Department of Biological Sciences, Columbia University, New York, NY, USA.
Department of Biomedical Engineering, Columbia University, New York, NY, USA.
Nat Biomed Eng. 2024 Sep;8(9):1109-1123. doi: 10.1038/s41551-024-01249-9. Epub 2024 Aug 29.
Light-sheet fluorescence microscopy (LSFM) is a widely used technique for imaging cleared tissue and living samples. However, high-performance LSFM systems are typically expensive and not easily scalable. Here we introduce a low-cost, scalable and versatile LSFM framework, which we named 'projected light-sheet microscopy' (pLSM), with high imaging performance and small device and computational footprints. We characterized the capabilities of pLSM, which repurposes readily available consumer-grade components, optimized optics, over-network control architecture and software-driven light-sheet modulation, by performing high-resolution mapping of cleared mouse brains and of post-mortem pathological human brain samples, and via the molecular phenotyping of brain and blood-vessel organoids derived from human induced pluripotent stem cells. We also report a method that leverages pLSM for the live imaging of the dynamics of sparsely labelled multi-layered bacterial pellicle biofilms at an air-liquid interface. pLSM can make high-resolution LSFM for biomedical applications more accessible, affordable and scalable.
光片荧光显微镜（LSFM）是一种广泛用于成像透明组织和活样本的技术。然而，高性能的 LSFM 系统通常价格昂贵且不易扩展。在这里，我们引入了一种低成本、可扩展且多功能的 LSFM 框架，我们称之为“投影光片显微镜”（pLSM），它具有高成像性能和小的设备和计算足迹。我们通过对清除的小鼠大脑和死后病理性人脑样本进行高分辨率映射，以及通过对源自人类诱导多能干细胞的脑和血管类器官进行分子表型分析，对 pLSM 进行了表征，该系统重新利用了现成的消费级组件、优化的光学器件、基于网络的控制架构和软件驱动的光片调制。我们还报告了一种利用 pLSM 对空气-液体界面上稀疏标记的多层细菌菌膜生物膜的动力学进行活细胞成像的方法。pLSM 可以使用于生物医学应用的高分辨率 LSFM 更容易获得、更经济实惠和更具扩展性。
