An intelligent detection method and device

Abstract

The present application provides an intelligent detection method and device, belonging to the field of flow cytometers, and is used to solve the problem that the accuracy of the detection results of flow cytometers in the related art is easily affected by various factors. In this method and device, a self-developed algorithm model is used to screen and analyze optimized microspheres and position-optimized microspheres, and the optimized microspheres are used to analyze and eliminate the detection results of single-cell liquid flow fluctuations, and the position-optimized microspheres are used to analyze and eliminate the optical system variation conditions. By combining the two, the final detection results that eliminate the influence of single-cell liquid flow fluctuations and optical system variations can be obtained, which is beneficial to making the final detection results accurate and reliable.

Description

Technical Field

[0001] This application relates to the field of flow cytometers, and particularly to an intelligent detection method and device. Background Art

[0002] The working principle of a flow cytometer is to make the cells or other biological particles to be measured pass through a liquid flow system to form a single-cell liquid flow, and through the irradiation of a laser beam, to generate scattered light and fluorescence. These optical signals will be captured by an optical system and converted into electrical signals, and then amplified and digitized by an electronic system. Finally, a data processing system will analyze these data to obtain information on the physical and biological characteristics of the cells or other biological particles, such as cell size, shape, DNA content, protein expression, etc.

[0003] During the application process of a flow cytometer, there may be many factors that affect the accuracy of its detection results. How to overcome these factors and ensure the accuracy of the detection results of the flow cytometer is a problem that those skilled in the art have been working hard to solve. Summary of the Invention

[0004] This application provides an intelligent detection method and device, which can overcome the influence of optical system variation and cell liquid flow fluctuation on the detection results, and is beneficial to improving the accuracy of the detection results.

[0005] In a first aspect, this application provides an intelligent detection method. The method includes:

[0006] Obtaining microsphere correlation information during the process of each microsphere passing through the light spot, where the microsphere correlation information includes one or more of forward scattered light, side scattered light, and microsphere fluorescence;

[0007] Analyzing the analysis standard degree and position standard degree of each microsphere based on the microsphere correlation information;

[0008] Determining an analysis preferred microsphere and a position preferred microsphere among all microspheres based on the analysis standard degree and the position standard degree;

[0009] Analyzing the optical variation rate according to the microsphere correlation information of the position preferred microsphere, where the optical variation rate reflects the variation direction and variation degree of the optical system in the flow cytometer;

[0010] Judging whether the absolute value of the optical variation rate is higher than the optical variation threshold;

[0011] If so, outputting an abnormal fault message, where the abnormal fault message reflects that an abnormal fault has occurred in the optical system of the flow cytometer;

[0012] If not, analyzing and obtaining a preferred detection result according to the analysis preferred microsphere and the optical variation rate.

[0013] By adopting the above technical solution, it is possible to select microspheres with less disturbance from the fluctuation of the cell fluid flow as the preferred microspheres to analyze the detection results, and the final preferred detection result is the detection result after being corrected by the calculated optical variation rate, so that the detection result finally output by the flow cytometer is relatively accurate, and an abnormal fault is prompted when the optical variation cannot be corrected by the algorithm, so that the abnormal fault of the flow cytometer can be repaired and maintained in time, which is beneficial to further ensuring the accuracy of the detection result of the flow cytometer.

[0014] Further, the analysis of the analysis standard degree and the position standard degree of each microsphere based on the microsphere correlation information includes:

[0015] Analyzing the first scattered light distribution data of the forward scattered light during the process of each microsphere passing through the light spot;

[0016] Determining the forward position sub-standard degree of the microsphere according to the first scattered light distribution data, the position standard degree is positively correlated with the forward position sub-standard degree, and the forward position sub-standard degree is positively correlated with the distribution stability and / or distribution symmetry of the first scattered light distribution data.

[0017] Further, the determination of the forward position sub-standard degree of the microsphere according to the first scattered light distribution data includes:

[0018] Determining the forward spectral peak value, forward spectral valley value, forward spectral peak width, forward spectral coefficient of variation and forward spectral distribution skewness of the forward scattered light according to the first scattered light distribution data;

[0019] Let the forward spectral peak value be , the forward spectral valley value be , the forward spectral peak width be , the forward spectral coefficient of variation be , the forward spectral distribution skewness be , and the forward position sub-standard degree be , then

[0020] ;

[0021] ;

[0022] ;

[0023] ;

[0024] In the formula, are all pre-acquired calculation weights.

[0025] Further, the analysis of the analysis standard degree and the position standard degree of each microsphere based on the microsphere correlation information further includes:

[0026] Analyze the second scattering distribution data of the lateral scattered light during the process of each microsphere passing through the light spot;

[0027] Determine the scattered light ratio distribution curve by combining the second scattered light distribution data and the first scattered light distribution data;

[0028] Analyze the ratio distribution curve to determine the comprehensive position sub-standard degree, the position standard degree is also positively correlated with the comprehensive position standard degree, and the comprehensive position sub-standard degree is positively correlated with the stability of the ratio distribution curve.

[0029] Further, the analyzing the ratio distribution curve to determine the comprehensive position sub-standard degree includes:

[0030] Let the coefficient of variation of the scattered light ratio distribution curve be and the maximum change rate be and the cumulative duration exceeding the preset ratio range be , and the comprehensive position sub-standard degree be , then

[0031] ;

[0032] In the formula, are all pre-obtained pre-designed calculation weights greater than zero.

[0033] Further, the analyzing the optical variation rate according to the microsphere correlation information for preferentially selecting microspheres includes:

[0034] Analyze the first scattered light distribution data of the forward scattered light and the fluorescence distribution data of the microsphere fluorescence during the process of each microsphere passing through the light spot;

[0035] Determine the forward spectral distribution mean value of the forward scattered light and the fluorescence distribution mean value according to the first scattered light distribution data;

[0036] Combine the forward spectral distribution mean value and the fluorescence distribution mean value, and the expected forward distribution mean value and the expected fluorescence distribution mean value of a plurality of pre-obtained preset coded microspheres, and the historical forward distribution mean value and the historical fluorescence distribution mean value of each expected coded microsphere in a plurality of pre-obtained historical microsphere groups;

[0037] Based on the expected forward distribution mean value and the expected fluorescence distribution mean value, perform clustering processing on the microspheres according to the forward spectral distribution mean value and according to the forward spectral distribution mean value and the fluorescence distribution mean value, and determine a forward spectral class mean value and a fluorescence distribution class mean value for each preset coded microsphere;

[0038] Let the forward spectral class mean value of the i-th preset coded microsphere be and the fluorescence distribution class mean value be and the expected forward distribution mean value be , the expected mean fluorescence distribution is , and the optical variation rate is G, then

[0039] ;

[0040] In the formula, are all pre - obtained pre - designed calculation weights greater than zero.

[0041] Furthermore, the analysis of the analysis standard degree and position standard degree of each microsphere based on the microsphere correlation information includes:

[0042] Analyze the first scattered light distribution data of the forward scattered light, the scattered light ratio distribution curve of the ratio of the lateral scattered light to the forward scattered light, and the fluorescence distribution data of the microsphere fluorescence during the process of each microsphere passing through the light spot;

[0043] Analyze the first scattered light distribution data, the ratio distribution curve, and the fluorescence distribution data respectively, and obtain the forward analysis sub - standard degree, the comprehensive analysis sub - standard degree, and the fluorescence analysis sub - standard degree respectively. The analysis standard degree is positively correlated with one or more of the forward analysis sub - standard degree, the comprehensive analysis sub - standard degree, and the fluorescence analysis sub - standard degree. The forward analysis sub - standard degree is positively correlated with the distribution stability, and / or distribution symmetry, and / or distribution expectancy of the first scattered light distribution data. The comprehensive analysis sub - standard degree is positively correlated with the stability of the ratio distribution curve. The fluorescence analysis sub - standard degree is positively correlated with the stability of the fluorescence distribution data.

[0044] Furthermore, the method for obtaining the analysis sub - standard degree includes:

[0045] Determine the forward spectral peak value, forward spectral valley value, forward spectral peak width, forward spectral coefficient of variation, forward spectral distribution skewness, forward spectral distribution mean, microsphere forward passing time, and forward spectral sudden change moment of the forward scattered light according to the first scattered light distribution data;

[0046] Let the forward spectral peak value be , the forward spectral valley value be , the forward spectral peak width be , the forward spectral coefficient of variation be , the forward spectral distribution skewness be , the forward spectral distribution mean be , the microsphere forward passing time be , the forward spectral sudden change moment be , and the forward analysis sub - standard degree be , then

[0047] ;

[0048] ;

[0049] ;

[0050] ;

[0051] ;

[0052] ;

[0053] ;

[0054] In the formula, is the expected forward spectral mean of the i-th preset coded microsphere, is the expected time for the i-th preset coded microsphere to pass through the light spot, are respectively the start time and the end time when the current microsphere passes through the light spot, are all pre-acquired pre-designed calculation weights greater than zero;

[0055] The method for obtaining the comprehensive analysis sub-standard degree includes:

[0056] Obtain the coefficient of variation, the maximum change rate, and the cumulative duration exceeding the preset ratio range of the scattered light ratio distribution curve;

[0057] Let the coefficient of variation of the scattered light ratio distribution curve be , the maximum change rate be , and the cumulative duration exceeding the preset ratio range be , and the comprehensive analysis sub-standard degree be , then

[0058] ;

[0059] In the formula, are all pre-acquired pre-designed calculation weights greater than zero;

[0060] The method for obtaining the fluorescence analysis sub-standard degree includes:

[0061] Obtain the coefficient of variation of the fluorescence distribution data;

[0062] Determine the fluorescence analysis sub-standard degree according to the coefficient of variation of the fluorescence distribution data, and the fluorescence analysis sub-standard degree is negatively correlated with the coefficient of variation of the fluorescence distribution data.

[0063] Furthermore, the obtaining of the preferred detection result according to the analysis of the preferred microspheres and the optical variation rate includes:

[0064] Let the value of the i-th dimension of the detection result data obtained by the analysis of the preferred microspheres be , the optical variation rate is G, and preferably the value of the i-th dimension in the detection result data is , then , where is the optical variation rate threshold, is a pre-designed calculation weight greater than zero.

[0065] In a second aspect, the present application provides an intelligent detection device. The device is used to execute any one of the methods described in the first aspect above.

[0066] In summary, the present application at least includes the following beneficial effects:

[0067] An intelligent detection method and device are provided, which can adjust the influence of single-cell fluid flow fluctuations and optical system variations in a flow cytometer on the detection result to a certain extent, and is beneficial to improving the accuracy of the detection result.

[0068] It should be understood that the content described in the invention content part is not intended to limit the key or important features of the embodiments of the present application, nor is it used to limit the scope of the present application. Other features of the present application will become easily understood through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0069] Combined with the drawings and referring to the following detailed description, the above and other features, advantages and aspects of the embodiments of the present application will become more obvious. In the drawings, the same or similar reference numerals represent the same or similar elements, where:

[0070] Figure 1 Shows a flowchart of an intelligent detection method in an embodiment of the present application. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0071] To make the objectives, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the drawings in the embodiments of the present application. Obviously, the described embodiments are some, but not all, of the embodiments of the present application. All other embodiments obtained by those of ordinary skill in the art without creative efforts based on the embodiments in the present application belong to the scope of protection of the present application.

[0072] In addition, the term "and / or" in this article is only a description of the association relationship of associated objects, indicating that there can be three relationships. For example, A and / or B can represent: A exists alone, A and B exist simultaneously, and B exists alone. In addition, the character " / " in this article generally represents an "or" relationship between the associated objects before and after.

[0073] The present application provides an intelligent detection method and device, which can correct the influence of single-cell fluid flow fluctuation and optical system variation on the detection results of a flow cytometer through algorithms, facilitating more accurate detection results of the flow cytometer.

[0074] In a first aspect, an embodiment of the present application discloses an intelligent detection method. This method can be executed by a flow cytometer and is directed to an entire microsphere group. Generally speaking, an entire microsphere group corresponds to one detection or experiment.

[0075] Figure 1 The flowchart of an intelligent detection method in an embodiment of the present application is shown.

[0076] Referring to Figure 1 , the method specifically includes the following steps:

[0077] S110: Obtain the microsphere correlation information during the process of each microsphere passing through the light spot.

[0078] In an embodiment of the present application, the light spot refers to the light spot formed by the focusing of the laser beam in the flow cytometer. The microsphere refers to the cell to be detected or other biological particles in the flow cytometer. The single-cell fluid flow can basically ensure that single microspheres pass through the light spot in sequence. After being excited by the light spot of the laser beam, forward scatter light, side scatter light, and fluorescence are generated. The forward scatter light, side scatter light, and fluorescence can be captured by the optical system of the flow cytometer and converted into electrical signals.

[0079] In the method of this step, the microsphere correlation information includes one or more of forward scatter light, side scatter light, and microsphere fluorescence. The forward scatter light, side scatter light, and microsphere fluorescence mentioned here are all converted electrical signals.

[0080] S120: Analyze the analysis standard degree and position standard degree of each microsphere based on the microsphere correlation information.

[0081] The analysis standard degree here is a quantitative index characterizing the preferred degree of the microsphere for analyzing the final detection result, and the position standard degree is a quantitative index characterizing the preferred degree of the microsphere for analyzing the optical system variation. The analysis standard degree and position standard degree are independently determined for each microsphere.

[0082] In the method of this step, the method for analyzing the analysis standard degree of microspheres based on microsphere correlation information specifically includes: analyzing the first scattered light distribution data of the forward scattered light, the scattered light ratio distribution curve of the ratio of the lateral scattered light to the forward scattered light, and the fluorescence distribution data of the microsphere fluorescence during the process of each microsphere passing through the light spot; respectively analyzing the first scattered light distribution data, the ratio distribution curve, and the fluorescence distribution data to obtain the forward analysis sub-standard degree, the comprehensive analysis sub-standard degree, and the fluorescence analysis sub-standard degree. The analysis standard degree is positively correlated with one or more of the forward analysis sub-standard degree, the comprehensive analysis sub-standard degree, and the fluorescence analysis sub-standard degree. The forward analysis sub-standard degree is positively correlated with the distribution stability, and / or distribution symmetry, and / or distribution expectancy of the first scattered light distribution data. The comprehensive analysis sub-standard degree is positively correlated with the stability of the ratio distribution curve. The fluorescence analysis sub-standard degree is positively correlated with the stability of the fluorescence distribution data.

[0083] Specifically, the method for obtaining the analysis sub-standard degree includes: determining the forward spectral peak value, forward spectral valley value, forward spectral peak width (width of the signal intensity distribution), forward spectral coefficient of variation, forward spectral distribution skewness, forward spectral distribution mean, microsphere forward passing time, and forward spectral sudden change moment of the forward scattered light according to the first scattered light distribution data; setting the forward spectral peak value as , the forward spectral valley value as , the forward spectral peak width as , the forward spectral coefficient of variation as , the forward spectral distribution skewness as , the front line spectral distribution mean as , the microsphere forward passing time as , the forward spectral sudden change moment as , and the forward analysis sub-standard degree as , then

[0084] ;

[0085] ;

[0086] ;

[0087] ;

[0088] ;

[0089] ;

[0090] ;

[0091] In the formula, is the expected forward spectral mean of the i-th preset coded microsphere, is the expected time for the i-th preset coded microsphere to pass through the light spot, are respectively the start time and the end time when the current microsphere passes through the light spot, are all pre-acquired pre-designed calculation weights greater than zero;

[0092] The method for obtaining the comprehensive analysis sub-standard degree includes: obtaining the coefficient of variation, the maximum change rate, and the cumulative duration exceeding the preset ratio range of the scattered light ratio distribution curve; let the coefficient of variation of the scattered light ratio distribution curve be , the maximum change rate be , the cumulative duration exceeding the preset ratio range be , and the comprehensive analysis sub-standard degree be , then

[0093] ;

[0094] In the formula, are all pre-acquired pre-designed calculation weights greater than zero;

[0095] The method for obtaining the fluorescence analysis sub-standard degree includes: obtaining the coefficient of variation of the fluorescence distribution data; determining the fluorescence analysis sub-standard degree according to the coefficient of variation of the fluorescence distribution data, and the fluorescence analysis sub-standard degree is negatively correlated with the coefficient of variation of the fluorescence distribution data. In a specific example, let the fluorescence analysis sub-standard degree be , and the coefficient of variation of the fluorescence distribution data be , then , in the formula, is a pre-acquired calculation weight greater than zero.

[0096] It should be understood that when determining the analysis standard degree in the foregoing content, the distribution stability of the forward scattered light of the microsphere (the relevant parameters are the forward spectral peak value, the forward spectral valley value, and the forward spectral coefficient of variation), the distribution symmetry (the relevant parameter is the forward spectral distribution skewness), and the distribution expectancy (the relevant parameters are the forward spectral distribution mean, the forward passing time of the microsphere, and the forward spectral sudden change moment) are comprehensively considered, as well as the distribution stability of the ratio of the lateral scattered light to the forward scattered light, and the distribution stability of the fluorescence value of the microsphere. For the above factors, only one or several of them can be considered in combination, or other parameters reflecting the distribution stability, distribution symmetry, and distribution expectancy of the forward scattered light, lateral scattered light, and fluorescence of the microsphere can also be considered. Other examples are not listed and introduced one by one here.

[0097] In the method of this step, the method for analyzing the position standard degree of microspheres based on microsphere correlation information specifically includes: analyzing the first scattered light distribution data of the forward scattered light during the process of each microsphere passing through the light spot; determining the forward position sub-standard degree of the microsphere according to the first scattered light distribution data, the position standard degree being positively correlated with the forward position sub-standard degree, and the forward position sub-standard degree being positively correlated with the distribution stability and / or distribution symmetry of the first scattered light distribution data.

[0098] The analysis of the analysis standard degree and position standard degree of each microsphere based on the microsphere correlation information may further include: analyzing the second scattered light distribution data of the lateral scattered light during the process of each microsphere passing through the light spot; combining the second scattered light distribution data and the first scattered light distribution data to determine the scattered light ratio distribution curve; analyzing the ratio distribution curve to determine the comprehensive position sub-standard degree, the position standard degree being positively correlated with the comprehensive position standard degree, and the comprehensive position sub-standard degree being positively correlated with the stability of the ratio distribution curve.

[0099] Specifically, the determination of the forward position sub-standard degree of the microsphere according to the first scattered light distribution data includes: determining the forward spectral peak value, forward spectral valley value, forward spectral peak width (width of the signal intensity distribution), forward spectral coefficient of variation, and forward spectral distribution skewness of the forward scattered light according to the first scattered light distribution data; setting the forward spectral peak value as , the forward spectral valley value as , the forward spectral peak width as , the forward spectral coefficient of variation as , the forward spectral distribution skewness as , and the forward position sub-standard degree as , then

[0100] ;

[0101] ;

[0102] ;

[0103] ;

[0104] In the formula, are all pre-obtained calculation weights.

[0105] The analysis of the ratio distribution curve to determine the comprehensive position sub-standard degree includes: setting the coefficient of variation of the scattered light ratio distribution curve as , the maximum change rate as , the cumulative duration exceeding the preset ratio range as , and the comprehensive position sub-standard degree as , then

[0106] ;

[0107] wherein are all pre-obtained pre-designed calculation weights greater than zero.

[0108] Similarly, the factors considered for the above calculation position standardization can also be analogously considered in the same way as when analyzing the standardization. Each factor can be applied independently or in any combination, or extended and supplemented to specific examples by analogy to similar factors, which will not be elaborated here.

[0109] When the considered factors are different, the requirements for the forward scattered light, side scattered light, and microsphere fluorescence applied in the foregoing steps are different, that is, the specific content of the microsphere correlation information matches the factors considered in this step.

[0110] S130: Based on the analysis standardization and position standardization, determine the analysis preferred microspheres and position preferred microspheres among all microspheres respectively.

[0111] In a specific example of the method in this step, it is possible to consider pre-setting an analysis standard threshold corresponding to the analysis standardization and a position standard threshold corresponding to the position standardization, and determining that the microspheres with an analysis standardization higher than the standard analysis threshold are analysis preferred microspheres, and determining that the microspheres with a position standardization higher than the position standard threshold are position preferred microspheres.

[0112] In another example, it is also possible to consider pre-setting a standard analysis ratio and a standard position ratio, and taking the number of microspheres with a higher analysis standardization in the microsphere group corresponding to the standard analysis ratio as the analysis preferred microspheres, and taking the positions of the number of microspheres with a higher position standardization in the microsphere group corresponding to the standard position ratio as the position preferred microspheres.

[0113] Other similar examples will not be listed one by one. It only needs to be able to determine the analysis preferred microspheres and position preferred microspheres based on the analysis standardization and position standardization.

[0114] S140: Analyze the optical variation rate according to the microsphere correlation information of the position preferred microspheres.

[0115] In the method of this step, the optical variation rate reflects the variation direction and degree of the optical system in the flow cytometer.

[0116] In one example, the method of this step specifically includes: analyzing the first scattered light distribution data of the forward scattered light and the fluorescence distribution data of the microsphere fluorescence during the process of each microsphere passing through the light spot; determining the forward spectral distribution mean value and the fluorescence distribution mean value of the forward scattered light according to the first scattered light distribution data; combining the forward spectral distribution mean value and the fluorescence distribution mean value, the expected forward distribution mean value and the expected fluorescence distribution mean value of a variety of preset coded microspheres obtained in advance, and the historical forward distribution mean value and the historical fluorescence distribution mean value of each expected coded microsphere in a plurality of historical microsphere groups obtained in advance; based on the expected forward distribution mean value and the expected fluorescence distribution mean value, clustering the microspheres according to the forward spectral distribution mean value and the forward spectral distribution mean value and the fluorescence distribution mean value, and determining a forward spectral class mean value and a fluorescence distribution class mean value for each preset coded microsphere.

[0117] Let the forward spectral class mean value of the i-th preset coded microsphere be , the fluorescence distribution class mean value be , the expected forward distribution mean value be , the expected fluorescence distribution mean value be , and the optical variation rate be G, then

[0118] ;

[0119] In the formula, are all pre-obtained pre-designed calculation weights greater than zero.

[0120] The sign of the optical variation rate reflects the variation direction of the optical system (that is, whether the analyzed fluorescence value is higher or lower than the actual fluorescence value), and the magnitude of the optical variation rate reflects the variation degree of the optical system (that is, the magnitude of the difference between the analyzed fluorescence value and the actual fluorescence value). It should be understood that considering that in the same environment, the variation direction and the variation degree of the excitation and acquisition of the optical system are basically the same, so it can be considered that the signs of the result of subtracting the expected forward distribution mean value from the forward spectral class mean value and the result of subtracting the expected fluorescence distribution mean value from the fluorescence distribution class mean value are the same and the magnitudes are basically the same.

[0121] S150: Judge whether the absolute value of the optical variation rate is higher than the optical variation threshold.

[0122] The optical variation threshold is a preset constant, and the principle of this step is the absolute value comparison process, which will not be elaborated.

[0123] S160: If the absolute value of the optical variation rate is higher than the optical variation threshold, output an abnormal fault message.

[0124] In the method of this step, the abnormal fault information reflects an abnormal fault in the optical system of the flow cytometer. The absolute value of the optical variation rate is higher than the optical variation threshold, indicating that the optical variation with this degree of variation cannot be corrected by algorithmic means, and it is necessary to repair and maintain the flow cytometer to correct the optical system. Therefore, the abnormal fault information is output, and the abnormal fault information can reach relevant personnel in ways such as direct display and triggering prompts on terminals such as mobile phones.

[0125] S170: If the absolute value of the optical variation rate is not higher than the optical variation threshold, then the optimal detection result is obtained based on the analysis of the selected microspheres and the optical variation rate.

[0126] If the absolute value of the optical variation rate is not higher than the optical variation threshold, it indicates that the optical variation with this degree of variation can be corrected by algorithmic means. At this time, the detection result data is first analyzed based on the selected microspheres for analysis. The analysis process here is the same as the analysis process using the entire microsphere group, so the analysis process will not be elaborated. Only the detection result data analyzed using the selected microspheres for analysis has statistical significance, can represent the entire microsphere group, and can eliminate the influence of single-cell fluid flow fluctuations on the detection result to a certain extent.

[0127] The detection result data is generally fluorescence values in multiple dimensions. After obtaining the detection result data, a specific example of the method for specifically determining the optimal detection result is as follows: Let the value of the i-th dimension of the detection result data obtained by analyzing the selected microspheres for analysis be , the optical variation rate be G, and the value of the i-th dimension in the optimal detection result data be , then , where is the optical variation rate threshold, and is a pre-designed calculation weight greater than zero. Using the optical variation rate to correct the detection result can eliminate the influence of optical system variation on the detection result to a certain extent, which is beneficial to further improving the accuracy of the detection result of the flow cytometer.

[0128] In summary, this method uses a self-developed microsphere screening model to respectively determine the selected microspheres for analysis and the selected microspheres for position, analyzes the variation of the optical system based on the selected microspheres for position, analyzes the detection result of eliminating single-cell fluid flow disturbance based on the selected microspheres for analysis, and then combines the variation of the optical system and the detection result of eliminating single-cell fluid flow disturbance to comprehensively determine the final detection result, so that the final detection result can overcome the two factors affecting the accuracy of the detection result in the flow cytometer, which is beneficial to improving the accuracy of the detection result.

[0129] It should be noted that, for the foregoing method embodiments, for the sake of simple description, they are all expressed as a series of action combinations. However, those skilled in the art should know that this application is not limited by the described action sequence, because according to the embodiments of this application, certain steps can be performed in other sequences or simultaneously. Secondly, those skilled in the art should also know that the embodiments described in the specification are all optional embodiments, and the actions and modules involved are not necessarily essential to this application.

[0130] In a second aspect, embodiments of the present application disclose an intelligent detection method and device. The device is used to execute any one of the methods disclosed in the first aspect above.

[0131] Those skilled in the art can clearly understand that for the convenience and brevity of description, the specific working process of the described device can refer to the corresponding process in the foregoing method embodiments, and will not be elaborated herein.

[0132] In summary, the present application at least includes the following beneficial effects:

[0133] An intelligent detection method and device are provided, which can, to a certain extent, adjust the influence of single-cell liquid flow fluctuations and optical system variations in a flow cytometer on the detection results, and is beneficial to improving the accuracy of the detection results.

[0134] The above description is only a preferred embodiment of the present application and an explanation of the applied technical principles. Those skilled in the art should understand that the scope of disclosure involved in the present application is not limited to the technical solutions formed by the specific combination of the above technical features, and should also cover other technical solutions formed by any combination of the above technical features or their equivalent features without departing from the foregoing disclosure concept. For example, the technical solutions formed by mutually replacing the above features with (but not limited to) technical features having similar functions disclosed in the present application.

Claims

1. An intelligent detection method, characterized in that: include: Acquire microsphere-related information of each microsphere when it passes through the light spot, wherein the microsphere-related information includes one or more of forward scattered light, side scattered light, and microsphere fluorescence; Analyzing the analysis standard and position standard of each microsphere based on the microsphere association information, the analysis standard is positively correlated to the distribution stability, distribution symmetry and distribution expectancy of the forward scattered light of the microsphere, and / or is used to characterize the distribution stability of the ratio of the side scattered light to the forward scattered light, and / or is used to characterize the distribution stability of the fluorescence value of the microsphere, the position standard is positively correlated to the distribution stability and / or distribution symmetry of the first scattered light distribution data, and / or is positively correlated to the stability of the ratio distribution curve of the second scattered light distribution data and the first scattered light distribution data; Based on the analysis standard and the position standard, respectively, analysis-preferred microspheres and position-preferred microspheres are determined among all microspheres; Analyzing the optical variation rate according to the microsphere association information of the position-preferred microsphere, wherein the optical variation rate reflects the variation direction and degree of variation of the optical system in the flow cytometer; Determining whether the absolute value of the optical variation rate is higher than the optical variation threshold; If yes, then outputting abnormal fault information, wherein the abnormal fault information reflects that an abnormal fault occurs in the optical system of the flow cytometer; If not, then the preferred test results are obtained based on the preferred microsphere and optical variability analysis according to the analysis.

2. The method according to claim 1, characterized in that The analyzing the analysis standard and position standard of each microsphere based on the microsphere association information includes: Analyze the first scattered light distribution data of the forward scattered light of each microsphere in the process of passing through the light spot; The forward position substandard of the microsphere is determined according to the first scattered light distribution data, wherein the position standard is positively correlated to the forward position substandard, and the forward position substandard is positively correlated to the distribution stability and / or distribution symmetry of the first scattered light distribution data.

3. The method according to claim 2, characterized in that Determining the forward position substandard of the microsphere according to the first scattered light distribution data comprises: Determining a forward spectrum peak value, a forward spectrum valley value, a forward spectrum peak width, a forward spectrum coefficient of variation, and a forward spectrum distribution skewness of the forward scattered light according to the first scattered light distribution data; Assume that the forward spectrum peak is The forward spectrum valley value is , the forward spectrum peak width is , the coefficient of variation of the forward spectrum is , the forward spectral distribution skewness is , the forward position substandard degree is ,but ； ； ； ； In the formula, These are all pre-acquired calculation weights.

4. The method according to claim 2, characterized in that: The analyzing the analysis standard and position standard of each microsphere based on the microsphere association information further includes: Analyze the second scattering distribution data of the side scattered light of each microsphere when it passes through the light spot; determining a scattered light ratio distribution curve by combining the second scattered light distribution data and the first scattered light distribution data; The ratio distribution curve is analyzed to determine a comprehensive position sub-standard, the position standard is also positively correlated to the comprehensive position standard, and the comprehensive position sub-standard is positively correlated to the stability of the ratio distribution curve.

5. The method according to claim 4, characterized in that The analyzing the ratio distribution curve to determine the comprehensive position sub-standard includes: Assume that the coefficient of variation of the scattered light ratio distribution curve is , the maximum rate of change is , the cumulative duration exceeding the preset ratio range is , the comprehensive position sub-standard is ,but ； In the formula, All are pre-acquired preset calculation weights greater than zero.

6. The method according to claim 1, characterized in that The analyzing the optical variation rate according to the microsphere association information of the position-preferred microspheres comprises: Analyze the first scattered light distribution data of the forward scattered light of each microsphere passing through the light spot and the fluorescence distribution data of the microsphere fluorescence; Determine a forward spectral distribution mean and a fluorescence distribution mean of the forward scattered light according to the first scattered light distribution data; Combining the forward spectral distribution mean and the fluorescence distribution mean, as well as the pre-acquired expected forward distribution means and expected fluorescence distribution means of a plurality of preset coded microspheres, and the pre-acquired historical forward distribution means and historical fluorescence distribution means of each expected coded microsphere in a plurality of historical microsphere groups; Based on the expected forward distribution mean and the expected fluorescence distribution mean, clustering the microspheres according to the forward spectral distribution mean and the forward spectral distribution mean and the fluorescence distribution mean, determining a forward spectral class mean and a fluorescence distribution class mean for each preset coded microsphere; Assume that the forward spectrum class mean of the i-th preset coded microsphere is , the mean value of fluorescence distribution is , the expected forward distribution mean is , the expected fluorescence distribution mean is , the optical variation rate is G, then ； In the formula, and All are pre-acquired preset calculation weights greater than zero.

7. The method according to claim 1, characterized in that The analyzing the analysis standard and position standard of each microsphere based on the microsphere association information includes: Analyze the first scattered light distribution data of the forward scattered light of each microsphere passing through the light spot, the scattered light ratio distribution curve of the ratio of the side scattered light to the forward scattered light, and the fluorescence distribution data of the microsphere fluorescence; The first scattered light distribution data, ratio distribution curve and fluorescence distribution data are analyzed respectively to obtain a forward analysis sub-standard, a comprehensive analysis sub-standard and a fluorescence analysis sub-standard respectively, the analysis standard is positively correlated to one or more of the forward analysis sub-standard, the comprehensive analysis sub-standard and the fluorescence analysis sub-standard, the forward analysis sub-standard is positively correlated to the distribution stability, and / or distribution symmetry, and / or distribution expectancy of the first scattered light distribution data, the comprehensive analysis sub-standard is positively correlated to the stability of the ratio distribution curve, and the fluorescence analysis sub-standard is positively correlated to the stability of the fluorescence distribution data.

8. The method according to claim 7, characterized in that The method for obtaining the analysis substandard degree includes: Determine the forward spectrum peak value, the forward spectrum valley value, the forward spectrum peak width, the forward spectrum coefficient of variation, the forward spectrum distribution skewness, the forward spectrum distribution mean, the microsphere forward passage time and the forward spectrum sudden change moment of the forward scattered light according to the first scattered light distribution data; Assume that the forward spectrum peak is The forward spectrum valley value is , the forward spectrum peak width is , the coefficient of variation of the forward spectrum is , the forward spectral distribution skewness is , the mean of the front spectral distribution is , the forward transit time of the microsphere is , the forward spectrum suddenly changes at , the forward analysis sub-criterion is ,but ； ； ； ； ； ； ； In the formula, is the expected forward spectrum mean of the i-th preset coded microsphere, is the expected time for the i-th preset coded microsphere to pass through the light spot, and are the start time and end time of the current microsphere passing through the light spot, All are pre-acquired preset calculation weights greater than zero; The method for obtaining the comprehensive analysis sub-standard degree includes: Obtain the coefficient of variation, maximum rate of change, and cumulative duration of the scattered light ratio distribution curve exceeding a preset ratio range; Assume that the coefficient of variation of the scattered light ratio distribution curve is , the maximum rate of change is , the cumulative duration exceeding the preset ratio range is , the comprehensive analysis sub-standard degree is ,but ； In the formula, All are pre-acquired preset calculation weights greater than zero; The method for obtaining the fluorescence analysis substandard degree includes: Obtain the coefficient of variation of fluorescence distribution data; The fluorescence analysis sub-standard is determined according to the coefficient of variation of the fluorescence distribution data, and the fluorescence analysis sub-standard is negatively correlated to the coefficient of variation of the fluorescence distribution data.

9. The method according to any one of claims 1 to 8, characterized in that: The preferred detection results obtained by analyzing the preferred microspheres and the optical variation rate analysis include: Suppose the value of the i-th dimension of the test result data obtained by analyzing the optimal microspheres is , the optical variation rate is G, and the value of the i-th dimension in the preferred test result data is ,but , where is the optical variation rate threshold, Calculates weights for presets that are greater than zero.

10. An intelligent detection device, characterized in that: Used to perform the method according to any one of claims 1 to 8.

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