Graduate School of Biomedical Engineering, Tohoku University, 6-6 Aoba, Aramaki, Aoba-ku, Sendai, 980-8579, Japan.
Langmuir. 2010 Feb 2;26(3):1949-52. doi: 10.1021/la902522j.
In the present study, we propose a method for preparing stable free-standing bilayer lipid membranes (BLMs). The BLMs were prepared in a microfabricated aperture with a smoothly tapered edge, which was prepared in a nanometer-thick Si(3)N(4) septum by the wet etching method. Owing to this structure, the stress on lipid bilayers at the contact with the septum was minimized, leading to remarkable membrane stability. The BLMs were not broken by applying a constant voltage of +/-1 V. The membrane lifetime was 15-45 h with and without an incorporated gramicidin channel. Gramicidin single-channel currents were recorded from the same BLM preparation when the aqueous solutions surrounding the BLM were repeatedly exchanged, demonstrating the tolerance of the present BLM to repetitive solution exchanges. Such stable membranes enable analysis of channel functions under various solution conditions from the same BLM, which will open up a variety of applications including a high throughput drug screening for ion channels.
在本研究中，我们提出了一种制备稳定的自由支撑双层脂质膜（BLM）的方法。BLM 是在具有平滑渐缩边缘的微制造孔中制备的，该孔是通过湿法刻蚀方法在纳米厚的 Si(3)N(4) 隔板上制备的。由于这种结构，与隔板接触的脂质双层上的应力最小化，导致膜的稳定性显著提高。通过施加+/-1 V 的恒定电压，BLM 不会破裂。在有和没有掺入的短杆菌肽通道的情况下，BLM 的寿命为 15-45 小时。当反复交换围绕 BLM 的水溶液时，从相同的 BLM 制备中记录了短杆菌肽单通道电流，证明了本 BLM 对重复溶液交换的耐受性。这种稳定的膜能够在相同的 BLM 下分析各种溶液条件下的通道功能，这将开辟各种应用，包括离子通道的高通量药物筛选。
