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Dual-color total internal reflection fluorescence cross-correlation spectroscopy
- Leutenegger, Marcel (author)
-
- Blom, Hans (author)
- KTH,Experimentell biomolekylär fysik
-
- Widengren, Jerker (author)
- KTH,Experimentell biomolekylär fysik
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- Eggeling, Christian (author)
- Goesch, Michael (author)
- Leitgeb, Rainer A. (author)
- Lasser, Theo (author)
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- (creator_code:org_t)
- SPIE-Intl Soc Optical Eng, 2006
- 2006
- English.
- In: Journal of Biomedical Optics. - : SPIE-Intl Soc Optical Eng. - 1083-3668 .- 1560-2281. ; 11:4
- Journal article (peer-reviewed)
Abstract
Subject headings
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- We present the development and first application of a novel dual-color total internal reflection (TIR) fluorescence system for single-molecule coincidence analysis and fluorescence cross-correlation spectroscopy (FCCS). As a performance analysis, we measured a synthetic DNA-binding assay, demonstrating this dual-color TIR-FCCS approach to be a suitable method for measuring coincidence assays such as biochemical binding, fusion, or signal transduction at solid/liquid interfaces. Due to the very high numerical aperture of the epi-illumination configuration, our setup provides a very high fluorescence collection efficiency resulting in a two- to three- fold increase in molecular brightness compared to conventional confocal FCCS. Further improvements have been achieved through global analysis of the spectroscopic data.
Keyword
- fluorescence spectroscopy
- laser-induced fluorescence
- correlation
- high count-rate
- microscopy
- molecule
- cell
- biomolecules
- kinetics
- surface
Publication and Content Type
- ref (subject category)
- art (subject category)
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