Super-resolution microscope method and device based on random fluorescence bleaching

Abstract

The invention discloses a super-resolution microscope method based on random fluorescence bleaching. The super-resolution microscope method comprises the following steps of: 1, focusing a laser beam on a to-be-detected sample with a fluorescence mark, exciting the to-be-detected sample by the laser beam to emit fluorescent; 2, collecting the fluorescent emitted by the to-be-detected sample to obtain fluorescent strength information; 3, arranging an area-of-interest on the to-be-detected sample, bleaching the area-of-interest, collecting the fluorescent strength information of the fluorescent emitted from the to-be-detected sample in a bleaching process; and 4, comparing and analyzing the fluorescent strength information by a computer to obtain position information of a fluorescent molecule in the area-of-interest, and obtaining a super-resolution image by using a restructing algorithm. The invention also discloses a super-resolution microscope device based on random fluorescence bleaching. The super-resolution microscope device has the advantages of simple structure, high transverse resolution and high signal to noise ratio.

Description

technical field [0001] The invention belongs to the field of optical super-resolution microscopy, in particular to a super-resolution microscopy method and device based on random fluorescence bleaching. Background technique [0002] With the development of science and technology, people are constantly pursuing smaller and smaller size structures and higher resolution capabilities, especially in the fields of microelectronics, aerospace, nanoprocessing, life sciences and materials engineering. demands are becoming increasingly urgent. [0003] Although electron microscopes, atomic force microscopes, scanning electron microscopes, and near-field scanning microscopes have high resolving power, their high requirements for samples or operating environments and their dependence on probes greatly limit their application range. Far-field optical microscopy has the characteristics of no contact and less damage to the sample, but according to Abbe's imaging theory, its resolution is ...

AI Technical Summary

Problems solved by technology

[0005] STED is one of the mainstream super-resolution methods at present, but this super-resolution method needs to integrate new modules or devices on the basis of traditional confocal microscopes, and these modules or devices are often more expensive

PALM and STORM use the randomness of fluorescent molecular light emission to achieve super-resolution by sparsely activating a small number of fluorescent molecules to emit light each time and then position fitting. The microscopic method based on single-molecule localization is the highest resolution among far-field optical super-resolution methods. , but both PALM and STORM need to be equipped with special fluorescent dyes and cannot be directly applied to common fluorescent dyes

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