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Time-Resolved Fluorescence Anisotropy Imaging

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Fluorescence Spectroscopy and Microscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1076))

Abstract

Fluorescence can be characterized by its intensity, position, wavelength, lifetime, and polarization. The more of these features are acquired in a single measurement, the more can be learned about the sample, i.e., the microenvironment of the fluorescence probe. Polarization-resolved fluorescence lifetime imaging—time-resolved fluorescence anisotropy imaging, TR-FAIM—allows mapping of viscosity or binding or of homo-FRET which can indicate dimerization or generally oligomerization.

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Notes

  1. 1.

    Only in the specific case of the cerulean fluorescent protein (CFP) the fluorescence lifetime has been reported to change due to homo-FRET [67].

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Acknowledgements

We would like to thank Dr Simon Ameer-Beg in the Randall Division for Cell and Molecular Biophysics at King’s College London for fruitful discussions and Dr Aleksandar Ivetic from the Membrane/Cytoskeleton Signalling Group in the Cardiovascular Division at King’s College London for the samples in Fig. 3. We would also like to thank Dr Steven Vogel from the National Institute on Alcohol Abuse and Alcoholism in Bethesda, USA, and editor Prof Yves Engelborghs for valuable comments on the manuscript. Finally, we would like to acknowledge the UK’s Medical Research Council (MRC) for funding.

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Authors and Affiliations

  1. Department of Physics, King’s College London, London, UK

    Klaus Suhling, James Levitt & Pei-Hua Chung

  2. Faculty of Life Sciences, University of Manchester, Manchester, UK

    Pei-Hua Chung

Authors
  1. Klaus Suhling
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  2. James Levitt
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  3. Pei-Hua Chung
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Editors and Affiliations

  1. KULeuven-Biomolecular Dynamics BioSCENTer, Leuven, Belgium

    Yves Engelborghs

  2. MicroSpectroscopy Centre, University of Wageningen Biochemistry, Wageningen, The Netherlands

    Antonie J.W.G. Visser

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Suhling, K., Levitt, J., Chung, PH. (2014). Time-Resolved Fluorescence Anisotropy Imaging. In: Engelborghs, Y., Visser, A. (eds) Fluorescence Spectroscopy and Microscopy. Methods in Molecular Biology, vol 1076. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-649-8_22

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