Fluorescence nanoscopy by ground-state depletion and single-molecule return

Nat Methods. 2008 Nov;5(11):943-5. doi: 10.1038/nmeth.1257. Epub 2008 Sep 15.


We introduce far-field fluorescence nanoscopy with ordinary fluorophores based on switching the majority of them to a metastable dark state, such as the triplet, and calculating the position of those left or those that spontaneously returned to the ground state. Continuous widefield illumination by a single laser and a continuously operating camera yielded dual-color images of rhodamine- and fluorescent protein-labeled (living) samples, proving a simple yet powerful super-resolution approach.

MeSH terms

  • Cell Line
  • Cell Survival
  • Humans
  • Microscopy, Fluorescence / methods*
  • Spectrometry, Fluorescence / methods*