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A versatile tiling light sheet microscope for cleared tissue imaging

By Yanlu Chen, Xiaoliang Li, Dongdong Zhang, Chunhui Wang, Ruili Feng, Xuzhao Li, Yao Wen, Hao Xu, Xinyi Shirley Zhang, Xiao Yang, Yongyi Chen, Yi Feng, Bo Zhou, Bi-Chang Chen, Kai Lei, Shang Cai, Jie-Min Jia, Liang Gao

Posted 04 Nov 2019
bioRxiv DOI: 10.1101/829267

We present a tiling light sheet microscope compatible with all tissue clearing methods for rapid multicolor 3D imaging of centimeter-scale cleared tissues with micron-scale (4x4x10 micron) to nanometer-scale (70x70x200 nm) spatial resolution. The microscope uses tiling light sheets to achieve a more advanced multicolor 3D imaging ability than conventional light sheet microscopes. The illumination light is phase modulated to adjust the 3D imaging ability of the microscope based on the tissue size and the desired spatial resolution and imaging speed, and also to keep the microscope aligned. The ability of the microscope to align via phase modulation alone also makes the microscope reliable and easy to operate. Here we describe the working principle and design of the microscope. We demonstrate its imaging ability by imaging various cleared tissues prepared by different tissue clearing and tissue expansion techniques. ### Competing Interest Statement A patent regarding the presented microscope was filed by the Westlake University on behalf of L.G..

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