CN115398209B

CN115398209B - Cell analysis method, cell analyzer, and computer-readable storage medium

Info

Publication number: CN115398209B
Application number: CN202080099809.8A
Authority: CN
Inventors: 阮楚良; 汪东生; 刘斌; 杨克勤
Original assignee: Shenzhen Mindray Bio Medical Electronics Co Ltd
Current assignee: Shenzhen Mindray Bio Medical Electronics Co Ltd
Priority date: 2020-04-13
Filing date: 2020-04-13
Publication date: 2025-08-26
Anticipated expiration: 2040-04-13
Also published as: WO2021207894A1; CN115398209A

Abstract

A cell analysis method, a cell analyzer and a computer readable storage medium comprise the steps of obtaining first original optical information and second original optical information of each cell to be tested in a sample liquid to be tested, taking the first original optical information as reference optical information to obtain a first group of effective detection data of each cell to be tested, taking the second original optical information as reference optical information to obtain a second group of effective detection data of each cell to be tested, and determining final effective detection data of each cell to be tested according to a preset rule by the first group of effective detection data and the second group of effective detection data to be used for identifying target cells in the sample liquid to be tested. According to the cell analysis method, the cell analyzer and the computer readable storage medium, cell characteristic information reflected by different optical signals is fully utilized, and the accuracy of cell classification, scatter diagram morphology and cell counting is improved.

Description

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

Technical Field

Embodiments of the present invention relate to medical device technology, and in particular, to a cell analysis method, a cell analyzer, and a computer-readable storage medium.

Background

Currently, a blood cell analyzer generally performs cell detection and analysis by using a laser scattering principle, and mainly performs cell detection and analysis by using a low-angle scattered light signal (forward scattered light signal FS), a side scattered light signal (SS) and a side fluorescent signal (FL), where FS represents cell volume information, FS represents cell surface complexity, and FL represents calculated cell inner content. The light scattering cell detection technology combines the optical characteristics and biological characteristics of cells, and has great development and wide application in cell identification by the advantages of rapidness, simplicity, accuracy, no damage, good repeatability and the like. When the light scattering principle is adopted for detection, the light beam irradiates the cell to be detected passing through the optical detection area, and scattered light signals are generated by the interaction of the light and the cell, wherein the scattered light signals contain information related to the size and distribution of the cell. In the case of fluorescent staining of the test cells, the test cells also generate a fluorescent signal that characterizes the intracellular nucleic acid content after being irradiated with light. When the cell to be detected passes through the optical detection area rapidly, the intensities of the generated three optical signals (namely FS, SS and FL) change instantaneously to generate optical pulses, the optical pulses enter the photoelectric sensor and the processing circuit to be converted into electric pulses, then the pulses generated by the cell to be detected are identified by taking the low-angle scattering signals as reference signals, the pulse amplitudes of the cell to be detected in the three optical signals are obtained, the pulse numbers are counted, a scatter diagram can be generated and the cell classification can be carried out by analyzing the pulse amplitude information, and the pulse numbers can represent the cell numbers. However, at present, only the low-angle scattering signals acquired in the low-angle direction are used for pulse recognition, and because the low-angle scattering signals represent the volume information of cells, the cells with larger volumes can be better recognized based on the low-angle scattering signals, however, the conditions of missing recognition or incapacity of recognition can occur when certain volumes are small, so that the problems of inaccurate cell classification, abnormal scatter diagram, inaccurate cell counting and the like are caused.

Disclosure of Invention

Embodiments of the present invention provide a cell analysis method, a cell analyzer, and a computer readable storage medium, which solve at least one of the above problems.

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

the original optical information acquisition step of acquiring first original optical information detected in a first optical detector and second original optical information detected in a second optical detector when each cell to be detected in the sample liquid to be detected passes through an optical detection zone of a cell analyzer within a preset time period;

A first group of effective detection data acquisition step of respectively determining first effective light pulse information and second effective light pulse information of each cell to be detected from the first original optical information and the second original optical information by taking the first original optical information as reference optical information for identifying effective light pulses generated by each cell to be detected through the optical detection area so as to acquire first group of effective detection data of each cell to be detected, wherein the first group of effective detection data comprises the first effective light pulse information and the second effective light pulse information;

A second group of effective detection data acquisition step of respectively determining third effective light pulse information and fourth effective light pulse information of each cell to be detected from the first original optical information and the second original optical information by taking the second original optical information as the reference optical information so as to acquire second group of effective detection data of each cell to be detected, including the third effective light pulse information and the fourth effective light pulse information;

And a cell identification step, namely determining final effective detection data of each cell to be detected according to a preset rule by using the first group of effective detection data and the second group of effective detection data of each cell to be detected, wherein the final effective detection data are used for identifying target cells in the sample liquid to be detected.

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

selecting the type of reference optical information according to the type of the target cells, wherein the reference optical information is used for identifying effective light pulses generated by each cell to be tested in the sample liquid to be tested passing through an optical detection area of a cell analyzer;

acquiring at least two kinds of optical information of each cell to be detected in the sample liquid to be detected passing through the optical detection area, wherein the at least two kinds of optical information comprise the reference optical information and at least one kind of non-reference optical information;

determining effective detection data corresponding to the effective light pulse from the at least two optical information according to the reference optical information;

And identifying target cells in the sample liquid to be detected according to the effective detection data.

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

Acquiring at least two kinds of optical information generated when each cell to be detected in the sample liquid to be detected passes through an optical detection area of a cell analyzer, wherein the at least two kinds of optical information comprise side scattering optical information or fluorescence information;

Determining first effective detection data corresponding to the effective light pulses from the at least two kinds of optical information by taking the side scattering light information or the fluorescence information as reference optical information for identifying the effective light pulses generated by each cell to be detected passing through the optical detection region;

and identifying platelets in the sample liquid to be detected according to the first effective detection data.

According to a fourth aspect of the present invention there is provided a cell analyser comprising:

a sampling device having a pipette with a pipette nozzle and having a driving device for driving the pipette to quantitatively aspirate a blood sample through the pipette nozzle;

a sample preparation device having a reaction cell for receiving a blood sample sucked by a sampling device and a reagent supply part for supplying a reagent to the reaction cell so that the blood sample sucked by the sampling device and the reagent supplied by the reagent supply part are mixed in the reaction cell to prepare a sample liquid to be measured;

An optical detection device comprising a light source, a flow cell, and a photodetector, wherein each cell to be tested of the sample liquid can flow in the flow cell, the light emitted by the light source irradiates the cells in the flow cell to generate optical information, the photodetector is used for collecting 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 the first to third aspects when the computer program is executed by the processor.

According to a fifth aspect of an embodiment of the present invention, there is provided a computer readable storage medium comprising a program executable by a processor to implement the method according to the first to third aspects.

According to the cell analysis method, the cell analyzer and the computer readable storage medium, a plurality of different types of optical signals generated by cells are respectively used as reference optical information for pulse identification, cell characteristic information reflected by the different optical signals is fully utilized, the condition that target cells are not identified or cannot be identified is avoided, the accuracy of cell classification, scatter diagram morphology and cell counting is improved, and the accuracy of the blood cell analyzer is further improved.

Drawings

In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is a schematic diagram of a cell analyzer according to an embodiment of the present invention;

FIG. 2 is a schematic diagram of a frame of an optical detection device according to an embodiment of the present invention;

FIG. 3 is a schematic diagram of a frame of a control device according to an embodiment of the present invention;

FIG. 4 is a logical schematic of a prior art method of cell analysis;

FIG. 5 is a schematic flow chart of a method of cell analysis according to an embodiment of the invention;

FIGS. 6 to 8 are schematic logic diagrams of methods of cell analysis according to various embodiments of the present invention;

FIG. 9 is a schematic flow chart diagram of another method of cell analysis according to an embodiment of the invention;

FIG. 10 is a logical schematic of another method of cell analysis according to an embodiment of the present invention.

Detailed Description

The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

According to an embodiment of the present invention, there is provided a cell analyzer including a sampling device, a sample preparation device, an optical detection device, and a data processing device.

The sampling device has a pipette with a pipette nozzle and has a drive device for driving the pipette to quantitatively aspirate a liquid sample to be measured through the pipette nozzle.

The sample preparation device is provided with a reaction tank and a reagent supply part, wherein the reaction tank is used for receiving the liquid sample to be tested, which is sucked by the sampling device, and the reagent supply part supplies at least one reagent to the reaction tank, so that the liquid sample to be tested sucked by the sampling device and the at least one reagent supplied by the reagent supply part are mixed in the reaction tank to prepare a sample liquid to be tested.

The optical detection device comprises a light source, a flow chamber and at least two detectors, wherein cells in the sample liquid to be detected can flow in the flow chamber, the cells in the flow chamber are irradiated by light emitted by the light source to generate original optical information, and the detectors are used for collecting the original optical information.

The data processing device is electrically connected to the optical detection device and comprises a processor and a computer readable storage medium storing a computer program, wherein the data processing device is configured to perform part or all of the steps of the cell analysis method according to an embodiment of the invention or any combination of the steps thereof, when the computer program is executed by the processor. The cell analysis method according to the present invention will be described in detail below.

Alternatively, the processor may be a CPU, GPU or other chip with computing capabilities. In practical applications, the Processor may be implemented by software, hardware, firmware, or a combination thereof, and may use at least one of a Circuit, a single or multiple Application Specific Integrated Circuits (ASIC), a digital signal Processor (DIGITAL SIGNAL Processor, DSP), a digital signal processing device (DIGITAL SIGNAL Processing Device, DSPD), a programmable logic device (Programmable Logic Device, PLD), a field programmable gate array (Field Programmable GATE ARRAY, FPGA), a central processing unit (Central Processing Unit, CPU), a controller, a microcontroller, a microprocessor, or any combination thereof, so that the Processor may perform part or all of the steps in the cell analysis method in various embodiments of the present application.

The computer readable storage medium may be volatile memory or nonvolatile memory, or may include both volatile and nonvolatile memory. The nonvolatile 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, a compact disc, or a read-only compact disc, and the magnetic surface memory may be a magnetic disk memory or a tape memory. The volatile memory may be a random access memory that acts as an external cache. By way of example, and not limitation, many forms of RAM are available, such as sram, dram, sdram, ddr, sdram with enhanced, sdram with synchronous connection, dram with direct memory bus, and RAM with direct memory bus. The memory described in embodiments of the present invention is intended to comprise these and any other suitable types of memory.

In one embodiment, FIG. 1 shows a schematic diagram of a cell analyzer according to an embodiment of the present invention. Referring to fig. 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 section 90. In practical applications, the output 90 may be a user interface. The optical detection device 50 and the data processing device 70 are disposed inside the second casing 200, and disposed on both sides of the second casing 200. The sample preparation device 30 is disposed inside the first housing 100, and the output section 90 and the sampling device 10 are disposed on the outer surface of the first housing 100.

It should be appreciated that the liquid sample to be tested may be any type of liquid sample containing cells, such as blood, body fluids, etc., without limitation. In this embodiment, a liquid sample to be measured is taken as a blood sample as an example.

Optionally, the sampling device 10 has a sampling needle for collecting a blood sample and delivering the collected blood sample to the sample preparation device 30. According to various embodiments, the sampling device may collect multiple blood samples, provide different chambers of the sample preparation device for different treatments, and then perform different tests.

Alternatively, the sample preparation device 30 has a reaction well and a reagent supply that stores reagents for reacting with the blood sample, e.g., at least a first reagent and/or a second reagent, and supplies the corresponding reagents to the reaction well as needed. The first reagent may be a diluent or a hemolyzing agent, wherein the hemolyzing agent is used for lysing erythrocytes in the liquid sample to be tested into fragments and keeping the morphology of leukocytes in the liquid sample to be tested substantially intact. Further, the second reagent may be a fluorescent dye for staining cells in the liquid sample to be measured.

Optionally, the optical detection device 50 includes at least two of a forward scatter detector, a side scatter detector, and a side fluorescence detector. The forward scatter detectors are typically arranged in a line with the light source and the flow cell, and the light source is arranged on each side of the flow cell to detect forward scatter light intensity or low angle scatter light intensity of cells flowing in the flow cell. The side scatter detectors are typically arranged at an angle to the line in which the light source and the flow cell are located to detect the side scatter light intensity of cells flowing in the flow cell. The fluorescence detector is typically arranged at an angle to the line in which the light source and the flow cell are located to detect the lateral fluorescence intensity of the cells flowing in the flow cell.

In one embodiment, referring to fig. 2, fig. 2 shows an example of an optical detection device according to an embodiment of the present invention. The optical detection device has a light source 101, a beam shaping assembly 102, a flow cell 103 and a forward scatter detector 104 arranged in sequence in a straight line. On one side of the flow chamber 103, a dichroic mirror 106 is arranged at an angle of 45 ° to the straight line. A part of the side light emitted by the cells in the flow cell 103 is transmitted through the dichroic mirror 106 and captured by the fluorescence detector 105 arranged behind the dichroic mirror 106 at an angle of 45 ° to the dichroic mirror 106, while another part of the side light is reflected by the dichroic mirror 106 and captured by the side scatter detector 107 arranged in front of the dichroic mirror 106 at an angle of 45 ° to the dichroic mirror 106.

It should be understood that the line of the light source and the flow chamber may be aligned with the forward scatter detector or may form an angle, which may be set as desired, without limitation, and that the angle between the line of the light source and the flow chamber and the dichroic mirror, and the angle between the dichroic mirror and the side scatter detector and the fluorescence detector, respectively, may be set as desired, without limitation.

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

The output section 90 is configured to output the detection result. The output unit 90 may be a display device (not shown) for displaying the detection result, and may be provided on the cell analyzer, or may be provided outside the cell analyzer and electrically connected to the data processing device 70. Further, the display device may be a touch display screen, a liquid crystal display screen, or the like, or may be a display screen on an electronic device such as a mobile phone, a tablet computer, or the like.

According to the embodiment of the invention, the cell analyzer can also provide a corresponding operation interface for an operator to operate, and the operation interface can comprise corresponding controls, such as a mark selection box or a menu bar, so that the operator can input an operation instruction on the operation interface according to actual use conditions to realize cell analysis through the cell analyzer. Further, the output section 90 provides the operation interface.

In one embodiment, as shown in FIG. 3, data processing device 70 includes at least a processor 71, RAM72, ROM73, a communication interface 74, memory 76, and I/O interface 75. The processor 71, RAM72, ROM73, communication interface 74, memory 76 and I/O interface 75 communicate via bus 77. The processor may be a CPU, GPU or other chip with computing capabilities. Various computer programs such as an operating system and application programs to be executed by the processor 71 and data necessary for executing the computer programs are stored in the memory 76. In addition, during cell detection, data stored locally may be stored in memory 76, if desired. The I/O interface 75 is constituted by a serial interface such as USB, IEEE1394, or RS-232C, a parallel interface such as SCSI, IDE, or IEEE1284, and an analog signal interface composed of a D/a converter, an a/D converter, and the like. An input device, such as a keyboard, mouse, touch screen or other control buttons, is connected to the I/O interface 75, and a user can directly input data to the data processing apparatus 70 using the input device. In addition, the I/O interface 75 may be connected to a display having a display function, such as a liquid crystal screen, a touch screen, an LED display screen, etc., and the data processing device 70 may output the processed data as image display data to the display for display, such as analysis data, instrument operation parameters, etc. The communication interface 74 is an interface that may be any communication protocol known to date. The communication interface 74 communicates with the outside through a network. The data processing device 70 may communicate data with any device connected via the network via a communication interface 74 in a communication protocol.

In the present practice, as shown in FIG. 4, one prior art cell analysis method is shown. In the prior art, in the process of analyzing cells in a liquid sample to be tested, forward scattered light information (FS signal) collected by a forward scattered light detector is used as reference optical information to perform pulse identification, so as to obtain pulse information of the forward scattered light information, where the pulse information includes pulse number, FS pulse amplitude H1, FS pulse position t (i.e. time corresponding to the identified pulse), and the like. Then, based on the pulse positions t corresponding to the respective pulses identified from the forward scattered light information, the pulse amplitudes H2, 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 the pulse positions t are respectively acquired, and then three pieces of FS-based amplitude data (H1, H2, H3) corresponding to the forward scattered light information, the side scattered light information and the side fluorescence information can be obtained for all the identified pulses, and then cell classification is performed based on the three pieces of FS-based amplitude data (H1, H2, H3) and a corresponding scatter diagram is obtained, and the number of target cells is obtained based on the number of pulses. However, as described above, FS represents cell volume information, and for some cells (e.g., platelets) whose cell surface complexity is high (i.e., SS signal intensity is high) or whose intracellular computational content is high (i.e., FL signal intensity is high), pulse recognition using only the FS signal as a reference signal may cause cases where recognition is missed or cannot be recognized, resulting in inaccurate cell classification, abnormal scatter diagram, and inaccurate cell count.

With the above in mind, a method of cell analysis is provided according to an embodiment of the present invention, see FIG. 5, in which a schematic flow chart of a method of cell analysis according to an embodiment of the present invention is shown. As shown in fig. 5, the method 500 includes:

a primary optical information acquisition step S510 of acquiring first primary optical information detected in a first optical detector and second primary optical information detected in a second optical detector when each cell to be measured in the sample liquid to be measured passes through an optical detection region of the cell analyzer within a preset period of time;

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

A second set of valid detection data acquisition step S530 of determining third valid light pulse information and fourth valid light pulse information of each cell to be measured from the first original optical information and the second original optical information with the second original optical information as the reference optical information, to acquire second set of valid detection data including the third valid light pulse information and the fourth valid light pulse information of each cell to be measured;

And S540, determining final effective detection data of each cell to be detected according to a preset rule by using the first group of effective detection data and the second group of effective detection data of each cell to be detected, and identifying target cells in the sample liquid to be detected.

In an embodiment of the invention, the original optical information may be a signal, such as an electrical signal, that is detected and output by the optical detector for a predetermined time. The reference optical information refers to original optical information in which pulse information of other original optical information is acquired based on the identified pulse position thereof, and pulse information acquired from the reference optical information and pulse information acquired from other original optical information based on the reference optical information can be recorded as valid optical pulse information.

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, n pulse information corresponding to the n pulses is obtained and is recorded as first effective optical pulse information, then pulse information corresponding to the n pulse positions is obtained from the second original optical information according to the n pulse positions of the n pulses and is recorded as second effective optical pulse information, and the first effective optical pulse information and the second effective optical pulse information are recorded as a first group of effective detection data. When the second original optical information is taken as reference optical information, m pulses are identified from the second original optical information, m pulse information corresponding to the m pulses is obtained and is recorded as fourth effective optical pulse information, then pulse information corresponding to the m pulse positions is obtained from the first original optical information according to the m pulse positions of the m pulses and is recorded as third effective optical pulse information, and the third effective optical pulse information and the fourth effective optical pulse information are recorded as second group of effective detection data. Finally, according to an intelligent screening algorithm, namely a preset rule, final effective detection data are obtained from the first group of effective data and the second group of effective data so as to identify the target cells.

According to the cell analysis method provided by the embodiment of the invention, the different optical information acquired by the at least two optical detectors is used as the reference optical information of pulse identification, so that the principle that the different optical information reflects different characteristics of cells is fully utilized, complementation is formed in pulse identification, and the condition that target cells are not identified or cannot be identified is avoided. Compared with the traditional method for analyzing the cells by only adopting forward scattered light information as reference optical information, the method overcomes the limitation of pulse identification based on single optical information and greatly improves the accuracy of cell analysis.

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.

In some embodiments, the first original optical information or the second original optical information is fluorescence information. In the case of fluorescence information as reference optical information, it is particularly suitable for identifying small-volume cells in a sample liquid to be measured, such as platelets, reticulocytes, and the like.

In some embodiments, the first set of valid detection data acquisition steps S520 include:

Identifying pulses generated by each test cell passing through the optical detection zone from the first original optical information based on a first amplitude threshold value and acquiring first effective light pulse information of each test cell, wherein the first effective light pulse information comprises first pulse amplitude information and first time information,

Acquiring second effective optical pulse information of each cell to be detected from the second original optical information according to the first time information, wherein the second effective optical pulse information comprises second pulse amplitude information;

accordingly, the second set of valid detection data acquisition step S530 includes:

identifying pulses generated by each cell to be tested passing through the optical detection zone from the second original optical information based on a second amplitude threshold value and acquiring fourth effective light pulse information of each cell to be tested, wherein the fourth effective light pulse information comprises fourth pulse amplitude information and second time information,

And acquiring third effective light pulse information of each cell to be detected from the first original optical information according to the second time information, wherein the third effective light pulse information comprises third pulse amplitude information.

Wherein different amplitude thresholds may be set for identifying pulses for different original optical information. 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 zone, and 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 to be a pulse generated by signal interference. In other words, when no cell passes through the optical detection zone, the first original optical information and the second original optical information are typically maintained under respective dc baseline voltages, and when a cell passes through the optical detection zone and generates optical information, the amplitude of the first original optical information and/or the second original optical information changes, and whether the cell is a valid pulse can be identified by judging the trend of the rising edge and the falling edge of the pulse, while recording the difference between the peak value of the pulse and the baseline as the pulse amplitude and recording the corresponding peak position (i.e., the time of pulse generation), and the number of cells can be calculated by accumulating the number of valid pulses.

As shown in fig. 6, 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. Taking a blood sample as an example, referring to fig. 1 and 2 together, a sampling device 10 in a cellular analyzer collects the blood sample through a sampling needle and conveys the collected blood sample to a sample preparation device 30. The reagent supply part in the sample preparation device 30 supplies the diluent and the fluorescent dye to the reaction tank according to the requirement, the blood sample reacts with the diluent and the fluorescent dye in the reaction tank in sequence to obtain a liquid sample to be tested containing a plurality of cells, and the cells in the liquid sample to be tested are queued one by one to 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 are excited by the light to generate different optical information. The forward scatter light detector or the side scatter light detector detects the forward scatter light intensity or the side scatter light intensity of the cells flowing in the flow chamber to obtain first original optical information, and the fluorescence detector detects the side fluorescence intensity of the cells flowing in the flow chamber to obtain second original optical information. The data processing device 70 performs cell analysis after acquiring the first and second original optical information. When forward scattered light information or side scattered light information (for example, forward scattered light information in fig. 6) is used as reference optical information, n pulses (for example, the number of pulses 1 in fig. 6) are identified from the forward scattered light information or side scattered light information and n pieces of first effective light pulse information corresponding to the n pulses are acquired, each piece of first effective light pulse information including first pulse amplitude information H11 (for example, FS pulse amplitude in fig. 6) and first time information t1 (for example, FS pulse position in fig. 6), and then second pulse amplitude information H12 corresponding to the vicinity of time t1 in the fluorescent information is acquired based on the first time information t1, whereby n pieces of second effective light pulse information including second pulse amplitude information H12 are obtained, and n pieces of first sets of effective detection data, that is, n pieces (t 1, H11, H12) (for example, FS-based two amplitude data in fig. 6) are obtained. When the fluorescence information is taken as the reference optical information, m pulses (for example, the number of pulses 3 in fig. 6) are identified from the fluorescence information and m pieces of fourth effective light pulse information corresponding to the m pulses are acquired, each piece of fourth effective light pulse information including fourth pulse amplitude information H24 (for example, FL pulse amplitude in fig. 6) and second time information t2 (for example, FL pulse position in fig. 6), and then third pulse amplitude information H23 corresponding to the vicinity of time t2 in the forward scattered light information or the side scattered light information is acquired based on the second time information t2, thereby obtaining m pieces of third effective light pulse information including third pulse amplitude information H23, and m pieces of second sets of effective detection data (for example, m pieces of (t 2, H23, H24) (for example, two amplitude data based on FL in fig. 6) are obtained. In other words, after two kinds of original optical information are received, pulse identification is performed by using the first original optical information, pulse amplitude information and position information are obtained, the number of pulses is counted, pulses at the same position are found on the second original optical information, and amplitude information is obtained, pulse identification is performed by using the second original optical information, pulse amplitude information and position information are obtained, the number of pulses is counted, and pulses at the same position are found on the first original optical information, and amplitude information is obtained. Finally, final valid detection data is determined from the n first and m second sets of valid detection data according to preset rules (e.g., the smart screening algorithm of fig. 6) for identifying and counting target cells and optionally for generating a scatter plot. wherein the pulse amplitude information in the final effective detection data is used for identifying the target cells, and the pulse amplitude information and the time information (pulse position information) in the final effective detection data are used for determining the number of the target cells.

In some embodiments, the first and second original optical information may also be forward and side scattered light information, respectively. Taking a blood sample as an example, referring again to fig. 1 and 2, a sampling device 10 in a cellular analyzer collects a blood sample through a sampling needle and conveys the collected blood sample to a sample preparation device 30. The reagent supplying section in the sample preparing apparatus 30 supplies the diluent to the reaction cell as needed, and the blood sample reacts with the diluent to obtain a liquid sample to be measured containing a plurality of cells, and causes the cells in the liquid sample to be measured to flow through the flow chamber of the optical detecting apparatus 50 in a line-by-line manner. 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 are excited by the light to generate different optical information. The forward scatter light detector and the side scatter light detector detect the forward scatter light intensity and the side scatter light intensity of the cell flowing in the flow chamber, respectively, to obtain first original optical information and second original optical information. The data processing device 70 performs cell analysis after acquiring the first and second original optical information. For specific cell analysis, reference is made to the embodiment described above in connection with fig. 6, and no further description is given here.

In some embodiments, the original optical information obtaining step S510 further includes obtaining a third original optical information detected by each cell to be tested in the third optical detector when each cell to be tested in the sample liquid passes through the optical detection zone within the preset time period, and correspondingly, the first set of valid detection data obtaining step S520 further includes determining a fifth valid optical pulse information of each cell to be tested from the third original optical information with the first original optical information as the reference optical information, the first set of valid detection data further includes the fifth valid optical pulse information, and correspondingly, the second set of valid detection data obtaining step S530 further includes determining a sixth valid optical pulse information of each cell to be tested from the third original optical information with the second original optical information as the reference optical information, and the second set of valid detection data further includes the sixth valid optical pulse information.

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.

As shown in fig. 7, in some embodiments, the first original optical information is forward scattered light information, the second original optical information is fluorescence information, and the third original optical information is side scattered light information. When forward scattered light information is taken as reference optical information, n pulses (i.e., the number of pulses 1 in fig. 7) are identified from the forward scattered light information and n pieces of first effective light pulse information corresponding to the n pulses are acquired, each of the first effective light pulse information including first pulse amplitude information H11 (i.e., FS pulse amplitude in fig. 7) and first time information t1 (i.e., FS pulse position in fig. 7), and then second pulse amplitude information H12 corresponding to the vicinity of time t1 in the fluorescent information and fifth pulse amplitude information H15 corresponding to the vicinity of time t1 in the side scattered light information are acquired based on the first time information t1, whereby n pieces of second effective light pulse information including second pulse amplitude information H12 and n pieces of fifth effective light pulse information including fifth pulse amplitude information H15 are respectively obtained, and n pieces of first sets of effective detection data (i.e., n pieces of (t 1, H11, H12, H15) (i.e., three pieces of FS-based data in fig. 7) are obtained. When the fluorescence information is taken as the reference optical information, m pulses (i.e., the number of pulses 3 in fig. 7) are identified from the fluorescence information and m pieces of fourth effective light pulse information corresponding to the m pulses are acquired, each of the fourth effective light pulse information including fourth pulse amplitude information H24 (i.e., FL pulse amplitude in fig. 7) and second time information t2 (i.e., FL pulse position in fig. 7), and then, based on the second time information t2, third pulse amplitude information H23 corresponding to the vicinity of time t2 in the forward scattered light information and sixth pulse amplitude information H26 corresponding to the vicinity of time t2 in the side scattered light information are acquired, whereby m pieces of third effective light pulse information including third pulse amplitude information H23 and m pieces of sixth effective light pulse information including sixth pulse amplitude information H26 are respectively obtained, and m pieces of second sets of effective detection data (i.e., m pieces of (t 2, H23, H24, H26) (i.e., three pieces of FL-based amplitude data in fig. 7) are obtained.

It should be understood that the first original optical information, the second original optical information, and the third original optical information are merely different original optical information, and there is no sequential relationship among the three. Therefore, the first original optical signal may be side scattered light information or fluorescence information, the second original optical signal may be forward scattered light information or fluorescence information, and the third original optical signal may be forward scattered light information or side scattered light information, which only needs to satisfy that the first original optical signal, the second original optical signal, and the third original optical signal are different from each other.

When two original optical information is used as the reference optical information identification pulse, compared with the single original optical signal used as the reference optical signal, the cell identification omission can be avoided, and the accuracy of cell analysis is improved. Further, when three pieces of original optical information are used as reference optical information, the accuracy of cell analysis can be further improved.

Thus, according to an embodiment of the present invention, the method further comprises a third set of valid detection data acquisition steps:

Using the third original optical information as the reference optical information, and respectively determining seventh effective light pulse information, eighth effective light pulse information and ninth effective light pulse information of each cell to be detected from the first original optical information, the second original optical information and the third original optical information to obtain a third group of effective detection data including the seventh effective light pulse information, the eighth effective light pulse information and the ninth effective light pulse information of each cell to be detected;

Correspondingly, the cell identification step S540 comprises determining final effective detection data of each cell to be detected from the first set of effective detection data, the second set of effective detection data and the third set of effective detection data according to a preset rule, wherein the final effective detection data is used for identifying target cells in the sample liquid to be detected.

Optionally, the first set of valid detection data acquiring step S520 includes:

Acquiring second effective light pulse information and fifth effective light pulse information of each cell to be detected from the second original optical information and the third original optical information according to the first time information, wherein the second optical information and the fifth effective light pulse information respectively comprise second pulse amplitude information and fifth pulse amplitude information;

a second set of valid detection data acquisition steps S520, including:

Acquiring third effective light pulse information and sixth effective light pulse information of each cell to be detected 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 comprise third pulse amplitude information and sixth pulse amplitude information;

A third set of valid detection data acquisition steps comprising:

Identifying pulses generated by each of the test cells passing through the optical detection zone from the third original optical information based on a third amplitude threshold value and acquiring ninth effective light pulse information of each of the test cells, the ninth effective light pulse information including ninth pulse amplitude information and third time information,

And respectively acquiring seventh effective light pulse information and eighth effective light pulse information of each cell to be detected from the first original optical information and the second original optical information according to the third time information, wherein the seventh effective light pulse information and the eighth effective light pulse information respectively comprise seventh pulse amplitude information and eighth pulse amplitude information.

Referring to fig. 8, when first original optical information is taken as reference optical information, n pulses (for example, the number of pulses 1 in fig. 8) are identified from the first original optical information based on a first amplitude threshold and n pieces of first effective light pulse information corresponding to the n pulses are acquired, each piece of first effective light pulse information including first pulse amplitude information H11 (for example, FS pulse amplitude in fig. 8) and first time information t1 (for example, FS pulse position in fig. 8), and then second pulse amplitude information H12 corresponding to the vicinity of time t1 in the second original optical information and fifth pulse amplitude information H15 corresponding to the vicinity of time t1 in the third original optical information are acquired based on the first time information t1, whereby n pieces of second effective light pulse information including second pulse amplitude information H12 and n pieces of fifth effective light pulse information including fifth pulse amplitude information H15 are respectively obtained, and n pieces of first sets of effective detection data are obtained, that is, n pieces of (t 1, H11, H12, H15) (for example, FS data based on three pieces of amplitude in fig. 8). When the second original optical information is taken as the reference optical information, m pulses (for example, the number of pulses 2 in fig. 8) are identified from the second original optical information based on the second amplitude threshold value and m pieces of fourth effective light pulse information corresponding to the m pulses are obtained, each piece of fourth effective light pulse information including fourth pulse amplitude information H24 (for example, SS pulse amplitude in fig. 8) and second time information t2 (for example, SS pulse position in fig. 8), then, according to the second time information t2, third pulse amplitude information H23 corresponding to the vicinity of time t2 in the first original optical information and sixth pulse amplitude information H26 corresponding to the vicinity of time t2 in the third original optical information are obtained, thereby obtaining m pieces of third effective light pulse information including third pulse amplitude information H23 and m pieces of sixth effective light pulse information including sixth pulse amplitude information H26, respectively, and m pieces of second sets of effective detection data (t 2, H23, H24, H26) (for example, SS data in fig. 8 are based on the three amplitudes) are obtained. When the third original optical information is taken as the reference optical information, i pulses (for example, the number of pulses 3 in fig. 8) are identified from the third original optical information based on the third amplitude threshold value and i pieces of ninth effective light pulse information corresponding to the i pulses are acquired, each of the ninth effective light pulse information including ninth pulse amplitude information H39 (for example, FL pulse amplitude in fig. 8) and third time information t3 (for example, FL pulse position in fig. 8), and then seventh pulse amplitude information H37 corresponding to the vicinity of time t3 in the first original optical information and eighth pulse amplitude information H38 corresponding to the vicinity of time t3 in the second original optical information are acquired based on the third time information t3, whereby i pieces of seventh effective light pulse information including seventh pulse amplitude information H37 and i pieces of eighth effective light pulse information including eighth pulse amplitude information H38 are respectively obtained, and i pieces of third sets of effective detection data, i pieces (t 3, H37, H38, FL, H39) (for example, three pieces of data based on the amplitude in fig. 8) are obtained. In other words, after three different kinds of original optical information are received, pulse identification is performed by using the first original optical information, pulse amplitude information and position information are obtained, the number of pulses is counted, pulses at the same position are found on the second original optical information and the third original optical information, and amplitude information are obtained, pulse identification is performed by using the second original optical information, pulse amplitude information and position information are obtained, pulse number is counted, pulses at the same position are found on the first original optical information and the third original optical information, and amplitude information are obtained, pulse identification is performed by using the third original optical information, pulse amplitude information and position information are obtained, pulse number is counted, 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, final valid detection data is determined from the n first, m second and i third sets of valid detection data according to a preset rule (e.g., the smart screening algorithm of fig. 8) for identifying and counting target cells, and optionally for generating a scatter plot.

In the embodiment of the present invention, the "acquisition amplitude" in fig. 6 to 8 indicates that, based on the pulse position obtained by pulse recognition based on one kind of original optical information, pulses are searched for on the other original optical information in the vicinity of the same position, and the difference between the peak value and the base line thereof is recorded as the pulse amplitude.

In some embodiments, the cell identification step S540 includes:

Judging whether time information in each group of effective detection data belongs to the same cell or not;

for the case of belonging to the same cell, selecting one of the groups of effective detection data corresponding to the same cell as final effective detection data of the same cell;

for the case of different cells, the effective detection data corresponding to the time information of the different cells is used as the final effective detection data of the different cells.

That is, the final valid detection data of each test cell is determined by comparing the time information in each set of valid detection data of each test cell. In other words, when a plurality of sets of valid detection data are acquired for the same cell, one set of valid detection data is selected as the final valid detection data of the cell, and when only one set of valid detection data is acquired for the same cell, the set of valid detection data is selected as the final valid detection data of the cell.

In some embodiments, where the pulses are identified with two different primary optical information as reference optical information, determining the final valid detection data by comparing time information in each set of valid detection data for each test cell includes:

judging whether the first time information t1 in each first group of effective detection data is the same as/basically the same as the second time information t2 in each second group of effective detection data, namely whether the first time information t1 and the second time information t2 belong to the same cell;

For the first time information t1 and the second time information t2 which are basically the same and belong to the same cell, selecting a first group of effective detection data or a second group of effective detection data corresponding to the same cell as final effective detection data of the same cell;

And for different first time information and second time information which belong to different cells, respectively taking a first group of effective detection data and a second group of effective detection data which correspond to the first time information and the second time information which belong to different cells as final effective detection data of the different cells.

In some embodiments, where the pulses are identified with three different primary optical information as reference optical information, determining the final valid detection data by comparing time information in each set of valid detection data for each test cell includes:

Judging whether the first time information t1 in each first group of effective detection data, the second time information t2 in each second group of effective detection data and the third time information t3 in the third group of effective detection data are the same or basically the same, namely whether the first time information t1 and the second time information t2 in each second group of effective detection data belong to the same cell;

for the first time information t1, the second time information t2 and the third time information t3 belonging to the same cell, selecting a first group of effective detection data or a second group of effective detection data or a third group of effective detection data corresponding to the same cell as final effective detection data of the same cell;

For the first time information t1 and the second time information t2 belonging to the same cell, selecting a first group of effective detection data or a second group of effective detection data corresponding to the same cell as final effective detection data of the same cell;

For the first time information t1 and the third time information t3 belonging to the same cell, selecting a first group of effective detection data or a third group of effective detection data corresponding to the same cell as final effective detection data of the same cell;

selecting a second set of valid detection data or a third set of valid detection data corresponding to the same cell as final valid detection data of the same cell for second time information t2 and third time information t3 belonging to the same cell;

For the first time information t1, the second time information t2 and the third time information t3 which do not belong to the same cell, the first group of effective detection data, the second group of effective detection data and the third group of effective detection data corresponding to the first time information, the second time information and the third time information which belong to different cells are respectively used as final effective detection data of the different cells.

That is, in general, the pulse generated by the cell passing through the optical detection region can be identified in two or three different kinds of original optical information, so that for the cell identified in the plurality of different kinds of original optical information, two or three sets of valid detection data obtained by taking the plurality of different kinds of original optical information as reference optical information are substantially identical, and one set of data can be taken as the final data of the cell identified in the plurality of different kinds of original optical information. However, pulses generated by some small-volume cells (e.g., platelets) cannot be identified in the forward scattered light information (the pulse amplitude of the forward scattered light information may be 0 or less than the amplitude threshold), but can only be identified in the fluorescence information or the side scattered light information, and the set of valid detection data obtained by using the fluorescence information or the side scattered light information as the reference optical information is used as the final valid detection data of the small-volume cells.

In some embodiments, the cell identification step S540 includes determining final valid detection data of the same cell by comparing amplitude information in each set of valid detection data corresponding to the same cell or according to the type of the target cell for the case that the time information in each set of valid detection data belongs to the same cell. In other words, for a plurality of sets of valid detection data obtained from the same cell, one of the sets of data is selected according to the type of the target cell, or one of the sets of data is selected according to the peak difference of each set of valid detection data.

For example, a group having the largest sum of the magnitudes of the sets of effective detection data corresponding to the same cell may be selected as the final effective detection data of the same cell. The sum of the amplitudes of a group of effective detection data refers to the sum of the pulse amplitudes corresponding to each original optical information of the same cell at the same pulse position.

In some embodiments, when different first and second original optical information are employed as reference optical information, if a first set of valid detection data and a second set of valid detection data are acquired for the same cell, wherein the first set of valid detection data comprises amplitude information (H11, H12) or (H11, H12, H15) and first time information t1, and the second set of valid detection data comprises amplitude information (H23, H24) or (H23, H24, H26) and second time information t2, t1 is substantially the same as t 2. At this time, the sum of the magnitudes of the first set of valid detection data is h11+h12 or h11+h12+h15, and the sum of the magnitudes of the second set of valid detection data is h23+h24 or h23+h24+h26, and valid detection data corresponding to a larger value of h11+h12 (or h11+h12+h15) and h23+h24 (or h23+h24+h26) may be selected as the final valid detection data of the cell. Further, when only the first or second set of valid detection data is acquired for the same cell, the first or second set of valid detection data is selected as the final valid detection data for that cell.

In some embodiments, when a different first original optical information is used, When the second original optical information and the 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 comprises amplitude information (H11, H12, H15) and first time information t1, the second set of valid detection data comprises amplitude information (H23, H24, H26) and second time information t2, t1 is substantially the same as t2, and at this time, the sum of the amplitudes of the first set of valid detection data is h11+h12+h15, the sum of the amplitudes of the second set of valid detection data is h23+h24+h26, and valid detection data corresponding to a larger value of h11+h12+h15 and h23+h24+h26 is selected as final valid detection data for the cell; (2) if a first set of valid detection data and a 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 third set of valid detection data includes amplitude information (H37, H38, H39) and third time information t3, t1 and t3 are substantially the same, and at this time, the sum of the amplitudes of the first set of valid detection data is H11 +H2, and the sum of the amplitudes of the third set of valid detection data is H37 +H2 +H39, valid detection data corresponding to a larger value of H11 +H2 +H15 and H37 +H2 +H39 can be selected as final valid detection data for the cell, (3) if a second set of valid detection data and a third set of valid detection data are obtained for the same cell, wherein the second 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 and t3 being substantially the same, where the sum of the amplitudes of the second set of valid detection data is H23+H24+H226 and the sum of the amplitudes of the third set of valid detection data is H237+H238+H239, and valid detection data corresponding to a larger value of H23+H224+H226 and H237+H238+H239 can be selected as final valid detection data of the cell, and (5) if the first set of valid detection data is obtained for the same cell, A second set of valid detection data and a third set of valid detection data, wherein the first set of valid detection data comprises amplitude information (H11, H12, H15) and first time information t1, the second set of valid detection data comprises amplitude information (H23, H24, H26) and second time information t2, the third set of valid detection data comprises amplitude information (H37, H38, H39) and third time information t3, t1, t2 and t3 are substantially the same, where the sum of the magnitudes of the first set of valid detection data is h11+h12+h15, the sum of the magnitudes of the second set of valid detection data is h23+h24+h26, and the sum of the magnitudes of the third set of valid detection data is h37+h38+h39, the valid detection data corresponding to the maximum value of h11+h12+h15, h23+h24+h26, and h37+h38+h39 may be selected as the final valid detection data of the cell, and (6) when only the first set of valid detection data, the second set of valid detection data, or the third set of valid detection data is obtained for the same cell, the first set of valid detection data, the second set of valid detection data, or the third set of valid detection data is selected as the final valid detection data of the cell.

Alternatively, the sum of the above-mentioned magnitudes may be replaced by the average magnitude, that is, a group having the largest average magnitude of the sets of valid detection data corresponding to the same cell may be selected as the final valid detection data of the same cell. The average amplitude of a set of valid data refers to the average of the amplitudes corresponding to the respective original optical information of the same cell at the same pulse position. For example, if a first set of valid detection data and a second set of valid detection data are acquired for the same cell, wherein the first set of valid detection data comprises amplitude information (H11, H12) and first time information t1, the second set of valid detection data comprises amplitude information (H23, H24) and second time information t2, t1 is substantially the same as t 2. At this time, the average amplitude of the first set of valid detection data is H2+H2/2, and the average amplitude of the second set of valid detection data is H2+H2/2.

In some embodiments, when multiple sets of valid detection data are obtained for the same cell, one of the sets of valid detection data is simply selected as the final valid detection data for that cell, depending on the type of target cell. For example, when the first original 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.

In some embodiments, the cell identification step S540 includes:

and selecting one group of the effective detection data as the final effective detection data according to the type of the target cells so as to identify the target cells in the sample liquid to be detected.

That is, in the cell identification step S540, it is possible to select one of the plurality of sets of valid detection data as the final valid detection data of each test cell directly and simply according to the type of the target cell without determining whether or not the time information of each set of valid detection data is the same.

Optionally, when the target cells are platelets and/or reticulocytes, the first set of valid detection data of each of the test cells is selected as final valid detection data of each of the test cells to identify the platelets and/or reticulocytes in the test sample fluid.

Because different types of target cells exhibit different characteristics in different optical detection directions, such as platelets may be better differentiated in fluorescence or side scatter light information than in forward scatter light information, the user may directly select the valid detection data detected with fluorescence or side scatter light information as reference optical information as final valid detection data.

In some embodiments, the cell identification step S540 includes identifying target cells in the sample fluid to be tested and determining the number of target cells in the sample fluid to be tested based on the amplitude information of the final valid detection data.

That is, the cells whose amplitude information satisfies a certain condition in the final effective detection data are target cells, and the number of cells whose amplitude information satisfies a certain condition in the final effective detection data is the number of target cells.

In some embodiments, since the characteristics of the cells in the different optical information are different, statistics of the pulse number based on any one optical information are inaccurate, and a manner of combining pulse counting with pulse position (i.e. time information) is adopted here, wherein the pulse information is associated with position information, and for repeated pulses identified by the same cell in the different optical information in the whole measurement process, only one valid position is reserved, and then the total number of pulse positions is the total number of pulses. For example, in the whole measurement process, three pulses are identified and recorded by using forward scattered light information as reference optical information, positions are (p 1, p2, p 3), four pulse positions are identified and recorded by using side scattered light information as reference optical information, four pulse positions are identified and recorded by using fluorescence information as reference optical information, the repetition positions are not counted repeatedly, and finally the pulse positions are (p 1, p2, p3, p4, p 5), namely the total number of obtained pulses is 5.

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

and generating a scatter diagram of the cells to be detected according to the amplitude information in the final effective data of each cell to be detected so as to identify the target cells in the sample liquid to be detected.

Specifically, the data processing device 70 in the cell analyzer according to the embodiment of the present invention can obtain a three-dimensional scatter diagram using fluorescence intensity, forward scattered light intensity, and side scattered light intensity at the same time, thereby distinguishing the target cell population from other cell populations, obtaining more accurately identified target cells, and calculating the number of target cells.

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 period of time, 50 ms. 1170 pulses are obtained using the forward scattered light information as reference optical information for pulse identification, and 1205 pulses are obtained using the fluorescence information as reference optical information for pulse identification. There are a total of 1162 pulses detected at the same position (time) in both modes, and 8 pulses are recognized in the forward scattered light information but are not recognized in the fluorescent information, and 43 pulses are recognized in the fluorescent information but are not recognized in the forward scattered light information. Among 1162 pulse data detected in both forward scattered light information and fluorescence information, a were platelets, among 8 pulse data unique to forward scattered light information, 4 were smaller in amplitude, which could be regarded as platelets empirically, and among 43 pulse data unique to fluorescence information, 11 were smaller in amplitude, which could be regarded as platelets empirically. Combining the above data, the following results can be obtained:

Reference to optical information	Total number of pulses	Platelet count
			Forward scattered light information	1170 Pieces	A+4
Fluorescence information	1205 Pieces of	A+11
			Combining the two	1213	A+15

It follows that the total number of pulses and the number of platelets identified by the pulse identification method with reference to the fluorescence information are greater than those with reference to the forward scattered light information, because the discrimination in the fluorescence information is better for small-volume cells of the type with respect to platelets. Furthermore, the total number of pulses and the number of platelets obtained by combining the fluorescence information and the forward scattered light information as reference optical information are increased. It is thus shown that the limitation of pulse recognition and cell analysis with reference to a single raw optical data can be overcome with reference to multiple raw optical data, which is superior to the case with reference to a single raw optical data.

There is also provided, in accordance with an embodiment of the present invention, as shown in fig. 9, a cell analysis method 900 including:

s910, selecting the type of reference optical information according to the type of the target cells, wherein the reference optical information is used for identifying that each cell to be tested in the sample liquid to be tested passes through an optical detection area of a cell analyzer to generate effective light pulses;

s920, obtaining at least two kinds of optical information of each cell to be detected in the sample liquid to be detected passing through the optical detection area, wherein the at least two kinds of optical information comprise the reference optical information and at least one kind of non-reference optical information;

s930, determining effective detection data corresponding to the effective light pulse from the at least two optical information according to the reference optical information;

S940, identifying target cells in the sample liquid to be detected according to the effective detection data.

For some types of target cells, the degree of distinction of the types of the cells in which original optical information is higher than that of other original optical information is known, so that the reference optical information can be directly determined according to the types of the target cells, and other collected original optical information is used as non-reference optical information, so that the accuracy is ensured, and meanwhile, the identification speed of the target cells is increased.

Optionally, determining the valid detection data according to the reference optical information includes:

Based on a preset amplitude threshold value, identifying pulses generated by each cell to be detected passing through the optical detection area from the reference optical information and acquiring effective light pulse data of each cell to be detected, wherein the effective light pulse data comprises first pulse amplitude information and time information;

Acquiring at least one non-reference optical data of each cell to be tested from at least one non-reference optical information according to the time information, wherein the non-reference optical data comprises second pulse amplitude information;

wherein the valid detection data comprises the valid light pulse data and the at least one non-reference optical data.

Optionally, when the target cell is a platelet and/or reticulocyte, side scattered light information or fluorescence information is selected as the reference optical information, and the at least two optical information includes at least two of forward scattered light information, side scattered light information, and fluorescence information.

In one embodiment, n reference effective light pulse information corresponding to n pulses is identified from reference optical information (e.g., fluorescence information), each reference effective light pulse information includes reference pulse amplitude information H and reference time information t, and then non-reference pulse amplitude information H1 corresponding to the vicinity of time t in at least one non-reference optical information (e.g., scattered light information) is acquired from the reference time information t, thereby obtaining non-reference effective light pulse information including non-reference pulse amplitude information, and effective detection data, i.e., (t, H1) is obtained.

Further, a scatter diagram of the test cells can be generated according to the amplitude information in the effective detection data to identify platelets in the test sample liquid, and the number of platelets is determined according to the reference time information in the effective detection data.

There is also provided, in accordance with an embodiment of the present invention, a method for detecting platelets and/or reticulocytes, including:

and identifying platelets and/or reticulocytes in the sample liquid to be detected according to the first effective detection data.

The method comprises the steps of determining first effective detection data corresponding to effective light pulses from at least two kinds of optical information, generating a scatter diagram of the to-be-detected cells according to amplitude information of the first effective detection data, and identifying the platelets and/or the reticulocytes in the to-be-detected sample liquid and determining the quantity of the platelets and/or the reticulocytes when the platelets and/or the reticulocytes in the to-be-detected sample liquid are identified by taking the side scattering light information or the fluorescence information as reference optical information.

Further, the at least two types of optical information include forward scattered optical information, the method further comprising:

determining second effective detection data corresponding to the effective light pulses from the at least two kinds of optical information by taking the forward scattering light information as reference optical information for identifying that each cell to be detected generates the effective light pulses through the optical detection area;

And identifying platelets and/or reticulocytes in the sample liquid to be tested according to the first effective detection data and the second effective detection data.

Embodiments of the present application also provide a computer readable storage medium storing a plurality of program instructions that, when invoked by a processor for execution, may perform part or all of the steps or any combination of the steps in the cell analysis method of the embodiments of the present application.

According to the cell analysis method, the cell analyzer and the computer readable storage medium, the plurality of different types of optical signals generated when cells pass through the flow chamber are respectively used as the reference signals for pulse identification, the principle that the different types of optical signals reflect different cell characteristic information is fully utilized, the condition that target cells are not identified or cannot be identified is avoided, the accuracy of cell classification, scatter diagram morphology and cell counting is improved, and the accuracy of the blood cell analyzer is further improved.

It will be understood by those skilled in the art that all of the features disclosed in this specification (including any accompanying claims, abstract and drawings), and all of the processes or units of any method or apparatus so disclosed, may be combined in any combination, except combinations where the features are mutually exclusive. Each feature disclosed in this specification (including any accompanying claims, abstract and drawings), may be replaced by alternative features serving the same, equivalent or similar purpose, unless expressly stated otherwise.

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. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. Further, as used in the specification, the terms "comprises" and/or "comprising" mean that there is a stated feature, integer, step, operation, element, and/or component, but does not preclude the presence or addition of one or more other features, integers, steps, operations, elements, and/or components.

The flow chart described in the present invention is merely one embodiment, and many modifications may be made to this illustration or the steps in the present invention without departing from the spirit of the invention. For example, the steps may be performed in a differing order, or steps may be added, deleted or modified. Those skilled in the art will recognize that the invention can be practiced with modification within the spirit and scope of the appended claims.

Claims

1. A method of cell analysis comprising:

The method comprises the steps of acquiring original optical information, namely acquiring first original optical information detected in a first optical detector and second original optical information detected in a second optical detector when each cell to be detected in a sample liquid to be detected passes through an optical detection zone of a cell analyzer within a preset time period, wherein the first original optical information and the second original optical information are different optical information;

2. The method of claim 1, wherein the first primary optical information and the second primary optical information are each selected from one of forward scattered light information, side scattered light information and fluorescence information.

3. The method of claim 2, wherein the first or second primary optical information is fluorescence information.

4. A method of cell analysis according to claim 3 wherein the target test cells are platelets and/or reticulocytes.

5. The method according to any one of claims 1 to 4, wherein the raw optical information obtaining step further comprises obtaining that each test cell in the test sample liquid detects third raw optical information in a third optical detector when passing through the optical detection zone within the preset time period;

The first group of effective detection data obtaining step further includes using the first original optical information as the reference optical information, determining fifth effective light pulse information of each cell to be detected from the third original optical information, the first group of effective detection data further including the fifth effective light pulse information;

The second set of valid detection data obtaining step further includes determining sixth valid light pulse information of each cell to be tested from the third original optical information with the second original optical information as the reference optical information, and the second set of valid detection data further includes the sixth valid light pulse information.

6. The method of claim 5, wherein the first set of valid detection data acquisition steps comprises:

the second set of valid detection data acquisition steps includes:

And acquiring third effective light pulse information and sixth effective light pulse information of each cell to be detected 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 comprise third pulse amplitude information and sixth pulse amplitude information.

7. The method of claim 5, further comprising a third set of valid detection data acquisition steps:

the cell identification step comprises the step of determining final effective detection data of each cell to be detected according to a preset rule by using the first group of effective detection data, the second group of effective detection data and the third group of effective detection data of each cell to be detected, and identifying target cells in the sample liquid to be detected.

8. The method of claim 7, wherein the first set of valid detection data acquisition steps comprises:

the second set of valid detection data acquisition steps includes:

the third set of valid detection data acquisition steps includes:

9. The method of claim 6 or 8, wherein the cell identification step comprises:

10. The method of claim 9, wherein the cell identification step comprises:

For the case of belonging to the same cell, determining a group from the sets of valid detection data corresponding to the same cell as final valid detection data of the same cell according to the amplitude information of the sets 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, wherein the cell detection step comprises:

and selecting one group from the effective detection data according to the type of the target cells as the final effective detection data so as to identify the target cells in the sample liquid to be detected.

12. The method of claim 11, wherein the first original optical information is fluorescence information or side scatter light information;

the cell detection step comprises the step of selecting the first set of valid detection data as the final valid detection data when the target cells are platelets and/or reticulocytes so as to identify the platelets and/or reticulocytes in the sample liquid to be detected.

13. The method according to any one of claims 6, 8 to 10, wherein the cell identification step comprises:

And identifying target cells in the sample liquid to be detected according to the amplitude information of the final effective detection data of each cell to be detected and determining the number of the target cells in the sample liquid to be detected.

14. The method according to any one of claims 6, 8 to 10, further comprising:

And generating a scatter diagram of the cells to be detected according to the amplitude information in the final effective detection data of each cell to be detected so as to identify the target cells in the sample liquid to be detected.

15. A method of cell analysis comprising:

16. The method of claim 15, wherein determining the valid detection data from the reference optical information comprises:

And acquiring at least one non-reference optical data of each cell to be detected from at least one non-reference optical information according to the time information, wherein the non-reference optical data comprises second pulse amplitude information, and the effective detection data comprises the effective light pulse data and the at least one non-reference optical data.

17. The method according to claim 15 or 16, wherein when the target cells are platelets and/or reticulocytes, side scatter light information or fluorescence information is selected as the reference optical information, said at least two optical information comprising at least two of forward scatter light information, side scatter light information and fluorescence information.

18. A method for detecting platelets and/or reticulocytes, comprising:

19. The method of claim 18, wherein the at least two types of optical information further comprise forward scattered optical information, the method further comprising:

Determining second effective detection data corresponding to the effective light pulse from the at least two kinds of optical information by taking the forward scattered light information as the reference optical information;

20. A cell analyzer, comprising:

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 of any one of claims 1 to 19 when the computer program is executed by the processor.

21. A computer readable storage medium comprising a program executable by a processor to implement the method of any one of claims 1 to 19.