Applied Physics B

, 93:725 | Cite as

Photoswitching microscopy with standard fluorophores

  • S. van de Linde
  • R. Kasper
  • M. Heilemann
  • M. Sauer
Open Access
Rapid communication

Abstract

We introduce far-field subdiffraction-resolution fluorescence imaging based on photoswitching of individual standard fluorophores in air-saturated solution. Here, photoswitching microscopy relies on the light-induced switching of organic fluorophores (ATTO 655 and ATTO 680) into long-lived metastable dark states and spontaneous repopulation of the fluorescent state. In the presence of low concentrations (2–10 mM) of reducing, thiol-containing compounds such as ß-mercaptoethylamine or glutathione, the density of fluorescent molecules can be adjusted to enable multiple localizations of individual fluorophores with an experimental accuracy of ∼20 nm. The method requires wide-field illumination with only a single laser beam for readout and photoswitching and provides superresolution fluorescence images of intracellular structures under live cell compatible conditions.

PACS

32.50.+d 61.05.-a 87.64.M- 87.80.Nj 

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Copyright information

© The Author(s) 2008

Authors and Affiliations

  • S. van de Linde
    • 1
  • R. Kasper
    • 1
  • M. Heilemann
    • 1
  • M. Sauer
    • 1
  1. 1.Applied Laser Physics and Laser Spectroscopy and Bielefeld Institute for Biophysics and NanoscienceBielefeld UniversityBielefeldGermany

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