Coaligned dual-channel STED nanoscopy and molecular diffusion analysis at 20 nm resolution
Authors: Göttfert F, Wurm CA, Mueller V, Berning S, Cordes VC, Honigmann A, Hell SW
CellNetworks People: Hell Stefan
Journal: Biophys J. 2013 Jul 2;105(1):L01-3. doi: 10.1016/j.bpj.2013.05.029

We report on a fiber laser-based stimulated emission-depletion microscope providing down to ∼20 nm resolution in raw data images as well as 15-19 nm diameter probing areas in fluorescence correlation spectroscopy. Stimulated emission depletion pulses of nanosecond duration and 775 nm wavelength are used to silence two fluorophores simultaneously, ensuring offset-free colocalization analysis. The versatility of this superresolution method is exemplified by revealing the octameric arrangement of Xenopus nuclear pore complexes and by quantifying the diffusion of labeled lipid molecules in artificial and living cell membranes.