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Quantum-Yield-Optimized Fluorophores for Site-Specific Labeling and Super-Resolution Imaging

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    Quantum-Yield-Optimized Fluorophores for Site-Specific Labeling and Super-Resolution Imaging
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    † Institute of Biochemistry, Goethe-University Frankfurt, Max-von-Laue-Str. 9, D-60438 Frankfurt/M., Germany
    ‡ University of Bordeaux, Interdisciplinary Institute for Neuroscience
    # CNRS UMR 5297, F-33000 Bordeaux, France
    § Laboratoire Photonique Numérique et Nanosciences, Université de Bordeaux, Institut d’Optique Graduate School and CNRS, 33405 Talence, France
    [email protected]
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    Journal of the American Chemical Society

    Cite this: J. Am. Chem. Soc. 2011, 133, 21, 8090–8093
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    https://doi.org/10.1021/ja200967z
    Published May 5, 2011
    Copyright © 2011 American Chemical Society
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    Abstract

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    Single-molecule applications, saturated pattern excitation microscopy, and stimulated emission depletion (STED) microscopy demand bright as well as highly stable fluorescent dyes. Here we describe the synthesis of quantum-yield-optimized fluorophores for reversible, site-specific labeling of proteins or macromolecular complexes. We used polyproline-II (PPII) helices as sufficiently rigid spacers with various lengths to improve the fluorescence signals of a set of different trisNTA–fluorophores. The improved quantum yields were demonstrated by steady-state and fluorescence lifetime analyses. As a proof of principle, we characterized the trisNTA–PPII–fluorophores with respect to in vivo protein labeling and super-resolution imaging at synapses of living neurons. The distribution of His-tagged AMPA receptors (GluA1) in spatially restricted synaptic clefts was imaged by confocal and STED microscopy. The comparison of fluorescence intensity profiles revealed the superior resolution of STED microscopy. These results highlight the advantages of biocompatible and, in particular, small and photostable trisNTA–PPII–fluorophores in super-resolution microscopy.

    Copyright © 2011 American Chemical Society

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    Detailed information regarding synthesis, spectroscopic characterization, specific protein binding, and in vivo visualization of trisNTA–Px–fluorophores in living neurons; complete ref 18. This material is available free of charge via the Internet at http://pubs.acs.org.

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    Journal of the American Chemical Society

    Cite this: J. Am. Chem. Soc. 2011, 133, 21, 8090–8093
    Click to copy citationCitation copied!
    https://doi.org/10.1021/ja200967z
    Published May 5, 2011
    Copyright © 2011 American Chemical Society
    Request reuse permissions

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