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

这项工作的目标是开发一种简单、可重复性的方法来检测平面脂质双层中功能离子通道的存在。改进脂质体离子通量分析和使用BLMS、贴片电极或全细胞的电测量将极大地促进生物物理和生化研究中离子通道活性的常规测试。由单层膜形成的膜片钳双层技术如果不是如此不可复制的话将会很好地发挥作用。我们建议测试一种形成稳定的平面双层膜的新方法，这种方法将很好地支持这样一个新的分析系统--这是在平面亲水表面上支撑双层膜，该表面由一个小的单孔穿孔，每个小孔包含一个Ag/AgCl电极。首先，我们将制作带有小亲水孔的平面硅和玻璃结构。然后，通过将该装置两次通过由具有或不具有离子通道的脂类单层覆盖的空气-水界面，将形成跨越这些孔的脂类双层。这种双层中的烟碱型乙酰胆碱受体的功能将用常规的电生理学技术进行测量。

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.