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Dual-color total internal reflection fluorescence cross-correlation spectroscopy
KTH, School of Engineering Sciences (SCI), Applied Physics, Experimental Biomolecular Physics.ORCID iD: 0000-0002-5584-9170
KTH, School of Engineering Sciences (SCI), Applied Physics, Experimental Biomolecular Physics.ORCID iD: 0000-0003-3200-0374
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2006 (English)In: Journal of Biomedical Optics, ISSN 1083-3668, E-ISSN 1560-2281, Vol. 11, no 4Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
2006. Vol. 11, no 4
Keyword [en]
fluorescence spectroscopy, laser-induced fluorescence, correlation, high count-rate, microscopy, molecule, cell, biomolecules, kinetics, surface
Identifiers
URN: urn:nbn:se:kth:diva-16046DOI: 10.1117/1.2221714ISI: 000241162000002Scopus ID: 2-s2.0-33751318060OAI: oai:DiVA.org:kth-16046DiVA: diva2:334088
Note
QC 20100525Available from: 2010-08-05 Created: 2010-08-05 Last updated: 2017-12-12Bibliographically approved

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Blom, Hans

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