Membrane Protein Array Chip using Micro/Nano Fabrication Technology

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We developed a highly reproducible method for planar lipid bilayer reconstitution using a micro-fluidic system made of a polymethyl methacrylate (PMMA) plastic substrate. Planar lipid bilayers are formed at apertures, 100 μm diameter, by flowing lipid solution and buffer alternately into an integrated micro-fluidic channel. Since the amount and distribution of the lipid solution at the aperture determines the state of the lipid bilayer, controlling them precisely is crucial. We designed the geometry of the fluidic system so that a constant amount of lipid solution is distributed at the aperture. Then, the layer of lipid solution is thinned by applying an external pressure, and finally becomes a bilayer when a pressure of 200-400 Pa is applied. The formation process can be simultaneously monitored with optical and electric recordings. The maximum yield for bilayer formation was 90%. Using this technique, four lipid bilayers are formed simultaneously in a single chip. Finally, a channel current through gramicidin peptide ion channels are recorded to prove the compatibility of the chip with single molecule electrophysiology.