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Updated: Aug 9, 2026

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination
Published on: May 13, 2017
Theory of measurement of Förster-type energy transfer in macromolecules
B Epe1, K G Steinhäuser, P Woolley
1Max-Planck-Institut für molekulare Genetik (Abteilung Wittmann), Ihnestrasse 63, D-1000 Berlin (West) 33.
Abstract:
We describe the theoretical basis of an unconventional method for the determination of the amount of energy transferred between two fluorophores by the Förster mechanism. The method involves an internal comparison made by separation of the fluorophores in situ (i.e., in the optical cell), for example, by means of enzymic digestion; it eliminates several important sources of error and it simplifies calculation while making maximal use of the information contained in the fluorescence spectra. The validity of the method is demonstrated by determination of the known distance between two modifiable sites on the transfer RNA molecule, and its usefulness is exemplified by its application to triangulation of the ribosome of Escherichia coli.
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