Real time observation of the photocleavage of single DNA molecules

A method for real time observation of photo-cleavage of stratched λDNA at single molecular level by a fluorescent microscope coupled with CCD is developed.DNA molecules stained with YOYO-1 are stretched by the mo-lecular combing technique and fixed on a modified slide.Then the Process of Photocleavage and relaxation of DNA under radiation of blue light is observed.We speculate that the conformation change of stretched DNA and the effect of water are likely to facilitate the effect of YOYO photocleav-age DNA molecules.The photocleavage effect of YOYO for stretched DNA may be useful to study DNA elasticity,cancer research as well as the interaction between DNA and dyes.     

Combined-dynamic mode “dip-pen” nanolithography and physically nanopatterning along single DNA molecules

Atomic force micriscope (AFM)-based dip-pen nanolithography (DPN) is an emerging approach for constructing nanostructures on material surfaces such as gold, silicon and silicon oxide. Although DPN is a powerful technique, it has not shown its ability of direct-writing and patterning of nanostructures on surfaces of soft materials, for example biomacromolecules. Direct depositing on soft surfaces becomes possible with the introduction of a combined-dynamic mode DPN rather than mostly used contact mode DPN or tapping mode DPN. In this report, the combined dynamic mode DPN is used for direct depositing protein ink on DNA molecules at the nanometer scale.     

Direct measurement of compression spring constant of single DNA molecule with AFM

The understanding of mechanical responses of individ-ual macromolecules under a small external load (usually F<0.5 nN) is of profound importance because the induced conformational transition can result in the change of bio-logical functions[1?3]. The advent of new instrumentations, especially the development of Pico-Newton force tech-nology, has made it possible to investigate the dynamic behavior of single biomolecule and manipulate it to make direct measurements of its elastic properties[4?…     

原子力显微镜观测DNA分子的表面沉积与扩展

原子力显微镜具有原子级分辨率,能够对生物样品进行观察.用原子力显微镜观察了DNA分子在Si、云母片及修饰过的云母片表面的沉积与扩展,对3种情况作了比较,用超声振荡方法可以有效地切断DNA分子链.     

DNA分子在云母表面拉直技术的研究

对DNA分子的生物、物理和化学性质的很多研究,都要求对单个DNA分子的性质进行深入探索,实现DNA分子在基底表面上的拉直是对其深入研究的物理基础。文章在APS溶液处理过的云母片表面,通过不同的方法完成了对DNA分子的拉直操作,利用原予力显微镜对拉直后的DNA分子进行了表征,获得了DNA分子拉直后的形貌,并对每种拉直方法进行了比较和讨论。     

Analysis of the Structure of Adsorbed Fibrinogen on Solid Surfaces and Its Dynamic Processes Using AFM

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Study on the specific interaction between angiogenin and aptamer by atomic force microscopy （AFM）

The specific interaction between angiogenin and aptamer has been investigated by using AFM. The specificity of the interaction is revealed by comparing the binding probability of aptamer to other elements in a series of control experiments. The results have shown that there is specific interaction force between angiogenin and aptamer. Moreover, the single molecular pull-off force between angiogenin and aptamer has also been determined using the Poisson statistical method to be 133.7±11.7 pN. These findings obtained are helpful to the better revelation of recognition mechanism between angiogenin and aptamer, which provided basis for further understanding the inhibition of the aptamer to angiogenic activity.     

壳聚糖与DNA相互作用的单分子方法研究

利用原子力显微镜与磁镊技术研究不同浓度的壳聚糖与多种DNA分子的作用,对其凝聚形态及力谱曲线进行观察。在恒力情况下,壳聚糖与DNA的电荷比k分别为3和4时,从DNA的力谱曲线上可以看见0.2~0.5μm左右的跳跃阶梯及线性增长区域,原因可能是壳聚糖-DNA聚合物中出现了环状、棒状及球状结构;利用原子力显微镜观察壳聚糖与λ-DNA及bpr322-DNA作用后的凝聚形态,发现随着壳聚糖与DNA电荷比k的增加,球状与棒状聚合物的比例明显减小,而且凝聚物的尺寸也整体减小。最后根据实验结果,建立了壳聚糖导致DNA构象变化的直观模型。     

Atomic force microscopic view of the fine topography on the tobacco stigma surface during its response to pollination

During compatible pollination in tobacco, an extracellular matrix （ECM） is secreted from the stigma surface; however, it is unknown whether the pattern of secretion across the stigma depends on the pollen source. In fact, technical limitations have prevented clear observation of ECM secretion. Here, we report the detailed topographic changes on the stigma surface that accompanies intraspecies and interspecies pollination in tobacco using contact mode atomic force microscopy （AFM）. Our results, which show the dynamics and time course of ECM secretion after pollination, indicate that a certain pattern of secretion already exists on the stigma prior to pollination. Intraspecies induced a two-step response, characterized by topographical changes on the stigma surface several hours after pollination, which was distinct from the pattern of ECM secretion induced by interspecies pollination. This difference was confirmed by root-mean-square analysis, which assessed the roughness of the stigma surface. Our findings indicate that compatible pollination not only induces ECM secretion from the stigma, but also results in a specific distribution of the ECM. Thus, this study demonstrates the powerful potential of AFM in studying the pollen-stigma interaction.     

黄原胶分子形貌的原子力显微镜研究

通过溶胶凝胶的方法制备了黄原胶水溶液,并采用轻敲模式原子力显微镜(AFM)对不同浓度的黄原胶分子溶液的表面微观结构形态进行了观察.实验结果表明,轻敲模式下的AFM能够观测到黄原胶分子的微观结构,浓度为0.1g/L时分子类似于球状体的颗粒,以多聚体形式存在.浓度达到0.01g/L时能够形成稳定的网状结构,当浓度达到0.001g/L的情况下形成单个纤维分子.利用AFM截面软件对网状结构和单个纤维进行分析,发现了云母表面出现的黄原胶分子分别为双链和单链.本文中提出了黄原胶分子从网状结构到单个纤维形成过程的假设模型,很好地解释了实验观察结果,对以后的实验研究有很大意义.     

Transmission electron microscopy and atomic force microscopy characterization of nickel deposition on bacterial cells

Recently bacterial cells have become attractive biological templates for the fabrication of metal nano- structures or nanomaterials due to their inherent small size, various standard geometrical shapes and abundant source. In this paper, nickel-coated bacterial cells (gram-negative bacteria of Escherichia coli) were fabricated via electroless chemical plating. Atomic force microscopy (AFM) and transmission electron microscopy (TEM) characterization results reveal evident morphological difference between bacterial cells before and after deposition with nickel. The bare cells with smooth surface presented transverse outspreading effect at mica surface. Great changes took place in surface roughness for those bacterial cells after metallization. A large number of nickel nanoparticles were observed to be equably distributed at bacterial surface after activation and subsequent metallization. Furthermore, ultra thin section analytic results validated the presence and uniformity of thin nickel coating at bacte- rial surface after metallization.     

有机溶剂对纳滤膜结构及透过特性的影响

观察了两种纳滤膜(NF-SH,MPF-44)的稳定性,并利用原子力电镜技术(AFM)观察膜的活性表面结构,发现有机溶剂导致膜结构发生显变化。观测在溶剂中的传递通量以及溶质截留率发现,影响NF-SH膜的通量的主要因素是膜本身的结构;对MPF-44膜,影响膜通量的主要因素是溶剂性质。对于这两种纳滤膜影响截留率的主要因素都是溶剂与溶质的相互作用。     

原子力显微镜针尖与样品间的材料转移

研究氮化硅针尖在十八烷基三甲氧基硅烷 (OTE) /云母表面的修饰过程。使用原子力 /摩擦力显微镜 ,以云母作为参考样品 ,研究了针尖在样品表面的修饰效应和修饰后针尖的清洁过程 ,并考察了湿度和载荷对针尖修饰效应的影响。修饰过程不是一个渐进的过程 ,在最初几次摩擦扫描中修饰较快 ,然后在 10～ 2 0次扫描后达到平衡态。在 OTE/云母表面修饰后的针尖在云母表面的摩擦力信号比修饰前针尖在云母表面的摩擦力信号小 ,并且大部分吸附在针尖表面的 OTE分子在云母表面的前 10次扫描中就被磨掉。相对湿度对针尖的修饰效应影响不大。在研究不同样品的摩擦性能时 ,尽量使用清洁针尖 ,并使用摩擦性能稳定的参考样品 (如云母 )来检测针尖的表面状态     

Si基底上DNA分子的自组装

利用自组装方法将DNA分子吸附于Si基底表面, 并通过原子力显微镜（AFM）观察不同质量浓度的DNA溶液在Si基底表面进行自组装后的结构： 当DNA溶液的质量浓度较低时, Si基底表面会吸附单个、 拉伸、 环状的分子; 当DNA溶液的质量浓度为60~160 ng/μL时, 在Si基底表面会形成网状结构; 当DNA溶液的质量浓度超过200 ng/μL时, 仅形成DNA薄膜. 实验结果表明, 可以利用该方法制作基于Si材料的DNA分子器件.     

DNA与CdS-COOH-EcoRI复合物相互作用的研究

采用荧光光谱、原子力显微镜、琼脂糖凝胶电泳等技术探究CdS-COOH-EcoRI复合物与DNA的相互作用.研究发现,小粒径的CdS-COOH-EcoRI复合物与DNA除了有特异性结合,还发生非特异性结合.DNA相对浓度比较大以及恒温孵育时间较短时,以特异性结合为主;而随着DNA相对浓度的减小以及恒温孵育时间的延长,会伴有非特异性结合.Ca2+存在时,可以通过延长孵育时间“激活”酶切反应,使得DNA被剪切.因此可以通过改变DNA相对浓度和孵育时间来调控EcoRI对DNA的剪切.     

金刚石膜在偏压电场作用下的成核行为

探讨了用微波等离子体化学气相沉积法(MPCVD)在Si(100)衬底上加偏压电场和不加偏压电场情况下金刚石膜的成核行为.并经用原子力显微镜(AFM)分析,偏压电场对金刚石成核有促进作用.文章也分析了偏压电场所以能促进金刚石成核的机制.     

Study on elastic modulus of individual ferritin

The mechanical property of individual ferritin was measured with force-volume mapping (FV) under contact mode of atomic force microscopy (AFM) in this work. The elastic modulus of individual ferritin was estimated by the Hertz mode. The estimated value of the elastic modulus of individual ferritin was about 250－800 MPa under a small deformation. In addition, the elastic modulus of individual ferritin was compared with that of the colloid gold nanoparticle.     

Micro-friction of diazoresin/polyacrylic acid self-assembly films in water with atomic force microscopy

Ultrathin films composed of diazoresin（DR）and polyacrylic acid（PAA）were fabricated.The surface morphology of the films in water was measured using an atomic force microscopy（AFM）.The self-assembly technique makes the surface rather flat and uniform.The friction force and its dependence on the velocity differ from the surface charge of the thin films.The friction force of repulsive DR/PAA film increases linearly with velocity and has lower values than that of attractive DR film over the full range of velocity.As the velocity increases,the attractive friction of DR film first decreases to a minimum at a velocity of 2 line/s and then increases all the way.When the surface is repulsive to the friction substrate,the friction of thin films that is determined by hydrated lubrication of polymer chains that is ultralubricated;when it is adhesive to the friction substrate,the friction is mainly contributed from the elastic deformation of adsorbed polymer chains in the low velocity region and from viscous sliding in the presence of hydrated-layer lubrication of the polymer chains in the higher velocity region.     

Application of image alignment and time averaging methods in AFM detection for single DNA molecules

Enhancement of the SNR (signal to noise ratio) in single-molecule imaging is significantly important for improving image resolu-tion and distinguishing the fine structures of single molecules at a higher precision level.Image processing techniques have dem-onstrated the remarkable capability to improve the SNR and the resolution level by breaking through some inherent limitations unresolved by instrument hardware optimization.In this paper, we focus on single-biomolecule imaging using atomic force mi-crosco...