Ultrafast, temporally stochastic STED nanoscopy of millisecond dynamics

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Data Interpretation, Statistical
  • Equipment Design
  • Equipment Failure Analysis
  • Image Enhancement / instrumentation*
  • Micro-Electrical-Mechanical Systems / instrumentation*
  • Microscopy, Fluorescence / instrumentation*
  • Molecular Imaging / instrumentation*
  • Nanotechnology / instrumentation*
  • Photometry / instrumentation*
  • Stochastic Processes