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Published September 2011 | Supplemental Material
Journal Article Open

Deep and fast live imaging with two-photon scanned light-sheet microscopy

Abstract

We implemented two-photon scanned light-sheet microscopy, combining nonlinear excitation with orthogonal illumination of light-sheet microscopy, and showed its excellent performance for in vivo, cellular-resolution, three-dimensional imaging of large biological samples. Live imaging of fruit fly and zebrafish embryos confirmed that the technique can be used to image up to twice deeper than with one-photon light-sheet microscopy and more than ten times faster than with point-scanning two-photon microscopy without compromising normal biology.

Additional Information

© 2011 Nature Publishing Group. Received 20 October 2010. Accepted 09 June 2011. Published online 17 July 2011. We thank P. Pantazis, W. Dempsey and L. Trinh for help with zebrafish sample preparations, and E. Beaurepaire for comments on the manuscript. This work was supported by grants to S.E.F. from the Caltech Beckman Institute and US National Institutes of Health (Center for Excellence in Genomic Science grant P50HG004071), and a fellowship to W.S. from the Caltech Biology Division. Author Contributions: T.V.T., W.S. and S.E.F. conceived and designed the project with consultation from D.S.K. and J.M.C.; T.V.T. designed and assembled the instrumentation; J.M.C. provided help with instrument software control. T.V.T. and W.S. imaged flies; T.V.T. imaged zebrafish; T.V.T. and D.S.K. imaged mice; W.S. and T.V.T. performed image reconstruction and analysis, and designed the figures; and T.V.T., W.S. and S.E.F. wrote the paper.

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August 22, 2023
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