Dresden 2017 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 2: Bioimaging and Spectroscopy I
BP 2.3: Vortrag
Montag, 20. März 2017, 10:15–10:30, HÜL 386
Live-cell super-resolution imaging of intrinsically fast moving flagellates — Marius Glogger1, Simone Stichler2, Ines Subota1, Sarah Bertlein2, Marie-Christin Spindler1, Jörg Teßmar2, Jürgen Groll2, Markus Engstler1, and •Susanne Fenz1 — 1Biocenter: Cell and Developmental Biology, University of Würzburg, Würzburg, Germany — 2Department of Functional Materials in Medicine and Dentistry, University of Würzburg, Würzburg, Germany
Recent developments in super-resolution microscopy make it possible to resolve structures in biological cells at a spatial resolution of a few nm. However, the optimal structural resolution requires repeated illumination cycles and is thus limited to chemically fixed cells. For live cell applications substantial improvement over classical Abbe-limited imaging can already be obtained in adherent or slow moving cells. Nonetheless, a large group of cells are fast moving and thus could not yet be addressed with live cell super-resolution microscopy. These include flagellate pathogens like African trypanosomes. Here, we present an embedding method based on an in situ forming cytocompatible UV-crosslinked hydrogel. The fast cross-linking hydrogel immobilizes trypanosomes efficiently to allow microscopy on the nanoscale. We characterized both the trypanosomes and the hydrogel with respect to their autofluorescence properties and found them suitable for single-molecule fluorescence microscopy (SMFM). As a proof of principle, SMFM was applied to super-resolve a structure inside the living trypanosome. We present an image of a flagellar axoneme component.