High-speed large-view-field digital scanning light sheet microimaging system

Abstract

The invention discloses a high-speed large-view-field digital scanning light sheet microscopic imaging system. The system comprises a light source module, a transverse view field expansion module, a scanning module, an illumination module and a detection module which are sequentially arranged according to a light path, light generated by the light source module passes through the transverse view field expansion module to obtain an effective view field for expanding a final illumination area. The scanning module scans a line beam received from the transverse view field expansion module into a surface beam, the illumination module performs fluorescence excitation by adopting a beam splitter, a first illumination objective lens and a second illumination objective lens, and the detection module realizes fluorescence detection imaging by adopting a line detector. The axial resolution is improved, the influence of phototoxicity and photobleaching on a sample is reduced, the imaging has higher resolution and contrast ratio by using the linear array detector, the imaging speed is higher, and the cost of the detector is saved.

Description

technical field [0001] The invention relates to the technical field of optical microscopic imaging, and specifically relates to a high-speed large-field digital scanning light-sheet microscopic imaging system. Background technique [0002] In optical microscopes, the resolution of the objective lens is limited to the cellular level (about 1 micron), and the diameter of the field of view is basically within 1 mm. In order to maintain the resolution while improving the imaging field of view, some related methods have been proposed. The most common and simple method is to fix the sample on a two-dimensional translation stage, and after imaging a specific area in a small field of view, move the sample, perform multiple imaging, and stitch multiple images to obtain a large field of view image. This method is slow, cannot be imaged in real time, and there are errors in field of view stitching and stacking of image edges is required, resulting in image distortion near the stitching...

AI Technical Summary

Problems solved by technology

The problem brought about by this is that the axial resolution is generally low, and the phototoxicity and photobleaching of the sample are obvious, which is not conducive to long-term imaging of the sample

[0007] Therefore, the main defects of the existing large-field fluorescence microscopes are: coaxial illumination and detection are used, the axial resolution is limited, the photobleaching and phototoxicity effects on the sample are obvious, the images in the mosaic imaging method are distorted, and for large-field Field surface detectors cannot balance high imaging speed with low cost

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