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WANG Peng, ZHOU Yao, ZHAO Yu-xuan, FEI Peng. Double-ring-modulated light sheet fluorescence microscopic technique for multi-scale high-resolution 3D imaging[J]. Chinese Optics. doi: 10.37188/CO.2022-0093
Citation: WANG Peng, ZHOU Yao, ZHAO Yu-xuan, FEI Peng. Double-ring-modulated light sheet fluorescence microscopic technique for multi-scale high-resolution 3D imaging[J]. Chinese Optics. doi: 10.37188/CO.2022-0093

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

doi: 10.37188/CO.2022-0093
Funds:  Supported by National Key Research and Development Program of China (No. 2017YFA0700501); National Natural Science Foundation of China (No. 21874052, No. 21927802)
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  • Corresponding author: feipeng@hust.edu.cn
  • Received Date: 09 May 2022
  • Accepted Date: 07 Jul 2022
  • Rev Recd Date: 31 May 2022
  • Available Online: 03 Aug 2022
  • In this paper, we propose a non-diffraction Light Sheet Fluorescence Microscopy (LSFM) technique, 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-diffraction Bessel LSFM, we invent a double-ring-modulated approach which can generate non-diffraction light sheets with ~0.4 to ~5 µm 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 various biomedical studies, such as cell biology, tissue pathology, and neuroscience.


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