Surface modification of AFM silicon probes for adhesion and wear reduction

Tip wear of silicon probes used for an atomic force microscope (AFM) is a critical issue. Wear can result in an increase of tip radius and adhesion between tip and sample, thus reducing the image resolution and introducing artifacts. In order to reduce adhesion, friction, and wear so as to reduce tip related artifacts, liquid lubricant (Z-TETRAOL), self-assembled monolayers (pentafluorophenyltriethoxysilane (PFPTES)), and fluorocarbon polymer (Fluorinert™) were applied on the silicon probe. A comprehensive investigation of adhesion, friction, and wear of the uncoated/coated tips in both ambient air and various humidity levels as well as the influence of the coatings on the image resolution was performed. Experiments showed that the coatings reduced the adhesion, friction, and wear of the silicon tip, improved the initial image resolution, and exhibited less deterioration as compared to that of uncoated tip in the long-term test.     

AFM picking-up manipulation of the metaphase chromosome fragment by using the tweezers-type probe

We have studied the development of a new procedure based on atomic force microscopy (AFM) for the analysis of metaphase chromosome. The aim of this study was to obtain detailed information about the specific locations of genes on the metaphase chromosome. In this research, we performed the manipulation of the metaphase chromosome by using novel AFM probes to obtain chromosome fragments of a smaller size than the ones obtained using the conventional methods, such as glass microneedles. We could pick up the fragment of the metaphase chromosome dissected by the knife-edged probe by using our tweezers-type probe.     

双探针原子力显微镜视觉对准系统

传统的原子力显微镜（AFM）受针尖形状和放置方式的影响很难测量线条的宽度和两个侧壁的形状,故本文提出采用双探针对顶测量方案来消除AFM针尖形状对测量结果的影响。介绍了一种基于机器视觉的双探针原子力显微镜对准系统,该系统将两个探针接触到一起,实现了双探针在三维方向上的对准。系统采用具有亚微米级分辨率的镜头,配合高分辨率的CCD来获得探针的清晰图像,用于在水平和垂直两个方向实时监控双探针的运动情况。采用基于石英音叉式的自传感自调节的原子力探针,无需外加光学探测系统,缩小了系统体积,避免了杂散光对视觉对准系统的干扰。最后对针尖进行了亚像素边缘提取,精确地获取了探针之间的相对位置,实现了亚微米级的双探针对准（1 μm以内）。该结论由探针之间距离与幅度/相位曲线得到了验证。     

纳结构的连续激光复合微纳探针刻划加工

为了加工形貌稳定且尺寸尽可能小的纳结构,建立了一套连续激光复合微纳探针的加工系统,并研究了光纤探针导光的连续激光辐照微纳探针的近场增强效应以及该系统的加工性能。首先,根据表面等离子体激元理论仿真分析了激光辐照原子力显微镜(AFM)探针的近场增强因子,并研究了微纳探针的针尖温度场和针尖热膨胀。接着,搭建了基于光纤探针导光的连续激光复合微纳探针的纳结构加工系统。最后,对聚乙烯片状材料样品进行了纳结构加工。结果显示:加工得到的纳米点尺度为200nm左右;纳米线的尺度为30~40nm。结果表明:光纤探针导光连续激光复合微纳探针系统避免了复杂的空间光路结构,是一种成本低廉,结构简单的系统,能够实现纳结构的加工。     

Modulus characterization of cells with submicron colloidal probes by atomic force microscope

Colloidal probes have been increasingly demanded for the characterization of cellular modulus in atomic force microscope because of their well-defined geometry and large contact area with cell. In this work, submicron colloidal probes are prepared by scanning electron microscope/focused ion beam and compared with sharp tip and micron colloidal probe, in conjunction with loading velocity and indentation depth on the apparent elastic modulus. NIM and cartilage cells are used as specimens. The results show that modulus value measured by sharp tip changes significantly with loading velocity while remains almost stable by colloidal probes. Also, submicron colloidal probe is superior in characterizing the modulus with increasing indentation depth, which could help reveal the mechanical details of cellular membrane and the modulus of the whole cell. To test the submicron colloidal probe further, the modulus distribution map of cell is scanned with submicron colloidal probe of 50 nm radius during small and large indentation depths with high spatial resolution. The outcome of this work will provide the effective submicron colloidal probe according to the effect of loading velocity and indentation depth, characterizing the mechanical properties of the cells.     

Mapping of enzyme activity by detection of enzymatic products during AFM imaging with integrated SECM-AFM probes

With the integration of submicro- and nanoelectrodes into atomic force microscopy (AFM) probes using microfabrication techniques, an elegant approach combining scanning electrochemical microscopy (SECM) with AFM has recently been introduced. Simultaneous contact mode imaging of a micropatterned sample with immobilized enzyme spots and imaging of enzyme activity is shown. In contrast to force spectroscopy the conversion of an enzymatic byproduct is directly detected during AFM imaging and correlated to the activity of the enzyme.     

Facilitating the pickup of individual DNA molecules by AFM nanomanipulation with tips mechanically worn on bare mica

The tip is one of the critical factors to improve the efficiency in picking up individual DNA molecules from solid substrates based on atomic force microscope (AFM) nanomanipulation. We found that wearing AFM tips on certain solid substrates in advance to nanomanipulation operation would largely improve the pickup efficiency, which was ascribed to the increasing affinity of the tip to the DNA molecules along with the increase of the tip radius after wearing. It was demonstrated that bare mica was superior to APTES-modified mica to keep the tip clean while wearing, which was crucial for DNA pickup during AFM nanomanipulation.     

金属薄膜导线的亚微米局域电导率精确测量技术

作为薄膜器件最重要物理量之一的局域电导率的定量测定,能在保证性能、提高成品率、完善制作工艺等方面起关键作用.利用基于原子力显微镜(Atomic force microscope,AFM)的4电极微探针局域电导率测量技术,精确测量厚度为350nm、宽度分别为50.0 μm、25.0μm、5.0 μm、2.0μm及600 ...     

Sensitivity and resonant frequency of an AFM with sidewall and top-surface probes for both flexural and torsional modes

The resonant frequencies and flexural sensitivities of an atomic force microscope (AFM) with assembled cantilever probe (ACP) are studied. This ACP comprises a horizontal cantilever, a vertical extension and two tips located at the free ends of the cantilever and the extension, which makes the AFM capable of simultaneous topography at top surface and sidewalls of microstructures especially microgears, which consequently leads to a time-saving swift scanning process. In this work, the effects of the sample surface contact stiffness and the geometrical parameters such as the ratio of the vertical extension length to the horizontal cantilever length and the distance of the vertical extension from clamped end of the horizontal cantilever on both flexural and torsional resonant frequencies and sensitivities are assessed. These geometrical effects are illustrated in some figures. The results show that the low-order vibration modes are more sensitive for low values of the contact stiffness, but the situation is reversed for high values.     

Analysis of photopolymerized acrylic films by AFM in pulsed force mode

This paper highlights the potential of atomic force microscopy in the pulsed force mode to investigate the photopatterning of acrylic‐based films. The pulsed force mode is a nonresonant mode designed to allow approach curves to be recorded along the scanning path. It thereby provides the topography of the sample and a direct and simple local characterization of adhesion and stiffness. This mode can be used either for imaging or for locally probing the mechanical properties of a surface. In particular, a correlation between stiffness and conversion of the monomer was established. The close examination of the pulsed force mode signal brought accurate information on the photoinduced modification of the film. Polymer films with submicron photopatterning generated by interferometric illumination were analyzed by pulsed force mode. It was established that the gradient of mechanical properties throughout the films was strongly dependant on the irradiation conditions.     

Phase imaging quality improvement by modification of AFM probes’ cantilever

Imaging of the surface of materials by atomic force microscopy under tapping and phase imaging mode, with use of modified probes is addressed. In this study, the circularly shaped holes located in varying distance from the probe base, were cut out by focused ion beam. Such modification was a consequence of the results of the previous experiments (probe tip sharpening and cantilever thinning) where significant improvement of image quality in tapping and phase imaging mode has been revealed. The solution proposed herein gives similar results, but is much simpler from the technological point of view. Shorter exposition time of the tip onto gallium ions during FIB processing allows to reduce material degradation. The aim of this modification was to change harmonic oscillators’ properties in the simplest and fastest way, to obtain stronger signal for higher resonant frequencies, which can be advantageous for improving the quality of imaging in PI mode. Probes shaped in that way were used for AFM investigations with Bruker AFM nanoscope 8. As a testing material, titanium roughness standard sample, supplied by Bruker, was used. The results have shown that the modifications performed within these studies influence the oscillation of the probes, which in some cases may result in deterioration of the imaging quality under tapping mode for one or both self‐resonant frequencies. However, phase imaging results obtained using modified probes are of higher quality. The numerical simulations performed by application of finite element method were used to explain the results obtained experimentally. Phenomenon described within this study allows to apply developed modelling methodology for prediction of effects of various modifications on the probes' tip, and as a result, to predict how proposed modifications will affect AFM imaging quality.     

Comparisons of the topographic characteristics and electrical charge distributions among Babesia‐infected erythrocytes and extraerythrocytic merozoites using AFM

Tick‐borne Babesia parasites are responsible for costly diseases worldwide. Improved control and prevention tools are urgently needed, but development of such tools is limited by numerous gaps in knowledge of the parasite–host relationships. We hereby used atomic force microscopy (AFM) and frequency‐modulated Kelvin probe potential microscopy (FM‐KPFM) techniques to compare size, texture, roughness and surface potential of normal and infected Babesia bovis, B. bigemina and B. caballi erythrocytes to better understand the physical properties of these parasites. In addition, AFM and FM‐KPFM allowed a detailed view of extraerythrocytic merozoites revealing shape, topography and surface potential of paired and single parasites. B. bovis‐infected erythrocytes display distinct surface texture and overall roughness compared to noninfected erythrocytes. Interestingly, B. caballi‐infected erythrocytes do not display the surface ridges typical in B. bovis parasites. Observations of extraerythrocytic B. bovis, B. bigemina and B. caballi merozoites using AFM revealed differences in size and shape between these three parasites. Finally, similar to what was previously observed for Plasmodium‐infected erythrocytes, FM‐KPFM images reveal an unequal electric charge distribution, with higher surface potential above the erythrocyte regions that are likely associated with Babesia parasites than over its remainder regions. In addition, the surface potential of paired extraerythrocytic B. bovis Mo7 merozoites revealed an asymmetric potential distribution. These observations may be important to better understand the unique cytoadhesive properties of B. bovis‐infected erythrocytes, and to speculate on the role of differences in the distribution of surface charges in the biology of the parasites.     

Kinetics of interaction of HIV fusion protein (gp41) with lipid membranes studied by real-time AFM imaging

One of the most important steps in the process of viral infection is a fusion between cell membrane and virus, which is mediated by the viral envelope glycoprotein. The study of activity of the glycoprotein in the post-fusion state is important for understanding the progression of infection. Here we present a first real-time kinetic study of the activity of gp41 (the viral envelope glycoprotein of human immunodeficiency virus—HIV) and its two mutants in the post-fusion state with nanometer resolution by atomic force microscopy (AFM). Tracking the changes in the phosphatidylcholine (PC) and phosphatidylcholine–phosphatidylserine (PC:PS) membrane integrity over one hour by a set of AFM images revealed differences in the interaction of the three types of protein with zwitterionic and negatively charged membranes. A quantitative analysis of the slow kinetics of hole formation in the negatively charged lipid bilayer is presented. Specifically, analysis of the rate of roughness change for the three types of proteins suggests that they exhibit different types of kinetic behavior.     

Quantitative temperature‐depending mapping of mechanical properties of bitumen at the nanoscale using the AFM operated with PeakForce TappingTM mode

The mechanical properties of bitumen, such as elasticity/Young's modulus, stickiness/adhesion, hardness and energy loss, and sample deformation were acquired quantitatively and simultaneously with the topology at the microscale, discriminating clearly two separate phases within the bitumen. Temperature‐dependent measurements revealed detailed and specific data about the changes of these properties with temperature, enabling the development of predictive models for the performance and durability of asphalt.     

Investigation of surface roughness values of various restorative materials after brushing with blue covarine containing whitening toothpaste by two different methods: AFM and profilometer

The aim of this study is to evaluate the effect of whitening toothpaste on the surface roughness of resin-based restorative materials by different measurement methods. Twenty four specimens from each of human enamel, a microhybrid composite and two nanohybrid composites discs (8.0 diameter × 4.0 mm thick) were divided into two groups (n = 12) according to toothbrushing solutıon and subjected to simulation toothbrushing (30,000 cycles) with both distilled water and whitening toothpaste containing blue covarine. Surface roughness was examined using atomic force microscopy (AFM), profilometer, and scanning electron microscopy (SEM), and the data obtained were subjected to analysis. Ra values of Tescera (TES) were significantly higher than Sonicfill 2 (SF2) when brushing both toothbrushing solutions for initial or 30,000 cycles. Roughness increased for SF2 and TES when brushed for 30,000 cycles and was higher than enamel and Herculite XRV Ultra (HXU). Human enamel was obtained lower surface roughness values brushed with toothpaste compared with distilled water. Evaluation of the surface roughness of control groups using the AFM revealed no statistically significant difference between the groups, but significant differences were found using a profilometer. The use of abrasive whitening toothpaste containing blue covarine and the number of brushing cycles affect the surface properties of human enamel and the restorative material, and also, the clinical success of the restoration. Toothbrushing for 30,000 cycles increased the surface roughness of all materials. The type of toothbrushing solution partially has affected surface roughness.     

The morphological and biomechanical changes of keratocytes cultured on modified p (HEMA‐MMA) hydrogel studied by AFM

The poor integration with host cornea tissue and the low mechanical properties of pHEMA hydrogel for artificial cornea remains a difficult problem to solve. A modified pHEMA hydrogel, MMA copolymerized and type‐I collagen and bFGF immobilized, was previously prepared in an attempt to solve the problems. In this study, the cytotoxicity of Col/bFGF‐p (HEMA‐MMA) and p (HEMA‐MMA) was studied by cell adhesion assay and atomic force microscopy (AFM). The results of cell adhesion assay show that the attachment of keratocytes on the modified membrane is much higher than that of the unmodified membrane. This indicates that the material after modification have better cell–material interaction. The AFM images reveal that the morphology of keratocytes cultured on different substrate is obviously different. The cell cultured on modified membrane presented a completely elongated and spindle‐shape morphology. The force?distance indicates that the biomechanical of keratocytes changes significantly after culturing on different substrates. The adhesion force (2328±523 pN) and Young's modulus (0.51±0.125 kPa) of the cell cultured on modified membrane are much higher, and the stiffness (0.08±0.022 mN/m) is lower than those of the cell cultured on unmodified membrane. These results show that the cytotoxicity of Col/bFGF‐p (HEMA‐MMA) for keratocytes is much improved. SCANNING 31: 246–252, 2009. © 2010 Wiley Periodicals, Inc.     

Nanoscale Fabrication of the Ferroelectric Polymer Poly(vinylidene Fluoride with Trifluoroethylene) P(VDF‐TrFE) 75:25 Thin Films by Atomic Force Microscope Nanolithography

Thin films of an organic ferroelectric system, poly(vinylidene fluoride with trifluoroethylene) P(VDF‐TrFE, Kureha Corporation, Tokyo, Japan) 75:25 layers, have been deposited on highly ordered pyrolytic graphite and silicon dioxide by the horizontal Schaefer method of Langmuir–Blodgett techniques. It is possible to “shave” or mechanically displace small regions of the polymer film by using atomic force microscope nanolithography techniques such as nanoshaving, leaving swaths of the surface cut to a depth of 4 nm and 12 nm exposing the substrate. The results of fabricating stripes by nanoshaving two holes close to each other show a limit to the material “stripe” widths of an average of 153.29 nm and 177.67 nm that can be produced. Due to the lack of adhesion between the substrates and the polymer P(VDF‐TrFE) film, smaller “stripes” of P(VDF‐TrFE) cannot be produced, and it can be shown by the sequencing of nanoshaved regions that “stripes” of thin films can be removed. SCANNING 34: 404‐409, 2012. © 2012 Wiley Periodicals, Inc.