Sapra K Tanuj
Department of Chemistry, University of Oxford, Oxford, UK.
Methods Mol Biol. 2013;974:73-110. doi: 10.1007/978-1-62703-275-9_5.
The atomic force microscope (AFM) has opened vast avenues hitherto inaccessible to the biological scientist. The high temporal (millisecond) and spatial (nanometer) resolutions of the AFM are suited for studying many biological processes in their native conditions. The AFM cantilever stylus is aptly termed as a "lab on a tip" owing to its versatility as an imaging tool as well as a handle to manipulate single bonds and proteins. Recent examples assert that the AFM can be used to study the mechanical properties and monitor processes of single proteins and single cells, thus affording insight into important mechanistic details. This chapter specifically focuses on practical and analytical protocols of single-molecule AFM methodologies related to high-resolution imaging and single-molecule force spectroscopy of membrane proteins. Both these techniques are operator oriented, and require specialized working knowledge of the instrument, theoretical, and practical skills.
原子力显微镜（AFM）为生物科学家开辟了迄今为止难以触及的广阔途径。AFM的高时间分辨率（毫秒级）和空间分辨率（纳米级）适合在自然条件下研究许多生物过程。AFM悬臂探针因其作为成像工具的多功能性以及用于操纵单键和蛋白质的手段，被恰当地称为“尖端实验室”。最近的例子表明，AFM可用于研究单个蛋白质和单个细胞的力学性质并监测其过程，从而深入了解重要的机制细节。本章特别关注与膜蛋白的高分辨率成像和单分子力谱相关的单分子AFM方法的实践和分析方案。这两种技术都以操作员为导向，需要仪器的专业操作知识、理论和实践技能。
