Adaptive scans allow targeted cell-ablations on curved cell sheets
Résumé
Tissue morphogenesis proceeds through the mechanical actuation of cells by active forces from the cytoskeleton. Multicellular laser ablation has emerged as an essential means to probe these active forces by severing the tissue locally and inferring the tension born by the severed domain from the recoil of the surrounding structures. The realm of applications of laser ablations is however limited by the 2D steering of the laser in most instrumental configurations, while embryos and developing tissues are intrinsically 3D structures. Here, we present a flexible near infrared (NIR) fs-pulsed laser ablation system in which ablation trajectories proceed in 3D and adapt to the curved surface of cell sheets, which are prominent structures in embryos. Trajectories are computed through an unsupervised search for the surface of interest. We demonstrate that, depending on the exact experimental setup, the surface estimation can rely on a high content 3D imaging with a combined confocal microscope, or alternatively on a rapid Lissajou scan of the sample space with a NIR stand-alone setup. We apply the developed strategy mapping tensions in the developing Drosophila wing imaginal disc. These targeted, adaptive scans could be applied to other forms of non-linear processes such as two-photon fluorescence imaging or opto-genetics.
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