Multiparameter single-molecule fluorescence measurements of DNA intercalating fluorophores

Benjamin P. Bowen, Jörg Enderlein, Neal Woodbury

Research output: Chapter in Book/Report/Conference proceedingConference contribution

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.

Original languageEnglish (US)
Title of host publicationProceedings of SPIE - The International Society for Optical Engineering
EditorsD.V. Nicolau, J Enderlein, R.C. Leif, D.L. Farkas
Pages22-26
Number of pages5
Volume4962
DOIs
StatePublished - 2003
EventManipulation and Analysis of Biomolecules, Cells , and Tissues - San Jose, CA, United States
Duration: Jan 28 2003Jan 29 2003

Other

OtherManipulation and Analysis of Biomolecules, Cells , and Tissues
CountryUnited States
CitySan Jose, CA
Period1/28/031/29/03

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Keywords

  • DNA
  • Single-molecule fluorescence
  • Thiazole Orange
  • Time-correlated single photon counting
  • TOTO

ASJC Scopus subject areas

  • Electrical and Electronic Engineering
  • Condensed Matter Physics

Cite this

Bowen, B. P., Enderlein, J., & Woodbury, N. (2003). Multiparameter single-molecule fluorescence measurements of DNA intercalating fluorophores. In D. V. Nicolau, J. Enderlein, R. C. Leif, & D. L. Farkas (Eds.), Proceedings of SPIE - The International Society for Optical Engineering (Vol. 4962, pp. 22-26) https://doi.org/10.1117/12.486348