Membrane Protein Array Chip using Micro/Nano Fabrication Technology

我们开发了一种高度可重复的方法，平面脂双层重建使用微流体系统的聚甲基丙烯酸甲酯（PMMA）塑料基板。通过使脂质溶液和缓冲液交替流入集成的微流体通道，在直径为100 μm的孔处形成平面脂质双层。由于孔径处脂质溶液的量和分布决定了脂质双层的状态，因此精确控制它们至关重要。我们设计了流体系统的几何形状，使得恒定量的脂质溶液分布在孔处。然后，通过施加外部压力使脂质溶液层变薄，并最终在施加200-400 Pa的压力时变成双层。形成过程可以用光学和电子记录同时监测。双层形成的最大产率为90%。使用这种技术，在单个芯片中同时形成四个脂质双层。最后，通过短杆菌肽离子通道记录通道电流，以证明芯片与单分子电生理学的兼容性。

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.