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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




Journal article


Methods mol biol

Publication Date





171 - 179


Calcein, Confocal microscopy, Fluorescence recovery after photobleaching, Fluorescence recovery curve, Gap junctions, In vitro, Intercellular communication, Mobile fraction percentage, Monolayer, Tenocytes, Cell Communication, Cell Culture Techniques, Cells, Cultured, Fluoresceins, Fluorescence Recovery After Photobleaching, Fluorescent Dyes, Gap Junctions, Humans, Microscopy, Confocal, Staining and Labeling, Tenocytes