CN115398209A

CN115398209A - Cell analysis method, cell analyzer, and computer-readable storage medium

Info

Publication number: CN115398209A
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: 2022-11-25
Also published as: WO2021207894A1

Abstract

A cell analysis method, a cell analyzer, and a computer-readable storage medium, the method comprising: acquiring first original optical information and second original optical information of each cell to be detected in a sample liquid to be detected; taking the first original optical information as reference optical information to obtain a first group of effective detection data of each cell to be detected; taking the second original optical information as reference optical information to obtain a second group of effective detection data of each cell to be detected; and determining the final effective detection data of each cell to be detected according to the first group of effective detection data and the second group of effective detection data according to a preset rule, wherein the final effective detection data is used for identifying the target cell in the sample liquid to be detected. According to the cell analysis method, the cell analyzer and the computer-readable storage medium, the 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 technologies, and in particular, to a cell analysis method, a cell analyzer, and a computer-readable storage medium.

Background

At present, a hemocyte analyzer generally uses a laser scattering principle to detect and analyze cells, and mainly uses a low-angle scattered light signal (forward scattered light signal FS), a side scattered light signal (SS) and a side fluorescent signal (FL) to detect and analyze cells, wherein FS represents cell volume information, FS represents cell surface complexity, and FL represents intracellular computational content. The light scattering cell detection technology combines the optical characteristic and the biological characteristic of the cell, and has the advantages of rapidness, simplicity, accuracy, no destructiveness, good repeatability and the like, and the light scattering cell detection technology is greatly developed and widely applied to cell identification. When the light scattering principle is adopted for detection, light beams irradiate cells to be detected passing through the optical detection area, light and the cells interact to generate scattered light signals, and the scattered light signals contain information related to the size and the distribution of the cells. Under the condition that the cell to be detected is stained by fluorescence, the cell to be detected also generates a fluorescence signal for representing the content of nucleic acid in the cell after being irradiated by light. When the cell to be detected rapidly passes through the optical detection area, the intensity of three generated optical signals (namely FS, SS and FL) is changed instantly 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 amplitude of the cell to be detected in the three optical signals is obtained, the pulse number is counted, a scatter diagram can be generated by analyzing pulse amplitude information, the cell is classified, and the pulse number can represent the cell number. However, at present, only the low-angle scattering signals acquired in the low-angle direction are used for pulse recognition, and the low-angle scattering signals represent volume information of cells, so that cells with large volumes can be well recognized based on the low-angle scattering signals, however, for some small volumes, recognition missing or recognition failure occurs, and thus 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 to solve at least one of the above problems.

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

an original optical information acquisition step: 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 area of a cell analyzer within a preset time period;

a first set of valid detection data acquisition steps: 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 passing through the optical detection area, so as to acquire a first set of effective detection data including the first effective light pulse information and the second effective light pulse information of each cell to be detected;

a second group of effective detection data acquisition steps: 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 obtain a second set of effective detection data of each cell to be detected, wherein the second set of effective detection data comprises the third effective light pulse information and the fourth effective light pulse information;

a cell recognition step: and determining the 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 is used for identifying the target cell in the sample liquid to be detected.

According to a second aspect of the present invention, there is provided a cell analysis method 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 detected in the sample liquid to be detected 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 when the cell to be detected passes 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 valid detection data corresponding to the valid light pulse from the at least two kinds of optical information according to the reference optical information;

and identifying the 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 light information or fluorescence information;

determining first effective detection data corresponding to the effective light pulse from the at least two kinds of optical information by using the side scattered light information or the fluorescence information as reference optical information for identifying the effective light pulse generated by each of the cells to be detected passing through the optical detection region;

and identifying the blood 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 analyzer comprising:

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

a sample preparation device having a reaction cell for receiving a blood sample drawn by a sampling device and a reagent supply portion for supplying a reagent to the reaction cell so that the blood sample drawn by the sampling device is mixed with the reagent supplied by the reagent supply portion in the reaction cell to prepare a sample solution to be measured;

the optical detection device comprises a light source, a flow chamber and a light detector, wherein each cell to be detected of the sample liquid to be detected can flow in the flow chamber, the light emitted by the light source irradiates the cell in the flow chamber to generate optical information, and the light detector is used for collecting the optical information; and

a data processing device electrically connected with 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 including: a program executable by a processor to implement a method as described in the first to third aspects.

According to the cell analysis method, the cell analyzer and the computer-readable storage medium provided by the embodiment of the invention, a plurality of different types of optical signals generated by cells are respectively used as reference optical information for pulse recognition, cell characteristic information reflected by different optical signals is fully utilized, missing recognition or incapability of recognition of target cells is avoided, and the accuracy of cell classification, scatter diagram form and cell counting is improved, so that the accuracy of the blood cell analyzer is improved.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.

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

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

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

FIG. 4 is a logic diagram of a prior art cell analysis method;

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

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

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

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

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

According to an embodiment of the present invention, there is provided a cellular 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 aspirate a liquid sample to be tested quantitatively 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 a liquid sample to be tested sucked by the sampling device, the reagent supply part is used for supplying at least one reagent to the reaction tank, and thus 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 a sample solution 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.

A 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 some or all of the steps or any combination of the steps of the cell analysis method according to an embodiment of the present invention when the computer program is executed by the processor. The cell analysis method practiced by 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 (ASICs), a Digital Signal Processor (DSP), a Digital Signal Processing Device (DSPD), a Programmable Logic Device (PLD), a Field Programmable Gate Array (FPGA), a Central Processing Unit (CPU), a controller, a microcontroller, and a microprocessor, so that the Processor may perform some or all of the steps of the cell analysis method in the embodiments of the present Application, or any combination of the steps thereof.

Alternatively, the computer readable storage medium may be either volatile memory or nonvolatile memory, and may include both volatile and nonvolatile memory. The nonvolatile memory can 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; the magnetic surface storage may be disk storage or tape storage. Volatile memory may be random access memory, which acts as external cache memory. By way of illustration and not limitation, many forms of RAM are available, such as SRAM, SDRAM, DRAM, SDRAM, DDR SDRAM, SSRAM, SDRAM, and DMA bus RAM. The memory described in connection with the embodiments of the invention is intended to encompass these and any other suitable types of memory.

In one embodiment, FIG. 1 shows a schematic structural diagram of a cellular 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 housing 200, and are disposed on both sides of the second housing 200. The sample preparation device 30 is disposed inside the first housing 100, and the output 90 and the sampling device 10 are disposed on an outer surface of the first housing 100.

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., without limitation. In this embodiment, a sample of a liquid to be tested is taken as a blood sample for illustration.

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

Alternatively, the sample preparation device 30 has a reaction cell and a reagent supply portion that stores reagents for reacting with the blood sample, for example, at least a first reagent and/or a second reagent, and supplies the respective reagents to the reaction cell as needed. The first reagent may be a diluent or a hemolytic agent, wherein the hemolytic agent is used for cracking red blood cells in the liquid sample to be tested into fragments and keeping the morphology of white blood cells 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 tested.

Optionally, the optical detection device 50 includes at least two of a forward scatter light detector, a side scatter light detector, and a side fluorescence detector. The forward scatter detectors are typically arranged in line with the light source and the flow cell, and the light source is arranged on each side of the flow cell to detect the forward scatter or low angle scatter intensity of the cells flowing in the flow cell. The side scatter detectors are typically arranged at an angle to a line along which the light source and the flow cell lie to detect the intensity of side scatter light from cells flowing in the flow cell. The fluorescence detector is typically disposed at an angle to a line in which the light source and the flow cell are located to detect lateral fluorescence intensity of 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 series. On one side of the flow cell 103, a dichroic mirror 106 is arranged at an angle of 45 ° to the straight line. A portion of the side light emitted by the cells in flow cell 103 is transmitted through dichroic mirror 106 and captured by fluorescence detector 105 arranged behind dichroic mirror 106 at an angle of 45 ° to dichroic mirror 106; while another portion of the side light is reflected by dichroic mirror 106 and captured by side scatter light detector 107 arranged at a 45 ° angle to dichroic mirror 106 in front of dichroic mirror 106.

It should be understood that the line where the light source and the flow cell are located may be in line with the forward scatter detector or may form an angle, which may be set as desired, and is not limited herein; similarly, the angle between the straight line where the light source and the flow cell are located 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 required, and are not limited herein.

The data processing device 70 is configured to detect the target cell flowing through the flow cell 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, 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 present invention, the cell analyzer may further provide a corresponding operation interface for an operator to operate, and the operation interface may include corresponding controls, such as an identification selection box or a menu bar, so that the operator may input an operation instruction on the operation interface according to an actual use condition, so as to implement cell analysis by 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 an I/O interface 75. The processor 71, RAM72, ROM73, communication interface 74, memory 76, and I/O interface 75 communicate over a bus 77. The processor may be a CPU, GPU or other chip with computing capabilities. The memory 76 stores various computer programs such as an operating system and an application program to be executed by the processor 71, and data necessary for executing the computer programs. In addition, data stored locally during the cell testing process, if desired, may be stored in memory 76. 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 and an a/D converter. The I/O interface 75 is connected to an input device comprising a keyboard, mouse, touch screen or other control buttons, which a user can use to input data directly to the data processing apparatus 70. In addition, 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 a display for displaying, for example: analytical data, instrument operating parameters, etc. The communication interface 74 is an interface that may be any communication protocol known today. The communication interface 74 communicates with the outside through a network. Data processing device 70 may communicate data with any device connected through the network via communication interface 74 in a communication protocol.

In the present practice, as shown in FIG. 4, a 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 recognition, so as to obtain pulse information of the forward scattered light information, including the number of pulses, FS pulse amplitude H1, FS pulse position t (i.e. time corresponding to the recognized pulse), and the like. Then, based on the pulse position t corresponding to each pulse identified according to the forward scattered light information, pulse amplitudes H2 and H3 corresponding to the pulse position t of 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 are respectively obtained, then three amplitude data (H1, H2, H3) based on FS 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 according to the three amplitude data (H1, H2, H3) based on FS and a corresponding scattergram 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 (for example, platelets) which are small in volume but have a large complexity on the cell surface (i.e., a large SS signal intensity) or a high intracellular core count (i.e., a large FL signal intensity), pulse recognition using only the FS signal as a reference signal may result in missed recognition or unrecognizable state, which may result in inaccurate cell classification, abnormal scatter diagram, and inaccurate cell count.

In view of the above, a cell analysis method is provided according to an embodiment of the present invention, and referring to fig. 5, a schematic flow chart of the cell analysis method according to an embodiment of the present invention is shown. As shown in fig. 5, the method 500 includes:

original optical information acquisition step S510: 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 area of a cell analyzer within a preset time period;

a first group valid detection data acquisition step S520: 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, and 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 so as to obtain a first set of effective detection data including the first effective light pulse information and the second effective light pulse information of each cell to be detected;

a second group valid detection data acquisition step S530: 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 obtain a second set of effective detection data of each cell to be detected, wherein the second set of effective detection data comprises the third effective light pulse information and the fourth effective light pulse information;

cell recognition step S540: and determining the 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 is used for identifying the target cell in the sample liquid to be detected.

In an embodiment of the present invention, the original optical information may be a signal, e.g., an electrical signal, which is detected and output by the optical detector continuously for a predetermined time. The reference optical information is original optical information in which pulse information of other original optical information is acquired with reference to the recognized pulse position, and the pulse information acquired from the reference optical information and the pulse information acquired from the other original optical information based on the reference optical information may be regarded as effective light pulse information.

For example, when first original optical information is used as reference optical information, n pulses are identified from the first original optical information and n pulse information corresponding to the n pulses are obtained and are marked as first effective light pulse information; then according to the n pulse positions of the n pulses, acquiring pulse information corresponding to the n pulse positions from second original optical information, and recording the pulse information as second effective optical pulse information; the first valid light pulse information and the second valid light pulse information are recorded as a first set of valid detection data. When the second original optical information is taken as the reference optical information, identifying m pulses from the second original optical information and obtaining m pulse information corresponding to the m pulses, and marking the m pulse information as fourth effective light pulse information; then according to the m pulse positions of the m pulses, acquiring pulse information corresponding to the m pulse positions from the first original optical information, and recording the pulse information as third effective optical pulse information; and the third effective light pulse information and the fourth effective light pulse information are marked as a second group of effective detection data. And finally, obtaining final effective detection data from the first group of effective data and the second group of effective data according to an intelligent screening algorithm, namely a preset rule so as to identify the target cells.

According to the cell analysis method provided by the embodiment of the invention, different optical information collected by at least two optical detectors is respectively used as reference optical information for pulse recognition, the principle that different optical information reflects different characteristics of cells is fully utilized, complementation is formed in pulse recognition, and missing recognition or incapability of recognition of target cells is avoided. Compared with the traditional method of analyzing cells by only adopting forward scattering light information as reference optical information, the method overcomes the limitation of pulse recognition 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 using fluorescence information as reference optical information, the method is particularly suitable for identifying small-sized cells, such as platelets, reticulocytes, and the like, in a sample solution to be tested.

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

based on a first amplitude threshold value, identifying pulses generated when each cell to be detected passes through the optical detection area from the first original optical information and acquiring first effective light pulse information of each cell to be detected, wherein the first effective light pulse information comprises first pulse amplitude information and first time information,

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

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

based on a second amplitude threshold value, identifying pulses generated when each cell to be detected passes through the optical detection area from the second original optical information and acquiring fourth effective optical pulse information of each cell to be detected, wherein the fourth effective optical 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 for different original optical information different amplitude thresholds can be set for identifying the 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 zone; if the peak of a pulse identified from within the original optical information is less than the amplitude threshold, the pulse is deemed not to be a pulse generated by the cell, but may be a pulse generated due to signal interference. In other words, when no cell passes through the optical detection region, the first original optical information and the second original optical information are generally maintained under respective dc baseline voltages, and when a cell passes through the optical detection region and optical information is generated, the amplitude of the first original optical information and/or the second original optical information changes, it is possible to identify whether or not the cell is a valid pulse by judging the trend of the rising edge and the falling edge of the pulse, simultaneously recording the difference between the peak value and the baseline of the pulse as the pulse amplitude and recording the corresponding peak position (i.e., the time at which the pulse is generated), 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 raw optical information is forward scattered light information or side scattered light information and the second raw optical information is fluorescence information. Taking a blood sample as an example, referring to fig. 1 and 2 together, the sampling device 10 in the cellular analyzer collects a blood sample through a sampling needle and transfers the collected blood sample to the sample preparation device 30. The reagent supply portion of the sample preparation device 30 supplies the diluent and the fluorochrome to the reaction cell as required, and the blood sample reacts with the diluent and the fluorochrome in the reaction cell in sequence to obtain a to-be-detected liquid sample containing a plurality of cells, and the cells in the to-be-detected liquid sample are lined 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 cell, and the cells passing through the optical detection area are excited by light to generate different optical information. A forward scatter light detector or a side scatter light detector detects a forward scatter light intensity or a side scatter light intensity of the cell flowing in the flow chamber to obtain first raw optical information, and a fluorescence detector detects a side fluorescence intensity of the cell flowing in the flow chamber to obtain second raw optical information. The data processing device 70 performs cell analysis after acquiring the first original optical information and the 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 the reference optical information, identifying n pulses (for example, the number of pulses 1 in fig. 6) from the forward scattered light information or side scattered light information and acquiring n first effective light pulse information corresponding to the n pulses, each 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); then, based on the first time information t1, second pulse amplitude information H12 corresponding to the fluorescence information in the vicinity of the time t1 is acquired, so that n pieces of second effective light pulse information including the second pulse amplitude information H12 are obtained, and n pieces of first group effective detection data, that is, n pieces of (t 1, H11, H12) (for example, two pieces of amplitude data based on FS in fig. 6) are obtained. When the fluorescent light information is taken as the reference optical information, identifying m pulses (e.g., pulse number 3 in fig. 6) from the fluorescent light information and acquiring m fourth effective light pulse information corresponding to the m pulses, each of the fourth effective light pulse information including fourth pulse amplitude information H24 (e.g., FL pulse amplitude in fig. 6) and second time information t2 (e.g., FL pulse position in fig. 6); then, according to the second time information t2, third pulse amplitude information H23 corresponding to the vicinity of the time t2 in the forward scattered light information or the side scattered light information is acquired, so that m pieces of third effective light pulse information including the third pulse amplitude information H23 are obtained, and m pieces of second group effective detection data, that is, m pieces (t 2, H23, H24) are obtained (for example, two pieces of amplitude data based on FL in fig. 6). In other words, after receiving two kinds of original optical information, using the first original optical information to perform pulse recognition and obtain pulse amplitude information and position information, and meanwhile counting the number of pulses, finding pulses at the same position on the second original optical information and obtaining amplitude information; and simultaneously, using the second original optical information to carry out pulse identification and obtain pulse amplitude information and position information, counting the number of pulses, finding out pulses at the same position on the first original optical information and obtaining the amplitude information. Finally, final valid detection data are determined from the n first sets of valid detection data and the m second sets of valid detection data according to preset rules (e.g., the intelligent screening algorithm in fig. 6) for identifying and counting the 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 scattered light information and side scattered light information, respectively. Taking a blood sample as an example, referring again to fig. 1 and 2, the sampling device 10 in the cell analyzer collects a blood sample through a sampling needle and transfers the collected blood sample to the sample preparation device 30. The reagent supply portion of the sample preparation device 30 supplies the diluent to the reaction cell as required, the blood sample reacts with the diluent to obtain a liquid sample to be tested containing a plurality of cells, and the cells in the liquid sample to be tested are lined 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 zone of the flow cell, and the cells passing through the optical detection zone are excited by light to generate different optical information. A forward scatter light detector and a side scatter light detector detect a forward scatter light intensity and a side scatter light intensity, respectively, of a cell flowing in the flow cell to obtain first original optical information and second original optical information. The data processing device 70 performs cell analysis after acquiring the first original optical information and the second original optical information. For a specific cell analysis process, reference may be made to the embodiment described above with reference to fig. 6, which is not described herein again.

In some embodiments, the original optical information acquiring step S510 further includes: acquiring third original optical information detected in a third optical detector when each cell to be detected in the sample liquid to be detected passes through the optical detection area within the preset time period; accordingly, the first set of valid detection data acquiring step S520 further includes: determining fifth effective light pulse information of each cell to be detected from the third original optical information by taking the first original optical information as the reference optical information, wherein the first set of effective detection data further comprises the fifth effective light pulse information; accordingly, the second set of valid detection data acquiring step S530 further includes: and determining sixth effective light pulse information of each cell to be detected from the third original optical information by taking the second original optical information as the reference optical information, wherein the second set of effective detection data further comprises the sixth effective light 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 the forward scattering light information is used as the reference optical information, n pulses (i.e., number of pulses 1 in fig. 7) are identified from the forward scattering light information and n first effective light pulse information corresponding to the n pulses is obtained, each 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); then, according to the first time information t1, second pulse amplitude information H12 corresponding to the vicinity of the time t1 in the fluorescence information and fifth pulse amplitude information H15 corresponding to the vicinity of the time t1 in the side scattered light information are acquired, so that n pieces of second effective light pulse information including the second pulse amplitude information H12 and n pieces of fifth effective light pulse information including the fifth pulse amplitude information H15 are respectively obtained, and n pieces of first group effective detection data, that is, n pieces of (t 1, H11, H12, H15) (that is, three pieces of amplitude data based on FS 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 piece of 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); 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, thereby obtaining m pieces of third effective light pulse information including the third pulse amplitude information H23 and m pieces of sixth effective light pulse information including the sixth pulse amplitude information H26, respectively, and further obtaining m pieces of second set of effective detection data, i.e., m pieces of (t 2, H23, H24, H26) (i.e., three pieces of amplitude data based on FL in fig. 7).

It should be understood that the first original optical information, the second original optical information, and the third original optical information are only used to distinguish different original optical information, and there is no sequential relationship between the three. Therefore, the first original optical signal may also be side scattered light information or fluorescence information, the second original optical information may also be forward scattered light information or fluorescence information, and the third original optical information may also be forward scattered light information or side scattered light information, as long as the first original optical information, the second original optical information, and the third original optical information are different from each other.

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

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

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 respectively by taking the third original optical information as the reference optical information so as to obtain a third group of effective detection data comprising 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;

accordingly, the cell recognition step S540 includes: and according to a preset rule, determining the final effective detection data of each cell to be detected according to the first group of effective detection data, the second group of effective detection data and the third group of effective detection data, and identifying the target cell in the sample liquid to be detected.

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

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

the second set of valid detection data acquiring step S520 includes:

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

a third set of valid detection data acquisition steps comprising:

based on a third amplitude threshold value, identifying pulses generated when each cell to be detected passes through the optical detection area from the third original optical information and acquiring ninth effective optical pulse information of each cell to be detected, wherein the ninth effective optical pulse information comprises ninth pulse amplitude information and third time information,

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

Referring to fig. 8, when the first original optical information is taken as the reference optical information, n pulses (e.g., number of pulses 1 in fig. 8) are identified from the first original optical information based on the first amplitude threshold and n first valid light pulse information corresponding to the n pulses is obtained, each first valid light pulse information including first pulse amplitude information H11 (e.g., FS pulse amplitude in fig. 8) and first time information t1 (e.g., FS pulse position in fig. 8); then, according to the first time information t1, 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 obtained, so that n pieces of second effective light pulse information including the second pulse amplitude information H12 and n pieces of fifth effective light pulse information including the fifth pulse amplitude information H15 are obtained, respectively, and n pieces of first group effective detection data, that is, n pieces of (t 1, H11, H12, H15) (for example, three amplitude data based on FS in fig. 8) are obtained. When the second original optical information is taken as the reference optical information, identifying m pulses (e.g., pulse number 2 in fig. 8) from the second original optical information based on the second amplitude threshold and obtaining m fourth effective light pulse information corresponding to the m pulses, each fourth effective light pulse information including fourth pulse amplitude information H24 (e.g., SS pulse amplitude in fig. 8) and second time information t2 (e.g., 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, so that m pieces of third effective light pulse information including the third pulse amplitude information H23 and m pieces of sixth effective light pulse information including the sixth pulse amplitude information H26 are obtained, respectively, and m pieces of second group effective detection data, that is, m pieces of (t 2, H23, H24, H26) (for example, three pieces of amplitude data based on SS in fig. 8) are obtained. When the third original optical information is taken as the reference optical information, identifying i pulses (e.g., pulse number 3 in fig. 8) from the third original optical information based on a third amplitude threshold value and acquiring i ninth effective optical pulse information corresponding to the i pulses, each of the ninth effective optical pulse information including ninth pulse amplitude information H39 (e.g., FL pulse amplitude in fig. 8) and third time information t3 (e.g., FL pulse position in fig. 8); then, according to the third time information t3, 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, so that i pieces of seventh effective optical pulse information including the seventh pulse amplitude information H37 and i pieces of eighth effective optical pulse information including the eighth pulse amplitude information H38 are respectively obtained, and then i pieces of third group effective detection data, i.e., i pieces of (t 3, H37, H38, H39) (for example, three pieces of amplitude data based on FL in fig. 8) are obtained. In other words, after receiving three different original optical information, using the first original optical information to perform pulse recognition and obtain pulse amplitude information and position information, and meanwhile counting the number of pulses, finding pulses at the same position on the second original optical information and the third original optical information and obtaining amplitude information; simultaneously, using the second original optical information to carry out pulse identification and obtain pulse amplitude information and position information, simultaneously counting the number of pulses, finding out pulses at the same position on the first original optical information and the third original optical information and obtaining amplitude information; and simultaneously, carrying out pulse identification by using the third original optical information to obtain pulse amplitude information and position information, counting the number of pulses, finding out pulses at the same position on the first original optical information and the second original optical information, and obtaining the amplitude information. 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 intelligent screening algorithm in fig. 8) for identifying and counting the target cells and optionally for generating a scatter plot.

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

In some embodiments, the cell identifying step S540 comprises:

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

selecting one of the sets of valid detection data corresponding to the same cell as the final valid detection data of the same cell when the cells belong to the same cell;

and regarding the condition of belonging to different cells, taking effective detection data corresponding to the time information of belonging to different cells as final effective detection data of the different cells.

That is, the final effective detection data of each cell to be detected is determined by comparing the time information in each set of effective detection data of each cell to be detected. In other words, when multiple 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; when only one set of valid detection data is acquired for the same cell, the set of valid detection data is used as the final valid detection data of the cell.

In some embodiments, in the case of identifying the pulse with two different original optical information as the reference optical information, determining the final valid detection data by comparing time information in each set of valid detection data of each cell to be detected includes:

judging whether the first time information t1 in each first group of effective detection data is the same/basically the same as the second time information t2 in each second group of effective detection data, namely whether the first time information and the second time information 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 regarding 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, in the case of identifying the pulse with three different kinds of original optical information as the reference optical information, determining the final valid detection data by comparing time information in each set of valid detection data of each cell to be detected 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/basically the same or not, namely whether the first time information, the second time information and the third time information belong to the same cell or not;

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

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

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

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

and regarding the first time information t1, the second time information t2 and the third time information t3 which do not belong to the same cell, respectively taking a first group of valid detection data, a second group of valid detection data and a third group of valid detection data which correspond to the first time information, the second time information and the third time information which belong to different cells as final valid 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 types of original optical information, so that two or three sets of valid detection data obtained by using the different types of original optical information as reference optical information are substantially identical for the cell identified in the different types of original optical information, and one set of data can be selected as the final data of the cell identified in the different types of original optical information. However, pulses generated by some cells (e.g., platelets) having a small volume cannot be identified in the forward scattered light information (the pulse amplitude of the forward scattered light information may be 0 or less than an amplitude threshold), and 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 cells having a small volume.

In some embodiments, the cell identifying step S540 comprises: and determining the final effective detection data of the same cell by comparing the amplitude information in each group of effective detection data corresponding to the same cell or according to the type of the target cell under the condition that the time information in each group of effective detection data belongs to the same cell. In other words, for a plurality of sets of valid detection data obtained for the same cell, one set of data is selected according to the type of the target cell, or one set of data is selected according to the peak difference of the valid detection data of each set.

For example, the group having the largest sum of the amplitudes of the respective groups of valid detection data corresponding to the same cell may be selected as the final valid detection data of the same cell. Wherein, the sum of the amplitudes of a set of valid detection data refers to the sum of the corresponding pulse amplitudes in each original optical information of the same cell at the same pulse position.

In some embodiments, when different first original optical information and second original optical information are used as the reference optical information, if a first set of valid detection data including amplitude information (H11, H12) or (H11, H12, H15) and first time information t1 and a second set of valid detection data including amplitude information (H23, H24) or (H23, H24, H26) and second time information t2 are acquired for the same cell, t1 is substantially the same as t 2. At this time, the sum of the amplitudes of the first set of valid detection data is H11+ H12 or H11+ H12+ H15, the sum of the amplitudes of the second set of valid detection data is H23+ H24 or H23+ H24+ H26, and the valid detection data corresponding to the 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 set of valid detection data or the second set of valid detection data is acquired 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.

In some embodiments, when different first original optical information, second original optical information, and third original optical information are employed as the reference optical information: (1) 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, H15) and first time information t1, the second set of valid detection data comprises amplitude information (H23, H24, H26) and second time information t2, and t1 is substantially the same as t2, 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 the larger value of H11+ H12+ H15 and H23+ H24+ H26 is selected as final valid detection data of the cell; (2) If a first group of valid detection data and a third group of valid detection data are acquired for the same cell, wherein the first group of valid detection data comprises amplitude information (H11, H12, H15) and first time information t1, the third group of valid detection data comprises amplitude information (H37, H38, H39) and third time information t3, and 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, the sum of the amplitudes of the third group of valid detection data is H37+ H38+ H39, and valid detection data corresponding to the larger value of H11+ H12+ H15 and H37+ H38+ H39 can be selected as final valid detection data of the cell; (3) If a second set of valid detection data and a third set of valid detection data are acquired 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, and 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, the sum of the amplitudes of the third set of valid detection data is H37+ H38+ H39, and valid detection data corresponding to the larger value of H23+ H24+ H26 and H37+ H38+ H39 can be selected as final valid detection data of the cell; (5) If a first set of valid detection data, a second set of valid detection data and a third set of valid detection data are acquired 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, the third set of valid detection data includes amplitude information (H37, H38, H39) and third time information t3, t1, t2 and t3 are substantially the same, 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 the sum of the amplitudes 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 can be selected as the final valid detection data for 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.

Alternatively, the sum of the amplitudes may be replaced by an average amplitude, that is, a group having the largest average amplitude of the groups 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 corresponding amplitudes of the original optical information of the same cell at the same pulse position. For example, if a first set of valid detection data comprising amplitude information (H11, H12) and first time information t1 and a second set of valid detection data comprising amplitude information (H23, H24) and second time information t2 are acquired for the same cell, t1 is substantially the same as t 2. At this time, the average amplitude of the first set of valid detection data is H11+ H12/2, and the average amplitude of the second set of valid detection data is H23+ H24/2.

In some embodiments, when multiple sets of valid assay data are acquired for the same cell, one of the sets of valid assay data is simply selected as the final valid assay data for that cell according to the type of target 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.

In some embodiments, the cell identifying step S540 comprises:

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

That is, in the cell identification step S540, one of the sets of valid detection data may be selected as the final valid detection data of each cell to be tested simply according to the type of the target cell without determining whether the time information of each set of valid detection data is the same.

Optionally, the first original optical information is fluorescence information or side scattered light information, and when the target cell is a platelet and/or a reticulocyte, the first set of valid detection data of each cell to be detected is selected as the final valid detection data of each cell to be detected, so as to identify the platelet and/or the reticulocyte in the sample liquid to be detected.

Since 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 scatter information than in forward scatter information, the user may directly select valid detection data detected with the fluorescence information or side scatter information as reference optical information as final valid detection data.

In some embodiments, the cell identifying step S540 comprises: and identifying the target cells in the sample liquid to be detected according to the amplitude information of the final effective detection data and determining the number of the target cells in the sample liquid to be detected.

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

In some embodiments, the statistics of the number of pulses based on any optical information are not accurate due to the different characteristics of the cell in different optical information, and a combination of pulse counting and pulse positions (i.e., time information) is used, wherein the pulse information is associated with the position information, and only one valid position is reserved for the repeated pulses identified in different optical information by the same cell in the whole measurement process, so that 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 with forward scattered light information as reference optical information, the positions are (p 1, p2, p 3), four pulse positions are identified and recorded with side scattered light information as reference optical information (p 1, p2, p3, p 4), four pulse positions are identified and recorded with fluorescence information as reference optical information (p 1, p2, p3, p 5), the repeated positions are not repeatedly counted, and finally the pulse positions are (p 1, p2, p3, p4, p 5), that is, the total number of pulses is 5.

Optionally, identifying the target cell in the sample solution to be detected 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 may obtain a three-dimensional scattergram using the fluorescence intensity, the forward scattered light intensity, and the side scattered light intensity at the same time, thereby distinguishing the target cell group from other cell groups, obtaining a more accurately identified target cell, 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 of 50 ms. Using the forward scattered light information as reference optical information for pulse identification, 1170 pulses are obtained; using the fluorescence information as reference optical information for pulse recognition, 1205 pulses were obtained. The total number of pulses detected at the same position (time) in the two methods is 1162, 8 pulses are recognized in the forward scattered light information and not recognized in the fluorescence information, and 43 pulses are recognized in the fluorescence information and not recognized in the forward scattered light information. Among 1162 pulse data detected in both forward scattered light information and fluorescence information, a is a platelet; of the pulse data unique to 8 forward scattered light information, 4 pulses were small in amplitude and empirically considered as platelets; of the pulse data unique to the 43 fluorescence information, 11 pulses were small in amplitude and were empirically considered as platelets. Combining the above data, the following results can be obtained:

referencing optical information	Total number of pulses	Number of platelets
Forward scattered light information	1170 devices	A+4
Fluorescence information	1205 by	A+11
Combine the two	1213 are provided with	A+15

It follows that the total number of pulses and the number of platelets identified by means of pulse identification with reference to fluorescence information is greater than in the case of forward scattered light information, since small-volume cells of this type are better differentiated in fluorescence information than platelets. In addition, the total number of pulses and the number of platelets obtained by combining the fluorescence information and the forward scattered light information as the reference optical information are more. It is thus shown that the limitation of pulse recognition and cell analysis with a single raw optical data as a reference can be overcome by using a plurality of raw optical data as the reference optical data, and the pulse recognition and cell analysis method with a plurality of raw optical data as a reference is superior to the case of using a single raw optical data as a reference.

According to an embodiment of the present invention, as shown in fig. 9, there is also provided a cell analysis method 900, including:

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

s920, acquiring at least two kinds of optical information of each cell to be detected in the sample liquid to be detected when the cell to be detected passes 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 valid detection data corresponding to the valid light pulse from the at least two kinds of optical information according to the reference optical information;

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

For some types of target cells, the discrimination of the type of 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 type of the target cells, and the acquired other original optical information is used as non-reference optical information, so that the accuracy is ensured and the recognition speed of the target cells is accelerated.

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

based on a preset amplitude threshold value, identifying pulses generated when each cell to be detected passes 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 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;

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 a reticulocyte, selecting side scattered light information or fluorescence information as the reference optical information, the at least two types of optical information including 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. fluorescent information), each reference effective light pulse information comprises reference pulse amplitude information H and reference time information t, and then non-reference pulse amplitude information H1 corresponding to time t in at least one type of non-reference optical information (e.g. scattered light information) is obtained according to the reference time information t, so that non-reference effective light pulse information comprising the non-reference pulse amplitude information is obtained, and further effective detection data, namely (t, H1) is obtained.

Further, a scatter plot of the test cells may be generated based on the amplitude information in the valid detection data to identify the platelets in the test sample fluid, and the number of platelets may be determined based on the number of reference time information in the valid 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 the blood platelets and/or reticulocytes in the sample liquid to be detected according to the first effective detection data.

When the platelets and/or the reticulocytes in the sample liquid to be detected are identified, the side scattered light information or the fluorescence information can be used as reference optical information for pulse identification, first effective detection data corresponding to effective light pulses are determined from the at least two kinds of optical information, and a scatter diagram of the cells to be detected can be generated according to amplitude information of the first effective detection data, so that the platelets and/or the reticulocytes in the sample liquid to be detected are identified and the number of the platelets and/or the reticulocytes is determined.

Further, the at least two types of light information include forward scattered light 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 with the forward scattered light information as reference optical information for identifying that each cell to be detected passes through the optical detection zone to generate the effective light pulse;

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

The embodiment of the present invention further provides a computer-readable storage medium, where multiple program instructions are stored, and after the multiple program instructions are called and executed by a processor, some or all of the steps in the cell analysis method in the embodiments of the present application, or any combination of the steps in the cell analysis method may be performed.

According to the cell analysis method, the cell analyzer and the computer-readable storage medium provided by the embodiment of the invention, a plurality of different types of optical signals generated when the cells pass through the flow chamber are respectively used as reference signals for pulse recognition, the principle that different types of optical signals reflect different cell characteristic information is fully utilized, missing recognition or incapability of recognition of target cells 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 elements of any method or apparatus so disclosed, may be combined in any combination, except combinations where such 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.

Technical terms used in the embodiments of the present invention are only used for illustrating 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, the use of "including" and/or "comprising" in the specification is intended to specify the presence of stated features, integers, steps, operations, elements, and/or components, 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 only an example, and various modifications can be made to the chart or the steps in the present invention without departing from the spirit of the present invention. For instance, the steps may be performed in a differing order, or steps may be added, deleted or modified. It will be understood by those skilled in the art that all or a portion of the above-described embodiments may be practiced and equivalents thereof may be resorted to as falling within the scope of the invention as claimed.

Claims

A method of analyzing cells, comprising:

original optical information acquisition: 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 area of a cell analyzer within a preset time period;

a first set of valid detection data acquisition steps: 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 passing through the optical detection area, so as to acquire a first set of effective detection data including the first effective light pulse information and the second effective light pulse information of each cell to be detected;

a second group of valid detection data acquisition steps: 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 obtain a second set of effective detection data of each cell to be detected, wherein the second set of effective detection data comprises the third effective light pulse information and the fourth effective light pulse information;

a cell recognition step: and determining the 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 is used for identifying the target cell in the sample liquid to be detected.
The method of claim 1, wherein the first and second original optical information are different optical information and are each selected from one of forward scattered light information, side scattered light information, and fluorescence information.
The method for cellular analysis of claim 2, wherein the first or second raw optical information is fluorescence information.
The method for analyzing cells according to claim 3, wherein the target test cell is a platelet and/or a reticulocyte.
The method according to any one of claims 1 to 4, wherein the original optical information obtaining step further comprises: acquiring third original optical information detected in a third optical detector when each cell to be detected in the sample liquid to be detected passes through the optical detection area within the preset time period;

the first set of valid detection data acquisition steps further comprises: determining fifth effective light pulse information of each cell to be detected from the third original optical information by taking the first original optical information as the reference optical information, wherein the first set of effective detection data further comprises the fifth effective light pulse information;

the second set of valid detection data acquisition steps further comprises: and determining sixth effective light pulse information of each cell to be detected from the third original optical information by taking the second original optical information as the reference optical information, wherein the second set of effective detection data further comprises the sixth effective light pulse information.
The method of claim 5, wherein the first set of valid detection data acquisition steps comprises:

based on a first amplitude threshold value, identifying pulses generated when each cell to be detected passes through the optical detection area from the first original optical information and acquiring first effective light pulse information of each cell to be detected, wherein the first effective light pulse information comprises first pulse amplitude information and first time information,

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

the second set of valid detection data acquisition steps comprises:

based on a second amplitude threshold value, identifying pulses generated when each cell to be detected passes through the optical detection area from the second original optical information and acquiring fourth effective optical pulse information of each cell to be detected, wherein the fourth effective optical pulse information comprises fourth pulse amplitude information and second time information,

and according to the second time information, obtaining 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, 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.
The method of claim 5, further comprising a third set of valid detection data acquisition steps:

determining seventh effective optical pulse information, eighth effective optical pulse information and ninth effective optical 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 respectively by taking the third original optical information as the reference optical information so as to obtain a third group of effective detection data of each cell to be detected, wherein the third group of effective detection data comprises the seventh effective optical pulse information, the eighth effective optical pulse information and the ninth effective optical pulse information;

the cell identification step comprises: and according to a preset rule, determining the final effective detection data of each cell to be detected according to 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 the target cell in the sample liquid to be detected.
The method of claim 7, wherein the first set of valid detection data acquisition steps comprises:

based on a first amplitude threshold value, identifying pulses generated when each cell to be detected passes through the optical detection region from the first original optical information and acquiring first effective light pulse information of each cell to be detected, wherein the first effective light pulse information comprises first pulse amplitude information and first time information,

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

the second set of valid detection data acquisition steps comprising:

based on a second amplitude threshold value, identifying pulses generated when each cell to be detected passes through the optical detection area from the second original optical information and acquiring fourth effective optical pulse information of each cell to be detected, wherein the fourth effective optical pulse information comprises fourth pulse amplitude information and second time information,

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

the third set of valid detection data acquisition steps comprising:

based on a third amplitude threshold value, identifying pulses generated when each cell to be detected passes through the optical detection area from the third original optical information and acquiring ninth effective optical pulse information of each cell to be detected, wherein the ninth effective optical pulse information comprises ninth pulse amplitude information and third time information,

according to the third time information, obtaining seventh effective optical pulse information and eighth effective optical pulse information of each cell to be detected from the first original optical information and the second original optical information respectively, wherein the seventh effective optical pulse information and the eighth effective optical pulse information respectively comprise seventh pulse amplitude information and eighth pulse amplitude information.
The method of claim 6 or 8, wherein the cell identification step comprises:

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

selecting one of the sets of valid detection data corresponding to the same cell as the final valid detection data of the same cell when the cells belong to the same cell;

and regarding the condition of belonging to different cells, taking effective detection data corresponding to the time information of belonging to different cells as final effective detection data of the different cells.
The method of claim 9, wherein the cell identification step comprises:

and determining one group from the groups of effective detection data corresponding to the same cell as the final effective detection data of the same cell according to the amplitude information of the groups of effective detection data corresponding to the same cell or the type of the target cell for the condition of belonging to the same cell.
The method of any one of claims 1 to 8, wherein the cell detection step comprises:

and selecting one group from the groups of 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.
The method of claim 11, wherein the first raw optical information is fluorescence information or side scatter information;

the cell detection step comprises: when the target cells are platelets and/or reticulocytes, the first set of valid detection data is selected as the final valid detection data to identify the platelets and/or reticulocytes in the test sample fluid.
The method according to any one of claims 6, 8 to 10, wherein the cell identification step comprises:

and identifying the 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.
The method of 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.
A method of analyzing cells, 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 detected in the sample liquid to be detected 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 when the cell passes 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 valid detection data corresponding to the valid light pulse from the at least two kinds of optical information according to the reference optical information;

and identifying the target cells in the sample liquid to be detected according to the effective detection data.
The method of claim 15, wherein determining the valid detection data from the reference optical information comprises:

based on a preset amplitude threshold value, identifying pulses generated when each cell to be detected passes 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 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;

wherein the valid detection data comprises the valid light pulse data and the at least one non-reference optical data.
The method according to claim 15 or 16, wherein when the target cell is a platelet and/or a reticulocyte, side scattered light information or fluorescence information is selected as the reference optical information, and the at least two types of optical information include at least two of forward scattered light information, side scattered light information, and fluorescence information.
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 light information or fluorescence information;

determining first effective detection data corresponding to the effective light pulse from the at least two kinds of optical information by using the side scattered light information or the fluorescence information as reference optical information for identifying the effective light pulse generated by each cell to be detected passing through the optical detection zone;

and identifying the blood platelets and/or the reticulocytes in the sample liquid to be detected according to the first effective detection data.
The method of claim 18, wherein the at least two types of light information further comprises forward scattered light information, the method further comprising:

determining second valid detection data corresponding to the valid light pulse from the at least two optical information with the forward scattered light information as the reference optical information;

and identifying the blood platelets and/or the reticulocytes in the sample liquid to be detected according to the first effective detection data and the second effective detection data.
A cellular analyzer, 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 drawn by a sampling device and a reagent supply portion for supplying a reagent to the reaction cell so that the blood sample drawn by the sampling device is mixed with the reagent supplied by the reagent supply portion in the reaction cell to prepare a sample solution to be measured;

an optical detection device including a light source, a flow chamber and a light detector, wherein each cell of the sample liquid to be detected can flow in the flow chamber, the light emitted by the light source irradiates the cell in the flow chamber to generate optical information, and the light detector is used for collecting the optical information; and

a data processing apparatus electrically connected with the optical detection apparatus and comprising a processor and a computer readable storage medium storing a computer program, wherein the data processing apparatus 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.
A computer-readable storage medium, characterized by comprising a program executable by a processor to implement the method of any one of claims 1 to 19.