19 June 2003 Multiparameter single-molecule fluorescence measurements of DNA intercalating fluorophores
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Abstract
Experiments using single-molecules of TOTO-1 intercalated into dsDNA were performed to investigate the DNA sequence dependence on the fluorescence detectable with single-molecule fluorescence spectroscopy. Previous work has shown that there is a difference in the fluorescence lifetime when TOTO-1 is intercalated in poly-AT DNA or in poly-GC DNA. The fluorescence detected from single-molecules in this work for poly-GC and poly-AT DNA showed fluorescence lifetimes of 2.1 and 1.8 nsec, respectively. Analysis of the fluorescence intensity detected from single-molecules of TOTO-1 was performed by fluorescence cross-correlation spectroscopy. TOTO-1 is shown to spend large amounts of time in dark states. These dark states reduce the detectable fluorescence intensity to approximately 10 photons per millisecond on average.
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Benjamin P. Bowen, Benjamin P. Bowen, Jorg Enderlein, Jorg Enderlein, Neal W. T. Woodbury, Neal W. T. Woodbury, } "Multiparameter single-molecule fluorescence measurements of DNA intercalating fluorophores", Proc. SPIE 4962, Manipulation and Analysis of Biomolecules, Cells, and Tissues, (19 June 2003); doi: 10.1117/12.486348; https://doi.org/10.1117/12.486348
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