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BP: Fachverband Biologische Physik
BP 8: Bioimaging and Spectroscopy I
BP 8.3: Talk
Monday, March 7, 2016, 12:15–12:30, H43
A novel membrane label for STED nanoscopy of living cardiomyocytes — •Elke Hebisch1, Stephan E. Lehnart2, and Stefan W. Hell1 — 1Max Planck Institute for Biophysical Chemistry, Department NanoBiophotonics, Am Fassberg 11, 37077 Goettingen, Germany — 2Research Unit for Cellular Biophysics and Translational Cardiology, Heart Research Center Goettingen, Robert-Koch-Str. 40, 37099 Goettingen, Germany
In heart muscle cells the fast and cell-wide propagation of rhythmic action potentials crucially depends on the architecture and composition of the plasma membrane (sarcolemma) and its extensive invaginations that form a transverse-axial tubular system (TATS). Here we report on the first application of the novel fluorescent membrane label cholesterol-PEG-KK114 (Chol-KK114) for STED nanoscopy of living mouse cardiomyocytes. Chol-KK114 enables fast and nontoxic in vivo labeling of cholesterol-rich cardiac membrane nanodomains. We could observe complex sarcolemmal and intracellular cholesterol signal patterns representing nanodomains sized far below the confocal resolution limit. These signal patterns are rich in detail and highly cell type specific since they could not be observed in HeLa or PtK2 cells. On the sarcolemma, we identified individual cholesterol-rich membrane nanodomains and higher order arrangements into ring structures and patches. Conclusively, we established a novel membrane label for superresolution microscopy of nanodomains in living primary cells.