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Nat Methods. 2015 Sep;12(9):827-30. doi: 10.1038/nmeth.3481. Epub 2015 Jul 27.

Ultrafast, temporally stochastic STED nanoscopy of millisecond dynamics.

Author information

1
German Cancer Research Center (DKFZ), Optical Nanoscopy Division, Heidelberg, Germany.
2
Bioquant Center, Heidelberg, Germany.
3
Institute for Biology and Genetics, Free University of Berlin, Berlin, Germany.
4
Department of Infectious Diseases, Virology, University of Heidelberg, Heidelberg, Germany.
5
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.

Abstract

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.

PMID:
26214129
DOI:
10.1038/nmeth.3481
[Indexed for MEDLINE]

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