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

In 2015, we launched the mesoSPIM initiative, 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 such microscopes. Here, we introduce the next-generation mesoSPIM ("Benchtop") with a significantly increased field of view, improved resolution, higher throughput, more affordable cost, and simpler assembly compared to the original version. We develop an optical method for testing detection objectives that enables us to select objectives optimal for light-sheet imaging with large-sensor cameras. The improved mesoSPIM achieves high spatial resolution (1.5 µm laterally, 3.3 µm axially) across the entire field of view, magnification up to 20x, and supports sample sizes ranging from sub-mm up to several centimeters while being compatible with multiple clearing techniques. The microscope serves a broad range of applications in neuroscience, developmental biology, pathology, and even physics.

Nikita Vladimirov, Fabian F. Voigt, Thomas Naert, Gabriela R. Araujo, Ruiyao Cai, Anna Maria Reuss, Shan Zhao, Patricia Schmid, Sven Hildebrand, Martina Schaettin, Dominik Groos, José María Mateos, Philipp Bethge, Taiyo Yamamoto, Valentino Aerne, Alard Roebroeck, Ali Ertürk, Adriano Aguzzi, Urs Ziegler, Esther Stoeckli, Laura Baudis, Soeren S. Lienkamp, Fritjof Helmchen