A flow analysis system signal processing method, device, equipment and storage medium

Abstract

The application provides a flow analysis system signal processing method and device, equipment and a storage medium, and the method comprises the following steps: acquiring a detection signal of scattered light emitted by a detected particle under laser irradiation by a scattered light detector, to obtain an electric signal; analyzing the electric signal, excluding interference through a preset threshold, to obtain a starting time and an ending time of the detected particle passing through a detection area; acquiring a detection signal of fluorescence emitted by the detected particle under laser irradiation by a fluorescence detector of each fluorescence channel, to obtain a light pulse signal; recording generation conditions of the light pulse signal in the starting and ending intervals and analyzing to obtain a peak value and an area of the channel as a detection result of the channel signal. For the detection result of the photon counting detector, the method provided by the application can simply and quickly extract the area and height parameters of the event signal, can be compared with the detection and processing result of the traditional flow cytometer using an analog detector, and the method has strong compatibility.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of signal processing, in particular to a flow analysis system signal processing method and device, equipment and storage medium. BACKGROUND

[0002] Flow cytometry is a basic general tool for cell quantification, biochemical analysis, cell sorting, etc., which realizes the detection of cell physicochemical characteristics through scattering and fluorescence signals. However, the traditional flow analysis system uses PMT or APD as a detector to detect relatively strong fluorescence signals, and the detection effect of weak light signals such as spontaneous fluorescence is not ideal.

[0003] In addition, an important development direction of the current flow is the spectral flow system, which needs to divide the already weak fluorescence into more channels, resulting in weaker fluorescence signals in each channel, which are easily overwhelmed by noise.

[0004] To solve this problem, using MPPC or SPAD as a detector, and using a photon counting method to identify weak fluorescence signals can obtain good results and provide better protection for the accuracy of weak fluorescence detection. However, there is still a lack of better methods for further effectively processing the photon pulse signals emitted after detection and combining with the flow system. SUMMARY

[0005] In order to achieve the above-mentioned purposes and other advantages of the present application, the first object of the present application is to provide a flow analysis system signal processing method, comprising the following steps:

[0006] Obtaining a detection signal of a scattering light detector for scattering light emitted by a detected particle under laser irradiation, to obtain an electrical signal;

[0007] Analyzing the electrical signal, excluding interference through a preset threshold, to obtain a starting time and an ending time of the detected particle passing through a detection area;

[0008] Obtaining a detection signal of a fluorescence detector for fluorescence emitted by the detected particle under laser irradiation in each fluorescence channel, to obtain a light pulse signal;

[0009] Recording the generation of the light pulse signal in the starting and ending intervals and analyzing to obtain a peak value and an area of the channel as a detection result of the channel signal.

[0010] Further, the step of analyzing the electrical signal, excluding interference through a preset threshold, to obtain a starting time and an ending time of the detected particle passing through a detection area comprises:

[0011] When the intensity of the light signal exceeds an event starting threshold, an event starting signal is sent out;

[0012] When the light signal intensity is detected to be lower than the event end threshold, an event end signal is sent out.

[0013] Further, the time number is recorded in a period t, and the amplitude A of the pulse is measured in a single-photon pulse intensity as a basic unit when the pulse signal arrives.

[0014] Further, the recording of the generation of the light pulse signal in the start and end interval and the analysis to obtain the peak value and the area of the channel as the detection result of the channel signal step includes:

[0015] The intensity of each pulse signal in the start and end interval is accumulated to obtain the total amount of pulses, which can be used as the area value of the event:

[0016] ;

[0017] When a new pulse signal arrives, the time number of the interval between the new pulse and the previous pulse is used as the dividend, and the intensity of the new pulse is used as the divisor, and the quotient obtained is used as the calculated interval time number of the current pulse:

[0018] ;

[0019] The number of pulse smoothing groups N is set, and the total sum of the time numbers of N pulses is calculated or smoothed as the modified calculation interval time number:

[0020] ;

[0021] wherein, is the weighting coefficient;

[0022] The reciprocal of is calculated to obtain the modified intensity value of each photon pulse after calculation The maximum value of the modified intensity of all pulses in the start and end interval is the peak value of the detection of the event of the channel.

[0023] Further, after the step of accumulating the intensity of each pulse signal in the start and end interval to obtain the total amount of pulses and using it as the area value of the event, the method further includes:

[0024] The start of noise measurement is waited for a set time after the end of the previous event, the total amount of pulses containing intensity information is recorded, and the end of noise measurement is set at a time before the start of the current event;

[0025] The time number between the start and end of noise measurement is , the total amplitude of the pulses in the interval is the total amount of baseline noise , and the ratio of the two is the average value of the baseline noise:

[0026] ;

[0027] The area value after baseline is:

[0028] .

[0029] Further, the weighting coefficient is defined as:

[0030] ;

[0031] Or, the weighting coefficient is defined as:

[0032] ;

[0033] Or, the weighting coefficient is defined as:

[0034] ;

[0035] Wherein, is the pulse intensity represented by .

[0036] The second object of the application is to provide a flow analysis system signal processing device, applying the above method, comprising a scattered light detector, a fluorescence detector, a filter, an analog-to-digital converter, a light splitting system, a pulse shaper, a pulse reading module, a data processing module; wherein,

[0037] When the detected particles pass through the detection area, they emit scattered light and fluorescence under laser irradiation;

[0038] The scattered light detector is used for collecting the scattered light signal emitted by the detected particles and converting it into an electrical signal;

[0039] The light splitting system is used for splitting the fluorescence into several channels according to the wavelength, and each channel corresponds to a fluorescence detector;

[0040] The fluorescence detector is used for detecting the fluorescence emitted by the detected particles;

[0041] The filter and the analog-to-digital converter sequentially filter and analog-to-digital convert the electrical signal output by the scattered light detector;

[0042] The pulse shaper is used for shaping the signal detected by the fluorescence detector;

[0043] The pulse reading module is used for judging the pulse signal intensity and converting the pulse into the number of photons;

[0044] The data processing module is used for processing the output of the analog-to-digital converter and the pulse reading module.

[0045] Further, the scattered light detector adopts a photodiode, an avalanche diode or a photomultiplier tube.

[0046] Further, the fluorescence detector adopts a silicon photomultiplier tube or a single-photon avalanche diode.

[0047] Further, the pulse reading module realizes an adjustable reference voltage through a digital-to-analog converter, realizes a predetermined number of voltage division through a resistance network, and each level of the reference voltage output after voltage division represents the intensity of the pulse signal formed by the corresponding number of photons, and the reference voltage of each level is compared with the intensity of the pulse signal through a comparator to determine the amplitude of the pulse signal, i.e., the number of photons represented by the pulse signal.

[0048] Alternatively, the pulse reading module adopts a high-speed analog-to-digital converter to collect the pulse signal in real time, and when the pulse signal arrives, the signal intensity is determined through a pulse peak value and the number of photons represented by the pulse is converted.

[0049] Further, when the pulse reading module adopts a high-speed analog-to-digital converter, a threshold value is formed through the intensity of the single-photon signal to suppress the interference of the bottom noise, and the signal is subjected to a smoothing filter processing to further suppress the interference of the noise on the amplitude determination.

[0050] A third object of the present application is to provide a computer device comprising a memory, a processor and a computer program stored on the memory and executable on the processor, wherein the processor implements the steps of the above method when executing the computer program.

[0051] A fourth object of the present application is to provide a computer-readable storage medium having a computer program stored thereon, wherein the computer program is executed by a processor to implement the steps of the above method.

[0052] Compared with the prior art, the present application has the following beneficial effects:

[0053] The present application provides a method for weak fluorescence signal acquisition and signal processing. For the detection results of a photon counting detector, the method provided by the present application can simply and quickly extract the area and height parameters of an event signal, can be compared with the detection and processing results of a traditional flow cytometer using an analog detector, is convenient for an operator to control the detection process and understand the detection results, and has strong compatibility.

[0054] The above description is only a summary of the technical solutions of the present application. In order to more clearly understand the technical means of the present application and can be implemented according to the content of the description, the following will be described in detail with the preferred embodiments of the present application and the accompanying drawings. The specific embodiments of the present application are given in detail by the following examples and the accompanying drawings. BRIEF DESCRIPTION OF DRAWINGS

[0055] The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this application, illustrate embodiments of the application and together with the description serve to explain the application. In the drawings:

[0056] Figure 1 Signal processing device schematic diagram for flow analysis system;

[0057] Figure 2 Signal processing flow diagram for flow analysis system;

[0058] Figure 3 Timing diagram for result correction by baseline calculation;

[0059] Figure 4 Pulse reading module implementation schematic diagram;

[0060] Figure 5 Signal processing method flow diagram for flow analysis system;

[0061] Figure 6 Flow diagram for determination of start time and end time of detected particle passing through detection area;

[0062] Figure 7 Flow diagram for channel signal detection of detected particle;

[0063] Figure 8 Computer device schematic diagram;

[0064] Figure 9 Computer readable storage medium schematic diagram. DETAILED DESCRIPTION

[0065] The application will be further described below in conjunction with the drawings and specific embodiments. Obviously, the described embodiments are only a part of the embodiments of the application, rather than all the embodiments. It should be noted that the following described embodiments or technical features can be combined with each other or other embodiments or technical features to form new embodiments, without conflict.

[0066] All other embodiments obtained by those skilled in the art based on the embodiments of the application without creative labor are within the scope of protection of the application.

[0067] The figure numbers in the present application are only used to distinguish each step in the scheme, and are not used to limit the execution order of each step. The specific execution order is subject to the description in the specification.

[0068] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used in the description herein is for describing particular embodiments only and is not intended to be limiting of the application.

[0069] The application provides a data processing method of photon counting signal and a corresponding device for analysis of flow fluorescence signal. The detection result obtained by the method can be combined with the detection parameters of the existing conventional flow system, and the operator can easily grasp the result. The specific scheme is as follows: Embodiment 1

[0070] A flow analysis system signal processing device, as shown in Figure 1 includes a scattered light detector, a fluorescence detector, a filter, an analog-to-digital converter, a light splitting system, a pulse shaper, a pulse reading module, and a data processing module. Wherein,

[0071] When the detected particles (such as cells) pass through the detection area, they will emit scattered light and fluorescence under laser irradiation.

[0072] The scattered light detector is used for collecting the scattered light signal emitted by the detected particles and converting it into an electrical signal.

[0073] Further, the scattered light detector uses a photodiode, avalanche diode or photomultiplier tube.

[0074] The scattered light has the same wavelength as the laser light, and the scattering angle and scattered light intensity are related to the consistency of the particle size and the particle structure. The light intensity of the scattered light is strong, and a PD, APD or PMT can be used as a detector to collect the light signal in the forward direction (forward scattering light, FSC), side direction or other angles (side scattering light, SSC) and convert it into an electrical signal.

[0075] The light splitting system is used to divide the fluorescence into several channels according to the wavelength, and each channel corresponds to a fluorescence detector.

[0076] The fluorescence detector is used for detecting the fluorescence emitted by the detected particles.

[0077] Further, the fluorescence detector uses a silicon photomultiplier tube or a single-photon avalanche diode.

[0078] The fluorescence signal is relatively weak and has spectral information. The fluorescence is divided into several channels by a spectral system (such as a filter array, a grating or a prism) according to the wavelength, and each channel corresponds to a fluorescence detector. The present application is aimed at weak fluorescence, and a silicon photomultiplier (MPPC / Si-PM) or a SPAD is used as a detector for fluorescence detection. The characteristic of this type of detector is that when it receives a photon, the photoelectric current generated under the action of a reverse bias will produce an avalanche amplification. The carriers generated by the photon will produce a large light pulse through multiplication, and the single photon can be detected by detecting the pulse. This is the working principle of the SPAD, and the MPPC further integrates multiple APDs together as pixels, so the MPPC can also respond to signals of multiple photons arriving at the same time to form pulses of different amplitudes.

[0079] The filter and the analog-to-digital converter sequentially filter and analog-to-digital convert the electrical signal output by the scattered light detector;

[0080] The pulse shaper is configured to shape the signal detected by the fluorescence detector;

[0081] The pulse reading module is configured to determine the intensity of the pulse signal and convert the number of photons represented by the pulse;

[0082] The data processing module is configured to process the outputs of the analog-to-digital converter and the pulse reading module.

[0083] In order to analyze the generation of the light pulse signal and facilitate the analysis of the data of the traditional flow cytometer, the present application sets a timer to record the number of time periods (intervals) t, and establishes an ADC or comparator group to measure the amplitude A of the pulse in units of single-photon pulse intensity when the pulse signal arrives.

[0084] In some embodiments, as shown in Figure 4 The pulse reading module realizes an adjustable reference voltage through a digital-to-analog converter, and the voltage is consistent with or slightly lower than the maximum output amplitude of the MPPC device (but the voltage difference should not exceed 1 / 16 of the maximum output). The number of pixels of the silicon photodiode is determined, such as 16-pixel MPPC, and a resistor network is used to realize a predetermined number of stages (i.e. 16 stages) of voltage division. The reference voltage output by each stage after voltage division represents the pulse signal intensity formed by the corresponding number of photons. By comparing the reference voltage of each stage with the intensity of the pulse signal through a comparator, the amplitude of the pulse signal, i.e. the number of photons it represents, can be determined.

[0085] In some other embodiments, the pulse reading module employs a high-speed analog-to-digital converter to collect the pulse signal in real time. When the pulse signal arrives, the signal strength is determined by the pulse peak value, and the number of photons represented by the pulse is calculated. Preferably, a threshold value can be formed by the intensity of the single-photon signal to suppress the interference of the bottom noise, and the signal can be subjected to smoothing filtering and other processing methods to further suppress the interference of the noise on the amplitude determination.

[0086] For detailed description of the flow cytometry signal processing method corresponding to the flow cytometry signal processing device, reference can be made to the corresponding description in the following method embodiments, which will not be repeated here. Embodiment 2

[0087] A flow cytometry signal processing method based on the above flow cytometry signal processing device. For detailed description of the flow cytometry signal processing device, reference can be made to the corresponding description in the above device embodiments, which will not be repeated here. As shown in Figure 2 、 Figure 5 The method comprises the following steps:

[0088] S100, obtaining the detection signal of the scattered light detector on the scattered light emitted by the detected particle under laser irradiation, to obtain an electrical signal;

[0089] When the detected particle (such as a cell) passes through the detection area, it will emit scattered light and fluorescence under laser irradiation. The scattered light is consistent with the wavelength of the laser, and the scattering angle and the scattered light intensity are related to the consistency of the particle size and the structure of the particle. The light intensity of the scattered light is strong, and a PD, APD or PMT can be used as a detector to collect and convert the light signal into an electrical signal in the forward direction (forward scattered light, FSC), side direction or other angles (side scattered light, SSC).

[0090] S200, analyzing the electrical signal, excluding noise or other small particle interference by a preset threshold, to obtain the start time and end time of the detected particle passing through the detection area;

[0091] Further, as shown in Figure 6 The step of analyzing the electrical signal and excluding interference by a preset threshold to obtain the start time and end time of the detected particle passing through the detection area comprises:

[0092] S210, when the light signal intensity is detected to exceed the event start threshold, an event start signal is sent out, such as a rising edge from low level to high level.

[0093] S220, when the light signal intensity is detected to be lower than the event end threshold, an event end signal is sent out, such as a falling edge from high level to low level.

[0094] S300, acquiring a detection signal of a fluorescence detector of each fluorescence channel for fluorescence emitted by the detected particle under laser irradiation, to obtain a light pulse signal;

[0095] The fluorescence signal is relatively weak and has spectral information. The fluorescence is divided into several channels by a light splitting system (such as a filter array, a grating or a prism) according to wavelength, and each channel corresponds to a fluorescence detector. The present application is aimed at weak fluorescence, and a silicon photomultiplier (MPPC / Si-PM) or a SPAD is used as a detector for fluorescence detection. The feature of this type of detector is that when it receives a photon, the photoelectric current generated under the action of a reverse bias will produce an avalanche amplification. The carrier generated by the photon will produce a large light pulse through multiplication, and the single photon can be detected by detecting the pulse. This is the working principle of SPAD, and MPPC further integrates multiple APDs as pixels, so MPPC can also respond to signals of multiple photons arriving at the same time to form pulses of different amplitudes.

[0096] S400, recording the generation of the light pulse signal in the start and end interval and analyzing to obtain the peak value and the area of the channel as the detection result of the channel signal.

[0097] In order to analyze the generation of the light pulse signal and facilitate the analysis of the data of the traditional flow cytometer, the present application sets a timer to record the number of time with a period (interval) t, and establishes an ADC or a comparator group to measure the amplitude A of the pulse with a single-photon pulse intensity as a basic unit when the pulse signal arrives.

[0098] The period t determines the resolution of the pulse detection result, and the smaller the period, the higher the resolution. The minimum period is related to the period of the MPPC responding to the photon pulse signal and the integration time of the sampling circuit. t generally does not need to be less than the duration of the photon pulse generated electrical signal.

[0099] Further, as shown in Figure 7 The step of recording the generation of the light pulse signal in the start and end interval and analyzing to obtain the peak value and the area of the channel as the detection result of the channel signal comprises:

[0100] The "area" parameter in the parameter of the traditional flow cytometer can be calculated by the following method: S410, accumulating the intensity of each pulse signal in the start and end interval to obtain a total amount of pulses , which can be used as the area value of the event:

[0101] ;

[0102] Further, the step of accumulating the intensity of each pulse signal in the start and end interval to obtain a total amount of pulses and taking it as the area value of the event further comprises:

[0103] To reduce the influence of noise on the result of area calculation, the result can be corrected by "baseline" calculation, as shown in Figure 3 .

[0104] Wait for a set time from the end of the previous event as the start of noise measurement, record the total number of pulses containing intensity information, and at a set time before the start of the current event as the end of noise measurement. The number of times between the start and end of noise measurement is , the total amplitude of the pulses in the interval is the total amount of baseline noise , and the ratio of the two is the average baseline noise:

[0105] ;

[0106] The area value after baseline removal is :

[0107] .

[0108] The "height" parameter in the traditional flow cytometer parameter can be calculated as follows: S420, when a new pulse signal arrives, the number of times between the previous pulse and the new pulse is taken as the dividend, the intensity A of the new pulse is taken as the divisor, and the quotient obtained is taken as the calculated interval time number of the current pulse:

[0109] ;

[0110] S430, set the number of pulse smoothing groups N, and calculate the weighted sum or smooth accumulation of the total number of times of N pulses as the corrected calculation interval time number:

[0111] ;

[0112] The weighting coefficient can be defined as:

[0113] ;

[0114] The average result of the calculation interval time number is obtained.

[0115] The weighting coefficient can also be defined as:

[0116] ;

[0117] Ignore the pulse intensity factor to suppress the influence of noise under very weak light signals.

[0118] The weighting coefficient It can also be defined as:

[0119]

[0120] By The pulse intensity represented by Determine the weight to get the average interval of the equivalent single photon pulse. Strengthen the weight of the multi-photon pulse, and adapt to the relatively strong signal.

[0121] S440, calculate The reciprocal of the modified intensity value of each photon pulse calculated The maximum value of the modified intensity of all pulses in the start and end interval That is, the peak value of the event detection of this channel.

[0122] The present application provides a method for collecting and processing weak fluorescent signals. For the detection results of the photon counting detector, the processing method can simply and quickly extract the area and height parameters of the event signal, and can be compared with the detection and processing results of the traditional flow cytometer using analog detectors. It is convenient for operators to control the detection process and understand the detection results. The method has strong compatibility. Example 3

[0123] A computer device 500, as shown in Figure 8 It includes a memory 510, a processor 520, and a computer program 530 stored on the memory and executable on the processor, and the processor executes the computer program to realize the steps of a flow analysis system signal processing method. For detailed description of the method, please refer to the corresponding description in the above method embodiment, which will not be repeated here. Example 4

[0124] A computer readable storage medium, as shown in Figure 9 It stores a computer program, and the computer program is executed by a processor to realize the steps of a flow analysis system signal processing method. For detailed description of the method, please refer to the corresponding description in the above method embodiment, which will not be repeated here.

[0125] The number of devices and the size of the processing described here are used to simplify the description of the present application. The application, modification and change of the present application are obvious to those skilled in the art.

[0126] ​Although embodiments of the present application have been disclosed in connection with the explicitly reited operating mechanisms of the application, it should be understood that they can not be limited to those precisely reited mechanisms, and that obvious modifications and / or coinations can be resorted to by those skilled in the art, and that such modifications and / or coinations fall within the scope of the application as reited in the claims and their equivalents.

[0127] The apparatus, computer device, and non-transitory computer storage medium provided by the embodiments of the present application are corresponding, and thus the apparatus, computer device, and non-transitory computer storage medium also have similar beneficial technical effects as the corresponding method. Since the beneficial technical effects of the method have been described in detail above, the beneficial technical effects of the corresponding apparatus, computer device, and non-transitory computer storage medium will not be described here again.

[0128] Those skilled in the art will appreciate that, in addition to implementing the controller in the form of a purely computer-readable program code, the controller can also be implemented in the form of logic gates, switches, application-specific integrated circuits, programmable logic controllers, and embedded microcontrollers, etc. by logically programming the method steps to achieve the same functions. Thus, the controller can be considered as a hardware component, and the means included therein for achieving various functions can also be considered as structures within the hardware component. Alternatively, the means for achieving various functions can even be considered as both software units implementing the method and structures within the hardware component.

[0129] The system, apparatus, or unit illustrated by the above embodiments can be specifically implemented by a computer chip or entity, or by a product with certain functions. For the convenience of description, the above apparatus is described by dividing various units according to functions during description. Of course, the functions of each unit can be implemented in the same or multiple software and / or hardware when implementing one or more embodiments of the present application.

[0130] Those skilled in the art will appreciate that the embodiments of the present application can be provided as a method, a system, or a computer program product. Thus, the embodiments of the present application can take the form of an entirely hardware embodiment, an entirely software embodiment, or an embodiment combining software and hardware aspects. Moreover, the embodiments of the present application can take the form of a computer program product implemented on one or more computer-usable storage media (including, but not limited to, disk memory, CD-ROM, optical memory, etc.) including computer-usable program code.

[0131] The specification is presented with reference to flow diagrams and / or block diagrams of methods, apparatus (systems) and computer program products according to embodiments of the specification. It will be understood that each block of the flow diagrams and / or block diagrams, and combinations of blocks in the flow diagrams and / or block diagrams, can be implemented by computer program instructions. These computer program instructions can be provided to a processor of a general purpose computer, special purpose computer, embedded processing system or other programmable data processing apparatus to produce a machine, such that the instructions, which execute via the processor of the computer or other programmable data processing apparatus, create means for implementing the functions specified in the flow diagrams and / or block diagrams block or blocks. Figure 1 The flow diagram and / or block diagram in the flow diagrams and / or block diagrams can also be implemented by the functional blocks of a computer system. Figure 1 The flow diagram and / or block diagram in the flow diagrams and / or block diagrams can also be implemented by the functional blocks of a computer system.

[0132] These computer program instructions can also be stored in a computer- readable memory that can direct a computer or other programmable data processing apparatus to function in a particular manner, such that the instructions stored in the computer-readable memory produce an article of manufacture including instructions which implement the flow diagrams and / or block diagrams block or blocks. Figure 1 The flow diagram and / or block diagram in the flow diagrams and / or block diagrams can also be implemented by the functional blocks of a computer system. Figure 1 The flow diagram and / or block diagram in the flow diagrams and / or block diagrams can also be implemented by the functional blocks of a computer system.

[0133] These computer program instructions can also be loaded onto a computer or other programmable data processing apparatus to cause a series of operational steps to be performed on the computer or other programmable apparatus to produce a computer-implemented process such that the instructions which execute on the computer or other programmable apparatus provide steps for implementing the flow diagrams and / or block diagrams block or blocks. Figure 1 The flow diagram and / or block diagram in the flow diagrams and / or block diagrams can also be implemented by the functional blocks of a computer system. Figure 1 The flow diagram and / or block diagram in the flow diagrams and / or block diagrams can also be implemented by the functional blocks of a computer system.

[0134] It should also be noted that the terms "comprising," "including," and any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but can also include other elements not expressly listed or inherent to such process, method, article, or apparatus. An element proceeded by "comprises... a" does not, without more constraints, exclude the existence of additional identical elements in the process, method, article, or apparatus that comprises the element.

[0135] The specification can be described in the general context of computer-executable instructions, such as program modules, being executed by a computer. Generally, program modules include routines, programs, objects, components, data structures, and the like, that perform particular tasks or implement particular abstract data types. Computer-executable instructions, associated data structures, and program modules represent examples of the program code means for executing steps of the methods disclosed herein. The particular sequence of steps and methods described in this specification is not the only manner in which the methods can be practiced. Likewise, the general description of the sequence of operations above applies to the methods in one or more embodiments.

[0136] Each of the embodiments in the specification is described in a progressive manner, and the same or similar parts between the embodiments can be referred to each other. Each of the embodiments focuses on the difference from other embodiments. In particular, the system embodiments are described simply because they are basically similar to the method embodiments, and the relevant parts can be referred to the description of the method embodiments.

[0137] The above only describes the embodiments of the specification and does not limit one or more embodiments of the specification. One or more embodiments of the specification can have various changes and variations for those skilled in the art. Any modification, equivalent replacement, improvement, etc. within the spirit and principle of one or more embodiments of the specification shall be included in the scope of claims of one or more embodiments of the specification.

Claims

1. A flow analysis system signal processing method characterized by, It comprises the following steps: a scattered light detector acquires a detection signal of scattered light emitted by the detected particle under laser irradiation to obtain an electrical signal; the electrical signal is analyzed to exclude interference by a preset threshold to obtain a starting time and an ending time of the detected particle passing through the detection area; a fluorescence detector acquires a detection signal of fluorescence emitted by the detected particle under laser irradiation to obtain a light pulse signal; the generation of the light pulse signal in the starting and ending intervals is recorded and analyzed to obtain a peak value and an area value of the channel as the detection result of the channel signal; a cycle t is recorded, and when the pulse signal arrives, the amplitude A of the pulse is measured as a basic unit of single-photon pulse intensity; the step of recording the generation of the light pulse signal in the starting and ending intervals and analyzing the peak value and the area value of the channel as the detection result of the channel signal comprises: the intensity of each pulse signal in the starting and ending intervals is accumulated to obtain a total pulse amount, which is used as the area value of the event; when a new pulse signal arrives, the time interval between the new pulse signal and the previous pulse signal is used as the divisor, and the intensity of the new pulse is used as the dividend to obtain a quotient as the calculated time interval number of the current pulse; a pulse smoothing group number N is set, and the total sum of the time numbers of N pulses is calculated as the modified calculated time interval number; wherein are weighting coefficients; The reciprocal of the calculation of the modified intensity value of each photon pulse after calculation The maximum value of the modified intensity of all pulses in the starting and ending intervals is the peak value of this event detection of the channel.

2. A signal processing method for a flow cytometric system as claimed in claim 1, characterized in that, the step of analyzing the electrical signal to exclude interference by a preset threshold to obtain a starting time and an ending time of the detected particle passing through the detection area comprises: when the intensity of the light signal is detected to be higher than an event starting threshold, an event starting signal is sent out; when the intensity of the light signal is detected to be lower than an event ending threshold, an event ending signal is sent out.

3. A signal processing method for a flow cytometric system as claimed in claim 1, characterized in that, after the step of accumulating the intensity of each pulse signal in the starting and ending intervals to obtain a total pulse amount and using the total pulse amount as the area value of the event, the following steps are further included: an event ending is taken as a starting point, and a pulse total amount containing intensity information is recorded after a set time as a starting point of noise measurement, and a set time before the starting of the current event is taken as an ending point of noise measurement; The number of times between the beginning and the end of the noise measurement is The total amplitude of the pulses in the interval is the total amount of baseline noise The ratio of the two is the average baseline noise: the area value after baseline removal is: 。 4. A signal processing method for a flow cytometric system as recited in claim 1, wherein, the weighting coefficient is defined as: or, the weighting coefficient is defined as: or, the weighting coefficient is defined as: wherein is the pulse intensity represented by I.

5. A signal processing device for a flow cytometric system, applying the method according to any one of claims 1 to 4, characterized in that: it comprises a scattered light detector, a fluorescence detector, a filter, an analog-to-digital converter, a light splitting system, a pulse shaper, a pulse reading module, and a data processing module; when the detected particle passes through the detection area, it emits scattered light and fluorescence under laser irradiation; the scattered light detector is used to collect light signals of the scattered light signal emitted by the detected particle and convert them into electrical signals; the light splitting system is used to divide the fluorescence into several channels according to the wavelength, and each channel corresponds to a fluorescence detector; the fluorescence detector is used to detect the fluorescence emitted by the detected particle; the filter and the analog-to-digital converter sequentially filter and analog-to-digital convert the electrical signal output by the scattered light detector; the pulse shaper is used to shape the signal detected by the fluorescence detector; the pulse reading module is used to judge the intensity of the pulse signal and convert the number of photons represented by the pulse; the data processing module is used to process the output of the analog-to-digital converter and the pulse reading module.

6. A signal processing apparatus for a flow analysis system as defined in claim 5, characterized in that: The scattered light detector adopts a photodiode, an avalanche diode or a photomultiplier tube.

7. A signal processing apparatus for a flow analysis system as defined in claim 5, characterized in that: The fluorescence detector adopts a silicon photomultiplier tube or a single-photon avalanche diode.

8. A signal processing apparatus for a flow analysis system as defined in claim 7, characterized in that: The pulse reading module realizes adjustable reference voltage through a digital-to-analog converter, realizes a predetermined number of voltage division through a resistance network, and each level output after voltage division represents the intensity of the pulse signal formed by the corresponding number of photons. The reference voltage of each level is compared with the intensity of the pulse signal through a comparator to determine the amplitude of the pulse signal, i.e., the number of photons it represents. Alternatively, the pulse reading module adopts a high-speed analog-to-digital converter to collect the pulse signal in real time. When the pulse signal arrives, the signal intensity is determined by the pulse peak value, and the number of photons represented by the pulse is converted.

9. A signal processing apparatus for a flow analysis system as defined in claim 8, characterized in that: When the pulse reading module adopts a high-speed analog-to-digital converter, a threshold value is formed by the intensity of the single-photon signal to suppress the interference of the bottom noise, and the signal is smoothed and filtered to further suppress the interference of the noise on the amplitude determination.

10. A computer device comprising a memory, a processor, and a computer program stored on the memory and executable on the processor, characterized in that, The processor executes the computer program to realize the steps of the method according to any one of claims 1-4.

11. A computer readable storage medium having stored thereon a computer program, characterized in that, The computer program is executed by the processor to realize the steps of the method according to any one of claims 1-4.

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