In vivo analysis of Drosophila embryo developmental dynamics by femtosecond pulse-induced ablation and multimodal nonlinear microscopy

Animal embryo development exhibits a complex ensemble of cell movements that are tightly regulated by developmental gene expression. It was proposed recently that mechanical factors may also play an important role during development. Investigating these dynamical processes is technically challenging and requires novel in vivo investigation methods. We show that multiphoton microscopy can be used for both perturbing and analyzing morphogenetic movements in vivo, (i) nonlinear microscopy is well adapted for the sustained imaging of early Drosophila embryos despite their highly scattering nature; (ii) femtosecond pulse-induced ablation can be used to process specific tissues in vivo. Combining this approach with multimodal microscopy (two-photon-excited fluorescence (2PEF) and third-harmonic generation (THG)), we report the successful quantitative modulation of morphogenetic movements in vivo. Our data provides insight to the issue of morphogenesis regulation.