A three-dimensional super-resolution method based on Bessel film imaging

Abstract

The invention discloses a three-dimensional super-resolution method based on Bessel film imaging, comprising the following steps: acquiring a three-dimensional image of a sample, wherein the three-dimensional image is obtained by quickly scanning a biological sample by using a piezoelectric ceramic displacement stage to collect fluorescent molecules emitted by stimulation in the sample; performinga radial fluctuation based super-resolution analysis on the three-dimensional image, and generating a first super-resolution image by analyzing a high-order variation of the fluorescent molecule withtime; guided by the first super-resolution image, the three-dimensional image is subjected to three-dimensional Bayesian analysis based on the scintillation and bleaching characteristics of fluorescent molecules to obtain a final super-resolution image. The invention extends the traditional two-dimensional super-resolution radial fluctuation analysis and two-dimensional Bayesian analysis to threedimensions for the first time, and realizes three-dimensional super-resolution; and the invention organically combines two super-resolution algorithms to improve the operation speed of the super-resolution algorithm and the spatial resolution of the super-resolution image.

Description

technical field [0001] The invention relates to the technical field of microscopic imaging, and more specifically, to a three-dimensional super-resolution method based on Bessel light sheet imaging. Background technique [0002] Light sheet fluorescence microscopy is a cutting-edge optical microscopic imaging method in the 21st century, which is of great significance to modern life science research. Traditional wide-field microscopy and confocal microscopy need to irradiate or scan the entire biological sample, which has high phototoxicity and low signal-to-noise images with poor axial resolution. The light sheet microscopy technology uses an excitation light sheet with a thickness of micron to excite the fluorescence of biological samples, and detects the fluorescence signal of the sample in a direction perpendicular to the direction of the excitation light sheet to form a two-dimensional image. Light sheet microscopy only excites fluorescent molecules near the focal plane...

AI Technical Summary

Problems solved by technology

[0004] In view of the defects of the prior art, the purpose of the present invention is to solve the disadvantages of the two existing super-resolution algorithms, the super-resolution image obtained by the super-resolution radial fluctuation analysis has reconstruction artifacts, and the blinking and bleaching based on fluorescent molecules The data analysis process of the characteristic Bayesian analysis is longer

At the same time, the current super-resolution technology cannot meet the requirements of both large field of view and high resolution, and the three-dimensional super-resolution technology for deep tissue and large-volume biological samples is a major technical bottleneck.

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