High-speed light-sheet microscopy for the in-situ acquisition of volumetric histological images of living tissue

作者：Kripa Patel, Wenxuan Liang, Malte Casper, Venkatakaushik Voleti, Wenze Li, Alexis J. Yagielski, Hanzhi Zhao, Citlali Pérez Campos, Grace Sooyeon Lee, Joyce M. Liu, Elizabeth Philipone, Angela J. Yoon, Kenneth P. Olive, Shana M. Coley, Elizabeth M. C. Hillman · 发表于：Nature Biomedical Engineering · 年份：2022 · DOI：10.1038/s41551-022-00849-7 · 被引用次数：80 · 研究领域：Cell Image Analysis Techniques、Advanced Fluorescence Microscopy Techniques、Digital Holography and Microscopy

Histological examinations typically require the excision of tissue, followed by its fixation, slicing, staining, mounting and imaging, with timeframes ranging from minutes to days. This process may remove functional tissue, may miss abnormalities through under-sampling, prevents rapid decision-making, and increases costs. Here, we report the feasibility of microscopes based on swept confocally aligned planar excitation technology for the volumetric histological imaging of intact living tissue in real time. The systems' single-objective, light-sheet geometry and 3D imaging speeds enable roving image acquisition, which combined with 3D stitching permits the contiguous analysis of large tissue areas, as well as the dynamic assessment of tissue perfusion and function. Implemented in benchtop and miniaturized form factors, the microscopes also have high sensitivity, even for weak intrinsic fluorescence, allowing for the label-free imaging of diagnostically relevant histoarchitectural structures, as we show for pancreatic disease in living mice, for chronic kidney disease in fresh human kidney tissues, and for oral mucosa in a healthy volunteer. Miniaturized high-speed light-sheet microscopes for in-situ volumetric histological imaging may facilitate the point-of-care detection of diverse cellular-level biomarkers.