Dual-color total internal reflection fluorescence cross-correlation spectroscopy
2006 | journal article
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- Authors
- Leutenegger, M. ; Blom, H.; Widengren, J.; Eggeling, C. ; Goesch, M.; Leitgeb, R. A.; Lasser, T.
- Abstract
- 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.
- Issue Date
- 2006
- Journal
- Journal of Biomedical Optics
- Language
- English