A method and system for washing-free image flow-like fluorescence detection

Abstract

This application provides a wash-free image flow cytometry fluorescence detection method and system. The method includes using a preset target fluorescent dye and a dyeing method to dye the target sample to ensure that a certain amount of free fluorescent dye is formed after the dyeing and incubation. All cells in the target sample can collect fluorescence intensity values ​​for density function cluster resolution analysis under a wash-free image-like flow fluorescence detection system; obtain microscopic fluorescence images of the stained target sample based on the microscopic imaging system ; Use computer image recognition technology to extract the gray value and pixel value of the target sample; calculate the fluorescence intensity value of the target sample based on the gray value and pixel value of the target sample, and output the microscopic fluorescence image and density function cluster analysis of the target sample the result of. It solves the problem that under the image flow cytometry fluorescence detection system, the fluorescence intensity value of unstained or slightly stained negative cells will be zero, or there will be an obvious cliff after detection after ordinary staining and washing.

Description

technical field [0001] The invention relates to the technical field of cell detection, in particular to a wash-free image-like flow fluorescence detection method and system. Background technique [0002] Flow cytometry is an important cell analysis technology. Its core principle is to perform fluorescent staining on cells or detection units with fluorescent dyes. After the fluorescently stained cells are irradiated with light of a specific wavelength (incident light), they will Another specific wavelength of light (emission light) is emitted, and a photomultiplier tube (pmt) is used to collect the voltage pulse signal corresponding to the emission light, and the voltage pulse signal is used to express the fluorescence intensity of the cell at a specific wavelength. Then, collect the size and fluorescence intensity values ​​of a large number of cells, use the density function, and use the difference in the fluorescence intensity values ​​of negative cells and positive cells t...

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Problems solved by technology

[0006] In view of this, an embodiment of the present invention provides a wash-free image-like flow-type fluorescence detection method and system to solve the problem that if the fluorescence intensity value of the measured sample is lower than that of the fluorescence imaging system in the prior art When the lower limit of the sensitivity is set, it cannot be used for image flow analysis to obtain correct results

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