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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.

Original publication

DOI

10.1007/978-1-4939-3664-9_12

Type

Journal

Methods Mol Biol

Publication Date

2016

Volume

1437

Pages

171 - 179

Keywords

Calcein, Confocal microscopy, Fluorescence recovery after photobleaching, Fluorescence recovery curve, Gap junctions, In vitro, Intercellular communication, Mobile fraction percentage, Monolayer, Tenocytes