Nanoscopy using localization and temporal separation of fluorescence from single molecules

Abstract

Since the development of the first techniques [1, 2] that bypass the diffraction limit postulated by Ernst Abbe in 1873, so called superresolution microscopies have spread rapidly diverging into different variants that are capable of resolving details smaller than 200 nm. This article focuses on those based on the subsequent localization of single molecules. Therein, most molecules are prepared in a non-fluorescent dark state, leaving only a few single molecules fluorescing, which can be localized by fitting a two dimensional Gaussian function to their respective point spread functions (PSF). © 2011 Springer Science+Business Media B.V.