Uchihashi Takayuki, Watanabe Hiroki, Kodera Noriyuki
Nagoya, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8601, Aichi, Japan.
Research Institute of Biomolecule Metrology Co., Ltd, Tsukuba, 305-0047, Ibaraki, Japan.
Methods Mol Biol. 2018;1814:159-179. doi: 10.1007/978-1-4939-8591-3_10.
Recent progresses in high-speed atomic force microscopy (HS-AFM) have enabled us to directly visualize dynamic processes of various proteins in liquid conditions. One of the key factors leading to successful HS-AFM observations is the selection of an appropriate substrate depending on molecules to be observed. For the HS-AFM imaging, a target molecule must be absorbed on a substrate by controlling its orientation without impairing the dynamics or physiological function of the molecule. In this chapter, we describe protocols for preparation of substrates that have been used for HS-AFM and then introduce observation examples on dynamic processes of biological molecules.
高速原子力显微镜(HS-AFM)的最新进展使我们能够直接观察液体条件下各种蛋白质的动态过程。成功进行HS-AFM观察的关键因素之一是根据要观察的分子选择合适的底物。对于HS-AFM成像,必须通过控制目标分子的方向将其吸附在底物上,同时又不损害分子的动力学或生理功能。在本章中,我们描述了用于HS-AFM的底物制备方案,然后介绍生物分子动态过程的观察实例。