Abstract
Sub–diffraction-limit imaging can be achieved by sequential localization of photoactivatable fluorophores, for which the image resolution depends on the number of photons detected per localization. We report a strategy for fluorophore caging that creates photoactivatable probes with high photon yields. Upon photoactivation, these probes can provide 104−106 photons per localization and allow imaging of fixed samples with resolutions of several nanometers. This strategy can be applied to many fluorophores across the visible spectrum.
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Acknowledgements
We thank T. Mitchison, M. Ericsson and W. Wang for help during this project. This work is supported in part by the US National Institutes of Health and a Collaborative Innovation Award from the Howard Hughes Medical Institute (to X.Z.). J.C.V. is supported in part by a Burroughs-Wellcome Career Award at the Scientific Interface. X.Z. is a Howard Hughes Medical Institute investigator.
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J.C.V. is primarily responsible for experimental design. J.C.V. and S.J. performed experiments and analysis. X.Z. supervised and guided the project. J.C.V. and X.Z. wrote the manuscript.
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Vaughan, J., Jia, S. & Zhuang, X. Ultrabright photoactivatable fluorophores created by reductive caging. Nat Methods 9, 1181–1184 (2012). https://doi.org/10.1038/nmeth.2214
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DOI: https://doi.org/10.1038/nmeth.2214
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