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Issue 6, 2014
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Single-molecule photophysics of dark quenchers as non-fluorescent FRET acceptors

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Abstract

Dark quencher chromophores are interesting alternatives to common single-molecule FRET acceptors. Due to their short excited state lifetime, they should be less prone to complex photophysics and bleaching. We find, however, that for common enzymatic oxygen scavenging systems and photoprotection strategies – the gold standard of single-molecule measurements – the quenchers BBQ650 and BHQ-2 induce frequent blinking of the donor molecule. They switch in a photoinduced process to what we identify as a radical anion state and back. We further make use of the broad absorption spectrum for selective bleaching of the quenchers in order to photoactivate the fluorescence of initially completely quenched dye molecules. This represents a general strategy to turn fluorescent dyes into photoactivatable probes.

Graphical abstract: Single-molecule photophysics of dark quenchers as non-fluorescent FRET acceptors

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Publication details

The article was received on 08 Aug 2013, accepted on 22 Sep 2013 and first published on 23 Sep 2013


Article type: Communication
DOI: 10.1039/C3PP50274K
Citation: Photochem. Photobiol. Sci., 2014,13, 853-858
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    Single-molecule photophysics of dark quenchers as non-fluorescent FRET acceptors

    P. Holzmeister, B. Wünsch, A. Gietl and P. Tinnefeld, Photochem. Photobiol. Sci., 2014, 13, 853
    DOI: 10.1039/C3PP50274K

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