F_oF₁-ATP synthases are membrane-embedded protein machines that catalyze the synthesis of adenosine triphosphate. Using photoactivation-based localization microscopy (PALM) in TIR-illumination as well as structured illumination microscopy (SIM), we explore the spatial distribution and track single F_oF₁-ATP synthases in living E. coli cells under physiological conditions at different temperatures. For quantitative diffusion analysis by mean-squared-displacement measurements, the limited size of the observation area in the membrane with its significant membrane curvature has to be considered. Therefore, we applied a 'sliding observation window' approach (M. Renz et al., Proc. SPIE 8225, 2012) and obtained the one-dimensional diffusion coefficient of F_oF₁-ATP synthase diffusing on the long axis in living E. coli cells.

Date Published: 9 March 2015
PDF: 12 pages
Proc. SPIE 9331, Single Molecule Spectroscopy and Superresolution Imaging VIII, 93310D (9 March 2015); doi: 10.1117/12.2080981

Published in SPIE Proceedings Vol. 9331:
Single Molecule Spectroscopy and Superresolution Imaging VIII
Jörg Enderlein; Ingo Gregor; Zygmunt Karol Gryczynski; Rainer Erdmann; Felix Koberling, Editor(s)