Multiphoton light-sheet microscopy using wavelength mixing: fast multicolor imaging of the beating Zebrafish heart with low photobleaching

Pierre Mahou; Julien Vermot; Emmanuel Beaurepaire; Willy Supatto

doi:10.1117/12.2079176

5 March 2015 Multiphoton light-sheet microscopy using wavelength mixing: fast multicolor imaging of the beating Zebrafish heart with low photobleaching

Pierre Mahou, Julien Vermot, Emmanuel Beaurepaire, Willy Supatto

Author Affiliations +

Proceedings Volume 9329, Multiphoton Microscopy in the Biomedical Sciences XV; 93290Z (2015) https://doi.org/10.1117/12.2079176
Event: SPIE BiOS, 2015, San Francisco, California, United States

Abstract

Two-photon laser scanning microscopy has become a standard to map thick and live tissues. However, its application for fast and multicolor imaging remains challenging. To address this issue, we report on the implementation of mixed wavelength excitation in a two-photon light-sheet microscope. We illustrate the potential of the technique by recording sustained multicolor two-photon movies of the beating heart in zebrafish embryos with negligible photobleaching at 28 million pixels/second. In particular, 3D reconstructions of the heart periodic motion are obtained with sufficient spatiotemporal resolution to track the fast movements of individual cells during a cardiac cycle.

Citation Download Citation

Pierre Mahou, Julien Vermot, Emmanuel Beaurepaire, and Willy Supatto "Multiphoton light-sheet microscopy using wavelength mixing: fast multicolor imaging of the beating Zebrafish heart with low photobleaching", Proc. SPIE 9329, Multiphoton Microscopy in the Biomedical Sciences XV, 93290Z (5 March 2015); https://doi.org/10.1117/12.2079176

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