Römer Winfried, Steinem Claudia
Institut für Analytische Chemie, Chemo- und Biosensorik, Universität Regensburg, 93040 Regensburg, Germany.
Biophys J. 2004 Feb;86(2):955-65. doi: 10.1016/S0006-3495(04)74171-5.
Ordered porous alumina substrates with pore diameters of 55 and 280 nm, respectively, were produced and utilized as a support to prepare membranes suspending the pores of the material. Highly ordered porous alumina was prepared by an anodization process followed by dissolution of the remaining aluminum and alumina at the backside of the pores. The dissolution process of Al(2)O(3) at the backside of the pores was monitored by electrical impedance spectroscopy ensuring the desired sieve-like structure of the porous alumina. One side of the porous material with an area of 7 mm(2) was coated with a thin gold layer followed by chemisorption of 1,2-dipalmitoyl-sn-glycero-3-phosphothioethanol. The hydrophobic monolayer on top of the upper surface was a prerequisite for the formation of suspending membranes, termed nano-black lipid membranes (nano-BLMs). The formation process, and long-term and mechanical stability of the nano-BLMs were followed by electrical impedance spectroscopy indicating the formation of lipid bilayers with typical specific membrane capacitances of (0.65 +/- 0.2) micro F/cm(2) and membrane resistances of up to 1.6 x 10(8) Omega cm(2). These high membrane resistances allowed for single-channel recordings. Gramicidin as well as alamethicin was successfully inserted into the nano-BLMs exhibiting characteristic conductance states.
分别制备了孔径为55纳米和280纳米的有序多孔氧化铝基底,并将其用作支撑体来制备填充材料孔隙的膜。通过阳极氧化工艺制备高度有序的多孔氧化铝,随后溶解孔隙背面剩余的铝和氧化铝。通过电阻抗光谱监测孔隙背面Al₂O₃的溶解过程,以确保多孔氧化铝具有所需的筛状结构。将面积为7平方毫米的多孔材料的一侧涂上一层薄金层,然后化学吸附1,2 - 二棕榈酰 - sn - 甘油 - 3 - 磷硫乙醇。上表面顶部的疏水单分子层是形成悬浮膜(称为纳米黑色脂质膜,nano - BLMs)的先决条件。通过电阻抗光谱跟踪nano - BLMs的形成过程、长期稳定性和机械稳定性,结果表明形成了具有典型比膜电容为(0.65±0.2)微法/平方厘米和膜电阻高达1.6×10⁸欧姆·平方厘米的脂质双层。这些高膜电阻允许进行单通道记录。短杆菌肽以及短杆菌酪肽成功插入nano - BLMs并表现出特征性的电导状态。