Background: Since the first morphological description of the gap junctions use electron microscopy, a considerable number of techniques has been introduced to evaluate gap junction channel functionality, many of which use dye transfer techniques, such as dye injection and fluorescent dye transfer, analyzed by flow cytometry. Methods: To analyze dye transfer, generally one population of cells is incubated with calcein-AM (0.5 lM) for 30 min at 37°C, and the other population was incubated with the lipophilic dye DiIC 18 (3) (10 lM) for 1 h at 37°C; after incubation, these cells were washed five times with PBS and cocultured for different times, and then the dye transfer was analyzed by flow cytometry.
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