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FRET imaging

Nature Biotechnology volume 21, pages 1387–1395 (2003)Cite this article

Abstract

Förster (or Fluorescence) Resonance Energy Transfer (FRET) is unique in generating fluorescence signals sensitive to molecular conformation, association, and separation in the 1–10 nm range. We introduce a revised photophysical framework for the phenomenon and provide a systematic catalog of FRET techniques adapted to imaging systems, including new approaches proposed as suitable prospects for implementation. Applications extending from a single molecule to live cells will benefit from multidimensional microscopy techniques, particularly those adapted for optical sectioning and incorporating new algorithms for resolving the component contributions to images of complex molecular systems.

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Figure 1: Parametric FRET functions.
Figure 2: The 'Michaelis-Menten' view of a fluorophore as a photon conversion catalyst or 'enzyme' (see Box 1).
Figure 3: Photochromic FRET (pcFRET).
Figure 4: Comparative sizes of common fluorophores and protein carriers used in FRET imaging.
Figure 5: Photophysical cycles including triplet state, FRET, and photobleaching.

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Acknowledgements

E.A.J.-E. is indebted to the Agencia Nacional de Promoción de la Ciencia y Tecnología (ANPCyT), Fundación Antorchas, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Secretaría de Ciencia, Tecnología e Innovación Productiva (SECyT) and the Universidad de Buenos Aires (UBA) for financial support. T.M.J. was supported by the Max Planck Society, European Union FP5 Projects QLG1-2000-01260 and QLG2-CT-2001-02278, and the Center of the Molecular Physiology of the Brain funded by the German Research Council (DFG). The authors were the recipients of a joint grant from the Volkswagen Foundation for their work on photochromic compounds and acknowledge the contribution of graduate student Luciana Giordano to the research depicted in Figure 4, as well as the efforts of many colleagues over the years in the general area represented by this review. They are also indebted professionally and personally for the inspiration offered by the late Gregorio Weber, the acknowledged father of fluorescence in biology. We thank Rainer Heintzmann, Pedro Aramendía, Carla Spagnuolo and Vinod Subramaniam for critical reading of the manuscript.

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  1. Departamento de Química Orgánica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, 1428, Argentina

    Elizabeth A Jares-Erijman

  2. Department of Molecular Biology, Max Planck Institute for Biophysical Chemistry, Göttingen, 37077, Germany

    Thomas M Jovin

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Jares-Erijman, E., Jovin, T. FRET imaging. Nat Biotechnol 21, 1387–1395 (2003). https://doi.org/10.1038/nbt896

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