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

Rapid 3D imaging of entire organs and organisms at cellular resolution is challenging. The authors present a computational light-sheet microscopy platform that combines dual-side confocally-scanned Bessel light-sheet illumination (thin, wide optical sectioning for deep tissues) with a content-aware compressed sensing (CACS) pipeline that improves contrast and resolution from a single acquisition. The method produces 3D images with high, isotropic resolution over two orders of magnitude faster than conventional 3D microscopy. They demonstrate whole mouse brain (~400 mm³) and entire gastrocnemius/tibialis muscles (~200 mm³) imaging at 5–10 minutes per sample with post-improved subcellular resolution of ~1.5 µm (0.5-µm iso-voxel). The approach enables system-level cellular analyses such as brain cell population mapping, whole-brain long-range neuron tracing, and whole-muscle neuromuscular junction occupancy quantification.

Chunyu Fang, Tingting Yu, Tingting Chu, Wenyang Feng, Fang Zhao, Xuechun Wang, Yujie Huang, Yusha Li, Peng Wan, Wei Mei, Dan Zhu, Peng Fei