CN-121988413-A - Micro-droplet detection method, apparatus, system, readable storage medium and program product

CN121988413ACN 121988413 ACN121988413 ACN 121988413ACN-121988413-A

Abstract

The application relates to a micro-droplet detection method, a device, a system, a computer device, a computer readable storage medium and a computer program product, the method comprises the steps of obtaining a sampling sequence of micro-droplets; the sampling sequence comprises a plurality of sampling signal values, wherein the plurality of sampling signal values comprise fluorescent signal values of a fluorescent sampling interval and background signal values of a background sampling interval, a trend curve of a background signal is fitted according to the background signal values of the background sampling interval, fitting signal values corresponding to the fluorescent signal values in the fluorescent sampling interval on the trend curve are determined, and the state result of the micro liquid drops is obtained by comparing the fluorescent signal values in the fluorescent sampling interval with the fitting signal values corresponding to the fluorescent signal values. The application can quantify the enrichment characteristic of the fluorescence signal, and further can reduce the influence of the enrichment of the fluorescence signal on the nature of the micro-droplets when the micro-droplets are nature-characterized, thereby realizing the accurate nature of the micro-droplets and further improving the accuracy of screening the micro-droplets.

Inventors

YAN XIANGWEN
Shen Mengzhe
ZHANG HUI
WANG BAOBAO
LI JING
XIE QINGQING
ZHENG YUE
ZHANG WENWEI

Assignees

深圳华大生命科学研究院

Dates

Publication Date: 20260508
Application Date: 20241105

Claims (15)

1. A method of detecting a microdroplet, comprising: The method comprises the steps of obtaining a sampling sequence of micro-droplets, wherein the sampling sequence comprises a plurality of sampling signal values, and the plurality of sampling signal values comprise a fluorescent signal value of a fluorescent sampling interval and a background signal value of a background sampling interval; Fitting a trend curve of the background signal according to the background signal value of the background sampling interval; Determining fitting signal values corresponding to the fluorescent signal values in the fluorescent sampling interval on the trend curve; And comparing each fluorescence signal value in the fluorescence sampling interval with the corresponding fitting signal value to obtain a state result of the micro liquid drop.
2. The method of claim 1, wherein each sampled signal value has a corresponding sampling sequence number describing a sampling order of the sampled signal values, and wherein the obtaining the sampling sequence of microdroplets comprises: acquiring a plurality of sampling signal values and sampling serial numbers corresponding to the sampling signal values; dividing a plurality of sampling serial numbers into a fluorescent sampling interval and a background sampling interval, taking a sampling signal value of the fluorescent sampling interval as a fluorescent signal value, and taking a sampling signal value of the background sampling interval as a background signal value.
3. The method of claim 2, wherein the dividing the plurality of sample sequence numbers into a fluorescence sample interval and a background sample interval comprises: Acquiring a maximum sampling serial number corresponding to a maximum sampling signal value; Dividing a fluorescence sampling interval containing the maximum sampling sequence number according to the position of the maximum sampling sequence number in the whole sampling sequence; and dividing the sampling sequence numbers outside the fluorescence sampling interval into background sampling intervals.
4. The method of claim 3, wherein the partitioning the fluorescence sample interval containing the largest sample sequence number based on the location of the largest sample sequence number in the entire sample sequence comprises: Calculating the sequence ratio of the maximum sampling sequence number to the total sampling sequence number; And determining a sequence starting point and a sequence end point of the fluorescence sampling interval according to the sequence ratio to obtain the fluorescence sampling interval containing the maximum sampling sequence number.
5. The method of claim 4, wherein determining the sequence start and sequence end of the fluorescence sampling interval based on the sequence ratio comprises: acquiring a preset radius proportion of a fluorescence sampling interval; If the sequence ratio is smaller than a first ratio, determining a difference value between the first ratio and a preset radius ratio as a starting point ratio of a fluorescence sampling interval, and determining a sum of the first ratio and the preset radius ratio as an ending point ratio of the fluorescence sampling interval; If the sequence ratio is larger than a second ratio, determining a difference value between the second ratio and a preset radius ratio as a starting point ratio of a fluorescence sampling interval, and determining a sum of the second ratio and the preset radius ratio as an end point ratio of the fluorescence sampling interval, wherein the second ratio is larger than the first ratio; If the sequence ratio is greater than or equal to the first ratio and less than or equal to the second ratio, determining a difference value between the sequence ratio and a preset radius ratio as a starting point ratio of a fluorescence sampling interval, and determining a sum of the sequence ratio and the preset radius ratio as an end point ratio of the fluorescence sampling interval; Taking the product of the starting point proportion of the fluorescence sampling interval and the total number of the serial numbers as the sequence starting point of the fluorescence sampling interval, and taking the product of the ending point proportion of the fluorescence sampling interval and the total number of the serial numbers as the sequence ending point of the fluorescence sampling interval.
6. The method of claim 1, wherein said obtaining a status result of said microdroplet by comparing each fluorescence signal value in said fluorescence sampling interval with its corresponding said fitted signal value comprises: determining a continuous interval in which the fluorescence signal value is larger than the corresponding fitting signal value from the fluorescence sampling interval; if the duty ratio of the continuous interval in the fluorescence sampling interval is smaller than a preset duty ratio, judging that the micro liquid drop is a negative micro liquid drop; And if the duty ratio of the continuous interval in the fluorescence sampling interval is larger than the preset duty ratio, obtaining the enhancement ratio corresponding to each fluorescence signal value in the fluorescence sampling interval, and obtaining the state result of the micro-droplet by counting the enhancement ratio, wherein the enhancement ratio is the ratio of the fluorescence signal value to the fitting signal value corresponding to the fluorescence signal value.
7. The method of claim 6, wherein the obtaining the status result of the microdroplet by counting the enhancement ratio comprises: determining the maximum enhancement ratio of the enhancement ratios and the average enhancement ratio of the enhancement ratios; If the average enhancement ratio reaches a preset average enhancement ratio threshold value and/or the maximum enhancement ratio reaches a preset maximum enhancement ratio threshold value, judging the micro-droplet to be a positive micro-droplet; Otherwise, judging the micro-droplet to be a negative micro-droplet.
8. The method of claim 1, wherein the plurality of sampled signal values of the microdroplet form a sampled dataset of the microdroplet, the sampled dataset being obtained by compressing an original dataset to a target data length, the acquiring a sampled sequence of microdroplets further comprising, prior to: acquiring an original data set, wherein the original data set comprises a plurality of original sampling signal values; Determining an original data length of the original data set; Determining a compression multiple s for compressing the original data set according to the original data length and the target data length, wherein the compression multiple s is a positive integer; The sampling sequence for obtaining micro-droplets comprises: And taking each s original sampling signal values as a group according to the sampling sequence of the original sampling signal values, taking the average value of each group of original sampling signal values as the sampling signal value, and obtaining a sampling sequence comprising the sampling signal values.
9. The method of claim 1, further comprising, after said comparing each fluorescence signal value in said fluorescence sampling interval with its corresponding said fitted signal value to obtain a status result of said microdroplet: if the micro-droplet is a positive micro-droplet, controlling a deflection electrode of the micro-fluidic chip to be opened, and controlling the flow direction of the micro-droplet to deflect through the deflection electrode; the duration of the deflection electrode is controlled so that the micro-droplets flow out of a preset outlet of the microfluidic chip.
10. A micro-droplet detection apparatus, comprising: The device comprises an acquisition module, a sampling module and a sampling module, wherein the acquisition module is used for acquiring a sampling sequence of micro-droplets, the sampling sequence comprises a plurality of sampling signal values, and the plurality of sampling signal values comprise a fluorescent signal value of a fluorescent sampling interval and a background signal value of a background sampling interval; The fitting module is used for fitting a trend curve of the background signal according to the background signal value of the background sampling interval; the determining module is used for determining fitting signal values corresponding to the fluorescent signal values in the fluorescent sampling interval on the trend curve; And the statistics module is used for obtaining the state result of the micro liquid drop by comparing each fluorescence signal value in the fluorescence sampling interval with the fitting signal value corresponding to each fluorescence signal value.
11. A micro-droplet detection system, comprising: the microfluidic chip is provided with a flow channel, the flow channel comprises at least two inlets and at least two outlets, and micro liquid drops are generated at the inlets and react in the flow channel to generate fluorescent signals; The micro-liquid droplet state detection device comprises a screening platform, a detection platform and a detection platform, wherein the screening platform is used for collecting fluorescent signals of the micro-liquid droplet to obtain a sampling sequence of the micro-liquid droplet, the sampling sequence comprises a plurality of sampling signal values, the sampling signal values comprise fluorescent signal values of a fluorescent sampling interval and background signal values of a background sampling interval, a trend curve of the background signal is fitted according to the background signal values of the background sampling interval, fitting signal values corresponding to the fluorescent signal values in the fluorescent sampling interval on the trend curve are determined, and the state result of the micro-liquid droplet is obtained by comparing the fluorescent signal values in the fluorescent sampling interval with the fitting signal values corresponding to the fluorescent signal values.
12. The system of claim 11, wherein a deflection electrode is disposed outside the flow channel of the microfluidic chip, the at least two outlets comprise preset outlets, and the screening platform is further configured to: if the micro-droplet is a positive micro-droplet, controlling a deflection electrode of the micro-fluidic chip to be opened, controlling the flow direction of the micro-droplet to deflect through the deflection electrode, and controlling the duration of the deflection electrode so as to enable the micro-droplet to flow out of a preset outlet of the micro-fluidic chip.
13. A computer device comprising a memory and a processor, the memory storing a computer program, characterized in that the processor implements the steps of the method of any one of claims 1 to 9 when the computer program is executed.
14. A computer readable storage medium, on which a computer program is stored, characterized in that the computer program, when being executed by a processor, implements the steps of the method of any of claims 1 to 9.
15. A computer program product comprising a computer program, characterized in that the computer program, when being executed by a processor, implements the steps of the method of any one of claims 1 to 9.

Description

Micro-droplet detection method, apparatus, system, readable storage medium and program product Technical Field The present application relates to the field of bioengineering, and in particular to a micro droplet detection method, apparatus, system, computer device, computer readable storage medium and computer program product. Background This section is intended to provide a background or context for the implementation of the embodiments of the invention that are set forth in the claims and the detailed description. The description herein is not admitted to be prior art by inclusion in this section. The micro-droplet technology has the application characteristics of microminiaturization, integration, easy preparation, mutual independence and mutual noninterference among the micro-droplets, small reagent consumption, high flux, capability of detecting secretory antibodies and the like, so that the research on the micro-droplet technology has strong practical significance and social benefit. With the development of biotechnology, the demand for antibodies has grown. The micro-droplet technology can judge the micro-droplet as negative or positive according to a fluorescent signal generated after the micro-droplet reaction, so as to control the flow direction of the micro-droplet, realize the screening of antigen-specific antibodies, and can achieve the ultra-high flux which cannot be achieved by the traditional flat plate screening technology with few reagents. However, the enrichment region generating fluorescent signals after the reaction of the micro-droplets has randomness, which easily leads to erroneous judgment of the negative and positive of the micro-droplets and influences the screening accuracy. Disclosure of Invention In view of the foregoing, it is desirable to provide a microdroplet detection method, apparatus, system, computer device, computer readable storage medium, and computer program product that can improve the accuracy of microdroplet screening. In a first aspect, the present application provides a method for detecting micro-droplets, comprising: The method comprises the steps of obtaining a sampling sequence of micro-droplets, wherein the sampling sequence comprises a plurality of sampling signal values, and the plurality of sampling signal values comprise a fluorescent signal value of a fluorescent sampling interval and a background signal value of a background sampling interval; Fitting a trend curve of the background signal according to the background signal value of the background sampling interval; Determining fitting signal values corresponding to the fluorescent signal values in the fluorescent sampling interval on the trend curve; And comparing each fluorescence signal value in the fluorescence sampling interval with the corresponding fitting signal value to obtain a state result of the micro liquid drop. In a second aspect, the present application also provides a micro-droplet detection apparatus, comprising: The device comprises an acquisition module, a sampling module and a sampling module, wherein the acquisition module is used for acquiring a sampling sequence of micro-droplets, the sampling sequence comprises a plurality of sampling signal values, and the plurality of sampling signal values comprise a fluorescent signal value of a fluorescent sampling interval and a background signal value of a background sampling interval; The fitting module is used for fitting a trend curve of the background signal according to the background signal value of the background sampling interval; the determining module is used for determining fitting signal values corresponding to the fluorescent signal values in the fluorescent sampling interval on the trend curve; And the statistics module is used for obtaining the state result of the micro liquid drop by comparing each fluorescence signal value in the fluorescence sampling interval with the fitting signal value corresponding to each fluorescence signal value. In a third aspect, the present application also provides a micro-droplet detection system, comprising: the microfluidic chip is provided with a flow channel, the flow channel comprises at least two inlets and at least two outlets, and micro liquid drops are generated at the inlets and react in the flow channel to generate fluorescent signals; The micro-liquid droplet state detection device comprises a screening platform, a detection platform and a detection platform, wherein the screening platform is used for collecting fluorescent signals of the micro-liquid droplet to obtain a sampling sequence of the micro-liquid droplet, the sampling sequence comprises a plurality of sampling signal values, the sampling signal values comprise fluorescent signal values of a fluorescent sampling interval and background signal values of a background sampling interval, a trend curve of the background signal is fitted according to the background signal values of the background sampling interval, fitting