Dresden 2020 – wissenschaftliches Programm

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BP: Fachverband Biologische Physik

BP 20: Poster VIII

BP 20.14: Poster

Dienstag, 17. März 2020, 14:00–16:00, P2/4OG

Femtosecond laser cell surgery and wound healing on Drosophila embryos — •Elena Ramela Ciobotea, Ruby van Dijk, Bastian Zielinski, Mostafa Aakhte, Cristian Sarpe, Arne Senftleben, H.Arno J. Müller, and Thomas Baumert — Heinrich-Plett-Straße 40, 34132 Kassel, Germany

In this work we combine fluorescence microscopy and fs laser tissue ablation on a living Drosophila melanogaster embryo to investigate the wound closure in different stages of development. In contrast to typical larger scale UV laser damage, single cells or membranes can be targeted. Pulses from an 800nm Ti:Sapphire high energy oscillator were selected by the Pockels cell and coupled into a Leica confocal fluorescent microscope with a 63x oil immersion objective (NA: 1.32). 3h old embryos were dechorionated, washed, mounted in halocarbon oil between a microscope slide and a cover slip and imaged with a 488 nm Ar laser for up to 30 minutes after fs treatment. Images were assembled and analyzed using ImageJ software and a preliminary LabVIEW program. Firsts experiments were performed on embryos with cell nucleus marker and plasma membrane. Enhancement of the setup lead to well defined control of the energy deposition on the target tissue by varying exposure time and burst sequence with the Pockels cell, as well as optimizing temporal pulse length of the ultrashort laser pulses. Recent experiments on Myosin-GFP-labeled embryo with ablation in different stages show a larger migration of the myosin towards the damage over time for early stages in comparison with late stages of development.