Journal
OPTICS EXPRESS
Volume 17, Issue 17, Pages 14567-14573Publisher
OPTICAL SOC AMER
DOI: 10.1364/OE.17.014567
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We report sub-diffraction resolution in two-photon excitation (TPE) fluorescence microscopy achieved by merging this technique with stimulated-emission depletion (STED). We demonstrate an easy-to-implement and promising laser combination based on a short-pulse laser source for two-photon excitation and a continuous-wave (CW) laser source for resolution enhancement. Images of fluorescent nanoparticles and the immunostained transcription regulator NF kappa B in mammalian cell nuclei exhibit resolutions of <50 nm and similar to 70 nm in the focal plane, respectively, corresponding to a 4-5.4-fold improvement over the diffraction barrier. (C) 2009 Optical Society of America
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