Ando Toshio
Department of Physics, Kanazawa University, Kakuma-machi, Kanazawa, Japan.
Microscopy (Oxf). 2013 Feb;62(1):81-93. doi: 10.1093/jmicro/dfs093. Epub 2013 Jan 4.
High-speed atomic force microscopy (HS-AFM) has been developed as a nano-dynamics visualization technique. This microscopy permits direct observation of structure dynamics and dynamic processes of biological molecules in physiological solutions, at a subsecond to sub-100 ms temporal resolution and an ∼2 nm lateral and a 0.1 nm vertical resolution. Importantly, tip-sample interactions do not disturb the biomolecules' functions. Various functioning proteins including myosin V walking on an actin filament and bacteriorhodopsin responding to light have been successfully visualized with HS-AFM. In the quest for understanding the functional mechanisms of proteins, inferences no longer have to be made from static snapshots of molecular structures and dynamic behavior of optical markers attached to proteins. High-resolution molecular movies obtained from HS-AFM observations reveal the details of molecules' dynamic behavior in action, without the need for intricate analyses and interpretations. In this review, I first describe the fundamentals behind the achieved high imaging rate and low invasiveness to samples, and then highlight recent imaging studies. Finally, future studies are briefly described.
高速原子力显微镜（HS-AFM）已发展成为一种纳米动力学可视化技术。这种显微镜能够在生理溶液中以亚秒至亚100毫秒的时间分辨率以及约2纳米的横向分辨率和0.1纳米的垂直分辨率，直接观察生物分子的结构动力学和动态过程。重要的是，针尖-样品相互作用不会干扰生物分子的功能。利用HS-AFM已成功可视化了各种功能蛋白，包括在肌动蛋白丝上行走的肌球蛋白V以及对光作出反应的细菌视紫红质。在探索蛋白质功能机制的过程中，不再需要从蛋白质分子结构的静态快照以及附着在蛋白质上的光学标记的动态行为来进行推断。从HS-AFM观察中获得的高分辨率分子电影揭示了分子动态行为的细节，而无需复杂的分析和解释。在这篇综述中，我首先描述实现高成像速率和对样品低侵入性背后的基本原理，然后重点介绍近期的成像研究。最后，简要描述未来的研究。
