Single-molecule super-resolution imaging by tryptophan-quenching-induced photoswitching of phalloidin-fluorophore conjugates

Authors

  • Siddharth Nanguneri,

    1. Institute of Anatomy and Cell Biology, Heidelberg University, Heidelberg, Germany
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  • Benjamin Flottmann,

    1. Institute of Anatomy and Cell Biology, Heidelberg University, Heidelberg, Germany
    2. BioQuant Centre, Heidelberg University, Heidelberg, Germany
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  • Frank Herrmannsdörfer,

    1. Institute of Anatomy and Cell Biology, Heidelberg University, Heidelberg, Germany
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  • Thomas Kuner,

    1. Institute of Anatomy and Cell Biology, Heidelberg University, Heidelberg, Germany
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  • Mike Heilemann

    Corresponding author
    1. Institute of Anatomy and Cell Biology, Heidelberg University, Heidelberg, Germany
    2. BioQuant Centre, Heidelberg University, Heidelberg, Germany
    3. Institute of Physical & Theoretical Chemistry, Goethe-University Frankfurt, Frankfurt/Main, Germany
    • Correspondence to: Mike Heilemann; Institute of Physical & Theoretical Chemistry, Goethe-University Frankfurt, Max-von-Laue Str. 7, 60438 Frankfurt/Main, Germany. E-mail: heilemann@chemie.uni-frankfurt.de

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  • REVIEW EDITOR: Dr. Francesca Cella Zanacchi

ABSTRACT

Photophysical properties of any fluorophore are governed by the chemical nanoenvironment. In the context of imaging biological samples, this translates to different photophysical properties for different labels and probes. Here, we demonstrate that the nanoenvironment of fluorophores within a probe can be advantageously used to induce particular properties such as light-induced photoswitching. We demonstrate efficient photoswitching and single-molecule super-resolution imaging for various fluorophore-phalloidin conjugates in aqueous buffer without the addition of further chemicals. We further demonstrate the utility of two-color imaging of fluorophore-phalloidin and a photoactivatable fluorescent protein in presynaptic nerve terminals. Microsc. Res. Tech. 77:510–516, 2014. © 2014 Wiley Periodicals, Inc.

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