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Two-photon excitation STED microscopy

opg.optica.org · 103 words · saved by 1 readers

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 NFκB in mammalian cell nuclei exhibit resolutions of <50 nm and ~70 nm in the focal plane, respectively, corresponding to a 4–5.4-fold improvement over the diffraction barrier. ©2009 Optical Society of America Far-field fluorescence microscopy is by far the most commonly used microscopy technique for biological investigations due to the non-invasiveness of focused light and the high molecular specificity of fluorescent labeling. Among the noteworthy developments that have been made in this field during the past two decades, one can cite the imp

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