Bilayer reconstitution of voltage-dependent ion channels using a microfabricated silicon chip
Bilayer reconstitution of voltage-dependent ion channels using a microfabricated silicon chip
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DOI：
10.1016/s0006-3495(01)75885-7
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发表时间：
2001-10-01
影响因子：
3.4
通讯作者：
Heath, JR
中科院分区：
文献类型：
--
作者：
Pantoja, R;Sigg, D;Heath, JR
含有重构离子通道的涂漆双层可作为通道结构和功能的电生理学研究的明确模型系统。通过在蚀刻到 200 微米厚的硅片上的圆柱形孔上涂上磷脂：癸烷混合物，可以获得水平取向的双层，溶液可以轻松进入两侧。 SiO2 层的硅烷化产生了疏水表面，促进了脂质混合物的粘附。使用标准光刻技术和各向异性深反应离子蚀刻来创建直径为 50 至 200 μm 的孔隙。隔板中孔的圆柱形结构和表面处理产生了稳定的双层。这些用于在直径为 100 微米和 200 微米的孔中重建 Maxi K 通道。悬浮在微芯片中的双层的电生理学特征与其他双层制剂的电生理学特征相当。水平方向和良好的电压钳位特性使微芯片双层方法成为与光学探针同时研究重组膜蛋白电特性的优秀系统。
Painted bilayers containing reconstituted ion channels serve as a well defined model system for electrophysiological investigations of channel structure and function. Horizontally oriented bilayers with easy solution access to both sides were obtained by painting a phospholipid:decane mixture across a cylindrical pore etched into a 200-mum thick silicon wafer. Silanization of the SiO2 layer produced a hydrophobic surface that promoted the adhesion of the lipid mixture. Standard lithographic techniques and anisotropic deep-reactive ion etching were used to create pores with diameters from 50 to 200 mum. The cylindrical structure of the pore in the partition and the surface treatment resulted in stable bilayers. These were used to reconstitute Maxi K channels in the 100- and 200-mum diameter pores. The electrophysiological characteristics of bilayers suspended in microchips were comparable with that of other bilayer preparations. The horizontal orientation and good voltage clamping properties make the microchip bilayer method an excellent system to study the electrical properties of reconstituted membrane proteins simultaneously with optical probes.