Double-ring-modulated light sheet fluorescence microscopic technique for multi-scale high-resolution 3D imaging

In this paper, we propose a non-diffraction Light Sheet Fluorescence Microscopy (LSFM) tech-nique, which readily enables multi-scale 3D fluorescence imaging of diverse biological samples with size ranging from microns to centimeters. To solve the problem of heavy sidelobes in conventional non -diffrac-tion Bessel LSFM, we invent a double-ring-modulated approach which can generate non-diffraction light sheets with-0.4 to-5 mu m tunable thickness and the ratio of the sidelobe was reduced to less than 30%. Then we built a multi-scale LSFM system based on this novel approach. The system showed versatile multi-scale imaging abilities, such as dual-color 3D dynamic imaging of single live cell, 3D super-resolution imaging of expansion cells and high-throughput 3D mapping of entire meso-scale organs. Therefore, we demonstrate that this multi-scale imaging modality can substantially improve the efficiency of LSFM for advancing vari-ous biomedical studies, such as cell biology, tissue pathology, and neuroscience.