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Journal of Biomedical Optics • Open Access

Dual-color total internal reflection fluorescence cross-correlation spectroscopy

Paper 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.

Paper Details

Date Published: 1 July 2006
PDF: 3 pages
J. Biomed. Opt. 11(4) 040502 doi: 10.1117/1.2221714
Published in: Journal of Biomedical Optics Volume 11, Issue 4
Show Author Affiliations
Marcel Leutenegger, École Polytechnique Fédérale de Lausanne (Switzerland)
Michael Gösch, École Polytechnique Fédérale de Lausanne (Switzerland)
Rainer Andreas Leitgeb, École Polytechnique Fédérale de Lausanne (Switzerland)
Theo Lasser, École Polytechnique Fédérale de Lausanne (Switzerland)
Hans Blom, École Polytechnique Fédérale de Lausanne (Switzerland)
Jerker Widengren, Kungliga Tekniska Högskolan (Sweden)
Christian Eggeling, Max Planck Institute for Biophysical Chemistry (Germany)

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