Mapping CD20 molecules on the lymphoma cell surface using atomic force microscopy

Atomic force microscopy (AFM) was used to locate CD20 molecules on the surface of lymphoma Raji cells. Rituximab (a monoclonal antibody against CD20) molecules were linked onto the AFM tip via a polyethylene glycol (PEG) linker. Raji cells were adsorbed onto glass slides coated with poly-L-lysine. First, the CD20 distribution in a local area of the cell surface was visualized using the AFM lift scan mode. Second, 16 × 16 force curves were obtained from the same cell area to construct the CD20-rituximab binding force map. Finally, free rituximab was added to block the CD20 molecules on the cell surface and the lift phase image and CD20-rituximab force map were obtained again. The experimental results indicated that when the lift height was greater than the length of the PEG linker, no recognition sites were observed in the lift phase image. However, as the lift height decreased to the length of the PEG linker, some recognition sites were observed in the lift phase image and these sites were generally consistent with the pixels in the force map. After blocking, both the recognition sites in the lift phase image and the gray pixels in the binding force map decreased markedly. These results can improve our understanding of the distribution of protein molecules on the cell surface and facilitate further investigations into cellular functions.     

橡胶混凝土改良之研究

探讨橡胶混凝土之改善,借由橡胶表面改质技术的开发,并应用原子力显微镜直接量测橡胶以水泥水化物间之作用力,以作为橡胶表面改质的参考.文中除做简单之文宪回顾外,并以三种表面改质技术为例,说明橡胶混凝土改进方法之研究方法.     

HCAP1基因对Raji细胞凋亡及凋亡相关蛋白Bax/Bcl - 2的作用

目的：研究外源HCAP1基因产物对Burkitt淋巴瘤细胞系Raji细胞凋亡及凋亡相关蛋白Bax／Bcl-2的作用。方法：用脂质体介导的基因转移方法，把外源HCAP1基因转染到Raji细胞中，用流式细胞术和Hochest 33258荧光染色检测细胞凋亡；用Western blot检测外源HCAP1基因对凋亡相关蛋白Bax和Bcl-2蛋白表达的调节。结果：外源HCAPl基因导入Raji细胞24、48和96h后，细胞凋亡率增加。Western blot显示Bax／Bel-2蛋白表达比值明显增高。结论：转染外源性HCAP1基因可诱导Raji细胞凋亡，使Bax／Bcl-2蛋白表达比例上调可能是HCAP1诱导凋亡的机制之一。     

Research progress in quantifying the mechanical properties of single living cells using atomic force microscopy

The advent of atomic force microscopy （AFM） provides a powerful tool for investigating the behaviors of single living cells under near physiological conditions. Besides acquiring the images of cellular ultra-microstruc- tures with nanometer resolution, the most remarkable advances are achieved on the use of AFM indenting tech- nique to quantify the mechanical properties of single living ceils. By indenting single living cells with AFM tip, we can obtain the mechanical properties of cells and monitor their dynamic changes during the biological processes （e.g., after the stimulation of drugs）. AFM indentation-based mechan- ical analysis of single cells provides a novel approach to characterize the behaviors of cells from the perspective of biomechanics, considerably complementing the traditional biological experimental methods. Now, AFM indentation technique has been widely used in the life sciences, yielding a large amount of novel information that is meaningful to our understanding of the underlying mechanisms that govern the cellular biological functions. Here, based on the authors＇ own researches on AFM measurement of cellular mechani- cal properties, the principle and method of AFM indentationtechnique was presented, the recent progress of measuring the cellular mechanical properties using AFM was summa- rized, and the challenges of AFM single-cell nanomechani- cal analysis were discussed.     

用原子力显微镜观察活海马区神经元三维超微结构

在液体环境中,利用原子力显微镜成功地对体外培养的大鼠海马区神经元进行了超微结构的成像研究。分别获取了活海马神经元胞体、轴突、树突以及生长锥等结构清晰的原子力显微图像,并对其三维结构进行了定量分析。为进一步利用原子力显微镜研究海马神经元打下了基础。     

利用原子力显微镜测量石英岩表面分子沉积膜的粘附力

对原子力显微镜（ＡＦＭ）探针测出的一条理想的力位移曲线进行了分析,推导出了根据实测的力位移曲线计算粘附力的公式。对表面分子沉积膜生长前后的石英岩和扫描探针之间的粘附力进行了实验研究,结果发现这种沉积膜可以降低石英岩表面的粘附力     

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.     

原子力显微镜在癌细胞观察和研究中的应用

原子力显微镜(AFM),能够在接近生理条件下以具有原子级的分辨率对活细胞进行表面成像和超微结构观察,同时可以研究细胞的生物过程、细胞与药物之间和细胞之间的相互作用,成为细胞生物学研究的一种有效工具.近年来AFM在细胞生物学研究中的应用进展很快,许多研究成果在生物医学和临床医学方面有良好的应用前景.本文分析了原子力显微镜的成像机理、工作模式和技术要点,综述了原子力显微镜在癌细胞的研究应用现状和前景.     

Progress of AFM single-cell and single-molecule morphology imaging

Atomic force microscopy (AFM) can probe single living cells and single native membrane proteins in natural fluid environments with label-free high spatial resolution. It has thus become an important tool for cellular and molecular biology that significantly complements traditional biochemical and biophysical techniques such as optical and electron microscopy and X-ray crystallog-raphy. Imaging surface topography is the primary application of AFM in the life sciences. Since the early 1990s, researchers have used AFM to investigate morphological features of living cells and native membrane proteins with impressive results. Steady improvements in AFM techniques for imaging soft biological samples have greatly expanded its applications. Based on the authors’ own research in AFM imaging of living cell morphologies, a review of sample preparation procedures for single-cell and single-molecule imaging experiments is presented, along with a summary of recent progress in AFM imaging of living cells and native membrane proteins. Finally, the challenges of AFM high-resolution imaging at the single-cell and single-molecule levels are discussed.     

间歇模原子力显微镜扫描探针的受迫振动解

提出了一个间隙模原子力显微镜全动态工作的模型 ,用参数变换和模式展开方法 ,得到了具有粘滞阻尼和分段线性相互作用力的扫描探针的受迫弯曲振动微分方程的分析解 ,间歇模原子力显微镜扫描探针的受迫振动解 ,由接触模解和非接触模解交替工作而构成     

影响电镀紫铜胎圈钢丝粘合力因素的工艺探索

对胎圈钢丝电镀紫铜生产过程中不同工艺参数下钢丝粘合力的变化进行研究和分析,探索了影响钢丝粘合力的工艺因素,获得了最佳生产工艺参数,达到了预期目标。     

Experimental Study on the Surface Modification of Ultra Thin DLC Films

Surface modification of Diamond-like carbon （DLC） films was carried out in order to estimate the reliability of the ultra thin DLC films. The wear resistance, conductivity and mechatronic reliability of the films were studied by contact atomic force microscope （AFM）, electric force microscope （EFM） and conductive AFM. The failure mechanism of pits formed and the reason for conductivity changed of DLC films were examined.     

石油钻机液力变矩器轴向力的分析与计算

对石油钻机液力变矩器各工作轮中液体的静压力分布进行了分析,由此得出了液力变矩器各工作轮轴向力的理论计算公式。实例计算发现,泵轮的轴向力拉动泵轮向涡轮靠近;涡轮的轴向力拉动涡轮向泵轮靠近;导轮的轴向力拉动导轮向泵轮靠近。三个工作轮的轴向力随着变矩器的工况变化,在起动工况时达到最大值。泵轮的轴向力最大,且变化起伏较大;其他两轮的轴向力变化较平稳,且数值也较小。轴向力除了与供油压力有关外,主要是与泵轮转速的二次方和循环圆有效直径的四次方有关     

多孔硅的电化学形成及微结构研究

采用自制的电解池用电化学腐蚀方法制备多孔硅，使用正交试验法设计阳极腐蚀参数。使用原子力显微镜研究多孔硅的微结构，分析并讨论了实验参数对多孔硅微结构的影响，确定了最佳制备工艺。     

A novel colloid probe preparation method based on chemical etching technique

Several fundamental problems in hydrophobic force measurements using atomic force microscope (AFM) are discussed in this paper. A novel method for colloid probe preparation based on chemical etching technology is proposed, which is specially fit for the unique demands of hydrophobic force measurements by AFM. The features of three different approaches for determining spring constants of rectangular cantilevers, including geometric dimension, Cleveland and Sader methods are compared. The influences of the sizes of the colloids on the measurements of the hydrophobic force curves are investigated. Our experimental results showed that by selecting colloid probe with proper spring constant and tip size, the hydrophobic force and the complete hydrophobic interaction force curve can be measured by using AFM.     

刀翼式PDC钻头的侧向力平衡设计

聚晶金刚石(PDC)钻头的侧向不平衡力是造成钻头涡动的主要原因之一,而侧向不平衡力取决于钻头的布齿结构,通过合理的布齿结构设计,可有效地控制侧向不平衡力的大小。在对PDC钻头进行受力分析的基础上,建立了PDC钻头受力计算模型,提出了通过调整刀翼周向位置使钻头的侧向不平衡力达到最小的优化设计方法。利用该方法进行了实例计算,结果表明,该设计方法能将PDC钻头的侧向不平衡力控制在钻压的1%以内。     

邻苯二甲酸二丁酯与胰蛋白酶的相互作用

在模拟人体生理条件下(pH值为7.4),应用荧光光谱法、紫外光谱法和圆二色谱法(CD)并结合原子力显微镜(AFM)和分子模拟技术,研究了邻苯二甲酸二丁酯(DBP)对胰蛋白酶(Trypsin)的光谱性质、结构及催化活性的影响.结果表明,DBP通过氢键和范德华力与胰蛋白酶形成基态复合物而猝灭胰蛋白酶的内源荧光,DBP在胰蛋白酶上只有1个结合位点.同步荧光、紫外和CD光谱研究发现,DBP与胰蛋白酶的结合诱导了酶的α-螺旋、β-折叠和β-转角含量的减少,而增加了无规卷曲的含量.原子力显微镜图像显示,DBP的存在引起胰蛋白酶的表面形态发生变化,蛋白质发生了聚集.分子模拟结果表明,DBP结合于胰蛋白酶S1疏水空腔附近,与氨基酸His 57,Ser 195和Gly 193形成氢键.酶活测定结果显示,DBP的存在导致胰蛋白酶活性被抑制.     

Assembling Zwitterionic, Water-Soluble Porphyrin and Its Biological Studies

A kind of zwitterionic water-soluble porphyrin 5-(N-trimethyl-4-aminiumphenyl)-10, 15, 20-tri (4-sulfonatephenyl) porphyrin trisammonium salt, has been synthesized. Its biological studies, interactions with DNA and bacteria cells, were investigated. The damage to the cell wall of bacteria E. coli were observed firstly by AFM (Atomic Force Microscope).     

Progress in measuring biophysical properties of membrane proteins with AFM single-molecule force spectroscopy

Membrane proteins are crucial in cell physiological activities and are the targets for most drugs.Thus,investigating the behaviors of membrane proteins not only provide deeper insights into cell function,but also help disease treatment and drug development.Atomic force microscopy is a unique tool for investigating the structure of membrane proteins.It can both image the morphology of single native membrane proteins with high resolution and,via single-molecule force spectroscopy(SMFS),directly measure their biophysical properties during molecular physiological activities such as ligand binding and protein unfolding.In the context of molecular biomechanics,SMFS has been successfully used to understand the structure and function of membrane proteins,complementing the static three-dimensional structures of proteins obtained by X-ray crystallography.Here,based on the authors’antigen-antibody binding force measurements in clinical tumor cells,the principle and method of SMFS is discussed,the progress in using SMFS to characterize membrane proteins is summarized,and challenges for SMFS are presented.     

基于局部摩擦因数模型的切削力预测建模

基于局部摩擦因数模型分别建立前刀面摩擦区、切削刃钝圆区、后刀面摩擦区的受力预测模型,进而获得切削力预测值.以钨钼系高速钢（W6Mo5Cr4V2Al）刀具和20Cr2Ni4合金钢为研究对象建立直角切削实验,通过三向测力仪测量直角切削主切削力和切深抗力,并与预测切削力进行对比,数值基本吻合.分析了切削参数以及刀具前角对切削力大小的影响规律.结果表明切削力随切削速度和刀具前角的增加有减小的趋势,随着切削深度的增加明显增大.