Multiparameter single-molecule fluorescence measurements of DNA intercalating fluorophores

Benjamin P. Bowen; Jorg Enderlein; Neal W. T. Woodbury

doi:10.1117/12.486348

19 June 2003 Multiparameter single-molecule fluorescence measurements of DNA intercalating fluorophores

Benjamin P. Bowen, Jorg Enderlein, Neal W. T. Woodbury

Proceedings Volume 4962, Manipulation and Analysis of Biomolecules, Cells, and Tissues; (2003) https://doi.org/10.1117/12.486348
Event: Biomedical Optics, 2003, San Jose, CA, United States

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.

Citation Download Citation

Benjamin P. Bowen, Jorg Enderlein, and 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); https://doi.org/10.1117/12.486348

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