STED microscopy with a supercontinuum laser source

Opt Express. 2008 Jun 23;16(13):9614-21. doi: 10.1364/oe.16.009614.

Abstract

We report on a straightforward yet powerful implementation of stimulated emission depletion (STED) fluorescence microscopy providing subdiffraction resolution in the far-field. Utilizing the same super-continuum pulsed laser source both for excitation and STED, this implementation of STED microscopy avoids elaborate preparations of laser pulses and conveniently provides multicolor imaging. Operating at pulse repetition rates around 1 MHz, it also affords reduced photobleaching rates by allowing the fluorophore to relax from excitable metastable dark states involved in photodegradation. The imaging of dense nanoparticles and of the microtubular network of mammalian cells evidences a spatial resolution of 30-50 nm in the focal plane, i.e. by a factor of 8-9 beyond the diffraction barrier.

MeSH terms

  • Equipment Design*
  • Equipment Failure Analysis
  • Lasers*
  • Lighting / instrumentation*
  • Microscopy, Confocal / instrumentation*
  • Microscopy, Fluorescence / instrumentation*
  • Reproducibility of Results
  • Sensitivity and Specificity