Issue 6, 2014

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

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

Article information

Article type
Communication
Submitted
08 Aug 2013
Accepted
22 Sep 2013
First published
23 Sep 2013

Photochem. Photobiol. Sci., 2014,13, 853-858

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

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Spotlight

Advertisements