Fluorescent staining method for target protein of DNA nano-structure labeled cell

Abstract

The invention belongs to the technical field of super-resolution microscopic imaging, and particularly relates to a fluorescent staining method for target protein of a DNA nano-structure labeled cell. Streptavidin is used as an intermediate, and a DNA nanostructure and an antibody of a biotin-modified target protein are connected together, so that the target protein is marked; and biotin, a group anchored on the hydrogel and more than two fluorescent dye molecules are simultaneously modified on the DNA nano structure. When the DNA nanostructure modifies the same dye, the brightness is very high, and the quality of the obtained microscopic picture can be greatly improved. When the dominant dye of the STED super-resolution microtechnique is modified, the expansion microtechnique can be well combined with the STED super-resolution technique. When a fluorescence resonance energy transfer dye pair is modified, the DNA nanostructure can have a flickering property, so that an expansion microtechnique is more conveniently combined with STORM and SOFI super-resolution techniques.

Description

technical field [0001] The invention belongs to the technical field of optical microscopy, and in particular relates to a fluorescent staining method for marking cell target proteins with DNA nanostructures. Background technique [0002] Expansion microscopy imaging technology is to specifically mark the target with fluorescence, such as protein, DNA, RNA, etc., and anchor the fluorescent group on the hydrogel. Fluorescent molecules that are indistinguishable from each other expand with the glue, increasing their distance from each other, allowing higher-resolution images to be acquired with common microscopy imaging equipment. Due to the simplicity of operation and readily available raw materials, expansion microscopy has a wide range of applications in single-celled animals, cell lines, tissues, and even animal body weight. [0003] The isotropic and uniform expansion of the sample is an important condition to ensure that the sample structure will not be distorted. To en...

AI Technical Summary

Problems solved by technology

This method uses streptavidin and biotin as the medium, and uses the advantages of DNA nanostructures with multiple terminals (free radicals) to modify multiple fluorescent dyes and the characteristics of not being digested by proteases to label the target protein, which can not only overcome In the existing expansion microscopy technology, the sample is digested by protease, which causes a large loss of sample fluorescence signal, and can greatly improve the brightness of the fluorescent label, so that the signal-to-noise ratio of the microscopic picture is higher and the picture quality is better.

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