Chen Y, Chauhan S, Gong C, Dayton H, Xu C, De La Cruz ED, Tsai YW, Datta MS, Rosoklija GB, Dwork AJ, Mann JJ, Boldrini M, Leong KW, Dietrich LEP, Tomer R. Low-cost and scalable projected light-sheet microscopy for the high-resolution imaging of cleared tissue and living samples. Nat Biomed Eng 2024;8(9):1109-1123. doi: 10.1038/s41551-024-01249-9.

 

Background: Light sheet fluorescence microscopy (LSFM) enables high-resolution imaging of large, living or optically cleared samples with minimal photodamage. However, high-performance LSFM systems are often expensive, complex and difficult to scale for high-throughput applications like organoid screening or whole-organ imaging.

 

Hypothesis: This study tested the hypothesis that a cost-effective, modular LSFM system built from consumer-grade components and controlled over a network can match the imaging performance of high-end commercial LSFM platforms while offering greater scalability and accessibility.

 

Methods: The authors developed projected Light Sheet Microscopy (pLSM), a novel LSFM framework using pocket laser projectors for multi-color illumination, a consumer-grade CMOS camera and 3D-printed immersion chambers. The system is controlled remotely via a Python interface on a Nvidia Jetson Nano board, allowing software-driven modulation of light sheet thickness and field of view. Its performance was evaluated through imaging of cleared mouse and human brains, iPSC-derived organoids and live bacterial biofilms.

Results: pLSM achieves ~5 µm axial resolution using single-pixel light sheets and delivers image quality comparable to a high-end, CLARITY-optimized LSFM (COLM) system. It enables multi-color imaging of brain vasculature, high-throughput organoid phenotyping and reveales subcellular features and heterogeneity. Live imaging of Pseudomonas aeruginosa pellicle biofilms captured distinct structural layers and dynamic cell behaviors not previously resolved.

 

Conclusions: pLSM is a robust, low-cost and scalable alternative to traditional LSFM systems. Its modular hardware, remote operability and flexible imaging capabilities make it well-suited for high-throughput and diverse biomedical imaging applications.