Membrane Protein Array Chip using Micro/Nano Fabrication Technology

采用微/纳米制造技术的膜蛋白阵列芯片

• 批准号: 16360119
• 负责人: SUZUKI Hiroaki
• 金额: $ 8.58万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16360119/
• 关键词: Mirco; Nano Fabrication Technology; Array Chip; Membrane Protein; Planar Lipid Bilayer

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.

我们开发了一种高重复性的平面脂质双层重构方法，使用由聚甲基丙烯酸甲酯（PMMA）塑料衬底制成的微流体系统。通过将脂质溶液和缓冲液交替注入集成微流体通道，在直径为100 μm的孔径处形成平面脂质双层膜。由于孔径处脂质溶液的数量和分布决定了脂质双分子层的状态，因此精确控制它们是至关重要的。我们设计了流体系统的几何形状，使油脂溶液在孔径处分布恒定。然后，通过施加外部压力使脂质溶液层变薄，最终在200- 400pa的压力下成为双层。地层过程可以通过光学和电子记录同时监测。双分子层形成的最大收率为90%。使用这种技术，四个脂质双分子层在一个芯片中同时形成。最后，记录了通过革兰杀菌肽离子通道的通道电流，证明了芯片与单分子电生理的兼容性。

项目成果

Highly reproducible method of planar lipid bilayer reconstitution in polymethyl methacrylate microfluidic chip

• DOI: 10.1021/la052534p
• 发表时间: 2006-02-14
• 期刊: LANGMUIR
• 影响因子: 3.9
• 作者: Suzuki, H;Tabata, KV;Takeuchi, S
• 通讯作者: Takeuchi, S