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) 是一种广泛使用的技术，用于对透明组织和活体样本进行成像。然而，高性能 LSFM 系统通常价格昂贵且不易扩展。在这里，我们介绍了一种低成本、可扩展且多功能的 LSFM 框架，我们将其命名为“投影光片显微镜”(pLSM)，具有高成像性能以及较小的设备和计算占用空间。我们描述了 pLSM 的功能，通过对清除的小鼠大脑和死后病理人类进行高分辨率映射，重新利用现成的消费级组件、优化的光学器件、网络控制架构和软件驱动的光片调制。大脑样本，并通过来自人类诱导多能干细胞的大脑和血管类器官的分子表型分析。我们还报告了一种利用 pLSM 对空气-液体界面处稀疏标记的多层细菌菌膜生物膜的动态进行实时成像的方法。 pLSM 可以使生物医学应用的高分辨率 LSFM 更容易获得、负担得起且可扩展。