Berlin 2015 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 13: Posters: Imaging and Superresolution Optical Microscopy
BP 13.6: Poster
Montag, 16. März 2015, 17:30–19:30, Poster A
Embedding of flagellate Trypanosoma brucei for single-molecule microscopy — •Marius Glogger1, Simone Stichler2, 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
The uniflagellate protozoa Trypanosoma brucei are the causal agents of African sleeping sickness. High-resolution microscopy of trypanosoma in vivo is challenging due to the high motility of the parasite. Here, we present an approach for complete cell immobilization suitable for single-molecule fluorescence microscopy (SMM) techniques. Immobilization of trypanosomes requires both efficient trapping to prevent cell motility and mild embedding conditions to ensure cell viability. Biopolymers like agarose or gelatine were used before to immobilize cells sufficiently for fluorescence recovery after photobleaching studies in vivo. However, complete inhibition of the flagellar beat was not achieved. Hence, novel gels are needed for sophisticated SMM methods, which require a high spatial accuracy. We use modified hyaluronic acid and a crosslinking reagent to generate gels that solidify quickly upon UV-illumination. The rigidity of these hydrogel can be adjusted easily to guarantee complete immobilization of cellular movement. At the same time we verify cell viability using a fluorescent marker. We aim to transfer our results on trypanosomes to flagellates in general.