原子力显微镜扫描成像DNA分子

采用Mg2+处理DNA、APTES或戊二醛修饰云母表面、DNA拉直方法制备了λ-DNA及DNA-组蛋白复合物样品.室温下原子力显微镜以轻敲模式在空气中扫描样品成像.实验结果表明:AFM扫描成像的效果与样品的制备方法有关,同时也受操作因素影响.     

人肝癌SMMC-7721细胞对低剂量γ射线辐射超敏感性的研究

研究了人肝癌细胞SMMC-7721在低剂量γ射线照射下超敏感性和增强的辐射抗性响应。选用对数生长期细胞接受0—6 Gy不同剂量的^60Coγ射线的照射。利用流式细胞仪对细胞进行分选计数,并用克隆形成法检测细胞存活率。发现SMMC-7721细胞存在低剂量辐射超敏感性和增强的辐射抗性响应,即在0—0．3Gy之间细胞表现出单位剂量杀伤增强现象,在0．3—1Gy细胞表现一定的辐射抗性,在1Gy以上,细胞的存活符合线性平方模型。     

原子力显微镜在分子细胞生物学研究中的应用

 1986年秉霖(G.Binning)等在扫描隧道显微镜的基础上发明了原子力显微镜(Atomicforcemicro-scope,AFM)。这种显微镜的放大倍数远远超过以往的任何显微镜,可以直接观察物质的分子和原子组成,这为微观世界的探索提供了理想的工具。AFM不仅可以以高分辨率表征样品表面形貌,分析研究与作用力相对应的各种表面性质,并可对样品的分子或原子进行纳米级力加工,也能对活的生命样品进行实时动态观测。这些特性使AFM在生命科学特别是在分子细胞生物学的研究中占据着独特的地位。一、AFM对细胞表面结构的研究AFM的样品制备简单,只需作一个渗涂片并在空气中干燥,且不需特殊的染色和固定;它的     

基于迁移学习的原子力显微镜成像恢复方法

受限于探针针尖结构尺寸,用原子力显微镜进行微纳测量时会产生图像边缘失真.提出了一种基于迁移学习的原子力显微镜成像恢复方法,通过迁移学习训练源模型和靶模型实现一维栅格成像恢复.该方法采用数学形态法中的腐蚀算法生成栅格点云数据,通过U-Net网络源模型从点云中提取针尖卷积效应的特征向量,将权重参数迁移至U-Net网络靶模型,靶模型在自适应正则化方法下进行监督学习.实验结果表明,该方法能有效恢复一维栅格的原子力显微镜测量图像,提高横向分辨力,可用于纳米栅格的线宽检测上.     

活细胞内单个大分子的行为

以往细胞内大分子活动的研究,多数是许多分子活动的一个平均结果,即一种集群平均（ensemble averaging）的结果,随着各种技术,特别是光学及荧光检测技术的成熟,实时视见活细胞内单个大分子的时代已经到来.在现时条件下,有许多方法可供选择,如荧光共振能量转移、原子力显微镜、全内反射显微技术、荧光相关光谱法等等.“活细胞内单个大分子的行为”的研究有可能为了解活细胞内单个分子的活动带来完全新的认识,但目前还存在不少方法学上的局限性,有待进一步提高,如有效的特异标记物、细胞深部荧光的检测、新型显微镜的开发等.     

细胞研究的光学显微成像

介绍了在细胞及亚细胞结构与功能研究中发挥至关重要作用的各种现代光学显微镜,包括近场显微镜、共焦扫描显微镜、双光子成像显微镜、图像恢复显微镜及时间相关单光子计数技术,着重描述了后两种系统的工作原理及先进功能。     

双波段全天空云量观测系统研制及数据分析

为实现全天时、全天空云量观测,提出并研制了一种双波段全天空云量观测系统。该系统在可见光波段通过内置遮光装置和高动态范围（HDR）图像合成技术,可采集清晰完整的全天空图像,同时,其在红外波段的一次成像可获取160°视场范围天空图像,较传统扫描拼接式红外观测模式更加简单快捷。主要阐述了该系统在不同波段的全天空成像原理以及云图分割方法。从双波段云量观测数据的一致性和准确性两方面进行分析,验证了该系统在云量观测上具有较高的准确性。     

利用扫描隧道显微镜在溶液中直接观测过渡族金属硫属化合物（英文）

本文利用自制的溶液扫描隧道显微镜在溶液环境下直接观测到TiSe2、MoTe_2和TaS_2样品的原子分辨率图像.通过将单晶样品在溶液中直接解理,可以保护解理过的新鲜样品表面在几个小时内不会被严重污染.利用自行搭建的溶液扫描隧道显微镜,首先观察到了TiSe2活泼样品的原子分辨率图像,并观察到了TiSe2表面所特有的点缺陷和三角形缺陷结构.此外,还观察到了MoTe2的原子分辨率超结构和TaS2表面的电荷密度波结构.结果表明:在室温、溶液环境下能更高效的研究过渡族金属硫属化合物等活泼样品的表面电子态结构,同样适用于溶液环境下的电催化和电化学研究.     

多光参量测量成像技术用于雾霾天气的实验研究

针对雾霾天气条件下目标对比度下降问题,研制成功了一套多光参量测量成像系统。通过多光参量成像的系统级辐射响应模型及模型参数分离式标定,实现雾霾天气情况下的多种场景目标成像实验观测。结果显示:楼房玻璃反射光中的偏振分量相对较强,通过线偏振、圆偏振、偏振角图像,可以清晰地分辨玻璃窗结构;普通的强度信息很难辨别雾霾天气下的路口车辆,在线偏振及偏振角图像中能够看到车辆,但是掺杂了树木的偏振信息,车辆信息较模糊;在圆偏振信息图像中,树木的圆偏振信息较少,车辆目标对比度提高20%。     

利用扫描近场红外显微镜对化学样品组分进行成像研究

介绍了用北京自由电子激光为光源的扫描近场红外显微镜对化学样品组分进行的成像研究，通过所得到的近场微区图像，可以对样品在微区范围内的成分组成，混合的均匀程度等作出比较清晰的判断. 关键词： 自由电子激光 近场光学 扫描近场红外显微镜     

Scanning ion conductance microscopy for imaging biological samples in liquid: a comparative study with atomic force microscopy and scanning electron microscopy

The present study was designed to show the applicability of scanning ion conductance microscopy (SICM) for imaging different types of biological samples. For this purpose, we first applied SICM to image collagen fibrils and showed the usefulness of the approach-retract scanning (ARS)/hopping mode for such samples with steep slopes. Comparison of SICM images with those obtained by AFM revealed that the ARS/hopping SICM mode can probe the surface topography of collagen fibrils and chromosomes at nanoscale resolution under liquid conditions. In addition, we successfully imaged cultured HeLa cells, with 15 μm in height by ARS/hopping SICM mode. Because SICM can obtain non-contact (or force-free) images, delicate cellular projections were visualized on the surface of the fixed cell. SICM imaging of live HeLa cells further demonstrated its applicability to study the morphological dynamics associated with biological processes on the time scale of minutes under liquid conditions. We further applied SICM for imaging the luminal surface of the trachea and succeeded in visualizing the surface of both ciliated and non-ciliated cells. These SICM images were comparable with those obtained by scanning electron microscopy. Although the dynamic mode of AFM provides better resolution than the ARS/hopping mode of SICM in some samples, only the latter can obtain contact-free images of samples with steep slopes, rendering it an important tool for observing live cells as well as unfixed or fixed soft samples with complicated shapes. Taken together, we demonstrate that SICM imaging, especially using an ARS/hopping mode, is a useful technique with unique capabilities for imaging the three-dimensional topography of a range of biological samples under physiologically relevant aqueous conditions.     

Scanning tunnelling microscopy imaging and modification of hydrogen-passivated Ge(100) surfaces

Scanning tunnelling microscopy (STM) study and modification of hydrogen (H)-passivated Ge(100) surfaces have been investigated. Thermal oxidation procedures were used to minimise surface roughness. Ge samples were passivated in HF solution after thermal oxidation. STM and atomic force microscope (AFM) imaging showed that, using HF etching after thermal oxidation, we can obtain a natural H-passivatedtopographically and chemically flat Ge(100) surface. The root-mean-square (rms) roughness ofa H-passivatedGe(100) surface measured both by STM and AFM is less than 2 ?. Electric properties of H-passivatedGe(100) surfaces were studied by scanning tunnelling spectroscopy (STS) in nitrogen ambient. STS showed that the H-passivated Ge surfaces were not pinned. Modification on H-passivated Ge(100) surfaces was carried out using STM by applying an electric voltage between the sample and tip in air. Modified features were characterised by STM and AFM imaging. On the H-passivated Ge(100) surfaces, stable, low-voltage, nanometer-scale modified features can be produced.     

Application of atomic force microscopy to microbial surfaces: from reconstituted cell surface layers to living cells.

The application of atomic force microscopy (AFM) to probe the ultrastructure and physical properties of microbial cell surfaces is reviewed. The unique capabilities of AFM can be summarized as follows: imaging surface topography with (sub)nanometer lateral resolution; examining biological specimens under physiological conditions; measuring local properties and interaction forces. AFM is being used increasingly for: (i) visualizing the surface ultrastructure of microbial cell surface layers, including bacterial S-layers, purple membranes, porin OmpF crystals and fungal rodlet layers; (ii) monitoring conformational changes of individual membrane proteins; (iii) examining the morphology of bacterial biofilms, (iv) revealing the nanoscale structure of living microbial cells, including fungi, yeasts and bacteria, (v) mapping interaction forces at microbial surfaces, such as van der Waals and electrostatic forces, solvation forces, and steric/bridging forces; and (vi) probing the local mechanical properties of cell surface layers and of single cells.     

High speed indentation measures by FV,QI and QNM introduce a new understanding of bionanomechanical experiments

Structural and mechanical mapping at the nanoscale by novel high-speed multiparametric Quantitative Imaging (QI) and PeakForce Quantitative Nanomechanical Mapping (PF-QNM) AFM modes was compared to the classical Force Volume (FV) mapping for the case of living Pseudomonas aeruginosa bacterial cells. QI and PF-QNM modes give results consistent with FV for the whole cells in terms of morphology and elastic modulus, while providing higher resolution and shorter acquisition time. As an important complement, the influence of scanning parameters on elastic modulus values was explored for small 0.2² μm² central area on top of cells. The modulus decreases with the indentation depth due to the effect of the hard cell wall, while it increases vs. tip oscillation frequency, displaying viscoelastic behaviour of the living bacterial cells. The ability of different AFM modes to follow correctly the bacteria viscoelastic behaviour at high oscillation frequency was tested.     

AFM imaging of fenestrated liver sinusoidal endothelial cells

Each microscope with its dedicated sample preparation technique provides the investigator with a specific set of data giving an instrument-determined (or restricted) insight into the structure and function of a tissue, a cell or parts thereof. Stepwise improvements in existing techniques, both instrumental and preparative, can sometimes cross barriers in resolution and image quality. Of course, investigators get really excited when completely new principles of microscopy and imaging are offered in promising new instruments, such as the AFM. The present paper summarizes a first phase of studies on the thin endothelial cells of the liver. It describes the preparation-dependent differences in AFM imaging of these cells after isolation. Special point of interest concerned the dynamics of the fenestrae, thought to filter lipid-carrying particles during their transport from the blood to the liver cells. It also describes the attempts to image the details of these cells when alive in cell cultures. It explains what physical conditions, mainly contributed to the scanning stylus, are thought to play a part in the limitations in imaging these cells. The AFM also offers promising specifications to those interested in cell surface details, such as membrane-associated structures, receptors, coated pits, cellular junctions and molecular aggregations or domains. The AFM also offers nano-manipulation possibilities, strengths and elasticity measurements, force interactions, affinity measurements, stiffness and other physical aspects of membranes and cytoskeleton. The potential for molecular approaches is there. New developments in cantilever construction and computer software promise to bring real time video imaging to the AFM. Home made accessories for the first generation of AFM are now commodities in commercial instruments and make the life of the AFM microscopist easier. Also, the combination of different microscopies, such as AFM and TEM, or AFM and SEM find their way to the market allowing comfortable correlative microscopy.     

Cortical rigidity of round cells in mitotic phase and suspended state

This paper describes the results of the analysis of cortical rigidity in two round cell states: mitotic round cells and detached round cells after trypsinization using atomic force microscopy (AFM). These two states are primary cell events with dynamic morphological alterations in vitro. The trypsinized detached cells were fixed on the substrate of membrane anchoring oleyl surface. Fluorescent images taken by confocal laser scanning microscopy revealed diverse cell surface protrusions and cortical actin development in the round cells under different conditions. Although the cortical actin of these cells seemed to develop similarly, cortical rigidity of the trypsinized round cells showed greater stiffness than that of mitotic round cells. The elasticity measurements by AFM may detect invisible information about the maturation or strength of F-actin structures and such measurements may indicate that the strength of the actomyosin cortex would be higher in trypsinized round cells compared to mitotic cells. The mechanical properties can help provide better insights into the characteristics of the actin cytoskeleton network in vicinity of cell surface during dynamic morphological alterations.     

Atomic force microscopy probing of cell elasticity

Atomic force microscopy (AFM) has recently provided the great progress in the study of micro- and nanostructures including living cells and cell organelles. Modern AFM techniques allow solving a number of problems of cell biomechanics due to simultaneous evaluation of the local mechanical properties and the topography of the living cells at a high spatial resolution and force sensitivity. Particularly, force spectroscopy is used for mapping mechanical properties of a single cell that provides information on cellular structures including cytoskeleton structure.

This entry is aimed to review the recent AFM applications for the study of dynamics and mechanical properties of intact cells associated with different cell events such as locomotion, differentiation and aging, physiological activation and electromotility, as well as cell pathology. Local mechanical characteristics of different cell types including muscle cells, endothelial and epithelial cells, neurons and glial cells, fibroblasts and osteoblasts, blood cells and sensory cells are analyzed in this paper.     

Atomic force microscopy studies of chemical–mechanical processes on silicon(100) surfaces

Atomic force microscopy (AFM) is used to examine chemical–mechanical processes on Si(100) surfaces. The AFM tip serves as a single asperity contact to exert tribological forces as well as an imaging tool. By scanning in chemically aggressive solutions, material removal can be observed directly. In the silicon system, high-force scans are used to remove oxide and initiate etching in selected locations, followed by low-force scans to image the resulting surfaces. Material removal rates were measured as a function of applied load, number of scans, solution composition, and time. In basic solution, places where the underlying silicon is exposed etch rapidly, producing structures 100 nm or less in size. Although the surface roughness initially increases during etching, the final surfaces are smooth. The oxide is extremely sensitive to applied stress: even very light scanning accelerates oxide dissolution. Once the oxide is removed, chemical etching proceeds through the underlying silicon with or without AFM scanning; but the silicon etches more rapidly if AFM scanning is continued, due to true chemical–mechanical (tribochemical) effects.     

Analyses of vibration responses on nanoscale processing in a liquid using tapping-mode atomic force microscopy

An analytical solution of the vibration responses of biological specimens using atomic force microscopy (AFM), which often requires operation in a liquid, is developed. In this study, the modal superposition method is employed to analyze the vibration responses of AFM cantilevers in tapping mode (TM) operated in a liquid and in air. The hydrodynamic force exerted by the fluid on AFM cantilevers is approximated by additional mass and hydrodynamic damping. The tip–sample interaction forces were transformed into axial, distributed transversal, and bending loading, and then applied to the end region of the AFM through the tip holder. The effects of transverse stress and bending stress were adopted to solve the dynamic model. With this model, a number of simulations were carried out to investigate the relationship between the transient responses of the cantilever in a liquid and the parameters considered in nanoscale processing. The simulations show that the vibration of AFM cantilevers in a liquid has dramatically different dynamic characteristics from these of that in air. The liquid reduces the magnitude of the transversal response and reduces the cantilever resonances. Moreover, the magnitudes of response become larger with increasing intermolecular distances and smaller with decreasing tip length. The cantilever vibration amplitudes significantly depend on the damping constant and the mass proportionality constant.     

拉曼光谱研究复方鹿仙草颗粒对SMMC-7721肝癌细胞的作用

应用拉曼光谱研究了不同浓度的鹿仙草溶液与肝癌细胞SMMC-7721的相互作用,通过对药物作用前后细胞的光谱变化进行分析,从而为阐明鹿仙草与肝癌细胞的作用方式提供重要的依据。拉曼光谱显示,加入鹿仙草后,细胞的许多峰都发生了变化。归属于磷酸骨架振动的785和1 092 cm-1的两个峰强度下降,对应碱基A和G的峰1 312和1 585 cm-1 等也有不同程度的降低,表明鹿仙草可能插入DNA碱基对之间,使DNA复制受到抑制,导致细胞DNA含量下降,而且会引起DNA单、双链的断裂。同时,研究还发现属于蛋白质的振动峰(1 005, 1 360, 1 656 cm-1)强度也有不同程度下降,说明蛋白质二级结构以及侧链氨基酸的环境均发生了改变。此外,鹿仙草对细胞的作用效果随浓度增加逐步增强。