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Updated: Aug 13, 2026

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination
Published on: May 13, 2017
Detection of three rotational correlation times for a rigid asymmetric molecule using frequency-domain fluorometry
I Gryczynski1, H Cherek, J R Lakowicz
1University of Maryland, School of Medicine, Department of Biological Chemistry, Baltimore 21201.
Abstract:
We measured the frequency response of the polarized emission of Yt-base in propylene glycol at 10 degrees C. Data were obtained for excitation wavelengths of 290, 312 and 346 nm, for which the fundamental anisotropies are 0.05, 0.19 and 0.32, respectively. Additionally, data were obtained using CCl4, to decrease the mean decay time from 9.1 to 4.2 ns. These nine sets of data were analyzed globally to recover the anisotropy decay law. Three correlation times were needed to fit the data, 0.8, 3.0 and 5.6 ns, a range of only 7-fold. We believe this is the first reported detection of three correlation times for a rigid molecule.
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