Curved light sheet microscopy for centimeter-scale cleared tissues imaging

Author:

Tang Lijuan,Wang Jiayu,Ding Jiayi,Sun Junyou,Chen Xing-jun,Shen Quqing,Song Ruiheng,Cao Peng,Gong Rong,Xu Fang,Ge Woo-ping,Sun Wenzhi,Zhao Hu,Wu JianglaiORCID

Abstract

Imaging large cleared tissues requires scaling the throughput of imaging techniques. Here we introduce curved light sheet microscope to perform optical sectioning with curved light sheets. This concept addresses the long-standing field curvature problem and lowers the barriers in designing high-throughput objectives. Leveraging a customized objective, the curved light sheet microscope achieves diffraction-limited resolution (1.0 μm laterally and 2.5 μm axially) and uniform contrast over a field of view of more than 1 × 1 cm2and is compatible with various tissue clearing techniques. Imaging an entire intact cleared mouse brain at 0.625 × 0.625 × 1.25 μm3voxel size takes less than 3 hours and image tiling is not required. We share full optics description of the objective and report imaging neuronal and vascular networks and tracing brain-wide long-distance axonal projections in intact mouse brains.

Publisher

Cold Spring Harbor Laboratory

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