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An Improved Substrate for Superior Imaging of Individual Biomacromolecules with Atomic Force Microscopy.
Colloids and Surfaces B: Biointerfaces ( IF 5.4 ) Pub Date : 2020-08-16 , DOI: 10.1016/j.colsurfb.2020.111321
Dmitry V Klinov 1 , Anna D Protopopova 2 , Dmitry S Andrianov 3 , Rustem I Litvinov 4 , John W Weisel 2
Affiliation  

High-resolution atomic force microscopy (AFM) of biomacromolecules is a valuable method for structural studies in biology. Traditionally, the surfaces used for AFM imaging of individual molecules are limited to mica, graphite, and glass. Because these substrates have certain shortcomings, new or modified surfaces that improve the quality of AFM imaging are highly desirable. Here, we describe an improved substrate for imaging of individual biomacromolecules with high-resolution AFM based on graphite surfaces coated by physical adsorption. We provide a detailed methodology, including the chemical structure, synthesis, characterization and the use of a substance that modifies the surface of freshly cleaved graphite, making it suitable for adsorption and AFM visualization of various biomacromolecules while minimizing spatial distortions. We illustrate the advantages of the modified graphite over regular surfaces with examples of high-resolution single-molecule imaging of proteins, polysaccharides, DNA and DNA-protein complexes. The proposed methodology is easy to use and helps to improve substantially AFM imaging of biomacromolecules of various natures, including flexible and/or unstructured sub-molecular regions that are not seen on other AFM substrates. The proposed technique has the potential to improve the use of AFM in structural biology for visualization and morphometric characterization of macromolecular objects.



中文翻译:

一种改进的基质,可利用原子力显微镜对单个生物大分子进行出色的成像。

生物大分子的高分辨率原子力显微镜(AFM)是生物学结构研究的一种有价值的方法。传统上,用于单个分子进行AFM成像的表面仅限于云母,石墨和玻璃。由于这些基材具有某些缺点,因此非常需要能够改善AFM成像质量的新表面或改性表面。在这里,我们描述了一种改进的基板,用于基于物理吸附涂覆的石墨表面的高分辨率AFM成像的单个生物大分子。我们提供了详细的方法,包括化学结构,合成,表征和使用能修饰新鲜裂解石墨表面的物质,使其适合各种生物大分子的吸附和AFM可视化,同时最大程度地减少空间扭曲。我们以蛋白质,多糖,DNA和DNA-蛋白质复合物的高分辨率单分子成像为例,说明了改性石墨在规则表面上的优势。所提出的方法易于使用,并且有助于实质上改善各种性质的生物大分子的AFM成像,包括在其他AFM基质上看不到的柔性和/或非结构化亚分子区域。所提出的技术有可能改善AFM在结构生物学中对大分子物体的可视化和形态计量表征的使用。所提出的方法易于使用,并且有助于实质上改善各种性质的生物大分子的AFM成像,包括在其他AFM基质上看不到的柔性和/或非结构化亚分子区域。所提出的技术有可能改善AFM在结构生物学中对大分子物体的可视化和形态计量表征的使用。所提出的方法易于使用,并且有助于实质上改善各种性质的生物大分子的AFM成像,包括在其他AFM基质上看不到的柔性和/或非结构化亚分子区域。所提出的技术有可能改善AFM在结构生物学中对大分子物体的可视化和形态计量表征的使用。

更新日期:2020-08-23
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