Nasrallah Hussein, Vial Anthony, Pocholle Nicolas, Soulier Jérémy, Costa Luca, Godefroy Cédric, Bourillot Eric, Lesniewska Eric, Milhiet Pierre-Emmanuel
INSERM, U1054, Montpellier, France.
Centre de Biochimie Structurale, Université de Montpellier, CNRS, UMR 5048, Montpellier, France.
Methods Mol Biol. 2019;1886:45-59. doi: 10.1007/978-1-4939-8894-5_3.
Supported lipid bilayers represent a very attractive way to mimic biological membranes, especially to investigate molecular mechanisms associated with the lateral segregation of membrane components. Observation of these model membranes with high-speed atomic force microscopy (HS-AFM) allows the capture of both topography and dynamics of membrane components, with a spatial resolution in the nanometer range and image capture time of less than 1 s. In this context, we have developed new protocols adapted for HS-AFM to form supported lipid bilayers on small mica disks using the vesicle fusion or Langmuir-Blodgett methods. In this chapter we describe in detail the protocols to fabricate supported artificial bilayers as well as the main guidelines for HS-AFM imaging of such samples.
支撑脂质双层是模拟生物膜的一种极具吸引力的方式,特别是用于研究与膜成分横向分离相关的分子机制。利用高速原子力显微镜(HS-AFM)观察这些模型膜,可以捕捉膜成分的形貌和动力学,空间分辨率在纳米范围内,图像捕获时间小于1秒。在此背景下,我们开发了适用于HS-AFM的新方案,通过囊泡融合或朗缪尔-布洛杰特方法在小云母盘上形成支撑脂质双层。在本章中,我们将详细描述制备支撑人工双层的方案以及此类样品的HS-AFM成像的主要指导原则。