Ostapchenko Valeriy, Gasset Maria, Baskakov Ilia V
Center for Biomedical Engineering and Technology, Department of Anatomy and Neurobiology, University of Maryland, Baltimore, MD, USA.
Methods Mol Biol. 2012;849:157-67. doi: 10.1007/978-1-61779-551-0_11.
Atomic force microscopy (AFM) has become a conventional tool for elucidation of the molecular mechanisms of protein aggregation and, specifically, for analysis of assembly pathways, architecture, aggregation state, and heterogeneity of oligomeric intermediates or mature fibrils. AFM imaging provides useful information about particle dimensions, shape, and substructure with nanometer resolution. Conventional AFM methods have been very helpful in the analysis of polymorphic assemblies formed in vitro from homogeneous proteins or peptides. However, AFM imaging on its own provides limited insight into conformation or composition of assemblies produced in the complex environment of a cell, or prepared from a mixture of proteins as a result of cross-seeding. In these cases, its combination with fluorescence microscopy (AFFM) increases its resolution.
原子力显微镜（AFM）已成为阐明蛋白质聚集分子机制的常规工具，特别是用于分析组装途径、结构、聚集状态以及寡聚中间体或成熟纤维的异质性。AFM成像以纳米分辨率提供有关颗粒尺寸、形状和亚结构的有用信息。传统的AFM方法在分析由同质蛋白质或肽体外形成的多晶型组装体方面非常有帮助。然而，仅AFM成像对在细胞复杂环境中产生的或因交叉接种由蛋白质混合物制备的组装体的构象或组成提供的见解有限。在这些情况下，将其与荧光显微镜（AFFM）结合可提高其分辨率。
