Henderson Robert M
University of Cambridge, Department of Pharmacology , Tennis Court Road, Cambridge CB2 1PD , UK +44 1223 334053 ;
Expert Opin Drug Discov. 2015 Mar;10(3):221-9. doi: 10.1517/17460441.2015.998195. Epub 2014 Dec 31.
Atomic force microscopy (AFM) is a scanning probe technique that has been in use in biology to generate sub-nanometre resolution images in near-physiological environments for over 20 years. Most AFM work uses instruments that take several minutes to generate each image but instruments that can produce real-time images have recently become available and there is now a reasonable body of work published on this technique. The importance of this high-speed AFM is that dynamic events of individual macromolecules can be studied.
This review focuses on specific examples that demonstrate the potential of the technique. It covers four areas in which high-speed AFM has been used to elucidate mechanisms that are either unstudied or not clearly understood. These areas are: protein-protein interactions; DNA-protein interactions; quantification of biological processes; the use of DNA origami scaffolds as nanostructures to build and study dynamic molecular events.
High-speed AFM shares advantages and disadvantages with conventional AFM, but it compares well in quality of data generated and in ease of use with other currently available techniques of high-resolution biological imaging. As the instruments become more widespread, the value of high-speed AFM and its potential to complement other techniques in molecular and cell biology should become more appreciated.
原子力显微镜(AFM)是一种扫描探针技术,在生物学领域已应用20多年,用于在近生理环境中生成亚纳米分辨率的图像。大多数原子力显微镜工作使用的仪器生成每张图像需要几分钟,但能够产生实时图像的仪器最近已问世,目前关于该技术也有了相当数量的已发表研究成果。这种高速原子力显微镜的重要性在于可以研究单个大分子的动态事件。
本综述聚焦于展示该技术潜力的具体实例。它涵盖了四个领域,高速原子力显微镜已被用于阐明那些尚未被研究或尚未被清楚理解的机制。这些领域包括:蛋白质-蛋白质相互作用;DNA-蛋白质相互作用;生物过程的量化;使用DNA折纸支架作为纳米结构来构建和研究动态分子事件。
高速原子力显微镜与传统原子力显微镜有共同的优缺点,但在生成的数据质量以及与其他当前可用的高分辨率生物成像技术相比的易用性方面表现良好。随着仪器的日益普及,高速原子力显微镜的价值及其在分子和细胞生物学中补充其他技术的潜力应该会得到更多认可。