Fast in vivo imaging using multiphoton light-sheet microscopy at optimal pulse frequency

Vincent Maioli; Antoine Boniface; Pierre Mahou; Júlia Ferrer Ortas; Lamiae Abdeladim; Emmanuel Beaurepaire; Willy Supatto

doi:10.1117/12.2578867

5 March 2021 Fast in vivo imaging using multiphoton light-sheet microscopy at optimal pulse frequency

Vincent Maioli, Antoine Boniface, Pierre Mahou, Júlia Ferrer Ortas, Lamiae Abdeladim, Emmanuel Beaurepaire, Willy Supatto

Author Affiliations +

Proceedings Volume 11654, High-Speed Biomedical Imaging and Spectroscopy VI; 116540E (2021) https://doi.org/10.1117/12.2578867
Event: SPIE BiOS, 2021, Online Only

Abstract

Light-sheet illumination enables major increase in multiphoton imaging speed for in vivo studies. However, photoperturbation in multiphoton light-sheet microscopy remains poorly investigated. We show here that the heart beat rate of zebrafish embryos is a sensitive probe of linear and nonlinear photoperturbations. By analyzing its behavior with respect to laser power, pulse frequency and wavelength, we derive guidelines to balance signal and photoperturbation. We then demonstrate one order-of-magnitude signal enhancement over previous implementations by optimizing the laser pulse frequency. These results open new opportunities for fast in vivo imaging.

Conference Presentation

Citation Download Citation

Vincent Maioli, Antoine Boniface, Pierre Mahou, Júlia Ferrer Ortas, Lamiae Abdeladim, Emmanuel Beaurepaire, and Willy Supatto "Fast in vivo imaging using multiphoton light-sheet microscopy at optimal pulse frequency", Proc. SPIE 11654, High-Speed Biomedical Imaging and Spectroscopy VI, 116540E (5 March 2021); https://doi.org/10.1117/12.2578867

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