Laboratory of Physics of Living Matter, École Polytechnique Fédérale de Lausanne (EPFL), CH-1015, Lausanne, Switzerland.
J Mol Recognit. 2012 Sep;25(9):486-93. doi: 10.1002/jmr.2212.
Interaction of the atomic force microscopy (AFM) tip with the sample can be invasive for soft samples. Frequency Modulation (FM) AFM is gentler because it allows scanning in the non-contact regime where only attractive forces exist between the tip and the sample, and there is no sample compression. Recently, FM-AFM was used to resolve the atomic structure of single molecules of pentacene and of carbon nanotubes. We are testing similar FM-AFM-based approaches to study biological samples. We present FM-AFM experiments on dsDNA deposited on 3-aminopropyltriethoxysilane modified mica in ultra high vacuum. With flexible samples such as DNA, the substrate flatness is a sub-molecular resolution limiting factor. Non-contact topographic images of DNA show variations that have the periodicity of the right handed helix of B-form DNA - this is an unexpected result as dehydrated DNA is thought to assume the A-form structure. Frequency shift maps at constant height allow working in the non-monotonic frequency shift range, show a rich contrast that changes significantly with the tip-sample separation, and show 0.2 to 0.4 nm size details on DNA. Frequency shift versus distance curves acquired on DNA molecules and converted in force curves show that for small molecules (height < 2.5 nm), there is a contribution to the interaction force from the substrate when the tip is on top of the molecules. Our data shine a new light on dehydrated and adsorbed DNA behavior. They show a longer tip-sample interaction distance. These experiments may have an impact on nanotechnological DNA applications in non-physiological environments such as DNA based nanoelectronics and nanotemplating.
原子力显微镜(AFM)针尖与样品的相互作用可能会对软样品造成侵入性损伤。调频(FM)AFM 则更为温和,因为它允许在非接触状态下进行扫描,此时针尖和样品之间只存在吸引力,不存在样品压缩。最近,FM-AFM 被用于解析并五苯和碳纳米管的单分子原子结构。我们正在测试类似的基于 FM-AFM 的方法来研究生物样品。我们展示了在超高真空下,在 3-氨丙基三乙氧基硅烷修饰云母上沉积的双链 DNA 的 FM-AFM 实验。对于柔性样品(如 DNA),基底平整度是亚分子分辨率的限制因素。非接触式形貌图像显示 DNA 存在周期性变化,具有 B 型 DNA 右手螺旋的周期性-这是一个意外的结果,因为脱水 DNA 被认为采用 A 型结构。在恒定高度下的频率偏移映射允许在非单调频率偏移范围内工作,显示出丰富的对比度,其随针尖-样品分离显著变化,并在 DNA 上显示出 0.2 到 0.4nm 的尺寸细节。在 DNA 分子上获取的频率偏移与距离曲线并转换为力曲线表明,对于小分子(高度<2.5nm),当针尖位于分子顶部时,相互作用力中存在来自基底的贡献。我们的数据为脱水和吸附 DNA 的行为提供了新的视角。它们显示出更长的针尖-样品相互作用距离。这些实验可能会对非生理环境中的基于 DNA 的纳米技术 DNA 应用产生影响,例如 DNA 基纳米电子学和纳米模板。