw optical information could also be forward scattered light information or side scattered light information. It only matters that the first, second, and third raw optical information are distinct from one another.

[0077] When two raw optical signals are used as reference optical information to identify pulses, compared to using a single raw optical signal as a reference optical signal, missed cell identification can be avoided, thereby improving the accuracy of cell analysis. Furthermore, when three raw optical signals are used as reference optical information, the accuracy of cell analysis can be further improved.

[0078] Therefore, according to an embodiment of the present invention, the method further includes a third set of steps for acquiring valid detection data:

[0079] Using the third original optical information as the reference optical information, determining seventh valid light pulse information, eighth valid light pulse information, and ninth valid light pulse information for each cell to be tested from the first original optical information, the second original optical information, and the third original optical information, respectively, to obtain a third set of valid detection data for each cell to be tested, including the seventh valid light pulse information, the eighth valid light pulse information, and the ninth valid light pulse information;

[0080] Accordingly, the cell identification step S540 includes: determining final valid detection data of each cell to be tested from the first set of valid detection data, the second set of valid detection data and the third set of valid detection data according to preset rules, so as to identify the target cells in the sample liquid to be tested.

[0081] Optionally, the first set of valid detection data acquisition step S520 includes:

[0082] Based on the first amplitude threshold, the pulses generated when each cell to be tested passes through the optical detection area are identified from the first original optical information and first effective light pulse information of each cell to be tested is obtained, wherein the first effective light pulse information includes first pulse amplitude information and first time information.

[0083] Acquire, according to the first time information, second effective light pulse information and fifth effective light pulse information of each cell to be tested from the second original optical information and the third original optical information, wherein the second optical information and the fifth effective light pulse information respectively include second pulse amplitude information and fifth pulse amplitude information;

[0084] The second set of valid detection data acquisition step S520 includes:

[0085] Based on the second amplitude threshold, the pulses generated when each cell to be tested passes through the optical detection area are identified from the second original optical information and fourth effective light pulse information of each cell to be tested is obtained, wherein the fourth effective light pulse information includes fourth pulse amplitude information and second time information.

[0086] Acquire third effective light pulse information and sixth effective light pulse information of each cell to be tested from the first original optical information and the third original optical information according to the second time information, wherein the third effective light pulse information and the sixth effective light pulse information respectively include third pulse amplitude information and sixth pulse amplitude information;

[0087] The third set of steps for obtaining effective detection data includes:

[0088] Based on the third amplitude threshold, the pulses generated when each cell to be tested passes through the optical detection area are identified from the third original optical information and ninth effective optical pulse information of each cell to be tested is obtained, wherein the ninth effective optical pulse information includes ninth pulse amplitude information and third time information.

[0089] According to the third time information, the seventh effective light pulse information and the eighth effective light pulse information of each cell to be tested are respectively obtained from the first original optical information and the second original optical information, wherein the seventh effective light pulse information and the eighth effective light pulse information respectively include the seventh pulse amplitude information and the eighth pulse amplitude information.

[0090] refer to Figure 8 , when the first original optical information is used as the reference optical information, n pulses (for example, Figure 8 The number of pulses in the image is 1) and n first effective optical pulse information corresponding to the n pulses is obtained, each first effective optical pulse information including first pulse amplitude information H11 (for example, Figure 8 FS pulse amplitude in ) and the first time information t1 (e.g., Figure 8 FS pulse position in the first time information); then, according to the first time information t1, the second pulse amplitude information H12 corresponding to the time t1 in the second original optical information and the fifth pulse amplitude information H15 corresponding to the time t1 in the third original optical information are obtained, thereby respectively obtaining n second valid optical pulse information including the second pulse amplitude information H12 and n fifth valid optical pulse information including the fifth pulse amplitude information H15, thereby obtaining n first groups of valid detection data, that is, n (t1, H11, H12, H15) (for example, Figure 8 When the second original optical information is used as the reference optical information, m pulses are identified from the second original optical information based on the second amplitude threshold (for example, Figure 8 2) and obtain m fourth effective optical pulse information corresponding to the m pulses, each fourth effective optical pulse information including fourth pulse amplitude information H24 (for example, Figure 8 SS pulse amplitude in) and second time information t2 (e.g., Figure 8SS pulse position in the first original optical information); then, according to the second time information t2, the third pulse amplitude information H23 corresponding to the time t2 in the first original optical information and the sixth pulse amplitude information H26 corresponding to the time t2 in the third original optical information are obtained, thereby respectively obtaining m third valid optical pulse information including the third pulse amplitude information H23 and m sixth valid optical pulse information including the sixth pulse amplitude information H26, thereby obtaining m second groups of valid detection data, i.e., m (t2, H23, H24, H26) (for example, Figure 8 When the third original optical information is used as the reference optical information, i pulses are identified from the third original optical information based on the third amplitude threshold (for example, Figure 8 The number of pulses in 3) and obtain i ninth effective optical pulse information corresponding to the i pulses, each ninth effective optical pulse information includes ninth pulse amplitude information H39 (for example, Figure 8 FL pulse amplitude in ) and third time information t3 (e.g., Figure 8 FL pulse position in the first original optical information); then, according to the third time information t3, the seventh pulse amplitude information H37 corresponding to the time t3 in the first original optical information and the eighth pulse amplitude information H38 corresponding to the time t3 in the second original optical information are obtained, thereby respectively obtaining i seventh valid optical pulse information including the seventh pulse amplitude information H37 and i eighth valid optical pulse information including the eighth pulse amplitude information H38, and further obtaining i third groups of valid detection data, i.e., i (t3, H37, H38, H39) (for example, Figure 8 In other words, after receiving three different kinds of original optical information, the first original optical information is used to perform pulse identification and obtain pulse amplitude information and position information, and the number of pulses is counted at the same time, and pulses at the same position are found on the second original optical information and the third original optical information and amplitude information is obtained; at the same time, the second original optical information is used to perform pulse identification and obtain pulse amplitude information and position information, and the number of pulses is counted at the same time, and pulses at the same position are found on the first original optical information and the third original optical information and amplitude information is obtained; at the same time, the third original optical information is used to perform pulse identification and obtain pulse amplitude information and position information, and the number of pulses is counted at the same time, and pulses at the same position are found on the first original optical information and the second original optical information and amplitude information is obtained. Finally, according to the preset rules (for example, Figure 8 ), the final valid detection data is determined from n first-group valid detection data, m second-group valid data, and i third-group valid data, for identifying and counting target cells, and optionally for generating a scatter plot.

[0091] In an embodiment of the present invention, Figures 6 to 8 The "acquisition amplitude" in the figure means that according to the pulse position obtained by pulse identification based on one original optical information, a pulse is searched near the same position on other original optical information, and the difference between its peak value and the baseline is recorded as the pulse amplitude.

[0092] In some embodiments, the cell identification step S540 includes:

[0093] Determine whether the time information in each group of valid detection data belongs to the same cell;

[0094] For the case of the same cell, one group of valid detection data corresponding to the same cell is selected as the final valid detection data of the same cell;

[0095] In the case of belonging to different cells, the valid detection data corresponding to the time information belonging to the different cells is used as the final valid detection data of the different cells.

[0096] In other words, the final valid detection data for each cell is determined by comparing the time information in each set of valid detection data for each cell. In other words, when multiple sets of valid detection data are obtained for the same cell, one set of valid detection data is selected as the final valid detection data for that cell; when only one set of valid detection data is obtained for the same cell, that set of valid detection data is used as the final valid detection data for that cell.

[0097] In some embodiments, when two different original optical information are used as reference optical information to identify pulses, the final valid detection data is determined by comparing the time information in each group of valid detection data of each cell to be tested, including:

[0098] Determine whether the first time information t1 in each first set of valid detection data is identical or substantially identical to the second time information t2 in each second set of valid detection data, that is, whether they belong to the same cell;

[0099] For substantially identical first time information t1 and second time information t2 belonging to the same cell, selecting the first set of valid detection data or the second set of valid detection data corresponding to the same cell as the final valid detection data of the same cell;

[0100] For different first time information and second time information belonging to different cells, the first group of valid detection data and the second group of valid detection data corresponding to the first time information and the second time information belonging to different cells are respectively used as the final valid detection data of the different cells.

[0101] In some embodiments, when three different original optical information are used as reference optical information to identify pulses, the final valid detection data is determined by comparing the time information in each group of valid detection data of each cell to be tested, including:

[0102] Determine whether the first time information t1 in each first set of valid detection data, the second time information t2 in each second set of valid detection data, and the third time information t3 in the third set of valid detection data are identical or substantially identical, that is, whether they belong to the same cell;

[0103] For the first time information t1, the second time information t2, and the third time information t3 belonging to the same cell, selecting the first set of valid detection data, the second set of valid detection data, or the third set of valid detection data corresponding to the same cell as the final valid detection data of the same cell;

[0104] For the first time information t1 and the second time information t2 belonging to the same cell, selecting the first set of valid detection data or the second set of valid detection data corresponding to the same cell as the final valid detection data of the same cell;

[0105] For the first time information t1 and the third time information t3 belonging to the same cell, selecting the first set of valid detection data or the third set of valid detection data corresponding to the same cell as the final valid detection data of the same cell;

[0106] For the second time information t2 and the third time information t3 belonging to the same cell, selecting the second set of valid detection data or the third set of valid detection data corresponding to the same cell as the final valid detection data of the same cell;

[0107] For the first time information t1, the second time information t2 and the third time information t3 that do not belong to the same cell, the first set of valid detection data, the second set of valid detection data and the third set of valid detection data corresponding to the first time information, the second time information and the third time information belonging to different cells are respectively used as the final valid detection data of the different cells.

[0108] That is, under normal circumstances, the pulses generated by cells passing through the optical detection zone can be identified in two or three different types of raw optical information. Therefore, for cells identified in multiple different types of raw optical information, the two or three sets of valid detection data obtained using the multiple different types of raw optical information as reference optical information are essentially consistent. In this case, any one set of data can be used as the final valid detection data for cells identified in multiple types of raw optical information. However, the pulses generated by certain very small cells (such as platelets) cannot be identified in forward scattered light information (the pulse amplitude of the forward scattered light information may be 0 or less than the amplitude threshold) and can only be identified in fluorescence information or side scattered light information. In this case, the set of valid detection data obtained using fluorescence information or side scattered light information as reference optical information is used as the final valid detection data for these very small cells.

[0109] In some embodiments, the cell identification step S540 includes determining final valid detection data for the same cell by comparing the amplitude information in the valid detection data sets corresponding to the same cell, or by comparing the amplitude information in the valid detection data sets corresponding to the same cell, or by comparing the amplitude information in the valid detection data sets corresponding to the same cell, or by comparing the amplitude information in the valid detection data sets based on the type of the target cell. In other words, for multiple sets of valid detection data obtained for the same cell, one set of data is selected based on the type of the target cell, or one set of data is selected based on the difference in peak values ​​between the valid detection data sets.

[0110] For example, the group of valid detection data corresponding to the same cell with the largest sum of amplitudes can be selected as the final valid detection data of the same cell. The sum of amplitudes of a group of valid detection data refers to the sum of pulse amplitudes corresponding to each of the original optical information at the same pulse position for the same cell.

[0111] In some embodiments, when different first and second original optical information are used as reference optical information, if a first set of valid detection data and a second set of valid detection data are obtained for the same cell, wherein the first set of valid detection data includes amplitude information (H11, H12) or (H11, H12, H15) and first time information t1, and the second set of valid detection data includes amplitude information (H23, H24) or (H23, H24, H26) and second time information t2, and t1 is substantially the same as t2. In this case, the sum of the amplitudes of the first set of valid detection data is H11+H12 or H11+H12+H15, and the sum of the amplitudes of the second set of valid detection data is H23+H24 or H23+H24+H26, then the valid detection data corresponding to the larger value of H11+H12 (or H11+H12+H15) and H23+H24 (or H23+H24+H26) can be selected as the final valid detection data of the cell. Further, when only the first set of valid detection data or the second set of valid detection data is obtained for the same cell, the first set of valid detection data or the second set of valid detection data is selected as the final valid detection data of the cell.

[0112] In some embodiments, when different first original optical information, second original optical information, and third original optical information are used as reference optical information: (1) if a first set of valid detection data and a second set of valid detection data are obtained for the same cell, wherein the first set of valid detection data includes amplitude information (H11, H12, H15) and first time information t1, and the second set of valid detection data includes amplitude information (H23, H24, H26) and second time information t2, and t1 is substantially the same as t2, then the amplitude of the first set of valid detection data is The sum of the amplitudes of the second group of valid detection data is H11+H12+H15, and the sum of the amplitudes of the second group of valid detection data is H23+H24+H26. The valid detection data corresponding to the larger value of H11+H12+H15 and H23+H24+H26 are selected as the final valid detection data of the cell; (2) If the first group of valid detection data and the third group of valid detection data are obtained for the same cell, wherein the first group of valid detection data includes amplitude information (H11, H12, H15) and the first time information t1, and the third group of valid detection data includes amplitude information (H37 , H38, H39) and the third time information t3, t1 is substantially the same as t3. At this time, the sum of the amplitudes of the first group of valid detection data is H11+H12+H15, and the sum of the amplitudes of the third group of valid detection data is H37+H38+H39. The valid detection data corresponding to the larger value of H11+H12+H15 and H37+H38+H39 can be selected as the final valid detection data of the cell; (3) If the second group of valid detection data and the third group of valid detection data are obtained for the same cell, among which the second group of valid detection data The third set of valid detection data includes amplitude information (H23, H24, H26) and first time information t2. The third set of valid detection data includes amplitude information (H37, H38, H39) and third time information t3. t2 is substantially the same as t3. At this time, the sum of the amplitudes of the second set of valid detection data is H23+H24+H26, and the sum of the amplitudes of the third set of valid detection data is H37+H38+H39. The valid detection data corresponding to the larger value of H23+H24+H26 and H37+H38+H39 can be selected as the final valid detection data of the cell.(5) If the first set of valid detection data, the second set of valid detection data and the third set of valid detection data are obtained for the same cell, wherein the first set of valid detection data includes amplitude information (H11, H12, H15) and first time information t1, the second set of valid detection data includes amplitude information (H23, H24, H26) and second time information t2, and the third set of valid detection data includes amplitude information (H37, H38, H39) and third time information t3, and t1, t2 and t3 are basically the same, then the sum of the amplitudes of the first set of valid detection data is H11+H12+H15, and the sum of the amplitudes of the second set is H11+H12+H15. The sum of the amplitudes of the valid detection data is H23+H24+H26, and the sum of the amplitudes of the third group of valid detection data is H37+H38+H39. The valid detection data corresponding to the maximum value among H11+H12+H15, H23+H24+H26 and H37+H38+H39 can be selected as the final valid detection data of the cell; (6) When only the first group of valid detection data, the second group of valid detection data or the third group of valid detection data is obtained for the same cell, the first group of valid detection data, the second group of valid detection data or the third group of valid detection data is selected as the final valid detection data of the cell.

[0113] Alternatively, the sum of the above amplitudes can be replaced by an average amplitude, that is, the group of valid detection data corresponding to the same cell with the largest average amplitude can be selected as the final valid detection data of the same cell. The average amplitude of a group of valid data refers to the average value of the amplitudes corresponding to the various original optical information of the same cell at the same pulse position. For example, if a first group of valid detection data and a second group of valid detection data are obtained for the same cell, wherein the first group of valid detection data includes amplitude information (H11, H12) and first time information t1, and the second group of valid detection data includes amplitude information (H23, H24) and second time information t2, t1 and t2 are substantially the same. At this time, the average amplitude of the first group of valid detection data is H11+H12 / 2, and the average amplitude of the second group of valid detection data is H23+H24 / 2.

[0114] In some embodiments, when multiple sets of valid detection data are acquired for the same cell, one set of valid detection data is simply selected based on the type of target cell as the final valid detection data for that cell. For example, when the first raw optical information is fluorescence information or side scattered light information, and the target cell is a platelet or reticulocyte, the first set of valid detection data is selected as the final valid detection data.

[0115] In some embodiments, the cell identification step S540 includes:

[0116] According to the type of the target cells, a group of the valid detection data is selected as the final valid detection data to identify the target cells in the sample liquid to be tested.

[0117] That is, in the cell identification step S540, it is not necessary to determine whether the time information of each group of valid detection data is the same. Instead, one group of the multiple groups of valid detection data is directly selected as the final valid detection data of each cell to be detected based on the type of the target cell.

[0118] Optionally, the first original optical information is fluorescence information or side scattered light information. When the target cells are platelets and / or reticulocytes, the first set of valid detection data of each cell to be tested is selected as the final valid detection data of each cell to be tested to identify the platelets and / or reticulocytes in the sample liquid to be tested.

[0119] Because different types of target cells exhibit different characteristics in different optical detection directions, for example, platelets may be better distinguished in fluorescence information or side scattered light information than in forward scattered light information. In this case, the user can directly select the effective detection data detected using fluorescence information or side scattered light information as the reference optical information as the final effective detection data.

[0120] In some embodiments, the cell identification step S540 includes: identifying target cells in the sample liquid to be tested according to the amplitude information of the final valid detection data and determining the number of target cells in the sample liquid to be tested.

[0121] That is, the cells whose amplitude information in the final effective detection data meets certain conditions are target cells, and the number of cells whose amplitude information in the final effective detection data meets certain conditions is the number of target cells.

[0122] In some embodiments, due to the different characteristics of cells in different optical information, pulse counts based on any one optical information are inaccurate. A method combining pulse counting and pulse position (i.e., time information) is employed herein, wherein the pulse information is associated with the position information. For repeated pulses identified in different optical information for the same cell throughout the measurement process, only one valid position is retained, and the total number of pulse positions is the total number of pulses. For example, during the entire measurement process, three pulses are identified and recorded at positions (p1, p2, p3) using forward scattered light information as the reference optical information, four pulses are identified and recorded at positions (p1, p2, p3, p4) using side scattered light information as the reference optical information, and four pulses are identified and recorded at positions (p1, p2, p3, p5) using fluorescence information as the reference optical information. The repeated positions are not counted repeatedly, resulting in pulse positions (p1, p2, p3, p4, p5), i.e., a total number of five pulses.

[0123] Optionally, identifying the target cells in the sample liquid to be tested according to the amplitude information of the final valid detection data includes:

[0124] A scatter plot of the cells to be tested is generated according to the amplitude information in the final valid data of each cell to be tested, so as to identify the target cells in the sample liquid to be tested.

[0125] Specifically, the data processing device 70 in the cell analyzer of the embodiment of the present invention can simultaneously use the fluorescence intensity, forward scattered light intensity and side scattered light intensity to obtain a three-dimensional scatter plot, thereby distinguishing the target cell population from other cell populations, obtaining more accurate identification of target cells and calculating the number of target cells.

[0126] In one embodiment, the optical detection device of the cell analyzer of the embodiment of the present invention collects forward scattered light information, side scattered light information and fluorescence information within a certain time period of 50ms. Using the forward scattered light information as the reference optical information for pulse identification, 1170 pulses are obtained; using the fluorescence information as the reference optical information for pulse identification, 1205 pulses are obtained. A total of 1162 pulses are detected at the same position (time) by the two methods, 8 pulses are identified in the forward scattered light information but not in the fluorescence information, and 43 pulses are identified in the fluorescence information but not in the forward scattered light information. Among them, among the 1162 pulse data detected in both the forward scattered light information and the fluorescence information, A are platelets; among the 8 pulse data unique to the forward scattered light information, 4 pulses have a small amplitude, which can be considered to be platelets based on experience; among the 43 pulse data unique to the fluorescence information, 11 pulses have a small amplitude, which can be considered to be platelets based on experience. Combining the above data, the following results can be obtained:

[0127] Reference optical information Total number of pulses platelet count Forward scattered light information 1,170 A+4 Fluorescence information 1205 A+11 Combining the two 1213 A+15

[0128] As can be seen, the pulse identification method using fluorescence information as a reference identified more total pulses and platelet counts than the method using forward scattered light information as a reference. This is because small cells such as platelets are better distinguishable in fluorescence information. Furthermore, the method using a combination of fluorescence and forward scattered light information as reference optical data yielded even higher total pulses and platelet counts. This demonstrates that using multiple raw optical data as reference data can overcome the limitations of using a single raw optical data source for pulse identification and cell analysis, and that pulse identification and cell analysis methods using multiple raw optical data sources are superior to those using a single raw optical data source as a reference.

[0129] According to an embodiment of the present invention, Figure 9As shown, a cell analysis method 900 is also provided, comprising:

[0130] S910, selecting a type of reference optical information according to the type of target cells, wherein the reference optical information is used to identify each test cell in the test sample solution that generates a valid light pulse when passing through the optical detection area of ​​the cell analyzer;

[0131] S920, acquiring at least two types of optical information of each cell to be tested in the sample liquid to be tested when the cell passes through the optical detection area, wherein the at least two types of optical information include the reference optical information and at least one type of non-reference optical information;

[0132] S930, determining valid detection data corresponding to the valid light pulse from the at least two types of optical information according to the reference optical information;

[0133] S940: Identify target cells in the sample solution according to the valid detection data.

[0134] Among them, for some types of target cells, it is already known which original optical information has a higher degree of discrimination for this type of cell than other original optical information. In this way, the reference optical information can be directly determined according to the type of target cell, and the other collected original optical information can be used as non-reference optical information, thereby speeding up the recognition of target cells while ensuring accuracy.

[0135] Optionally, determining the effective detection data according to the reference optical information includes:

[0136] Based on a preset amplitude threshold, identifying the pulses generated by each cell to be tested passing through the optical detection area from the reference optical information and obtaining effective light pulse data of each cell to be tested, the effective light pulse data including first pulse amplitude information and time information;

[0137] Acquire at least one non-reference optical data of each cell to be measured from at least one non-reference optical information according to the time information, wherein the non-reference optical data includes second pulse amplitude information;

[0138] The effective detection data includes the effective light pulse data and the at least one non-reference optical data.

[0139] Optionally, when the target cells are platelets and / or reticulocytes, side scattered light information or fluorescence information is selected as the reference optical information, and the at least two optical information include at least two of forward scattered light information, side scattered light information and fluorescence information.

[0140] In one embodiment, n pulses are identified from reference optical information (e.g., fluorescence information) and n reference effective light pulse information corresponding to the n pulses are obtained, each reference effective light pulse information including reference pulse amplitude information H and reference time information t. Then, based on the reference time information t, non-reference pulse amplitude information H1 corresponding to time t in at least one non-reference optical information (e.g., scattered light information) is obtained, thereby obtaining non-reference effective light pulse information including the non-reference pulse amplitude information, and further obtaining effective detection data, i.e., (t, H, H1).

[0141] Furthermore, a scatter plot of the cells to be tested may be generated based on the amplitude information in the valid detection data to identify platelets in the sample liquid to be tested, and the number of platelets may be determined based on the number of reference time information in the valid detection data.

[0142] According to an embodiment of the present invention, a method for detecting platelets and / or reticulocytes is also provided, comprising:

[0143] Acquiring at least two types of optical information generated when each cell to be tested in the sample liquid to be tested passes through the optical detection area of ​​the cell analyzer, wherein the at least two types of optical information include side scattered light information or fluorescence information;

[0144] Using the side scattered light information or the fluorescence information as reference optical information for identifying effective light pulses generated by each of the cells to be detected passing through the optical detection area, and determining first effective detection data corresponding to the effective light pulse from the at least two types of optical information;

[0145] The platelets and / or reticulocytes in the sample liquid to be tested are identified according to the first valid detection data.

[0146] Among them, the discrimination degree of platelets and / or reticulocytes in side scattered light information and fluorescence information is better than that in forward scattered light information. When identifying platelets and / or reticulocytes in the sample liquid to be tested, the side scattered light information or fluorescence information can be used as reference optical information for pulse identification, and then the first effective detection data corresponding to the effective light pulse is determined from the at least two optical information, and a scatter plot of the cells to be tested can be generated based on the amplitude information of the first effective detection data to identify the platelets and / or reticulocytes in the sample liquid to be tested and determine the number of platelets and / or reticulocytes.

[0147] Furthermore, the at least two types of light information include forward scattered light information, and the method further comprises:

[0148] Using the forward scattered light information as reference optical information for identifying effective light pulses generated by each of the cells to be detected passing through the optical detection area, and determining second effective detection data corresponding to the effective light pulses from the at least two types of optical information;

[0149] Platelets and / or reticulocytes in the sample liquid to be tested are identified according to the first valid detection data and the second valid detection data.

[0150] An embodiment of the present invention further provides a computer-readable storage medium storing a plurality of program instructions. After the plurality of program instructions are called and executed by a processor, some or all of the steps or any combination of the steps in the cell analysis method in each embodiment of the present application can be executed.

[0151] According to the cell analysis method, cell analyzer, and computer-readable storage medium of the embodiments of the present invention, multiple different types of optical signals generated when cells pass through the flow chamber are used as reference signals for pulse identification. This fully utilizes the principle that different types of optical signals reflect different cell characteristic information, avoids missed identification or failure to identify target cells, and improves the accuracy of cell classification, scatter plot morphology, and cell counting, thereby improving the accuracy of the hematology analyzer.

[0152] It will be understood by those skilled in the art that, except where mutually exclusive, all features disclosed in this specification (including the accompanying claims, abstract, and drawings) and all processes or units of any method or apparatus disclosed herein may be combined in any combination. Unless expressly stated otherwise, each feature disclosed in this specification (including the accompanying claims, abstract, and drawings) may be replaced by an alternative feature providing the same, equivalent, or similar purpose.

[0153] The technical terms used in the embodiments of the present invention are only used to illustrate specific embodiments and are not intended to limit the present invention. In this document, the singular forms "a," "the," and "said" are used to include the plural forms, unless the context clearly indicates otherwise. Furthermore, the terms "including" and / or "comprising" used in this specification refer to the presence of the described features, integers, steps, operations, elements, and / or components, but do not exclude the presence or addition of one or more other features, integers, steps, operations, elements, and / or components.

[0154] The flowchart described in this invention is merely an embodiment. Various modifications and variations may be made to this diagram or the steps described herein without departing from the spirit of the invention. For example, the steps may be performed in a different order, or certain steps may be added, deleted, or modified. Those skilled in the art will appreciate that equivalent variations that implement all or part of the above-described embodiment and are made in accordance with the claims of the present invention are still within the scope of the invention.

Claims

1. A cell analysis method, characterized in that: include: The step of acquiring original optical information comprises acquiring first original optical information detected by a first optical detector and second original optical information detected by a second optical detector when each cell to be tested in the sample liquid to be tested passes through an optical detection area of ​​the cell analyzer within a preset time period, wherein the first original optical information and the second original optical information are different optical information; A first set of valid detection data acquisition step: using the first original optical information as reference optical information for identifying valid light pulses generated by each cell to be detected passing through the optical detection area, and determining first valid light pulse information and second valid light pulse information of each cell to be detected from the first original optical information and the second original optical information, respectively, to acquire a first set of valid detection data for each cell to be detected, including the first valid light pulse information and the second valid light pulse information; A second set of valid detection data acquisition step: using the second original optical information as the reference optical information, determining third valid light pulse information and fourth valid light pulse information of each cell to be detected from the first original optical information and the second original optical information, respectively, to acquire a second set of valid detection data for each cell to be detected, including the third valid light pulse information and the fourth valid light pulse information; Cell identification step: According to preset rules, the final valid detection data of each cell to be tested is determined from the first set of valid detection data and the second set of valid detection data of each cell to be tested, so as to identify the target cells in the sample liquid to be tested.

2. The cell analysis method according to claim 1, wherein The first original optical information and the second original optical information are respectively selected from one of forward scattered light information, side scattered light information and fluorescence information.

3. The cell analysis method according to claim 2, characterized in that The first original optical information or the second original optical information is fluorescence information.

4. The cell analysis method according to claim 3, characterized in that The target cells to be detected are platelets and / or reticulocytes.

5. The method according to any one of claims 1 to 4, characterized in that The step of acquiring original optical information further includes: acquiring third original optical information detected in a third optical detector when each cell to be tested in the sample liquid to be tested passes through the optical detection area within the preset time period; The first set of valid detection data acquisition step further includes: using the first original optical information as the reference optical information, determining fifth valid light pulse information of each cell to be detected from the third original optical information, the first set of valid detection data further including the fifth valid light pulse information; The step of acquiring the second set of valid detection data further includes: using the second original optical information as the reference optical information, determining the sixth valid light pulse information of each cell to be tested from the third original optical information, and the second set of valid detection data further includes the sixth valid light pulse information.

6. The method according to claim 5, characterized in that The first set of steps for obtaining effective detection data includes: Based on the first amplitude threshold, the pulses generated when each cell to be tested passes through the optical detection area are identified from the first original optical information and first effective light pulse information of each cell to be tested is obtained, wherein the first effective light pulse information includes first pulse amplitude information and first time information. Acquire, according to the first time information, second effective light pulse information and fifth effective light pulse information of each cell to be tested from the second original optical information and the third original optical information, wherein the second optical information and the fifth effective light pulse information respectively include second pulse amplitude information and fifth pulse amplitude information; The second set of steps for obtaining effective detection data includes: Based on the second amplitude threshold, the pulses generated when each cell to be tested passes through the optical detection area are identified from the second original optical information and fourth effective light pulse information of each cell to be tested is obtained, wherein the fourth effective light pulse information includes fourth pulse amplitude information and second time information. According to the second time information, third effective light pulse information and sixth effective light pulse information of each cell to be tested are obtained from the first original optical information and the third original optical information, wherein the third effective light pulse information and the sixth effective light pulse information respectively include third pulse amplitude information and sixth pulse amplitude information.

7. The method according to claim 5, characterized in that Further comprising a third set of steps for obtaining effective detection data: Using the third original optical information as the reference optical information, determining seventh valid light pulse information, eighth valid light pulse information, and ninth valid light pulse information for each cell to be tested from the first original optical information, the second original optical information, and the third original optical information, respectively, to obtain a third set of valid detection data for each cell to be tested, including the seventh valid light pulse information, the eighth valid light pulse information, and the ninth valid light pulse information; The cell identification step includes: determining final valid detection data of each cell to be tested from the first set of valid detection data, the second set of valid detection data and the third set of valid detection data of each cell to be tested according to preset rules, so as to identify the target cells in the sample liquid to be tested.

8. The method according to claim 7, characterized in that The first set of steps for obtaining effective detection data includes: Based on the first amplitude threshold, the pulses generated when each cell to be tested passes through the optical detection area are identified from the first original optical information and first effective light pulse information of each cell to be tested is obtained, wherein the first effective light pulse information includes first pulse amplitude information and first time information. Acquire, according to the first time information, second effective light pulse information and fifth effective light pulse information of each cell to be tested from the second original optical information and the third original optical information, wherein the second optical information and the fifth effective light pulse information respectively include second pulse amplitude information and fifth pulse amplitude information; The second set of steps for obtaining effective detection data includes: Based on the second amplitude threshold, the pulses generated when each cell to be tested passes through the optical detection area are identified from the second original optical information and fourth effective light pulse information of each cell to be tested is obtained, wherein the fourth effective light pulse information includes fourth pulse amplitude information and second time information. Acquire third effective light pulse information and sixth effective light pulse information of each cell to be tested from the first original optical information and the third original optical information according to the second time information, wherein the third effective light pulse information and the sixth effective light pulse information respectively include third pulse amplitude information and sixth pulse amplitude information; The third set of steps for obtaining effective detection data includes: Based on the third amplitude threshold, the pulses generated when each cell to be tested passes through the optical detection area are identified from the third original optical information and ninth effective optical pulse information of each cell to be tested is obtained, wherein the ninth effective optical pulse information includes ninth pulse amplitude information and third time information. According to the third time information, the seventh effective light pulse information and the eighth effective light pulse information of each cell to be tested are respectively obtained from the first original optical information and the second original optical information, wherein the seventh effective light pulse information and the eighth effective light pulse information respectively include the seventh pulse amplitude information and the eighth pulse amplitude information.

9. The method according to claim 6 or 8, characterized in that The cell identification step comprises: Determine whether the time information in each group of valid detection data belongs to the same cell; For the case of the same cell, one group of valid detection data corresponding to the same cell is selected as the final valid detection data of the same cell; In the case of belonging to different cells, the valid detection data corresponding to the time information belonging to the different cells is used as the final valid detection data of the different cells.

10. The method according to claim 9, characterized in that The cell identification step comprises: For the case of the same cell, one group of valid detection data corresponding to the same cell is determined as the final valid detection data of the same cell according to the amplitude information of each group of valid detection data corresponding to the same cell or according to the type of the target cell.

11. The method according to any one of claims 1 to 8, characterized in that The cell detection step comprises: According to the type of the target cells, one group is selected from each group of the valid detection data as the final valid detection data to identify the target cells in the sample liquid to be tested.

12. The method according to claim 11, characterized in that The first original optical information is fluorescence information or side scattered light information; The cell detection step includes: when the target cells are platelets and / or reticulocytes, selecting the first set of valid detection data as the final valid detection data to identify the platelets and / or reticulocytes in the sample liquid to be tested.

13. The method according to any one of claims 6, 8 to 10, characterized in that The cell identification step comprises: The target cells in the sample liquid to be tested are identified according to the amplitude information of the final effective detection data of each cell to be tested, and the number of the target cells in the sample liquid to be tested is determined.

14. The method according to any one of claims 6, 8 to 10, characterized in that Also includes: A scatter plot of the cells to be tested is generated according to the amplitude information in the final valid detection data of each cell to be tested, so as to identify the target cells in the sample liquid to be tested.

15. A cell analysis method, characterized in that: include: Selecting the type of reference optical information according to the type of target cells, wherein the reference optical information is used to identify effective light pulses generated by each cell to be tested in the sample liquid to be tested passing through the optical detection area of ​​the cell analyzer; Acquiring at least two types of optical information of each cell to be tested in the sample liquid to be tested when the cell passes through the optical detection area, wherein the at least two types of optical information include the reference optical information and at least one type of non-reference optical information; determining, based on the reference optical information, valid detection data corresponding to the valid light pulse from the at least two types of optical information; The target cells in the sample liquid to be tested are identified according to the effective detection data.

16. The method according to claim 15, characterized in that Determining the effective detection data according to the reference optical information includes: Based on a preset amplitude threshold, identifying the pulses generated by each cell to be tested passing through the optical detection area from the reference optical information and obtaining effective light pulse data of each cell to be tested, the effective light pulse data including first pulse amplitude information and time information; According to the time information, at least one non-reference optical data of each cell to be tested is obtained from at least one non-reference optical information, wherein the non-reference optical data includes second pulse amplitude information; wherein the effective detection data includes the effective light pulse data and the at least one non-reference optical data.

17. The method according to claim 15 or 16, characterized in that When the target cells are platelets and / or reticulocytes, side scattered light information or fluorescence information is selected as the reference optical information, and the at least two optical information include at least two of forward scattered light information, side scattered light information and fluorescence information.

18. A method for detecting platelets and / or reticulocytes, characterized in that: include: Acquiring at least two types of optical information generated when each cell to be tested in the sample liquid to be tested passes through the optical detection area of ​​the cell analyzer, wherein the at least two types of optical information include side scattered light information or fluorescence information; Using the side scattered light information or the fluorescence information as reference optical information for identifying effective light pulses generated by each of the cells to be detected passing through the optical detection area, and determining first effective detection data corresponding to the effective light pulse from the at least two types of optical information; The platelets and / or reticulocytes in the sample liquid to be tested are identified according to the first valid detection data.

19. The method according to claim 18, characterized in that The at least two types of light information further include forward scattered light information, and the method further includes: Using the forward scattered light information as the reference optical information, determining second valid detection data corresponding to the valid light pulse from the at least two types of optical information; Platelets and / or reticulocytes in the sample liquid to be tested are identified according to the first valid detection data and the second valid detection data.

20. A cell analyzer comprising: a sampling device having a pipette with a pipette nozzle and a driving device for driving the pipette to quantitatively draw a blood sample through the pipette nozzle; A sample preparation device comprising a reaction pool and a reagent supply unit, wherein the reaction pool is used to receive a blood sample drawn by a sampling device, and the reagent supply unit provides a reagent to the reaction pool, so that the blood sample drawn by the sampling device and the reagent provided by the reagent supply unit are mixed in the reaction pool to prepare a sample liquid to be tested; An optical detection device, comprising a light source, a flow chamber, and a light detector, wherein each cell to be tested in the sample liquid to be tested can flow in the flow chamber, the light emitted by the light source illuminates the cells in the flow chamber to generate optical information, and the light detector is used to collect the optical information; and A data processing device electrically connected to the optical detection device and comprising a processor and a computer-readable storage medium storing a computer program, wherein the data processing device is configured to perform the steps of the method according to any one of claims 1 to 19 when the computer program is executed by the processor.

21. A computer-readable storage medium, characterized in that The method comprises a program which can be executed by a processor to implement the method according to any one of claims 1 to 19.

Images