Mica functionalization for imaging of DNA and protein-DNA complexes with atomic force microscopy.
Mica functionalization for imaging of DNA and protein-DNA complexes with atomic force microscopy.
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DOI：
10.1007/978-1-62703-056-4_14
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发表时间：
2013
期刊：
影响因子：
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通讯作者：
Lyubchenko, Yuri L
中科院分区：
文献类型：
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作者：
Shlyakhtenko, Luda S;Gall, Alexander A;Lyubchenko, Yuri L
表面处理是利用原子力显微镜(AFM)对DNA和蛋白质-DNA复合体进行可靠和重复性成像的关键步骤。本文介绍了云母表面化学功能化的方法。一种方法是使用3-氨基丙基三乙氧基硅烷(APTES)，使人们能够获得一种称为AP-云母的光滑表面。在不含二价阳离子的大范围pH范围内，该表面结合了多种离子强度的核酸和核蛋白复合体。另一种方法使用氨基丙基硅烷(APS)来制备APS-云母表面。与AP云母相比，APS云母的优势在于能够在水溶液中获得可靠且可重现的时间推移图像。本章介绍了AP-云母和APS-云母表面的制备方法，以及用于AFM成像的样品的制备。还提供了APS的合成和纯化方案。用大量的例子说明了这些应用。
Surface preparation is a key step for reliable and reproducible imaging of DNA and protein-DNA complexes with atomic force microscopy (AFM). This article describes the approaches for chemical functionalization of the mica surface. One approach utilizes 3-aminopropyl-trietoxy silane (APTES), enabling one to obtain a smooth surface termed AP-mica. This surface binds nucleic acids and nucleoprotein complexes in a wide range of ionic strengths, in the absence of divalent cations and in a broad range of pH. Another method utilizes aminopropyl silatrane (APS) to yield an APS-mica surface. The advantage of APS-mica compared with AP-mica is the ability to obtain reliable and reproducible time-lapse images in aqueous solutions. The chapter describes the methodologies for the preparation of AP-mica and APS-mica surfaces and the preparation of samples for AFM imaging. The protocol for synthesis and purification of APS is also provided. The applications are illustrated with a number of examples.