Development of the epithelium-like complex membrane system

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To develop the epithelium-like complex membrane system, we start to establish a single-membrane system with a reproducible way. Compared to the conventional droplet interface bilayer method, the size of the droplet was decreased to one tenth, and the background noise was substantially attenuated. The asymmetric membrane was formed by adding different liposomes composed of distinct lipid compositions to the droplets. We revealed that the gating of the KcsA potassium channel was regulated by the acidic lipid in the inner leaflet of the membrane. The rapid perfusion system was introduced into the bilayer system, in which two glass pipettes were inserted into one of the droplet. The solution was exchanged within several tens of ms, and the pH-dependent activation of the KcsA channel was readily measured.