title: High throughput assay of diffusion through Cx43 gap junction channels with a microfluidic chip.
abstract: This paper describes a microfluidic-based assay capable of measuring gap-junction mediated dye diffusion in cultured cells. The technique exploits multistream laminar flow to selectively expose cells to different environments, enabling continuous loading of cells in one compartment while monitoring, in real time, dye diffusion into cells of a neighboring compartment. A simple one-dimensional diffusion model fit to the data extracted the diffusion coefficient of four different dyes, 5-(6)-carboxyfluorescein, 5-chloromethylfluorescein, Oregon green 488 carboxylic acid, and calcein. Different inhibitors were assayed for their ability to reduce dye coupling. The chip can screen multiple inhibitors in parallel in the same cell preparation, demonstrating its potential for high throughput. The technique provides a convenient method to measure gap junction mediated diffusion and a screen for drugs that affect gap junction communication.
authors.affiliation: Department of Mechanical and Aerospace Engineering, SUNY-Buffalo, Buffalo, New York 14260, USA.
authors.forename: Cédric
authors.initials: C
authors.lastname: Bathany
authors.forename: Derek
authors.initials: D
authors.lastname: Beahm
authors.forename: James D
authors.initials: JD
authors.lastname: Felske
authors.forename: Frederick
authors.initials: F
authors.lastname: Sachs
authors.forename: Susan Z
authors.initials: SZ
authors.lastname: Hua
mesh_terms: D000818:Animals; D002460:Cell Line; D018031:Connexin 43 / Q000032:analysis*; D004058:Diffusion; D017629:Gap Junctions / Q000737:chemistry*; D044085:Microfluidics / Q000379:methods*; D051381:Rats
chemical_list: D018031:Connexin 43