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Photobleaching and Sensitized Emission-Based Methods for the Detection of Förster Resonance Energy Transfer

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Computer Optimized Microscopy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2040))

Abstract

Förster resonance energy transfer (FRET) is a non-radiative interaction between two molecules that happens at distances in the range of a few nanometers. Using FRET interactions between suitably selected fluorophores allows to study molecular interactions or conformational changes of single molecules on fluorescence microscopes even though the optical resolution of the microscope is limited to distances that are almost two orders of magnitude higher.

In this chapter several variants of FRET detection methods are described that are based either on the targeted photobleaching of one of the participating molecule species or on the direct detection of the fluorescence signal that is created as a result of the FRET interactions.

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Acknowledgments

The fitting macros “linear fit acceptor photobleaching.ijm” and “exponential fit donor photobleaching.ijm” were kindly provided by Raul Gomez, Centre for Genomic Regulation, Barcelona.

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Authors and Affiliations

  1. Advanced Light Microscopy Unit, Centre for Genomic Regulation, Barcelona, Spain

    Timo Zimmermann

Authors
  1. Timo Zimmermann
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Correspondence to Timo Zimmermann .

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Editors and Affiliations

  1. Molecular Imaging Platform, Molecular Biology Institute of Barcelona (CSIC), Barcelona, Spain

    Elena Rebollo

  2. Scientific and Technological Centers (CCiTUB), Universitat de Barcelona, Barcelona, Spain

    Manel Bosch

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Zimmermann, T. (2019). Photobleaching and Sensitized Emission-Based Methods for the Detection of Förster Resonance Energy Transfer. In: Rebollo, E., Bosch, M. (eds) Computer Optimized Microscopy. Methods in Molecular Biology, vol 2040. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9686-5_12

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  • DOI: https://doi.org/10.1007/978-1-4939-9686-5_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9685-8

  • Online ISBN: 978-1-4939-9686-5

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