1 July 2006 Dual-color total internal reflection fluorescence cross-correlation spectroscopy
Author Affiliations +
J. of Biomedical Optics, 11(4), 040502 (2006). doi:10.1117/1.2221714
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.
Marcel Leutenegger, Michael Gösch, Rainer Andreas Leitgeb, Theo Lasser, Hans Blom, Jerker Widengren, Christian Eggeling, "Dual-color total internal reflection fluorescence cross-correlation spectroscopy," Journal of Biomedical Optics 11(4), 040502 (1 July 2006). http://dx.doi.org/10.1117/1.2221714
Submission: Received ; Accepted
JOURNAL ARTICLE
3 PAGES


SHARE
KEYWORDS
Luminescence

Fluorescence spectroscopy

Spectroscopy

Molecules

Reflectance spectroscopy

Interfaces

Diffusion

Back to Top