Minutes-timescale 3D isotropic imaging of entire organs at subcellular resolution by content-aware compressed-sensing light-sheet microscopy

Chunyu Fang, Tingting Yu, Tingting Chu, Wenyang Feng, Fang Zhao, Xuechun Wang, Yujie Huang, Yusha Li, Peng Wan, Wei Mei (), Dan Zhu () and Peng Fei ()
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Chunyu Fang: Huazhong University of Science and Technology
Tingting Yu: Huazhong University of Science and Technology
Tingting Chu: Huazhong University of Science and Technology
Wenyang Feng: Huazhong University of Science and Technology
Fang Zhao: Huazhong University of Science and Technology
Xuechun Wang: Huazhong University of Science and Technology
Yujie Huang: Huazhong University of Science and Technology
Yusha Li: Huazhong University of Science and Technology
Peng Wan: Huazhong University of Science and Technology
Wei Mei: Huazhong University of Science and Technology
Dan Zhu: Huazhong University of Science and Technology
Peng Fei: Huazhong University of Science and Technology

Nature Communications, 2021, vol. 12, issue 1, 1-13

Abstract: Abstract Rapid 3D imaging of entire organs and organisms at cellular resolution is a recurring challenge in life science. Here we report on a computational light-sheet microscopy able to achieve minute-timescale high-resolution mapping of entire macro-scale organs. Through combining a dual-side confocally-scanned Bessel light-sheet illumination which provides thinner-and-wider optical sectioning of deep tissues, with a content-aware compressed sensing (CACS) computation pipeline which further improves the contrast and resolution based on a single acquisition, our approach yields 3D images with high, isotropic spatial resolution and rapid acquisition over two-order-of-magnitude faster than conventional 3D microscopy implementations. We demonstrate the imaging of whole brain (~400 mm3), entire gastrocnemius and tibialis muscles (~200 mm3) of mouse at ultra-high throughput of 5~10 min per sample and post-improved subcellular resolution of ~ 1.5 μm (0.5-μm iso-voxel size). Various system-level cellular analyses, such as mapping cell populations at different brain sub-regions, tracing long-distance projection neurons over the entire brain, and calculating neuromuscular junction occupancy across whole muscle, are also readily accomplished by our method.

Date: 2021
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DOI: 10.1038/s41467-020-20329-3

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