Sharper low-power STED nanoscopy by time gating

in: Nature Methods (2011)
Vicidomini, Giuseppe; Moneron, Gael; Han, Kyu Young; Westphal, Volker; Ta, Haisen; Reuss, Matthias ; Engelhardt, Johann ; Eggeling, Christian; Hell, Stefan W.
Applying pulsed excitation together with time-gated detection improves the fluorescence on-off contrast in continuous-wave stimulated emission depletion (CW-STED) microscopy, thus revealing finer details in fixed and living cells using moderate light intensities. This method also enables super-resolution fluorescence correlation spectroscopy with CW-STED beams, as demonstrated by quantifying the dynamics of labeled lipid molecules in the plasma membrane of living cells.

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