The Benchtop mesoSPIM: a next-generation open-source light-sheet microscope for large cleared samples

In 2015, we launched the mesoSPIM initiative (www.mesospim.org), an open-source project for making light-sheet microscopy of large cleared tissues more accessible. Meanwhile, the demand for imaging larger samples at higher speed and resolution has increased, requiring major improvements in the capabilities of light-sheet microscopy. Here, we introduce the next-generation mesoSPIM ("Benchtop") with significantly increased field of view, improved resolution, higher throughput, more affordable cost and simpler assembly compared to the original version. We developed a new method for testing objectives, enabling us to select detection objectives optimal for light-sheet imaging with large-sensor sCMOS cameras. The new mesoSPIM achieves high spatial resolution (1.5 µm laterally, 3.3 µm axially) across the entire field of view, a magnification up to 20x, and supports sample sizes ranging from sub-mm up to several centimetres, while being compatible with multiple clearing techniques. The new microscope serves a broad range of applications in neuroscience, developmental biology, and even physics.

2015年，我们启动了mesoSPIM计划（www.mesospim.org），这是一个旨在提升大型透明组织光片显微镜成像可及性的开源项目。与此同时，学界对以更高速度与分辨率成像更大尺寸样本的需求与日俱增，这要求光片显微镜的性能实现重大升级。在此基础上，我们推出了新一代mesoSPIM“台式型”设备，相较于初代版本，其视场范围显著扩大、分辨率得以优化、通量更高、成本更为亲民且组装流程更为简便。我们开发了一种全新的物镜测试方法，能够筛选出适配大传感器sCMOS（sCMOS）相机的光片成像最优检测物镜。这款新一代mesoSPIM可在整个视场内实现高空间分辨率（横向1.5 µm，轴向3.3 µm），最大放大倍率可达20倍，支持从亚毫米至数厘米的样本尺寸，同时兼容多种组织透明化技术。该新型显微镜可广泛应用于神经科学、发育生物学乃至物理学等诸多研究领域。