Multiparameter single-molecule fluorescence measurements of DNA intercalating fluorophores

Benjamin P. Bowen; Jörg Enderlein; Neal Woodbury

doi:10.1117/12.486348

Multiparameter single-molecule fluorescence measurements of DNA intercalating fluorophores

Benjamin P. Bowen, Jörg Enderlein, Neal Woodbury

Research output: Chapter in Book/Report/Conference proceeding › Conference 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 language	English (US)
Title of host publication	Proceedings of SPIE - The International Society for Optical Engineering
Editors	D.V. Nicolau, J Enderlein, R.C. Leif, D.L. Farkas
Pages	22-26
Number of pages	5
Volume	4962
DOIs	https://doi.org/10.1117/12.486348
State	Published - 2003
Event	Manipulation and Analysis of Biomolecules, Cells , and Tissues - San Jose, CA, United States Duration: Jan 28 2003 → Jan 29 2003

Other

Other	Manipulation and Analysis of Biomolecules, Cells , and Tissues
Country/Territory	United States
City	San Jose, CA
Period	1/28/03 → 1/29/03

Keywords

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

ASJC Scopus subject areas

Electrical and Electronic Engineering
Condensed Matter Physics

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10.1117/12.486348

Cite this

Multiparameter single-molecule fluorescence measurements of DNA intercalating fluorophores. / Bowen, Benjamin P.; Enderlein, Jörg; Woodbury, Neal.
Proceedings of SPIE - The International Society for Optical Engineering. ed. / D.V. Nicolau; J Enderlein; R.C. Leif; D.L. Farkas. Vol. 4962 2003. p. 22-26.

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

Bowen, BP, Enderlein, J & Woodbury, N 2003, Multiparameter single-molecule fluorescence measurements of DNA intercalating fluorophores. in DV Nicolau, J Enderlein, RC Leif & DL Farkas (eds), Proceedings of SPIE - The International Society for Optical Engineering. vol. 4962, pp. 22-26, Manipulation and Analysis of Biomolecules, Cells , and Tissues, San Jose, CA, United States, 1/28/03. https://doi.org/10.1117/12.486348

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title = "Multiparameter single-molecule fluorescence measurements of DNA intercalating fluorophores",

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.",

keywords = "DNA, Single-molecule fluorescence, Thiazole Orange, Time-correlated single photon counting, TOTO",

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N2 - 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.

AB - 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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Multiparameter single-molecule fluorescence measurements of DNA intercalating fluorophores

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Keywords

ASJC Scopus subject areas

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