The goal of the work is development of a simple reproducible method for assaying for the presence of functional ion channels in planar lipid bilayers. Improving upon liposomal ion flux assays and electrical measurements employing BLMs, patch electrodes, or whole cells would greatly facilitate routine testing for ion channel activity in biophysical and biochemical research. The technique of patch clamping bilayers formed from monolayers would serve well if it were not so irreproducible. We propose to test a new approach at forming stable planar bilayers that will lend itself well to such a new assay system--this is to support bilayers on planar hydrophilic surfaces perforated by small single orifices containing one Ag/AgCl electrode each. First we will fabricate planar silicon and glass structures with small hydrophilic holes. Lipid bilayers will then be formed across those holes by passing the device twice through an air-water interface covered by a lipid monolayer with or without ion channels. Measurement of the function of the nicotinic acetylcholine receptor in such bilayers will be made with conventional electrophysiological techniques.
