Blood cell pulse signal identification method, flow cytometer and readable storage medium

Abstract

The present application provides a method for identifying blood cell pulse signals, which first obtains each optical signal formed after laser irradiation to blood cells; then pre-processes each optical signal to obtain a reference signal and at least one synchronization signal; then extracts the reference characteristic parameters of the reference signal, and locates the reference position of the reference characteristic parameters; subsequently locates the position range of each synchronization signal based on a preset phase difference and the reference position; finally, extracts the synchronization characteristic parameters of the synchronization signal within the position range, and outputs the test result. The method of the present application locates the position range of the synchronization signal by a preset phase difference, which reduces the amount of data processing for the synchronization signal while ensuring its accuracy. The present application also relates to a flow cytometer and a computer-readable storage medium.

Description

Technical Field

[0001] The present application relates to the field of in vitro diagnostic analysis, and in particular to a blood cell pulse signal recognition method, a flow cytometer and a computer-readable storage medium. Background Art

[0002] At present, flow cytometers represented by blood cell analyzers mostly use optical methods for blood cell detection. By irradiating the blood cells flowing through the irradiation area with lasers of a specific wavelength, the blood cells produce scattered light or fluorescence, and then the light signals are collected and converted into electrical pulse signals. Finally, by extracting and analyzing the characteristics of the pulse signals, the test results such as the size, granularity, expression of antigen molecules, and the content of genetic material in the cells are obtained, thereby obtaining valuable reference information from the perspective of clinical medicine or biology.

[0003] Blood cell pulse signals usually have three channels, including the forward signal FS, the side signal SS and the fluorescence signal. In the existing blood cell detection technology, the side signal SS and the fluorescence signal are usually weak and easily interfered, which affects the accuracy of pulse recognition; at the same time, the number of blood cell particles is large and the flow rate is fast, and the processing speed of blood cell pulse signals usually needs to reach 20MHz or above, which requires high-performance hardware to meet the test requirements, and the cost is relatively high. Summary of the invention

[0004] The present application provides an optimized blood cell pulse signal recognition method, a flow cytometer, and a computer-readable storage medium, which can reduce the amount of data processed by pulse signals and improve the accuracy of pulse recognition. The present application specifically includes the following solutions:

[0005] In a first aspect, the present application provides a method for identifying a blood cell pulse signal, comprising the following steps:

[0006] Acquire various optical signals formed after the laser is irradiated to the blood cells, wherein the optical signals include a reference optical signal and at least one synchronous optical signal;

[0007] Pre-processing each optical signal to obtain a reference signal and at least one synchronization signal, wherein the reference signal is generated by processing the reference optical signal, and each synchronization signal is generated by processing the corresponding synchronization optical signal;

[0008] Extracting a reference characteristic parameter of the reference signal and locating a reference position of the reference characteristic parameter;

[0009] Based on a preset phase difference of each synchronization signal relative to the reference signal and the reference position, locating a position range of each synchronization signal;

[0010] Extracting synchronization characteristic parameters of the synchronization signal within the position range of each synchronization signal;

[0011] The reference characteristic parameter and each of the synchronous characteristic parameters are output as a test result.

[0012] The optical signals generated after the laser is irradiated on the blood cells include at least one of a forward light signal, a side light signal and a fluorescent light signal.

[0013] The reference light signal is one of the forward light signal, the side light signal and the fluorescent light signal.

[0014] The pre-processing of each optical signal includes:

[0015] The optical signals of each channel are subjected to photoelectric conversion processing, amplification and filtering processing, and analog-to-digital conversion processing respectively.

[0016] The preset phase difference of the synchronization signal relative to the reference signal specifically includes:

[0017] Respectively calculating a first circuit delay of each synchronization signal in the photoelectric conversion process, a second circuit delay in the amplification and filtering process, and a third circuit delay in the analog-to-digital conversion process compared to the reference signal;

[0018] The sum of the first circuit delay, the second circuit delay and the third circuit delay is calculated as a phase difference of the synchronization signal relative to the reference signal.

[0019] The extracting of the reference characteristic parameters of the reference signal and locating the reference position of the reference characteristic parameters comprises:

[0020] Extracting the reference characteristic parameter within a signal cycle of the reference signal, and locating a reference position of the reference characteristic parameter within the signal cycle;

[0021] The preset phase difference of the synchronization signal relative to the reference signal also includes:

[0022] The preset phase difference of the synchronization signal relative to the reference signal is controlled to be within the signal period.

[0023] The preset phase difference of the synchronization signal relative to the reference signal also includes:

[0024] The preset phase difference of the synchronization signal relative to the reference signal is adjusted based on actual test results and / or historical test results.

[0025] The extracting of the reference characteristic parameters of the reference signal and locating the reference position of the reference characteristic parameters comprises:

[0026] A pulse recognition algorithm is used to extract the reference characteristic parameters of the reference signal, and the reference position of the reference characteristic parameters is located.

[0027] The reference characteristic parameters extracted from the reference signal include one or more of a peak parameter, an area parameter and a width parameter.

[0028] The extracting of the reference characteristic parameters of the reference signal and locating the reference position of the reference characteristic parameters comprises:

[0029] A peak parameter of the reference signal is extracted, and a reference position of the peak parameter is located.

[0030] Wherein, the outputting the reference characteristic parameter and each of the synchronization characteristic parameters as the test result comprises:

[0031] The reference characteristic parameter and each of the synchronous characteristic parameters are outputted as test results in a graphical manner.

[0032] In the blood cell pulse signal recognition method of the first aspect of the present application, by acquiring each optical signal formed after the laser is irradiated to the blood cells and pre-processing each optical signal, a reference signal and at least one synchronization signal can be obtained. Then, by extracting the characteristic parameters of the reference signal, the reference position of the reference characteristic parameter can be located. Subsequently, each synchronization signal is preset with a phase difference relative to the reference signal, and the position range of each synchronization signal can be located by the phase difference and the reference position. Finally, the synchronization characteristic parameters of the synchronization signal are extracted within the position range without parsing the entire synchronization signal to extract the characteristic parameters. The method of the present application reduces the data processing amount of the synchronization signal, and can avoid the influence of noise interference on the synchronization characteristic parameters, thereby improving the recognition accuracy and recognition speed of the blood cell pulse signal.

[0033] In a second aspect, the present application provides a flow cytometer, comprising:

[0034] A flow chamber, for allowing blood cells in the sample to be tested to pass through one by one;

[0035] a laser emitter for irradiating the blood cells passing through the flow chamber;

[0036] A receiving component, used for acquiring various optical signals formed after the laser emitted by the laser transmitter irradiates the blood cells, wherein the optical signals include a reference optical signal and at least one synchronous optical signal;

[0037] A pre-processing component, used for pre-processing each optical signal to obtain a reference signal and at least one synchronization signal, wherein the reference signal is generated by processing the reference optical signal, and each synchronization signal is generated by processing the corresponding synchronization optical signal;

[0038] A pulse identification component is used to extract the reference characteristic parameters of the reference signal and locate the reference position of the reference characteristic parameters; and locate the position range of each synchronization signal based on the reference position and the preset phase difference of each synchronization signal relative to the reference signal; and finally extract the synchronization characteristic parameters of the synchronization signal within the position range of each synchronization signal;

[0039] An output component is used to output the reference characteristic parameter and each of the synchronous characteristic parameters as a test result.

[0040] Wherein, the receiving component includes one or more of a forward light receiving component, a side light receiving component and a fluorescent light receiving component.

[0041] The pre-processing component includes a photoelectric conversion unit, an amplification and filtering unit, and an analog-to-digital conversion unit.

[0042] In a third aspect, the present application provides a computer-readable storage medium storing executable instructions and configured to implement the above-mentioned blood cell pulse signal recognition method when causing a processor to execute the executable instructions.

[0043] In the second and third aspects of the present application, the principles of application are similar to the blood cell pulse signal recognition method of the first aspect of the present application, and both of them extract the synchronization characteristic parameters within the position range corresponding to the synchronization signal by locating the reference signal and combining the phase difference of each synchronization signal relative to the reference signal, thereby reducing the amount of data processing and improving the recognition accuracy and recognition speed of each synchronization signal. BRIEF DESCRIPTION OF THE DRAWINGS

[0044] Figure 1 is a schematic diagram of the internal framework of the flow cytometer provided in the embodiment of the present application;

[0045] Figure 2 is a schematic diagram of the structure of some components of the flow cytometer provided in an embodiment of the present application;

[0046] Figure 3 It is a schematic diagram of the internal framework of the pre-processing component in the flow cytometer provided in the embodiment of the present application;

[0047] Figure 4 is a flow chart of a blood cell pulse signal recognition method provided in an embodiment of the present application;

[0048] Figure 5 It is a parameter recognition schematic diagram of the blood cell pulse signal recognition method provided in an embodiment of the present application;

[0049] Figure 6 is a flow chart of a blood cell pulse signal recognition method provided by another embodiment of the present application;

[0050] Figure 7 It is a schematic diagram of a computer-readable storage medium provided in an embodiment of the present application. DETAILED DESCRIPTION

[0051] The following will be combined with the drawings in the embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of this application.

[0052] The serial numbers assigned to the components herein, such as "first", "second", etc., are only used to distinguish the objects described and do not have any order or technical meaning. The "connection" and "coupling" mentioned in this application, unless otherwise specified, include direct and indirect connections (couplings). In the description of this application, it should be understood that the orientation or position relationship indicated by the terms "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", etc. are based on the orientation or position relationship shown in the accompanying drawings, which is only for the convenience of describing this application and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation to this application.

[0053] In the present application, unless otherwise clearly specified and limited, a first feature being “above” or “below” a second feature may mean that the first and second features are in direct contact, or the first and second features are in indirect contact through an intermediate medium. Moreover, a first feature being “above”, “above”, and “above” a second feature may mean that the first feature is directly above or obliquely above the second feature, or simply means that the first feature is higher in level than the second feature. A first feature being “below”, “below”, and “below” a second feature may mean that the first feature is directly below or obliquely below the second feature, or simply means that the first feature is lower in level than the second feature.

[0054] Please see this application first Figure 1The flow cytometer 100 shown in the figure includes a flow chamber 10, a laser transmitter 20, a receiving component 30, a pre-processing component 40, a pulse recognition component 50 and an output component 60. Figure 2 The schematic diagram shows the specific structure of the flow chamber 10, the laser transmitter 20, and the receiving component 30. The receiving component 30 includes at least a first receiving sub-component 31 and a second receiving sub-component 32. The flow chamber 10 is located between the laser transmitter 20 and the first receiving sub-component 31, and the second receiving sub-component 32 is located at another position of the flow chamber 10 different from the first receiving sub-component 31. Figure 2 In the schematic diagram of FIG. 1 , the second sub-receiving component 32 is disposed at a vertical position of a line connecting the flow chamber 10 and the first sub-receiving component 31, so that the second sub-receiving component 32 is disposed obliquely above the flow chamber 10. It can be understood that in other embodiments, the second sub-receiving component 32 can also be disposed at other positions of the flow chamber 10 that are different from the first sub-receiving component 31.

[0055] The flow chamber 10 is used to allow blood cells in the sample to be tested to pass through one by one. The sheath flow technology can be used to make the blood cells in the sample to be tested flow through the flow chamber 10 in an orderly manner under the wrapping of the sheath liquid. The laser emitter 20 can be implemented by a helium-neon laser or a semiconductor laser. The laser emitter 20 is used to emit a laser of a specific wavelength to irradiate the blood cells flowing through the flow chamber 10 one by one. The laser irradiated on the blood cells will generate scattered light and fluorescence to the surroundings, and the first sub-receiving component 31 and the second sub-receiving component 32 located at different positions of the flow chamber 10 can receive scattered light signals or fluorescence signals in different directions, which is also described as the first sub-receiving component 31 receiving one light signal and the second sub-receiving component 32 receiving another light signal, wherein the light signal can be a scattered light signal or a fluorescence signal.

[0056] Generally, by setting the respective optical components in the first sub-receiving component 31 and the second sub-receiving component 32, the angle ranges of the optical signals received by the first sub-receiving component 31 and the second sub-receiving component 32 can also be controlled respectively. For example, the first sub-receiving component 31 is configured to receive optical signals within a first angle range, wherein the first angle range is 0 to 10 degrees or 1 to 10 degrees; the second sub-receiving component 32 is configured to receive optical signals within a second angle range, wherein the second angle range is 10 to 20 degrees. In one embodiment, the first angle range and the second angle range are continuous ranges.

[0057] The optical signals received by the receiving component 30 of the present application are not limited to two channels. Based on the different detection items of the flow cytometer 100, the receiving component 30 can also be provided with a third receiving component (not shown) and a fourth receiving component (not shown) to receive more optical signals. It can be understood that the types of optical signals between the multiple optical signals can be different, the receiving directions can be different, and even the angle ranges between the multiple optical signals can be different, which needs to be specifically matched and set according to the detection item requirements of the flow cytometer 100.

[0058] After receiving multiple optical signals through the receiving component 30, the flow cytometer 100 needs to perform pre-processing on each optical signal through the pre-processing component 40 to convert the optical signal into an electrical signal. Then, the pulse recognition component 50 is used to identify and extract the characteristic parameters of each electrical signal, and finally the output component 60 outputs the test results to the outside world. Figure 3 As shown in FIG. 4 , the pre-processing component 40 includes a photoelectric conversion unit 41, an amplifying and filtering unit 42, and an analog-to-digital conversion unit 43. The photoelectric conversion unit 41 is used to convert each optical signal into an analog electrical signal, and then transmit the analog electrical signal to the amplifying and filtering unit 42 for filtering and noise reduction. Finally, the analog-to-digital conversion unit 43 converts the filtered analog electrical signal into an electrical signal corresponding to each optical signal. The electrical signal here can be understood as a digital electrical signal.

[0059] In some embodiments, each sub-receiving component is provided with a corresponding pre-processing component 40, and the pre-processing component 40 only performs pre-processing on the optical signal, which can improve the speed of pre-processing; in other embodiments, a pre-processing component 40 can also be provided behind two or even multiple sub-receiving components to reduce the size and cost of the flow cytometer 100.

[0060] Typically, the receiving component 30 may have three paths, including a forward light receiving component, a side light receiving component, and a fluorescence receiving component. Therefore, the receiving component 30 usually includes at least one of the above three paths. In some embodiments, the receiving component 30 includes the above three paths of receiving components at the same time. The optical signal received by the forward light receiving component is a forward light signal, which forms a forward signal FS after being processed by the pre-processing component 40; the optical signal received by the side light receiving component is a side light signal, which forms a side signal SS after being processed by the pre-processing component 40; the optical signal received by the fluorescence receiving component is a fluorescent light signal, which forms a fluorescent signal FL after being processed by the pre-processing component 40. The forward signal FS is usually a low-angle signal, which can be used to distinguish the size of blood cells. The side signal SS and the fluorescent signal FL are used to distinguish blood cells in combination with the forward signal FS.

[0061] See also Figure 4The blood cell pulse signal identification method of the present application is implemented based on the above-mentioned flow cytometer 100. The method specifically includes the following steps:

[0062] S10, acquiring various optical signals formed after the laser is irradiated onto the blood cells, wherein the optical signals include a reference optical signal and at least one synchronous optical signal;

[0063] Specifically, the laser emitter 20 emits a laser of a specific wavelength to irradiate the blood cells flowing through the flow chamber 10, so that the receiving component 30 can receive at least two optical signals. One of the at least two optical signals is a reference optical signal, and the optical signals other than the reference optical signal are synchronization optical signals.

[0064] It is understandable that because the laser of a specific wavelength emitted by the laser transmitter 20 needs to irradiate the blood cells flowing through the flow chamber 10 to form scattered light or fluorescence, when there are no blood cells flowing through the flow chamber 10, the receiving component 30 will not receive the optical signal, or the optical signal received by the receiving component 30 is a signal with stable values. When blood cells flow through the flow chamber 10, the scattered light or fluorescence formed by the movement of the blood cells in the flow chamber 10 will cause the numerical fluctuations of each optical signal. And because the fluctuations of each optical signal are all based on the movement of the same blood cell in the flow chamber 10, for each optical signal, the starting time of the fluctuation, the duration of the fluctuation, the time when the peak appears, the time when the fluctuation ends, etc. are all consistent. Therefore, the optical signals received by the receiving component 30 can be regarded as optical signals synchronized with each other.

[0065] However, since the orientations of the sub-receiving components relative to the flow chamber 10 are different, the distances between the sub-receiving components and the flow chamber 10 may also be different. Therefore, there are still slight differences in the start time, duration, peak appearance time, and end time of the fluctuations formed by the optical signals. In this method, one of the multiple optical signals is selected as the reference optical signal, and the remaining optical signals except the reference signal are defined as multiple synchronous optical signals.

[0066] S20, pre-processing each optical signal to obtain a reference signal and at least one synchronization signal, wherein the reference signal is generated by processing the reference optical signal, and each synchronization signal is generated by processing its corresponding synchronization optical signal;

[0067] Specifically, the pre-processing component 40 is used to pre-process the multiple optical signals acquired by the receiving component 30. Because the multiple optical signals include one reference optical signal and multiple synchronization optical signals, the reference optical signal processed by the pre-processing component 40 can generate one reference signal, and each of the remaining synchronization optical signals is processed by the pre-processing component 40 to obtain its corresponding synchronization signal.

[0068] S30, extracting a reference characteristic parameter of a reference signal, and locating a reference position of the reference characteristic parameter;

[0069] For details, see Figure 5 Schematic diagram of each signal shown. For any signal processed by the pre-processing component 40, when a blood cell flows through the flow chamber 10, the fluctuation of the signal will form a pulse shape of unidirectional increase-unidirectional decrease. By extracting the characteristic parameters of the signal of this pulse shape, the peak value (P), area (A), width (W) and other characteristic parameters of the signal can be obtained. The parameters such as P, A, W of these scattered light or fluorescent signals can reflect the size, granularity and antigen molecule expression of blood cells, the content of genetic material in cells, etc., and thus obtain valuable reference information from the perspective of clinical medicine or biology.

[0070] In the method of the present application, after the reference light signal is determined, the reference signal obtained by pre-processing it needs to use the pulse recognition component 50 to perform feature recognition on the reference signal within the full cycle corresponding to a blood cell flowing through the flow chamber 10, so as to obtain the reference characteristic parameters of the reference signal. It can be understood that the reference characteristic parameters may include at least one or more of the peak parameter, area parameter and width parameter of the reference signal. Further, in combination with the time axis of the reference signal, the extraction position of the reference characteristic parameter can also be located, that is, the reference position corresponding to the reference characteristic parameter. For example, when the extracted reference characteristic parameter is the peak value of the reference signal, the peak position of the peak parameter in the reference signal can be located as the reference position of the reference signal. For area parameters, width parameters, etc., the reference position of the reference signal can be located by identifying the position of the average dividing line of the area parameter or the width parameter.

[0071] S40, locating the position range of each synchronization signal based on a preset phase difference of each synchronization signal relative to a reference signal and a reference position;

[0072] Specifically, it is mentioned above that there will be slight differences in the start time, duration, peak time, end time, etc. of the fluctuations formed between the optical signals. Therefore, after completing the pre-processing of the optical signals and obtaining a reference signal and multiple synchronization signals, there will be a time difference within a certain range between each synchronization signal and the reference signal, and between any two synchronization signals.

[0073] Furthermore, for an embodiment in which a set of pre-processing components 40 is provided corresponding to each receiving component, there will be differences in the circuit delays generated by each set of pre-processing components 40 during the process of converting each synchronization optical signal into a synchronization signal, and during the process of converting the reference optical signal into a reference signal, due to factors such as device selection and computing power.

[0074] Therefore, by accurately calculating the distance difference between each receiving component and the flow chamber 10, and the circuit delay of each receiving component and its corresponding pre-processing component 40, the phase difference between each optical signal after being converted into an electrical signal can be derived. Because in the design and production process of the flow cytometer 100 of the present application, the position of each receiving component relative to the flow chamber 10, and the device selection of each receiving component and its corresponding pre-processing component 40 have been determined, the phase difference between any two electrical signals can be derived by accurate calculation. After a reference optical signal is determined in the multi-channel optical signal, the phase difference between each processed synchronization signal and the reference signal corresponding to the reference optical signal can be derived. The phase difference is the preset phase difference of the synchronization signal relative to the reference signal.

[0075] The phase difference is usually a range value. Subsequently, after the reference position is located in step S30, the pulse identification component 50 can be used to locate the position range of each synchronization signal based on the phase difference preset by calculation between each synchronization signal and the reference signal.

[0076] S50, extracting synchronization characteristic parameters of the synchronization signal within the position range of each synchronization signal;

[0077] Specifically, after the position range of each synchronization signal is determined, the synchronization characteristic parameters of each synchronization signal can be extracted within the position range corresponding to the synchronization signal.

[0078] Because the reference signal and the synchronization signal can be roughly regarded as mutually synchronized optical signals, after the reference characteristic parameter of the reference synchronization signal is formed at the reference position, the synchronization characteristic parameter of the synchronization signal should also be obtained within a certain range compared to the reference position. The certain range is the range of the phase difference of the synchronization signal relative to the reference signal.

[0079] As mentioned above, in the process of identifying the reference signal and extracting its reference characteristic parameters, the pulse recognition component 50 recognizes the reference signal corresponding to the full cycle of a blood cell flowing through the flow chamber 10, and then obtains the reference characteristic parameters of the reference signal. However, due to the high processing speed requirement of the pulse recognition component 50, it usually needs to reach 20MHz or above to meet the performance requirements. Therefore, if the pulse recognition component 50 is used to recognize each synchronous signal within the full cycle, the data volume load of the pulse recognition component 50 is further increased.

[0080] The method of the present application presets the phase difference between each synchronization signal and the reference signal, and combines the reference position characteristics actually identified, so that the pulse identification component 50 only needs to identify each synchronization signal within the position range corresponding to the synchronization signal, and extract the synchronization characteristic parameters of the synchronization signal. It can be understood that the synchronization characteristic parameters and the reference characteristic parameters are preferably the same type of parameters, for example, the synchronization characteristic parameters and the reference characteristic parameters are both peak parameters. The setting of the position range reduces the data processing amount of the pulse identification component 50, and the determination of the position range of multiple synchronization signals can greatly reduce the data processing amount of the pulse identification component 50, thereby reducing the performance requirements of the flow cytometer 100 on the pulse identification component 50, while ensuring the normal testing of the flow cytometer 100.

[0081] Furthermore, for the lateral signal SS or the fluorescent signal FL mentioned above, which is weak in signal, because of its susceptibility to interference, the pulse recognition component 50 may mistakenly recognize the peak data formed by interference as the peak parameter of the lateral signal SS or the fluorescent signal FL. However, through the blood cell pulse signal recognition method of the present application, because the phase difference between each synchronization signal and the reference signal is preset, the pulse recognition component 50 only extracts the synchronization characteristic parameters within the position range of the synchronization signal. As for the noise interference phenomenon that may appear outside the position range, because it does not appear within the position range, even if the lateral signal SS or the fluorescent signal FL is interfered, it will not affect the extraction of the synchronization characteristic parameters, ensuring that the pulse recognition component 50 can accurately extract the synchronization characteristic parameters within the position range, thereby improving the test accuracy of the flow cytometer 100.

[0082] S60: Output the reference characteristic parameter and each synchronous characteristic parameter as the test result.

[0083] Specifically, after the pulse identification component 50 completes the identification of the reference characteristic parameters and each synchronization characteristic parameter in each optical signal, the output component 60 can output and display the reference characteristic parameters and each synchronization characteristic parameter to the user as a test result.

[0084] Therefore, the blood cell pulse signal recognition method of the first aspect of the present application determines a reference light signal and processes and recognizes it, and then determines the position range of the synchronization signal based on the reference signal, so that the pulse recognition component 50 can recognize the synchronization characteristic parameters within a more accurate range, thereby reducing the data processing amount of the pulse recognition component 50, while avoiding the phenomenon of insufficient test accuracy caused by noise interference, and improving the data processing accuracy and processing speed of the flow cytometer 100.

[0085] As mentioned above, in step S10, the light signals generated after the laser irradiates the blood cells include at least one of the forward light signal, the side light signal and the fluorescent light signal. Figure 5 In the embodiment, the optical signal acquired by the flow cytometer 100 includes a forward light signal, a side light signal and a fluorescent light signal at the same time. Therefore, after being processed by the pre-processing component 40, the forward signal FS, the side signal SS and the fluorescent signal FL can be obtained at the same time.

[0086] Among the above three light signals, any one light signal can be selected as the reference light signal. That is, the reference light signal can be one of the forward light signal, the side light signal and the fluorescent light signal. Figure 5 In the embodiment, the forward light signal is selected as the reference light signal. Because the forward light signal has a relatively stronger anti-interference ability than the side light signal and the fluorescent light signal, the reference characteristic parameter identified based on the forward light signal has a higher accuracy. Therefore, in this embodiment, the forward signal FS is defined as the reference signal.

[0087] Furthermore, Figure 5 The embodiment selects the peak parameter P of the forward signal FS FS As the reference characteristic parameter. In other embodiments, the area parameter or the width parameter may also be used as the reference characteristic parameter. FS The corresponding point P1 of the forward signal FS is the reference position of the peak value. The phase difference between the side signal SS and the forward signal FS is calculated as: Δt SS-FS =[t 1 ,t 2 ]. Thus, the position range of the lateral signal SS can be derived: [P1-t 1 ,P1+t 2 Therefore, the pulse identification component 50 is calculated by [P1-t 1 ,P1+t 2 ] can be identified within the position range, and the peak parameter P of the lateral signal SS can be extracted. SS .

[0088] Correspondingly, the phase difference between the fluorescence signal FL and the forward signal FS can be obtained by calculation: ΔtFL-FS =[t 3 ,t 4 ], the position range of the fluorescence signal FL can be deduced: [P1-t 3 ,P1+t 4 Then, the pulse identification component 50 is in [P1-t 3 ,P1+t 4 ] can be identified within the position range, and the peak parameter P of the fluorescence signal FL can be extracted. FL .

[0089] It needs to be pointed out that Figure 5 Middle pair t 1 ,t 2 ,t 3 and t 4 The schematic diagram is only used to express the phase relationship between the lateral signal SS and the fluorescent signal FL and the forward signal FS as the reference signal. The method of the present application does not strictly limit the peak parameter P of the lateral signal SS. SS The peak parameter P of the forward signal FS FS After that, the peak parameter P of the fluorescence signal FL is not limited. FL The peak parameter P of the forward signal FS FS In addition, this method does not limit the peak parameter P of the forward signal FS. FS The value must be greater than the peak parameter P of the lateral signal SS SS Or the peak parameter P of the fluorescence signal FL FL The numerical value of .

[0090] In one embodiment, when pre-processing is performed on each optical signal in step S20 of the method, because the pre-processing component 40 may include a photoelectric conversion unit 41, an amplification and filtering unit 42, and an analog-to-digital conversion unit 43, the pre-processing performed on each optical signal may include photoelectric conversion processing, amplification and filtering processing, and analog-to-digital conversion processing.

[0091] In one embodiment, when calculating the phase difference of a preset synchronization signal relative to a reference signal, the following steps may be specifically included:

[0092] Calculate the first circuit delay of each synchronization signal in the photoelectric conversion process, the second circuit delay in the amplification and filtering process, and the third circuit delay in the analog-to-digital conversion process compared to the reference signal respectively;

[0093] The sum of the first circuit delay, the second circuit delay and the third circuit delay is calculated as the phase difference of the synchronization signal relative to the reference signal.

[0094] Specifically, according to the differences in device selection and computing power between the pre-processing component 40 of each synchronous optical signal and the pre-processing component 40 of the reference optical signal, the circuit delay of each sub-step in the pre-processing of the synchronous optical signal can be calculated, and then the circuit delays of each sub-step are added together to obtain the total delay of the synchronous optical signal in the pre-processing process relative to the pre-processing of the reference optical signal. Then, the total delay is used as the phase difference of the synchronous signal relative to the reference signal, which can be used to determine the position range of the synchronous signal.

[0095] As mentioned above, the different orientations and distances of each receiving component relative to the flow chamber 10 will also affect the phase difference. However, due to the high propagation speed of light, the effect of distance on the phase difference is much smaller than the effect of circuit delay. Therefore, in this embodiment, only the effect of circuit delay on the phase difference can be considered, which can also improve the accuracy of the method.

[0096] For an example, see Figure 6 , for step S30 "extracting a reference characteristic parameter of a reference signal and locating a reference position of the reference characteristic parameter", it includes:

[0097] S32a, extracting a reference characteristic parameter within a signal cycle T of the reference signal, and locating a reference position of the reference characteristic parameter within the signal cycle T;

[0098] Then, in step S40, the "preset phase difference of the synchronization signal relative to the reference signal" also includes:

[0099] S41a, controlling the preset phase difference of the synchronization signal relative to the reference signal to be within the signal period T.

[0100] Specifically, in this embodiment, through the circuit delay calculation of the synchronization signal and the reference signal, there may be a phenomenon that the circuit delay is too large and the phase difference exceeds the signal period T of the same blood cell flowing through the flow chamber 10. That is, the speed of the blood cell flowing through the flow chamber 10 is too fast, causing the phase difference of one or more synchronization signals relative to the reference signal to exceed the time period of the blood cell flowing through the flow chamber 10. At this time, determining the position range of the synchronization signal based on the preset phase difference may cause the position range to exceed the signal period T of the current blood cell, which in turn increases the calculation amount of the pulse identification component 50.

[0101] It can be understood that when the position range exceeds the current signal cycle T, the increased computational effort of the pulse identification component 50 is an invalid computational effort. Therefore, it is necessary to define the phase difference of any synchronization signal relative to the reference signal to be within the same signal cycle T. When the phase difference of the synchronization signal relative to the reference signal exceeds the current signal cycle T at either end, the boundary of the position range needs to be replaced by the boundary of the current signal cycle T to avoid adding unnecessary computational load.

[0102] It should be noted that the speed of blood cells is not uniform when they flow through the flow chamber 10 one by one, so the signal period T for each blood cell may be different. Therefore, during the implementation of this method, the phase difference of the synchronization signal relative to the reference signal may change with the change of the signal period T.

[0103] In other embodiments, only the phase difference of one side of the synchronization signal relative to the reference signal may be calculated, and the phase difference boundary of the other side of the synchronization signal may be directly relaxed to the boundary position of the signal period T. In such a setting, when the phase difference range of the synchronization signal relative to the reference signal is large and some synchronization characteristic parameters fluctuate greatly within the position range, appropriately expanding the range of the phase difference is conducive to ensuring that the synchronization characteristic parameters are accurately located within the position range. At the same time, because the value of the phase difference is determined on one side of the synchronization signal, the operation of identifying the synchronization signal within the entire signal period T is also avoided, which can also have a certain effect of reducing the amount of data processing.

[0104] In one embodiment, in step S40, “presetting the phase difference of the synchronization signal relative to the reference signal” may further include:

[0105] S42a, adjusting a preset phase difference of the synchronization signal relative to the reference signal based on actual test results and / or historical test results.

[0106] Specifically, the phase difference preset by calculating the circuit delay may have a calculation deviation, resulting in the preset phase difference not being consistent with the actual test result. Therefore, when using this method for testing, the preset phase difference can also be corrected according to the actual test results so that the preset phase difference can be matched to the position range where the synchronization characteristic parameter is located. It is understandable that when calculating and presetting the phase difference, historical test results can also be called to combine the set phase difference or historical test results can be called alone to correct the preset phase difference, which can also improve the accuracy of the preset phase difference.

[0107] In one embodiment, in step S30 of "extracting a reference characteristic parameter of a reference signal and locating a reference position of the reference characteristic parameter", the following may also be included:

[0108] S31a, extracting the reference characteristic parameters of the reference signal by using a pulse recognition algorithm, and locating the reference position of the reference characteristic parameters.

[0109] Because the waveform of each optical signal after being converted into an electrical signal is a pulse-shaped waveform, the pulse recognition component 50 extracts the reference characteristic parameters of the reference signal through a pulse recognition algorithm, and its recognition accuracy is higher and the recognition result is more reliable. It can be understood that because each synchronization signal is also a pulse-shaped waveform, the pulse recognition component 50 can also use a pulse recognition algorithm when extracting the synchronization characteristic parameters of each synchronization signal.

[0110] In one embodiment, step S60 of "outputting the reference characteristic parameter and each synchronization characteristic parameter as the test result" further includes:

[0111] S60a, outputting the reference characteristic parameters and each synchronous characteristic parameter as a test result in a graphical manner.

[0112] Specifically, the test results output by the output component 60 can be as follows: Figure 5 As shown, when displaying the reference characteristic parameters and the synchronous characteristic parameters, the reference signal and the synchronous signal are displayed to the user in a graphical manner. The graphical display method also makes it easier for the user to intuitively judge the size and proportion of each signal waveform, as well as the size difference between the characteristic parameters.

[0113] Please look back Figure 1 The flow cytometer 100 provided in the second aspect of the present application is shown. The flow cytometer 100 can be used to implement the above-mentioned blood cell pulse signal recognition method. Therefore, in the flow cytometer 100 of the present application, after the laser emitter 20 emits a laser of a specific wavelength toward the blood cells flowing through the flow chamber 10 one by one, the receiving component 30 is also used to obtain the various optical signals formed after the laser emitted by the laser emitter 20 is irradiated to the blood cells, wherein the optical signals include a reference optical signal and at least one synchronous optical signal.

[0114] The pre-processing component 40 is used to perform pre-processing on each optical signal to obtain a reference signal and at least one synchronization signal, wherein the reference signal is generated by processing the reference optical signal, and each synchronization signal is generated by processing the corresponding synchronization optical signal.

[0115] The pulse identification component 50 is used to extract the reference characteristic parameters of the reference signal and locate the reference position of the reference characteristic parameters; then, based on the reference position and the phase difference of each synchronization signal relative to the reference signal, the position range of each synchronization signal is located; finally, the synchronization characteristic parameters of the synchronization signal are extracted within the position range of each synchronization signal.

[0116] The output component 60 is used to output the reference characteristic parameters and various synchronous characteristic parameters as test results.

[0117] It can be understood that, due to the above-mentioned configuration, the flow cytometer 100 of the present application reduces the data processing amount of the synchronization signal and can effectively avoid the interference of noise on the synchronization signal, thereby improving the test accuracy and test speed of the flow cytometer 100 of the present application.

[0118] See also Figure 7 The third aspect of the present application provides a computer-readable storage medium 200, which includes a storage device 202 storing executable program instructions and is configured to cause a processor 201 to execute the executable program instructions to implement the blood cell pulse signal recognition method provided in the first aspect of the present application.

[0119] Specifically, in one embodiment, the processor 201 calls the program instructions stored in the storage device 202 to perform the following operations:

[0120] Acquire various optical signals formed after the laser is irradiated onto the blood cells, wherein the optical signals include a reference optical signal and at least one synchronous optical signal;

[0121] Pre-processing each optical signal to obtain a reference signal and at least one synchronization signal, wherein the reference signal is generated by processing the reference optical signal, and each synchronization signal is generated by processing the corresponding synchronization optical signal;

[0122] Extracting a reference characteristic parameter of a reference signal and locating a reference position of the reference characteristic parameter;

[0123] Based on the preset phase difference of each synchronization signal relative to the reference signal and the reference position, the position range of each synchronization signal is located;

[0124] Extracting synchronization characteristic parameters of the synchronization signal within the position range of each synchronization signal;

[0125] The benchmark characteristic parameters and each synchronous characteristic parameter are output as the test results.

[0126] The storage device 202 may include a volatile memory device (volatile memory), such as a random-access memory device (random-access memory, RAM); the storage device 202 may also include a non-volatile memory device (non-volatile memory), such as a flash memory device (flash memory), a solid-state drive (solid-state drive, SSD), etc.; the storage device 202 may also include a combination of the above-mentioned types of storage devices.

[0127] The processor 201 may be a central processing unit (CPU). The processor 201 may also be other general-purpose processors, digital signal processors (DSP), application-specific integrated circuits (ASIC), field-programmable gate arrays (FPGA) or other programmable logic devices, discrete gate or transistor logic devices, discrete hardware components, etc. A general-purpose processor may be a microprocessor or the processor may also be any conventional processor, etc.

[0128] It should be pointed out that in the second and third aspects of the present application, because the principles applied are similar to those of the blood cell pulse signal identification method of the first aspect of the present application, one channel is extracted from multiple optical signals as a reference optical signal, and then the phase difference of each preset synchronization signal relative to the reference signal is calculated, and the synchronization characteristic parameters are identified within the position range corresponding to each synchronization signal, and finally the test results are output to the user, thereby reducing the amount of data processing for the synchronization signal and avoiding noise interference of the synchronization signal. Therefore, the development of each embodiment in the above two aspects can also be carried out based on the various embodiments of the blood cell pulse signal identification method of the first aspect of the present application, and this specification will not be repeated here.

[0129] The above-described implementation methods do not constitute a limitation on the protection scope of the technical solution. Any modification, equivalent replacement and improvement made within the spirit and principle of the above-described implementation methods shall be included in the protection scope of the technical solution.

Claims

1. A blood cell pulse signal recognition method, characterized in that: The steps include: Acquire various optical signals formed after the laser is irradiated to the blood cells, wherein the optical signals include a reference optical signal and at least one synchronization optical signal, and the various optical signals formed after the laser is irradiated to the blood cells include at least one of a forward optical signal, a side optical signal, and a fluorescent optical signal; Pre-processing each optical signal to obtain a reference signal and at least one synchronization signal, wherein the reference signal is generated by processing the reference optical signal, and each synchronization signal is generated by processing the corresponding synchronization optical signal; Extracting a reference characteristic parameter of the reference signal and locating a reference position of the reference characteristic parameter; Based on a preset phase difference of each synchronization signal relative to the reference signal and the reference position, locating a position range of each synchronization signal; Extracting synchronization characteristic parameters of the synchronization signal within the position range of each synchronization signal; Outputting the reference characteristic parameter and each of the synchronization characteristic parameters as a test result; The pre-processing of each optical signal includes: Performing photoelectric conversion, amplification and filtering, and analog-to-digital conversion on the optical signals of each channel; The extracting of the reference characteristic parameters of the reference signal and locating the reference position of the reference characteristic parameters comprises: Extracting the reference characteristic parameter within a signal cycle of the reference signal, and locating a reference position of the reference characteristic parameter within the signal cycle; The preset phase difference of the synchronization signal relative to the reference signal includes: Respectively calculating a first circuit delay of each synchronization signal in the photoelectric conversion process, a second circuit delay in the amplification and filtering process, and a third circuit delay in the analog-to-digital conversion process compared to the reference signal; The sum of the first circuit delay, the second circuit delay and the third circuit delay is calculated as a phase difference of the synchronization signal relative to the reference signal.

2. The blood cell pulse signal recognition method according to claim 1, characterized in that: The reference light signal is one of the forward light signal, the side light signal, and the fluorescent light signal.

3. The blood cell pulse signal recognition method according to claim 1, characterized in that: The method further comprises: The preset phase difference of the synchronization signal relative to the reference signal is controlled to be within the signal period.

4. The blood cell pulse signal recognition method according to claim 1, characterized in that: The method further comprises: The preset phase difference of the synchronization signal relative to the reference signal is adjusted based on actual test results and / or historical test results.

5. The blood cell pulse signal recognition method according to claim 1, characterized in that: The method further comprises: A pulse recognition algorithm is used to extract the reference characteristic parameters of the reference signal, and the reference position of the reference characteristic parameters is located.

6. The blood cell pulse signal recognition method according to any one of claims 1 to 4, characterized in that: The reference characteristic parameters of the reference signal extracted include one or more of a peak parameter, an area parameter, and a width parameter.

7. The blood cell pulse signal recognition method according to claim 6, characterized in that: The extracting the reference characteristic parameter of the reference signal and locating the reference position of the reference characteristic parameter comprises: A peak parameter of the reference signal is extracted, and a reference position of the peak parameter is located.

8. The blood cell pulse signal recognition method according to any one of claims 1 to 4, characterized in that: Outputting the reference characteristic parameter and each of the synchronization characteristic parameters as a test result comprises: The reference characteristic parameter and each of the synchronous characteristic parameters are outputted as test results in a graphical manner.

9. A flow cytometer, characterized in that: include: A flow chamber, for allowing blood cells in the sample to be tested to pass through one by one; a laser emitter for irradiating the blood cells passing through the flow chamber; A receiving component, used for acquiring various optical signals formed after the laser emitted by the laser transmitter irradiates the blood cells, wherein the optical signals include a reference optical signal and at least one synchronous optical signal; A pre-processing component, used for pre-processing each optical signal to obtain a reference signal and at least one synchronization signal, wherein the reference signal is generated by processing the reference optical signal, and each synchronization signal is generated by processing the corresponding synchronization optical signal; A pulse identification component is used to extract the reference characteristic parameters of the reference signal and locate the reference position of the reference characteristic parameters; and locate the position range of each synchronization signal based on the reference position and the preset phase difference of each synchronization signal relative to the reference signal; and finally extract the synchronization characteristic parameters of the synchronization signal within the position range of each synchronization signal; An output component, used for outputting the reference characteristic parameter and each of the synchronous characteristic parameters as a test result; Wherein, the receiving component includes one or more of a forward light receiving component, a side light receiving component and a fluorescence receiving component; The pre-processing component includes a photoelectric conversion unit, an amplification and filtering unit, and an analog-to-digital conversion unit; The pulse identification component is specifically used to extract the reference characteristic parameters within a signal cycle of the reference signal and locate the reference position of the reference characteristic parameters within the signal cycle; and respectively calculate the first circuit delay of each synchronization signal in the photoelectric conversion process, the second circuit delay in the amplification and filtering process, and the third circuit delay in the analog-to-digital conversion process compared to the reference signal; and the sum of the first circuit delay, the second circuit delay, and the third circuit delay is counted as the phase difference of the synchronization signal relative to the reference signal.

10. A computer-readable storage medium, characterized in that: Executable instructions are stored and configured to cause a processor to execute the executable instructions to implement the blood cell pulse signal recognition method according to any one of claims 1 to 8.

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