Abstract
Fluorescence recovery after photobleaching (FRAP) is a microscopy-based technique to study the movement of fluorescent molecules inside a cell. Although initially developed to investigate intracellular mobility, FRAP can be also used to measure intercellular dynamics. This chapter describes how to perform FRAP experiment to study gap junctional communication in living cells. The procedures described here can be carried out with a laser-scanning confocal microscope and any in vitro cultured cells known to communicate via gap junctions. In addition, the method can be easily adjusted to measure gap junction function in 3D cell cultures as well as ex vivo tissue.
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Acknowledgements
Our studies to develop the published methods were supported by Orthopaedic Research UK, Arthritis Research UK, the Oxford NIHR Musculoskeletal Biomedical Research Unit and the Botnar Research Centre Bioimaging Small Research Facility, University of Oxford.
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Kuzma-Kuzniarska, M., Yapp, C., Hulley, P.A. (2016). Using Fluorescence Recovery After Photobleaching to Study Gap Junctional Communication In Vitro. In: Vinken, M., Johnstone, S. (eds) Gap Junction Protocols. Methods in Molecular Biology, vol 1437. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3664-9_12
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DOI: https://doi.org/10.1007/978-1-4939-3664-9_12
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-3664-9
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