Stetter Frank W S, Kienle Sandra, Krysiak Stefanie, Hugel Thorsten
Physik-Department E22a, Technische Universität München; IMETUM, Technische Universität München.
Physik-Department E22a, Technische Universität München; IMETUM, Technische Universität München;
J Vis Exp. 2015 Feb 27(96):e52456. doi: 10.3791/52456.
Atomic force spectroscopy is an ideal tool to study molecules at surfaces and interfaces. An experimental protocol to couple a large variety of single molecules covalently onto an AFM tip is presented. At the same time the AFM tip is passivated to prevent unspecific interactions between the tip and the substrate, which is a prerequisite to study single molecules attached to the AFM tip. Analyses to determine the adhesion force, the adhesion length, and the free energy of these molecules on solid surfaces and bio-interfaces are shortly presented and external references for further reading are provided. Example molecules are the poly(amino acid) polytyrosine, the graft polymer PI-g-PS and the phospholipid POPE (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine). These molecules are desorbed from different surfaces like CH3-SAMs, hydrogen terminated diamond and supported lipid bilayers under various solvent conditions. Finally, the advantages of force spectroscopic single molecule experiments are discussed including means to decide if truly a single molecule has been studied in the experiment.
原子力光谱法是研究表面和界面分子的理想工具。本文介绍了一种将多种单分子共价连接到原子力显微镜(AFM)针尖上的实验方案。同时,对AFM针尖进行钝化处理,以防止针尖与基底之间的非特异性相互作用,这是研究连接到AFM针尖上的单分子的前提条件。本文简要介绍了用于确定这些分子在固体表面和生物界面上的粘附力、粘附长度和自由能的分析方法,并提供了进一步阅读的外部参考文献。示例分子包括聚氨基酸聚酪氨酸、接枝聚合物PI-g-PS和磷脂POPE(1-棕榈酰-2-油酰-sn-甘油-3-磷酸乙醇胺)。这些分子在各种溶剂条件下从不同表面(如CH3-SAMs、氢终止金刚石和支撑脂质双层)解吸。最后,讨论了力谱单分子实验的优点,包括判断实验中是否真正研究了单个分子的方法。