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Recording Intracellular cAMP Levels with EPAC-Based FRET Sensors by Fluorescence Lifetime Imaging

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cAMP Signaling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1294))

Abstract

Eukaryotic cells use second messengers such as Ca2+, IP3, cGMP, and cAMP to transduce extracellular signals like hormones, via membrane receptors to downstream cellular effectors. FRET-based sensors are ideal to visualize and measure these rapid changes of second messenger concentrations in time and place. Here, we describe the use of EPAC-based FRET sensors to measure cAMP with spatiotemporal resolution in cells by fluorescence lifetime imaging (FLIM).

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Authors and Affiliations

  1. Department of Cell Biology, The Netherlands Cancer Institute, Amsterdam, The Netherlands

    Jeffrey Klarenbeek & Kees Jalink

  2. Department of Cell Biology and Biochemistry, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX, Amsterdam, The Netherlands

    Marcel Raspe

Authors
  1. Marcel Raspe
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  2. Jeffrey Klarenbeek
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Corresponding author

Correspondence to Kees Jalink .

Editor information

Editors and Affiliations

  1. Department of Physiology, Anatomy, and Genetics, University of Oxford, Oxford, United Kingdom

    Manuela Zaccolo

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Raspe, M., Klarenbeek, J., Jalink, K. (2015). Recording Intracellular cAMP Levels with EPAC-Based FRET Sensors by Fluorescence Lifetime Imaging. In: Zaccolo, M. (eds) cAMP Signaling. Methods in Molecular Biology, vol 1294. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2537-7_2

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  • DOI: https://doi.org/10.1007/978-1-4939-2537-7_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2536-0

  • Online ISBN: 978-1-4939-2537-7

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