STED microscopy with a supercontinuum laser source.
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.
Department of NanoBiophotonics, Max Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Göttingen, Germany.
Dominik
D
Wildanger
Eva
E
Rittweger
Lars
L
Kastrup
Stefan W
SW
Hell
D004867:Equipment Design; D019544:Equipment Failure Analysis; D007834:Lasers; D008029:Lighting / Q000295:instrumentation*; D018613:Microscopy, Confocal / Q000295:instrumentation*; D008856:Microscopy, Fluorescence / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity