Hochauflösende Fluoreszenz-Messstation mit konfokaler Mikroskopie und TIRF-Mikroskopie

点击翻译按钮获取中文摘要

We apply for a fluorescence work-station for both TIRF- and confocal microscopy as part of our start-up funding. The main application of this work-station will be studies of the dynamics of the acto-myosin cytoskeleton, both on isolated protein complexes in vitro, but also in mammalian cell lines (e.g. HeLa and Cos-7) and in parasites. We will focus on the function of myosin-VI in different stages of clathrin-mediated endocytosis and on the role of myosin-XXI found in Leishmania parasites. We will study formation and localisation of motor protein complexes at high positional and time resolution on isolated single proteins in vitro and also in intact cells near the cell membrane in TIRF-mode. Dynamics of complex formation and motility deeper inside the cell will be studied using confocal microscopy with a fast resonant scanner. The resonant scanner will enable us to perform FRAP experiments, to study the dynamic distribution of protein complexes, and to carry out time resolved FRET studies for co-localisation of motor proteins and their binding partners. A spectral imaging detector combined with image analysis will help us to segregate mixed fluorescent signals and to improve signal-to-noise in detecting individual fluorophores in multi-colour fluorescence imaging.