On the resolution capabilities and limits of fluorescence lifetime correlation spectroscopy (FLCS) measurements.
Quantitative tests were performed in order to explore the practical limits of FLCS. We demonstrate that: a) FLCS yields precise and correct concentration values from as low as picomolar to micromolar concentrations; b) it is possible to separate four signal components in a single detector setup; c) diffusion times differing only 25% from each other can be resolved by separating a two component mixture based on the different fluorescence lifetimes of both components; d) most of the inherent technical limitations of conventional FCS are easily overcome by FLCS employing a single detector channel confocal detection scheme.
Rüttinger
Steffen
S
Physikalisch-Technische Bundesanstalt, Abbestr. 2-12, 10587, Berlin, Germany. ruettinger@gmail.com
Kapusta
Peter
P
Patting
Matthias
M
Wahl
Michael
M
Macdonald
Rainer
R
D002232:Carbocyanines / Q000737:chemistry; D004058:Diffusion; D005456:Fluorescent Dyes / Q000737:chemistry; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013997:Time Factors
D002232:Carbocyanines; D005456:Fluorescent Dyes; C085321:cyanine dye 5
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