Structured-illumination optical section fluorescence microscopic imaging method and device thereof

Abstract

The invention discloses a structured-illumination optical section fluorescence microscopic imaging method and a device thereof, and belongs to the technical field of optical imaging. The device comprises: a light source; a sample stage for carrying a fluorescent sample; a detection system for detecting the fluorescence emitted by the fluorescent sample; a polarizer for changing a light beam into radically polarized lights and a radial polarization converter, which are sequentially arranged between the light source and the sample stage along a light path; a phase mask plate for carrying out phase modulation on the radially polarized lights; a diffraction grating for diffracting the modulated light beam; a galvanometer for moving an illumination pattern on the fluorescent sample, wherein thelight beam emitted by the galvanometer is irradiated onto the fluorescent sample from the Y direction, and the detection system collects the fluorescence from the Z direction; and a processor for controlling the galvanometer to move the illumination pattern from the X, controlling the sample stage to move in the Z direction, and processing and reconstructing a light intensity distribution image collected by the detection system in order to generate a three-dimensional image of the sample. The light beam formed by the device and the method has a small main lube and a large view field.

Description

technical field [0001] The invention relates to the technical field of optical imaging, in particular to a method and device for structured illumination optical section fluorescence microscopic imaging. Background technique [0002] The rapid development of biotechnology in recent years requires us to have a deeper understanding of physiological processes, which also puts forward higher requirements for 3D real-time imaging technology. However, the development of 3D real-time imaging technology also faces many challenges—successful 3D real-time imaging technology needs to simultaneously achieve high spatial resolution, high imaging speed, good optical sectioning ability, low photodamage and photobleaching ability. These characteristics are often contradictory. For example, if you want to obtain an image with a higher spatial resolution, it may take more time, which sacrifices the imaging speed; and you need to add higher pump energy , which increases the photodamage and pho...

AI Technical Summary

Problems solved by technology

[0005] In order to improve the resolution of light-slicing fluorescence microscopy, researchers have made many attempts, combining super-resolution techniques such as stimulated emission depletion microscopy, structured light illumination, and single-molecule localization with light-slicing fluorescence microscopy. Combined with each other, trying to improve the resolving power of light sectioning fluorescence microscopy, but the results are not satisfactory

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