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Inverse-Fluorescence Cross-Correlation Spectroscopy
KTH, School of Engineering Sciences (SCI), Applied Physics, Experimental Biomolecular Physics.ORCID iD: 0000-0002-1850-5440
KTH, School of Engineering Sciences (SCI), Applied Physics, Experimental Biomolecular Physics.
2010 (English)In: Analytical Chemistry, ISSN 0003-2700, E-ISSN 1520-6882, Vol. 82, no 13, 5646-5651 p.Article in journal (Refereed) Published
Abstract [en]

Inverse-fluorescence correlation spectroscopy (iFCS) was recently introduced as an alternative version of FCS that does not require labeling of the analyzed particles or biomolecules. In iFCS, the signal from a medium surrounding the particles is analyzed, as opposed to a signal from the studied particles themselves. As unlabeled particles diffuse through the detection volume, they displace a fraction of the fluorescent medium, causing transient dips in the detected signal which give information about the mobility and concentration of the analyzed particles. Here inverse-fluorescence cross-correlation spectroscopy (iFCCS) is introduced as an extension of iFCS. In iFCCS, labeled particles/biomolecules are analyzed and their fluorescence signal is cross-correlated with the signal from the surrounding medium. When labeled particles are analyzed, a direct estimate of the volume of the particles is obtained or, alternatively, an estimate of the size of the detection volume. Another possibility is to analyze the interaction of small, labeled molecules with unlabeled particles, resulting in cross-correlation as an indication of binding, even though only one binding partner is labeled. This also enables accurate estimation of the degree of labeling, since the amounts of labeled and unlabeled particles are estimated independently.

Place, publisher, year, edition, pages
2010. Vol. 82, no 13, 5646-5651 p.
National Category
Analytical Chemistry
URN: urn:nbn:se:kth:diva-27252DOI: 10.1021/ac1006409ISI: 000279253300034ScopusID: 2-s2.0-77954210488OAI: diva2:379398
Knut and Alice Wallenberg Foundation

QC 20150721

Available from: 2010-12-17 Created: 2010-12-09 Last updated: 2015-07-21Bibliographically approved

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Wennmalm, StefanWidengren, Jerker
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