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Nat Methods. 2008 Nov;5(11):943-5. doi: 10.1038/nmeth.1257. Epub 2008 Sep 15.

Fluorescence nanoscopy by ground-state depletion and single-molecule return.

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  • 1Max Planck Institute for Biophysical Chemistry, Department of NanoBiophotonics, Am Fassberg 11, 37077 Göttingen, Germany.


We introduce far-field fluorescence nanoscopy with ordinary fluorophores based on switching the majority of them to a metastable dark state, such as the triplet, and calculating the position of those left or those that spontaneously returned to the ground state. Continuous widefield illumination by a single laser and a continuously operating camera yielded dual-color images of rhodamine- and fluorescent protein-labeled (living) samples, proving a simple yet powerful super-resolution approach.

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