Curved light sheet microscopy for centimeter-scale cleared tissues imaging

作者：Lijuan Tang, Jiayu Wang, Jiayi Ding, Junyou Sun, Xing-jun Chen, Quqing Shen, Ruiheng Song, Peng Cao, Rong Gong, Fang Xu, Woo‐Ping Ge, Wenzhi Sun, Hu Zhao, Jianglai Wu · 发表于：bioRxiv (Cold Spring Harbor Laboratory) · 年份：2024 · DOI：10.1101/2024.04.28.591483 · 被引用次数：9 · 研究领域：Advanced Fluorescence Microscopy Techniques、Cell Image Analysis Techniques、Optical Coherence Tomography Applications

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 cm 2 and is compatible with various tissue clearing techniques. Imaging an entire intact cleared mouse brain at 0.625 × 0.625 × 1.25 μm 3 voxel 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.