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The Contribution of Reactive Oxygen Species to the Photobleaching of Organic Fluorophores
Author(s) -
Zheng Qinsi,
Jockusch Steffen,
Zhou Zhou,
Blanchard Scott C.
Publication year - 2013
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/php.12204
Subject(s) - photobleaching , singlet oxygen , photochemistry , fluorophore , chemistry , radical , fluorescence , cyclooctatetraene , trolox , reactive oxygen species , oxygen , molecule , organic chemistry , biochemistry , physics , quantum mechanics , dpph , antioxidant
Photoexcitation of fluorophores commonly used for biological imaging applications generates reactive oxygen species ( ROS ) which can cause bleaching of the fluorophore and damage to the biological system under investigation. In this study, we show that singlet oxygen contributes relatively little to Cy5 and ATTO 647N photobleaching at low concentrations in aqueous solution. We also show that Cy5 generates significantly less ROS when covalently linked to the protective agents, cyclooctatetraene ( COT ), nitrobenzyl alcohol ( NBA ) or Trolox. Such fluorophores exhibit enhanced photostability both in bulk solutions and in single‐molecule fluorescence measurements. While the fluorophores ATTO 647N and ATTO 655 showed greater photostability than Cy5 and the protective–agent‐linked Cy5 derivatives investigated here, both of ATTO 647N and ATTO 655 generated singlet oxygen and hydroxyl radicals at relatively rapid rates, suggesting that they may be substantially more phototoxic than Cy5 and its derivatives.