Reduced Dyes Enhance Single-Molecule Localization Density for Live Superresolution Imaging

Authors

  • Lina Carlini,

    1. Laboratory of Experimental Biophysics, Institute of Physics of Biological Systems, École Polytechnique Fédérale de Lausanne (EPFL), National Centre of Competence in Research (NCCR) in Chemical Biology, Lausanne (Switzerland)
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  • Alexander Benke,

    1. Laboratory of Experimental Biophysics, Institute of Physics of Biological Systems, École Polytechnique Fédérale de Lausanne (EPFL), National Centre of Competence in Research (NCCR) in Chemical Biology, Lausanne (Switzerland)
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  • Dr. Luc Reymond,

    1. Institute of Chemical Sciences and Engineering, École Polytechnique Fédérale de Lausanne (EPFL), National Centre of Competence in Research (NCCR) in Chemical Biology, Lausanne (Switzerland)
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  • Dr. Gražvydas Lukinavičius,

    1. Institute of Chemical Sciences and Engineering, École Polytechnique Fédérale de Lausanne (EPFL), National Centre of Competence in Research (NCCR) in Chemical Biology, Lausanne (Switzerland)
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  • Prof. Suliana Manley

    Corresponding author
    1. Laboratory of Experimental Biophysics, Institute of Physics of Biological Systems, École Polytechnique Fédérale de Lausanne (EPFL), National Centre of Competence in Research (NCCR) in Chemical Biology, Lausanne (Switzerland)
    • Laboratory of Experimental Biophysics, Institute of Physics of Biological Systems, École Polytechnique Fédérale de Lausanne (EPFL), National Centre of Competence in Research (NCCR) in Chemical Biology, Lausanne (Switzerland)

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Abstract

Cell-permeable rhodamine dyes are reductively quenched by NaBH4 into a non-fluorescent leuco-rhodamine form. Quenching is reversible, and their fluorescence is recovered when the dyes are oxidized. In living cells, oxidation occurs spontaneously, and can result in up to ten-fold higher densities of single molecule localizations, and more photons per localization as compared with unmodified dyes. These two parameters directly impact the achievable resolution, and we see a significant improvement in the quality of live-cell point-localization super-resolution images taken with reduced dyes. These improvements carry over to increase the density of trajectories for single-molecule tracking experiments.

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