Non-wash image flow fluorescence detection method and system thereof

Abstract

The invention provides a non-wash image flow cytometry fluorescence detection method and a system thereof. The method includes the following steps: dyeing a target sample with a preset target fluorescent dye through a dyeing technology, and ensuring the formation of a certain free fluorescent dye after dyeing and incubation to make all cells in the target sample acquire the fluorescence intensity value used for density function clustering resolution analysis under the non-wash flow fluorescence detection system; acquiring the microscopic fluorescence image of the dyed target sample based on a microscopic imaging system; extracting the gray value and the pixel value of the target sample by adopting a computer image recognition technology; and calculating the fluorescence intensity value of the target sample based on the gray value and the pixel value of the target sample, and outputting the microscopic fluorescence image of the target sample and the density function clustering analysis result. A problem that the detected fluorescence intensity value of undyed or dyed negative cells is zero or obviously sharps declines under the image flow fluorescence detection system after common dyeing washing is solved in the invention.

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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