Stripping Material from a Supported Lipid Bilayer with High Speed Buffer Flow

Michael Ornstead, Ruth Hunter, Mason L Valentine, Cameron Cooper, Stephen Kent Smith, Christopher F. Monson
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Abstract

A microfluidic device was created and used to demonstrate that supported lipid bilayers can be deposited on clean glass slides and removed using high velocity buffer flow (1-4 m/s linear velocity). This was accomplished by forcing the flow through a microfluidic channel covering an annealed glass coverslip bearing a supported lipid bilayer (SLB). The removal of bilayer material was monitored via fluorescence microscopy, and two basic regimes were observed: at 1-2 m/s smaller areas were stripped, while at 3-4 m/s larger areas were stripped. SLB removal was verified by two means. First, lipid vesicles labeled with a different fluorescent dye were added to the device and filled in holes left by the removal of the original SLB, allowing stripping to be verified visually. Second, the solutions obtained from stripping were concentrated and the fluorescence in the concentrates was measured. The ability to strip SLB from glass provides a relatively gentle method of creating spatially inhomogeneous SLB, which could be a useful tool in the continued investigation of membrane properties and components. KEYWORDS: Supported Lipid Bilayer; Membrane Vesicle; Microfluidic Device
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剥离材料从支持脂双分子层与高速缓冲流
创建了一个微流体装置,并用于证明支持的脂质双层可以沉积在干净的玻璃载玻片上,并使用高速缓冲流(1-4 m/s线速度)去除。这是通过迫使流动通过覆盖有支持脂质双分子层(SLB)的退火玻璃盖盖的微流体通道来完成的。通过荧光显微镜监测双层材料的去除,并观察到两种基本机制:在1-2 m/s下剥离较小区域,而在3-4 m/s下剥离较大区域。通过两种方法验证了SLB的去除。首先,将用不同荧光染料标记的脂质囊泡添加到设备中,并填充去除原始SLB后留下的孔,从而可以直观地验证剥离。其次,对剥离所得溶液进行浓缩,测定浓缩液中的荧光。从玻璃中剥离SLB的能力提供了一种相对温和的方法来创建空间不均匀的SLB,这可能是继续研究膜性质和成分的有用工具。关键词:负载脂双分子层;膜泡;微流体装置
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