A light-sheet illumination microscopic imaging method and system for simultaneous multi-layer imaging

Abstract

The invention discloses an optical plate lighting micro-imaging system for simultaneous multi-layer imaging. An optical plate lighting micro-imaging method comprises the steps that after light beam output by a laser light source is expanded, the light beam is irradiated to an intensity modulator to shape to generate multiple parallel lamellar light beams with different Z-axis positions; the lightbeam is projected into a sample cavity by an optical path relay system, and meanwhile, fluorescence signals of different layers of samples are excited; a point diffusion function of an imaging objective lens is elongated by the refractive index difference of a medium between the imaging objective lens and the samples, so that the fluorescence signals from the different layers can be collected simultaneously; a 3D image reconstruction algorithm is used for performing three-dimensional reconstruction on obtained projection images of the different layers of fluorescence signals, and finally, a real three-dimensional structure of the samples is restored. According to the optical plate lighting micro-imaging system for the simultaneous multi-layer imaging, mechanical scanning in the system is avoided by simultaneous multi-layer lighting and signals of different depths of imaging samples, and the 3D reconstruction algorithm of later images is utilized to improve the quantitative scale of time resolution in 3D imaging.

Description

technical field [0001] The invention relates to the technical field of microscopic imaging, and more particularly, to a method and system for simultaneous multi-layer imaging of light sheet illumination microscopic imaging. Background technique [0002] Light-sheet illumination microscopy utilizes a side-illuminated sheet-like beam to activate a sample to a preset depth, with micron-scale tomographic capabilities. Perpendicular to the direction of the light sheet, the wide-field imaging method is used to quickly obtain the three-dimensional space information of the sample. Since the light sheet illumination microscope only activates the layer to be imaged each time, it has less photobleaching and phototoxicity than the traditional point scanning imaging method, and is suitable for long-term imaging of living biological samples. [0003] However, traditional light-sheet illumination microscopes only generate a single light sheet at a time, and use mechanical scanning to axia...

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Problems solved by technology

[0004] Aiming at the defects of the prior art, the purpose of the present invention is to solve the technical problem that the three-dimensional imaging speed of the traditional light sheet illumination microscope is slow

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