Ultrafast, temporally stochastic STED nanoscopy of millisecond dynamics
Authors: Schneider J, Zahn J, Maglione M, Sigrist SJ, Marquard J, Chojnacki J, Kräusslich HG, Sahl SJ, Engelhardt J, Hell SW
CellNetworks People: Hell Stefan, Kräusslich Hans-Georg
Journal: Nat Methods. 2015 Sep;12(9):827-30. doi: 10.1038/nmeth.3481

Electro-optical scanning (>1,000 frames/s) with pixel dwell times on the order of the lifetime of the fluorescent molecular state renders stimulated emission depletion (STED) nanoscopy temporally stochastic. Photon detection from a molecule occurs stochastically in one of several scanning frames, and the spatial origin of the photon is known with subdiffraction precision. Images are built up by binning consecutive frames, making the time resolution freely adjustable. We demonstrated nanoscopy of vesicle motions in living Drosophila larvae and the cellular uptake of viral particles with 5- to 10-ms temporal resolution.