Abstract
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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Acknowledgements
We thank A. Egner and A. Schönle for support with the instrumentation and data evaluation software and for valuable discussions, R.Y. Tsien (University of California, San Diego) for providing the plasmid coding for fluorescent proteins, V.N. Belov (Max Planck Institute for Biophysical Chemistry, Göttingen) for supplying the rhodamine dyes Rh-sart3b and Rh-sart3f, R. Schmitz-Salue and S. Löbermann for excellent technical assistance, S. Sahl for help with the experiments, and B. Rankin and J. Jethwa for carefully reading the manuscript.
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Fölling, J., Bossi, M., Bock, H. et al. Fluorescence nanoscopy by ground-state depletion and single-molecule return. Nat Methods 5, 943–945 (2008). https://doi.org/10.1038/nmeth.1257
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DOI: https://doi.org/10.1038/nmeth.1257
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