Kelvin probe force microscopy in application to biomolecular films: Frequency modulation,amplitude modulation,and lift mode

Kelvin probe force microscopy (KPFM) is a powerful technique to visualize the differences of work function in metals and lateral surface potential distribution in thin organic films. Earlier we have shown that frequency modulated-Kelvin probe force microscopy has significant advantages in both sensitivity and resolution when applied to metal and inorganic interfaces in vacuum. High resolution, high sensitivity, and performance in ambient conditions are required in order to study biologically relevant samples. In this work we compared the resolution of frequency modulation (FM-KPFM), amplitude modulation (AM-KPFM), and lift modes KPFM for imaging the local electrical surface potential of complex biomolecular films and demonstrated that FM-KPFM mode has superior resolution for biological applications. The power of the method was illustrated on pulmonary surfactant films, revealing nm spatial resolution and mV potential sensitivity in ambient air. At this level of resolution this method can provide critical insight into the molecular arrangement and function of complex biosystems in a way that other KPFM modes cannot do. Based on the observed changes in the local surface potential we discovered that excess cholesterol produces nm size electrostatic domains and change the electric fields.     

The comparison between force volume and peakforce quantitative nanomechanical mode of atomic force microscope in detecting cell's mechanical properties

Atomic force microscope (AFM) has been widely used in the biological field owing to its high sensitivity (subnanonewton), high spatial resolution (nanometer), and adaptability to physiological environments. Nowadays, force volume (FV) and peakforce quantitative nanomechanical (QNM) are two distinct modes of AFM used in biomechanical research. However, numerous studies have revealed an extremely confusing phenomenon that FV mode has a significant difference with QNM in determining the mechanical properties of the same samples. In this article, for the case of human benign prostatic hyperplasia cells (BPH) and two cancerous prostate cells with different grades of malignancy (PC3 and DU145), the differences were compared between FV and QNM modes in detecting mechanical properties. The results show measured Young's modulus of the same cells in FV mode was much lower than that obtained by QNM mode. Combining experimental results with working principles of two modes, it is indicated that surface adhesion is highly suspected to be a critical factor resulting in the measurement difference between two modes. To further confirm this conjecture, various weight ratios of polydimethylsiloxane (PDMS) were assessed by two modes, respectively. The results show that the difference of Young's modulus measured by two modes increases with the surface adhesion of PDMS, confirming that adhesion is one of the significant elements that lead to the measurement difference between FV and QNM modes.     

Characterisation of electronic and structural properties of thin silicon dioxide films by scanned probe microscopy

This paper concerns the application of scanned probe microscopy to the study of thin silicon dioxide films. We show how the formation of 7 nm diameter silicon carbide particles on a silicon surface during high temperature processing affects the quality of subsequently grown oxide. To measure the local dielectric properties of thin oxides we have developed a new type of probe measurement which allows high resolution surface imaging, based on an atomic force microscope, combined with local electrical measurements. The spatial resolution of the electrical measurements was shown to be < 40 nm. Applying the technique to 500 nm capacitors fabricated on device quality oxide it was shown that some capacitors broke down during imaging while others remained stable at electric fields up to 30 MVcm^?1. This higher breakdown strength may have substantial impact on future electronic device reliablity and performance.     

Interaction forces and conduction properties between multi wall carbon nanotube tips and Au(111)

We have studied the interaction forces and electrical conduction properties arising between multiwall carbon nanotube tips and the Au(111) surface in air, by means of amplitude modulation scanning force microscopy, also called intermittent contact. We have centered our work on tips with metallic electronic structure and for the specific parameters used we have found a preliminary interaction range where there is no contact between tip and surface. Stable imaging in this non-contact range is possible with multiwall carbon nanotube tips. These tips have also been used to obtain simultaneous topographic and current maps of the surface. They show excellent properties as tips due to their high aspect ratio and durability, as a result of their elastic and non-reactive properties. Correspondingly, multiwall carbon nanotube tips allow high resolution local analysis of electrical conductivity on a nanometer scale.     

Magnetooptical Analysis of the Subsurface Region in a Bearing Ring Subjected to Rolling Contact Fatigue

It has been known for decades that high contact load in bearings leads to fatigue alterations of subsurface material, which can be optically visualized on etched samples. In the literature, these alterations are termed dark etching regions and white etching bands, due to their peculiar patterns and contrast. In the current work, it is proposed to study these alterations by the magnetooptical Kerr effect, expecting that the material changes associated with rolling contact fatigue also affect the magnetic properties of bearing steel. The advantage of this method is that it has good spatial resolution, allowing recording of magnetic properties on microscopic scales. Samples containing dark etching regions and white etching bands were compared in the magnetooptical test with the reference sample containing no optically visible changes. It was found that the coercive force of material reduces as result of microstructural changes caused by rolling contact fatigue.     

Topography and nanomechanical properties of tribochemical films derived from zinc dialkyl and diaryl dithiophosphates

In this paper we present recent results from an on‐going effort to characterize the nanomechanical properties of a variety of tribochemical, antiwear films through the use of modern scanning probe techniques. The two types of antiwear wear films studied, derived from zinc dialkyl dithiophosphate (alkyl ZDDP) and zinc diaryl dithiophosphate (aryl ZDDP), were chosen because they possess significantly different wear characteristics. High resolution AFM topographic images showed significant differences between the two types of films. More interestingly, high resolution imaging and quantitative mechanical properties testing using the interfacial force microscope (IFM), revealed different elastic and plastic properties between the two types of films; in addition each type of film possessed several distinct regions with respect to mechanical properties. The maximum values for elastic modulus and hardness were located on the highly loaded regions of the alkyl ZDDP films which exhibited the best tribological performance. In contrast, the aryl ZDDP films, which exhibited poorer antiwear behaviour, were devoid of such resilient regions. Our results correlate the macroscopic wear behavior of the tribochemical films to differences in the mechanical properties on a nanometer scale. This revised version was published online in July 2006 with corrections to the Cover Date.     

Sample preparation procedures for biological atomic force microscopy

Since the late 1980s, atomic force microscopy (AFM) has been increasingly used in biological sciences and it is now established as a versatile tool to address the structure, properties and functions of biological specimens. AFM is unique in that it provides three-dimensional images of biological structures, including biomolecules, lipid films, 2D protein crystals and cells, under physiological conditions and with unprecedented resolution. A crucial prerequisite for successful, reliable biological AFM is that the samples need to be well attached to a solid substrate using appropriate, nondestructive methods. In this review, we discuss common techniques for immobilizing biological specimens for AFM studies.     

Structure-Property Relationships in Thin Solid Poly(methyl methacrylate) Boundary Films

The interface, properties of boundary films are commonly characterized try the measurement of the interface shear stress, τ, which is defined as the, fictional force per unit area of contact. The parameter τ has been observed to be a strong function of several contact mechanical variables which include the contact pressure, the temperature, the sliding velocity and the contact time, τ is also found to be a strong function of the processing methods used in the preparation of polymer films. It is this facet of the functionality of τ that is pursued in the present paper with an emphasis upon the apparent ductile or brittle lubrication modes, the locus of the shear zone and the molecular architectures which are responsible for these resultant lubrication modes.     

Polysaccharide properties probed with atomic force microscopy

In recent years, polysaccharides have been extensively studied using atomic force microscopy (AFM). Owing to its high lateral and vertical resolutions and ability to measure interaction forces in liquids at pico‐ or nano‐Newton level, the AFM is an excellent tool for characterizing biopolymers. The first imaging studies showed the morphology of polysaccharides, but gradually more quantitative image analysis techniques were developed as the AFM grew easier to use in aqueous liquids and in non‐contact modes. Recently, AFM has been used to stretch polysaccharides and characterize their physicochemical properties by application of appropriate polymer stretching models, using a technique called single‐molecule force spectroscopy. From application of such models as the wormlike chain, freely jointed chain, extensible‐freely jointed chain, etc., properties such as the contour length, persistence length and segment elasticity or spring constant can be calculated for polysaccharides. The adhesion between polysaccharides and surfaces has been quantified with AFM, and this application is particularly useful for studying polysaccharides on microbial and other types of cells, because their adhesion is controlled by biopolymer characteristics. This review presents a synthesis of the theory and techniques currently in use to probe the physicochemical properties of polysaccharides with AFM.     

Application of atomic force microscopy in bacterial research

The atomic force microscope (AFM) has evolved from an imaging device into a multifunctional and powerful toolkit for probing the nanostructures and surface components on the exterior of bacterial cells. Currently, the area of application spans a broad range of interesting fields from materials sciences, in which AFM has been used to deposit patterns of thiol‐functionalized molecules onto gold substrates, to biological sciences, in which AFM has been employed to study the undesirable bacterial adhesion to implants and catheters or the essential bacterial adhesion to contaminated soil or aquifers. The unique attribute of AFM is the ability to image bacterial surface features, to measure interaction forces of functionalized probes with these features, and to manipulate these features, for example, by measuring elongation forces under physiological conditions and at high lateral resolution (<1 Å). The first imaging studies showed the morphology of various biomolecules followed by rapid progress in visualizing whole bacterial cells. The AFM technique gradually developed into a lab‐on‐a‐tip allowing more quantitative analysis of bacterial samples in aqueous liquids and non‐contact modes. Recently, force spectroscopy modes, such as chemical force microscopy, single‐cell force spectroscopy, and single‐molecule force spectroscopy, have been used to map the spatial arrangement of chemical groups and electrical charges on bacterial surfaces, to measure cell–cell interactions, and to stretch biomolecules. In this review, we present the fascinating options offered by the rapid advances in AFM with emphasizes on bacterial research and provide a background for the exciting research articles to follow. SCANNING 32: 74–96, 2010. © 2010 Wiley Periodicals, Inc.     

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.     

X-ray microscopy of plant cells by using LiF crystal as a detector

A lithium fluoride (LiF) crystal has been utilized as a new soft X-ray detector to image different biological samples at a high spatial resolution. This new type of image detector for X-ray microscopy has many interesting properties: high resolution (nanometer scale), permanent storage of images, the ability to clear the image and reuse the LiF crystal, and high contrast with greater dynamic range. Cells of the unicellular green algae Chlamydomonas dysosmos and Chlorella sorokiniana, and pollen grains of Olea europea have been used as biological materials for imaging. The biological samples were imaged on LiF crystals by using the soft X-ray contact microscopy and contact micro-radiography techniques. The laser plasma soft X-ray source was generated using a Nd:YAG/Glass laser focused on a solid target. The X-ray energy range for image acquisition was in the water-window spectral range for single shot contact microscopy of very thin biological samples (single cells) and around 1 keV for multishots microradiography. The main aim of this article is to highlight the possibility of using a LiF crystal as a detector for the biological imaging using soft X-ray radiation and to demonstrate its ability to visualize the microstructure within living cells.     

基于介质层耦合的柔性触觉传感器空间分辨率补偿方法研究

传统柔性压力传感器采用离散电容结构,传感器空间分辨率受到最小离散单元的限制.对于作用面积较小的外力接触,现有柔性传感器仅能通过减小离散单元面积确定接触位置,而离散单元过小则会造成制作困难、接线复杂、成本升高.为解决微小接触面积下空间分辨率偏低问题,提出了一种基于整体介质层耦合结构柔性传感器,通过建立传感器接触及扩散电容...     

Morphology and Nanomechanical Properties of ZDDP Antiwear Films as a Function of Tribological Contact Time

We have employed scanning force microscopy (SFM) and nanoindentation analysis to track the evolution of tribologically generated antiwear films derived from zinc dialkyldithiophosphate (ZDDP) as a function of rubbing time. The SFM images reveal that film morphology evolves with time through a growth mechanism consisting of three stages. In the first stage nucleation on active sites at the steel surface leads to the growth of distinct segregated islands. In the second stage the islands coalesce causing the film to spread over a larger fraction of the surface. In the final stage continuous rubbing induces the large islands to divide into smaller, densely packed structures. In contrast to the observed morphological changes, rubbing time does not strongly influence the nanomechanical properties of the films. This highlights the importance of film morphology in determining the effectiveness of ZDDP antiwear films. We also observe large variation in both the morphology and nanomechanical properties that are likely due to the heterogeneity in contact pressure at the pin-sample interface of the wear rig.     

Combined AFM and confocal fluorescence microscope for applications in bio-nanotechnology

We present a custom-designed atomic force fluorescence microscope (AFFM), which can perform simultaneous optical and topographic measurements with single molecule sensitivity throughout the whole visible to near-infrared spectral region. Integration of atomic force microscopy (AFM) and confocal fluorescence microscopy combines the high-resolution topographical imaging of AFM with the reliable (bio)-chemical identification capability of optical methods. The AFFM is equipped with a spectrograph enabling combined topographic and fluorescence spectral imaging, which significantly enhances discrimination of spectroscopically distinct objects. The modular design allows easy switching between different modes of operation such as tip-scanning, sample-scanning or mechanical manipulation, all of which are combined with synchronous optical detection. We demonstrate that coupling the AFM with the fluorescence microscope does not compromise its ability to image with a high spatial resolution. Examples of several modes of operation of the AFFM are shown using two-dimensional crystals and membranes containing light-harvesting complexes from the photosynthetic bacterium Rhodobacter sphaeroides.     

Use of phase imaging in atomic force microscopy for measurement of viscoelastic contrast in polymer nanocomposites and molecularly thick lubricant films

Phase contrast microscopy, using an atomic force microscope, is used to detect and quantify changes in composition across polymer nanocomposites and molecularly thick lubricated surfaces. The technique takes advantage of the contrast in viscoelastic (viscous energy dissipation) properties of the different materials across the surface. Some materials, especially polymers, are found to display viscoelastic behavior. For such materials, the strain response lags the stress by a phase angle that is characteristic of the material. In tapping (or intermittent contact) mode, phase angle contrast is found to be highly dependent on vibration amplitude and mean tip-to-sample distance (setpoint). Phase angle contrast seems to be a stronger function of viscoelastic properties at relatively high vibration amplitude and low mean tip-to-sample distance. In this regime the effects of sample deformation, and thus viscoelastic properties, are dominant. In these contrast images, low phase angle corresponds to materials with low viscoelastic properties. This technique was used to find fairly reproducible phase angle contrast for polyethylene terephthalate (PET) films with embedded ceramic particles, metal particle (MP) magnetic tape, and Si(100) with a nonuniform Z-15 lubricant film. Very little correlation is found between phase angle images and friction force images for PET films with embedded ceramic particles and MP tape; phase angle images give information that cannot be obtained from topography or friction images. A numerical vibration model verifies that viscoelastic properties are dominant for high vibration amplitude and low mean tip-to-sample distance. For these conditions, the model also verifies that low phase angle corresponds to low viscoelastic properties.     

Magnetic refinement of tips for magnetic force microscopy

Silicon cantilever probes with monolithically integrated tips are commercially available and are routinely used for atomic force microscopy (AFM). For such probes, amagnetic refinement of the silicon tip has been developed and results in a deposition of ferromagnetic material such as nickel or CrCoTa in the top area of the tip. The method consists of essentially three steps: (1) A broad-area sputter deposition of a ferromagnetic material; (2) a selective electron beam-induced carbon deposition at the top of the tip; (3) a broad-area ion-beam sputter etching, which removes the magnetic layer everywhere except underneath the carbon cap. The method allows to control the total amount and extension of the magnetic material left at the tip. It is applicable to all kinds of ferromagnetic materials which can be deposited as a thin layer by sputter deposition or evaporation. Experiments indicate that the method is reliable and improves the resolution of magnetic force microscopy (MFM). With such magnetically refined tips on silicon cantilevers, MFM measurements have been performed in contact mode as well as in dynamic and static noncontact modes. In this paper, the method for magnetic tip refinement is described and MFM measurements with these tips are presented.     

High-resolution force microscopy of in-plane magnetization

We have shown for the first time, in previous studies, that the force microscope fitted with a magnetic tip could be used to image magnetic domains in TbFe thin films. In this report we show that the information provided by the magnetic force microscope can also include a measurement of the component of magnetization in the sample which is parallel to the surface. Measurements were taken with a magnetized tip tilted at 45° with respect to the surface normal. In a first experiment, we imaged 1 μm diameter domains thermomagnetically written in a TbFe thin film. In a second measurement, we imaged a series of alternating domains in a thin film Co-alloy disc which was decorated with small magnetized particles, allowing unambiguous identification of the domain boundaries. In both cases we interpret the asymmetric portion of the images to the in-plane component of the sample magnetization.     

Friction force microscopy investigations of potassium halide surfaces in ultrahigh vacuum: structure,friction and surface modification

Friction force microscopy measurements on the vacuum‐cleaved (001) surfaces of KF, KCl and KBr have been carried out. All surfaces exhibit atomically flat terraces with monatomic steps aligned preferentially along low‐index lattice directions. Stick‐slip lateral forces with the lattice periodicity are observed on all surfaces. Tip‐sample contact creates higher friction domains on the terraces of all three materials. The structure, topography and degree of friction force contrast of these domains is material dependent. The dependence of friction upon load generally does not coincide with the behavior expected for an elastic contact. We propose that the observed domains result from surface structural changes created by low load tip‐sample contact on these relatively soft materials. This revised version was published online in June 2006 with corrections to the Cover Date.     

Near-field second harmonic imaging of the c/a/c/a polydomain structure of epitaxial PbZrxTi1–xO3 thin films

Near-field optical second harmonic microscopy has been applied to imaging of the c/a/c/a polydomain structure of epitaxial PbZr _x Ti_{1– x} O₃ thin films in the 0 <  x  < 0.4 range. Comparison of the near-field optical images and the results of atomic force microscopy and X-ray diffraction studies show that an optical resolution of the order of 100 nm is achieved. Symmetry properties of the near-field second harmonic signal allow us to obtain good optical contrast between the local second harmonic generation in c- and a-domains. Experimentally measured near-field second harmonic images have been compared with the results of theoretical calculations. Good agreement between theory and experiment is demonstrated.