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

Vladimirov, N.; Voigt, F.; Naert, T.; Araujo, G. R.; Cai, R.; Reuss, A. M.; Zhao, S.; Schmid, P.; Hildebrand, S.; Schaettin, M.; Groos, D.; Mateos, J. M.; Bethge, P.; Yamamoto, T.; Aerne, V.; Roebroeck, A.; Ertürk, A.; Aguzzi, A.; Ziegler, U.; Stoeckli, E. T.; Baudis, L.; Lienkamp, S. S.; Helmchen, F.

doi:10.1101/2023.06.16.545256

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bioRxiv · 10.1101/2023.06.16.545256

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

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Abstract

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 {micro}m laterally, 3.3 {micro}m 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.

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Vladimirov, N., Voigt, F., Naert, T., Araujo, G. R., Cai, R., Reuss, A. M., Zhao, S., Schmid, P., Hildebrand, S., Schaettin, M., Groos, D., Mateos, J. M., Bethge, P., Yamamoto, T., Aerne, V., Roebroeck, A., Ertürk, A., Aguzzi, A., Ziegler, U., Stoeckli, E. T., Baudis, L., Lienkamp, S. S., Helmchen, F.. 2023-06-21. The Benchtop mesoSPIM: a next-generation open-source light-sheet microscope for large cleared samples. https://doi.org/10.1101/2023.06.16.545256

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