Toward a Practical Assay for Ion Channel Activity: Can a Silicon Device Support Planar Lipid Bilayers?

这项工作的目标是发展一个简单的可重复的 测定功能性离子通道存在的方法 在平面脂质双层中。 改善脂质体离子通量 使用BLM、贴片 电极或整个细胞将大大促进常规的 生物物理和生物化学中离子通道活性的测试 research. 膜片钳技术形成的双层， 如果不是如此不可再现，单层将很好地起作用。 我们 我建议测试一种形成稳定平面双层的新方法 这将很好地适用于这样一种新的分析系统--这将 在平面亲水表面上支撑双层， 每个孔含有一个Ag/AgCl电极。 首先，我们将制作平面硅和玻璃结构， 小的亲水孔。 脂质双分子层就会形成 穿过这些孔， 空气-水界面被脂质单层覆盖，有或没有 离子通道 烟碱功能的测量 在这种双层中的乙酰胆碱受体将用 常规电生理技术。

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.