title: Superresolution microscopy on the basis of engineered dark states.
abstract: New concepts for superresolution fluorescence microscopy by subsequent localization of single molecules using photoswitchable or photoactivatable fluorophores are rapidly emerging and provide new ways to resolve structures beyond the diffraction limit. Here, we demonstrate that superresolution imaging can be carried out with practically every single-molecule compatible, synthetic fluorophore by controlling their emission properties. We prepare dark states by removing oxygen that extends the triplet state lifetime to several milliseconds. We further increase the duration of the off-states using electron transfer reactions to create radical ion states of severalfold longer lifetimes. Imaging single molecules, actin filaments, and microtubules in fixed cells as well as simulations demonstrate that the thus created dark states are sufficiently long for resolution of approximately 50 nm.
authors.affiliation: Angewandte Physik-Biophysik, and Center for NanoScience, Ludwig-Maximilians-Universitat, Amalienstrasse 54, 80799 Munchen, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Steinhauer
authors.forename: Carsten
authors.initials: C
authors.lastname: Forthmann
authors.forename: Jan
authors.initials: J
authors.lastname: Vogelsang
authors.forename: Philip
authors.initials: P
authors.lastname: Tinnefeld
mesh_terms: D000199:Actins / Q000737:chemistry; D008856:Microscopy, Fluorescence / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D000199:Actins