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Photobleaching of asymmetric cyanines used for fluorescence imaging of single DNA molecules

Lookup NU author(s): Dr Eimer TuiteORCiD

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Abstract

The photobleaching of the cyanine dyes YO and YOYO has been investigated for both free and DNA-bound dyes, using absorption and fluorescence spectroscopy coupled with fluorescence microscopy. For the free dyes, the nature of the reactive species involved in the photodegradation process is different for the monomer and the dimer, as shown by scavenger studies. For DNA-bound dyes, photoinduced fading of the visible absorption band occurs by different pathways depending on the drug binding mode and can be attenuated by appropriate scavengers. However, none of these scavengers were found to have any significant effect on the photobleaching of dye fluorescence. It appears that the reduction of fluorescence intensity comes from a quenching of the dye fluorescence by modified DNA bases, possibly 8-oxo-7,8-dihydro-2 ' -deoxyguanosine.

Publication metadata

Author(s): Tuite E; Kanony C; Akerman B

Publication type: Article

Publication status: Published

Journal: Journal of the American Chemical Society

Year: 2001

Volume: 123

Issue: 33

Pages: 7985-7995

ISSN (print): 0002-7863

ISSN (electronic): 1520-5126

Publisher: American Chemical Society

URL: http://dx.doi.org/10.1021/ja001047n

DOI: 10.1021/ja001047n

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