Axial single-molecule high-accuracy location method and device

Abstract

The invention belongs to the technical field of super-resolution location fluorescence imaging and relates to an axial single-molecule high-accuracy fluorescence location method device and a use method thereof. According to the axial single-molecule high-accuracy fluorescence location method device disclosed by the invention, the high-accuracy characteristic of single-molecule location is combined, the characteristic that the slope of a single-molecule axial gauss light intensity distribution response curve is the maximum at the focal point position is utilized, and single-molecule axial accurate location is achieved; thus, super-resolution fluorescence imaging is achieved. The axial single-molecule high-accuracy fluorescence location method device disclosed by the invention comprises a laser A, a laser B, an objective lens, a three-dimensional sample displacement bench, an axial location system and the like, wherein the laser A and the laser B are used for stimulating and activating fluorescence molecules; the axial location system is composed of a beam splitter prism, a lens and an EMCCD; after fluorescence enters the axial location system, the fluorescence can be imaged at the EMCCD position after passing through the lens; the EMCCD position is symmetrical relative to the focus point.

Description

Technical field [0001] The invention belongs to the technical field of super-resolution positioning fluorescence imaging, and relates to a fluorescence positioning method device for axial single-molecule high-precision positioning and a use method thereof. Background technique [0002] In modern biomedical research, in order to better understand the process of life and the mechanism of disease, it is often necessary to observe the precise positioning and distribution of the organelles, viruses, parasites, etc. in the cell in the three-dimensional cell space. At the same time, protein science research also requires the clarification of the relationship between protein structure, location and function, and the temporal and spatial sequence of protein-protein interactions; biological macromolecules, mainly structural proteins and RNA and their complexes, constitute cells Basic structural system; how important active factors regulate the main life activities in cells, such as cell pr...

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Problems solved by technology

[0008] The purpose of the present invention is to solve the problem of low axial resolution of the single-molecule positioning system, and propose a method and device for accurately positioning the axial position of fluorescent molecules by using focal plane symmetric receiving information

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