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STED nanoscopy in living cells using Fluorogen Activating Proteins.

James A J Fitzpatrick, Qi Yan, Jochen J Sieber, Marcus Dyba, Ulf Schwarz, Chris Szent-Gyorgyi, Carol A Woolford, Peter B Berget, Alan S Waggoner, Marcel P Bruchez
Bioconjugate Chemistry 2009
We demonstrate the effectiveness of a genetically encoded Malachite Green (MG) binding fluorogen activating protein (FAP) for live cell stimulated emission depletion nanoscopy (STED). Both extracellular and intracellular FAPs were tested in living cells using fluorogens with either membrane expressed FAP or as an intracellular FAP-actin fusion. Structures with FWHM of 110-122nm were observed. Depletion data however suggests a resolution of 70nm with the given instrument.

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