Vesenka J, Manne S, Yang G, Bustamante C J, Henderson E
Department of Zoology and Genetics, Iowa State University, Ames 50011.
Scanning Microsc. 1993 Sep;7(3):781-8.
Recent work in atomic force microscopy (AFM) of deoxyribonucleic acid (DNA) has relied on immobilizing DNA molecules by drying a small volume of buffered DNA solution onto cleaved mica. When imaging in air, relative humidity has been known to affect both the resolution and measured height of the DNA strands. We present data of measured height versus humidity for DNA and attached gold labels, and we propose a model for this data based on swelling of coadsorbed buffer salts upon exposure to moisture. In this model, small particles (e.g., DNA) stay near the top of the swelling salt layer, whereas larger particles (e.g., gold spheres) tend to be anchored down to the substrate until a moderate humidity is reached. At high humidity (around 65%), the salt layer becomes fluid-like and susceptible to tip-induced motion; the salts are either removed from the scan area or aggregate into island structures, depending on initial salt concentration on the surface.
近期关于脱氧核糖核酸(DNA)的原子力显微镜(AFM)研究工作,一直依赖于将少量缓冲DNA溶液干燥在劈开的云母上以固定DNA分子。在空气中成像时,已知相对湿度会影响DNA链的分辨率和测量高度。我们展示了DNA及附着的金标记物的测量高度与湿度的数据,并基于吸附的缓冲盐在接触水分时的膨胀现象,为该数据提出了一个模型。在这个模型中,小颗粒(如DNA)停留在膨胀盐层的顶部附近,而较大颗粒(如金纳米球)往往会固定在基底上,直到达到适度湿度。在高湿度(约65%)时,盐层变得类似流体,容易受到针尖诱导的移动影响;盐要么从扫描区域被移除,要么聚集成岛状结构,这取决于表面的初始盐浓度。