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BP: Fachverband Biologische Physik

BP 34: New Technologies

BP 34.1: Hauptvortrag

Freitag, 18. März 2011, 10:15–10:45, ZEU 260

Super-resolution fluorescence imaging of cellular structure and dynamics — •Markus Sauer, Sebastian van de Linde, Teresa Klein, Anna Löschberger, Thorge Holm, and Sven Proppert — Biotechnology and Biophysics, Julius-Maximilians-University Würzburg, Germany

We introduce direct stochastic optical reconstruction microscopy (dSTORM), a new general approach for multicolor super-resolution fluorescence imaging based on reversible photoswitching of standard small organic fluorophores. Photoswitching of organic rhodamine and oxazine fluorophores, i.e. the reversible transition from a fluorescent to a non-fluorescent state in aqueous buffers exploits the formation of long-lived radical anions through reaction with thiol compounds and repopulation of the singlet ground state by reaction with molecular oxygen. We unravel the underlying switching mechanism, investigate the importance of labeling strategies and densities, and demonstrate super-resolution imaging with different commercially available organic fluorophores with high spatial and temporal resolution. Finally, we demonstrate that dSTORM in combination with suited chemical tags can be advantageously used for dynamic live cell imaging at resolutions ~ 20 nm.