Balancing signal and photoperturbation in multiphoton light-sheet microscopy by optimizing laser pulse frequency [article]

Vincent Maioli, Antoine Boniface, Pierre Mahou, Júlia Ferrer Ortas, Lamiae Abdeladim, Emmanuel Beaurepaire, Willy Supatto
2020 biorxiv/medrxiv   pre-print
Improving the imaging speed of multiphoton microscopy is an active research field. Among recent strategies, light-sheet illumination holds distinctive advantages for achieving fast imaging in vivo. 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
more » ... quency 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 live tissue imaging.
doi:10.1101/2020.06.02.130377 fatcat:mu7ildtxfzdo5cdl3qfdxwuttq