Simultaneous multi-lifetime multi-color STED imaging for colocalization analyses

Opt Express. 2011 Feb 14;19(4):3130-43. doi: 10.1364/OE.19.003130.


We describe a STED microscope optimized for colocalization experiments with up to three colors. Two fluorescence labels are separated by their fluorescence lifetime whereas a third channel is discriminated by the wavelength of fluorescence emission. Since it does not require a second STED beam, separating by lifetime is insensitive to drift and thus optimally suited for colocalization analyses. Furthermore, we propose a setup having a second STED beam for long duration multicolor recording.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Color
  • Imaging, Three-Dimensional / methods*
  • Lamins / metabolism
  • Microscopy, Fluorescence / methods*
  • Time Factors
  • Tubulin / metabolism


  • Lamins
  • Tubulin