Random optical reconstruction and structured light illumination composite super-resolution imaging system

Abstract

The invention discloses a random optical reconstruction and structured light illumination composite super-resolution imaging system, and the system comprises a light source module which is used for providing a plurality of combined illumination light rays with different wavelengths, controlling the illumination time sequence of the light rays, and controlling the illumination of a single-wavelength light ray, or the alternate illumination of a plurality of wavelength light rays, or the simultaneous illumination of a plurality of wavelength light rays; a composite light field regulation and control module which comprises a first optical regulation and control device used for regulating and controlling the incident light field into a cosine structure illumination light field and a second optical regulation and control device used for regulating and controlling the incident light field into a uniform illumination light field, wherein the two optical regulation and control devices can work independently, alternately or simultaneously; and a fluorescence imaging module which is used for collecting a plurality of original fluorescence images of the sample and reconstructing a super-resolution image by a computer. According to the invention, two super-resolution imaging technologies of random optical reconstruction and structured light illumination are combined on one set of optical imaging platform, and multi-mode and cross-resolution scale simultaneous imaging of a complex biological system can be realized.

Description

Technical field[0001]The present invention belongs to the field of microscopic imaging, particularly a random optical reconstruction and structural light illumination composite super-resolution imaging system.Background technique[0002]Fluorescence super-resolution imaging has a wide range of application prospects, there is already a variety of super-resolution imaging techniques, including random optical reconstruction super-resolution imaging techniques, structural light illumination super resolution imaging techniques, surreal radiation loss super resolution imaging techniques. While improving the resolution, these techniques are accompanied by the decrease in imaging speed, increased light power, increased light toxicity, and imaging.[0003]In the prior art, the technical principle of the random optical reconstruction super-resolution imaging method is to randomly activate a single molecule within the diffraction limit range from different times by optical switching fluorescent pr...

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Problems solved by technology

However, the imaging resolution of structured light illumination can be up to 2 times higher than that of traditional fluorescence microscopes, reaching the level of 100nm. It is difficult for fine biochemical reactions such as the movement of motor proteins along the cytoskeleton and the process of pathogen particles invading cells, which limits its application in ultramicrobiology, Applications in fields such as medicine

[0005] The demand for high-end imaging technology in the fields of life sciences and basic medicine covers ultra-high spatial resolution, fast imaging speed, low illumination power, low phototoxicity, and large imaging field of view. These requirements are difficult to reconcile from a technical perspective. Contradictions. At present, there is no single technical means that can meet these needs at the same time. The combination of different types of technologies is an important breakthrough direction to break through related difficulties.

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