Tunneling/shear force microscopy using piezoelectric tuning forks for characterization of topography and local electric surface properties

Characterization of novel nanoelectronic structures and materials requires advanced and high-resolution diagnostic methods. In this article new approach for high sensitivity measurements of electric surface properties using scanning probe microscopy is presented. In this procedure topography and tunneling current flowing between the metallic tip and the surface are observed simultaneously. In our design piezoelectric tuning fork equipped with metallic tip in shear force microscope is used.     

Force-gradient-induced mechanical dissipation of quartz tuning fork force sensors used in atomic force microscopy

We have studied the dynamics of quartz tuning fork resonators used in atomic force microscopy taking into account the mechanical energy dissipation through the attachment of the tuning fork base. We find that the tuning fork resonator quality factor changes even in the case of a purely elastic sensor-sample interaction. This is due to the effective mechanical imbalance of the tuning fork prongs induced by the sensor-sample force gradient, which in turn has an impact on dissipation through the attachment of the resonator base. This effect may yield a measured dissipation signal that can be different from the one exclusively related to the dissipation between the sensor and the sample. We also find that there is a second-order term in addition to the linear relationship between the sensor-sample force gradient and the resonance frequency shift of the tuning fork that is significant even for force gradients usually present in atomic force microscopy, which are in the range of tens of N/m.     

Time-lapse imaging of In Vitro myogenesis using atomic force microscopy

Myoblast therapy relies on the integration of skeletal muscle stem cells into distinct muscular compartments for the prevention of clinical conditions such as heart failure, or bladder dysfunction. Understanding the fundamentals of myogenesis is hence crucial for the success of these potential medical therapies. In this report, we followed the rearrangement of the surface membrane structure and the actin cytoskeletal organization in C2C12 myoblasts at different stages of myogenesis using atomic force microscopy (AFM) and confocal laser scanning microscopy (CLSM). AFM imaging of living myoblasts undergoing fusion unveiled that within minutes of making cell–cell contact, membrane tubules appear that unite the myoblasts and increase in girth as fusion proceeds. CLSM identified these membrane tubules as built on scaffolds of actin filaments that nucleate at points of contact between fusing myoblasts. In contrast, similarly behaving membrane tubules are absent during cytokinesis. The results from our study in combination with recent findings in literature further expand the understanding of the biochemical and membrane structural rearrangements involved in the two fundamental cellular processes of division and fusion.     

原子力显微镜原理与应用技术

本文简述原子力显微镜的工作原理,对比说明敲击模式的优越性,指出针尖-样品卷积效应和假象产生的原因,并例证其应用领域及其测试效果。     

Phase imaging with intermodulation atomic force microscopy

Intermodulation atomic force microscopy (IMAFM) is a dynamic mode of atomic force microscopy (AFM) with two-tone excitation. The oscillating AFM cantilever in close proximity to a surface experiences the nonlinear tip-sample force which mixes the drive tones and generates new frequency components in the cantilever response known as intermodulation products (IMPs). We present a procedure for extracting the phase at each IMP and demonstrate phase images made by recording this phase while scanning. Amplitude and phase images at intermodulation frequencies exhibit enhanced topographic and material contrast.     

Phase contrast in Simultaneous Topography and Recognition imaging

The operation of a force microscope in Simultaneous Topography and Recognition (TREC) imaging mode is analyzed by means of numerical simulations. Both topography and recognition signals are analyzed by using a worm-like chain force as the specific interaction between the functionalized tip probe and the sample. The special feedback mechanism in this mode is shown to couple the phase signal to the presence of molecular recognition interactions even in absence of dissipation.     

Molecular dynamics of DNA and nucleosomes in solution studied by fast-scanning atomic force microscopy

Nucleosome is a fundamental structural unit of chromatin, and the exposure from or occlusion into chromatin of genomic DNA is closely related to the regulation of gene expression. In this study, we analyzed the molecular dynamics of poly-nucleosomal arrays in solution by fast-scanning atomic force microscopy (AFM) to obtain a visual glimpse of nucleosome dynamics on chromatin fiber at single molecule level. The influence of the high-speed scanning probe on nucleosome dynamics can be neglected since bending elastic energy of DNA molecule showed similar probability distributions at different scan rates. In the sequential images of poly-nucleosomal arrays, the sliding of the nucleosome core particle and the dissociation of histone particle were visualized. The sliding showed limited fluctuation within ∼50 nm along the DNA strand. The histone dissociation occurs by at least two distinct ways: a dissociation of histone octamer or sequential dissociations of tetramers. These observations help us to develop the molecular mechanisms of nucleosome dynamics and also demonstrate the ability of fast-scanning AFM for the analysis of dynamic protein–DNA interaction in sub-seconds time scale.     

Investigating ultra-thin lubricant layers using resonant friction force microscopy

The ultrasonic friction mode of an atomic force microscope is a scanning probe technique allowing one to analyze the load and velocity dependence of friction. The technique is based on evaluation of the resonance behavior of an AFM cantilever when in contact with a vibrating sample surface. The effect of load and lateral displacement of the sample surface on the shape of the torsional resonance spectra of the AFM cantilever is evaluated under dry and lubricated sliding conditions. A characteristic flattening of the torsional resonance curve has been observed at large surface displacements, resulting from the onset of sliding friction in the AFM cantilever–sample surface contact. An analytical model describing torsional cantilever vibrations in Hertzian contact with a sample surface is presented, and numerical simulations have been carried out in order to confirm that the flattening of the resonance curve occurs with the onset of the sliding friction in the contact.     

Adhesion artefacts in atomic force microscopy imaging

Artefacts that affect contrast and arise from adhesion forces in atomic force microscopy images of aramid fibres (both fresh and plasma-treated) are investigated. It is demonstrated that these stem not only from variations in the chemical composition of the surface but also from certain topographical features (which may appear hidden or enhanced in the images), resulting in changes in the lateral forces that are detected by the cantilever and are comparable to the vertical forces. It is also shown that both types of contribution to the forces can be uncoupled to yield images free from these artefacts, thus allowing more accurate quantitative measurements. These artefactual effects are also generally applicable to many other materials.     

A symmetrical stretching stage for electrical atomic force microscopy

We present a remotely-controlled device for sample stretching, designed for use with atomic force microscopy (AFM) and providing electrical connection to the sample. Such a device enables nanoscale investigation of electrical properties of thin gold films deposited on polydimethylsiloxane (PDMS) substrate as a function of the elongation of the structure. Stretching and releasing is remotely controlled with use of a dc actuator. Moreover, the sample is stretched symmetrically, which gives an opportunity to perform AFM scans in the same site without a time-consuming finding procedure. Electrical connections to the sample are also provided, enabling Kelvin probe force microscopy (KPFM) investigations. Additionally, we present results of AFM imaging using the stretching stage.     

Second harmonic atomic force microscopy imaging of live and fixed mammalian cells

Higher harmonic contributions in the movement of an oscillating atomic force microscopy (AFM) cantilever are generated by nonlinear tip–sample interactions, yielding additional information on structure and physical properties such as sample stiffness. Higher harmonic amplitudes are strongly enhanced in liquid compared to the operation in air, and were previously reported to result in better structural resolution in highly organized lattices of proteins in bacterial S-layers and viral capsids [J. Preiner, J. Tang, V. Pastushenko, P. Hinterdorfer, Phys. Rev. Lett. 99 (2007) 046102]. We compared first and second harmonics AFM imaging of live and fixed human lung epithelial cells, and microvascular endothelial cells from mouse myocardium (MyEnd). Phase–distance cycles revealed that the second harmonic phase is 8 times more sensitive than the first harmonic phase with respect to variations in the distance between cantilever and sample surface. Frequency spectra were acquired at different positions on living and fixed cells with second harmonic amplitude values correlating with the sample stiffness. We conclude that variations in sample stiffness and corresponding changes in the cantilever–sample distance, latter effect caused by the finite feedback response, result in second harmonic images with improved contrast and information that is not attainable in the fundamental frequency of an oscillating cantilever.     

Measurement of thermal expansion coefficient of INVAR foil using atomic force microscopy

The production of organic light emitting diode (OLED) displays depends on the use of the low coefficient of thermal expansion (CTE) of INVAR foils as the shadow mask. The high-resolution of the smartphone displays requires increasingly thin INVAR in a two-step etching process. However, it is difficult to measure CTE for very thin metal foils. A simple method is developed to measure the linear CTE of the INVAR foil using atomic force microscopy (AFM). This method uses a focus-ion-beam (FIB) to etch a 5000 μm trench on the INVAR foil. The thermal drift of the system is calibrated from AFM images, and the average linear CTE of the INVAR foil is then calculated from the displacements of two side end points of the trench on the foil during temperature variation. The linear CTE obtained by the proposed method is quite close to the value of the bulk INVAR provided by the manufacturer.     

Atomic force microscopy for ultrafiltration membrane imaging

Atomic force microscopy (AFM) can reveal nanometer-scale structure of samples without the sample preparation techniques that involve dehydration. This is particularly important for hydrophilic organic materials. An asymmetric polysulfone ultrafiltration membrane (molecular weight cutoff rated at 10 kg/mol) was imaged by AFM. Sample mounting methods tried include cyanoacrylate, double-sided tape, and paraffin. Wax and tape bonding did not lead to usable images. Cyanoacrylate bonding resulted in images that appear to show 2.8° 10⁹ pores/m² approximately 3 nm in diameter, creating a porosity of 2%. This is consistent with estimates of molecular sizes for 10 kg/mol proteins, but not with the results of other AFM studies of similar membranes. The discrepancies can be explained largely by differences in sample preparation techniques.     

Atomic force microscopy imaging of fragments from the Martian meteorite ALH84001

A combination of scanning electron microscopy (SEM) and environmental scanning electron microscopy (ESEM) techniques, as well as atomic force microscopy (AFM) methods has been used to study fragments of the Martian meteorite ALH84001. Images of the same areas on the meteorite were obtained prior to and following gold/palladium coating by mapping the surface of the fragment using ESEM coupled with energy-dispersive X-ray analysis. Viewing of the fragments demonstrated the presence of structures, previously described as nanofossils by McKay et al . (Search for past life on Mars — possible relic biogenic activity in martian meteorite ALH84001. Science , 1996, pp. 924–930) of NASA who used SEM imaging of gold-coated meteorite samples. Careful imaging of the fragments revealed that the observed structures were not an artefact introduced by the coating procedure.     

An improved dynamic model of tuning fork probe for scanning probe microscopy

This paper presents a two coupled oscillators model to describe the dynamics of a tuning fork with a probe attached. The two coupled oscillators are unbalanced only in their effective masses and the damping ratios. By applying a frequency domain system identification approach in experimental investigation of various probe attachment cases, a good accuracy of the model is demonstrated. The effectiveness of the model is further demonstrated in quantitative analysis of the noise performance and the sensitivity of force sensing with a tuning fork probe. Compared with existing models, the proposed model can more accurately characterize the dynamics of a tuning fork probe.     

Ribosome substructure investigated by scanning force microscopy and image processing

Scanning force microscopy was used to study the ultrastructure of eukaryotic ribosomes from Chironomus pallidivittatus in the polysomal complex. Positively stained polysomes were imaged, and the resulting three-dimensional ribosomal structures were further processed by statistical analyses of virtual cross-sections parallel to the substrate plane. Structural investigations were based on parameters which are minimally influenced by the tip geometry, like section plane centre or axis ratio. In the lower part of the structure a shift of the section centres was observed, suggesting an attached structure. The geometry of the sections revealed an elliptical shape in the upper part (axis ratio 1.52 ± 0.22), with a less elongated shape in the lower region (axis ratio 1.41 ± 0.18). A model for the surface topography of a positively stained ribosome exhibiting a small subunit attached along the long side of an elliptical large structure is proposed.     

Intermittency in amplitude modulated dynamic atomic force microscopy

From a mathematical point of view, the atomic force microscope (AFM) belongs to a special class of continuous time dynamical systems with intermittent impact collisions. Discontinuities of the velocity result from the collisions of the tip with the surface. Transition to chaos in non-linear systems can occur via the following four routes: bifurcation cascade, crisis, quasi-periodicity, and intermittency. For the AFM period doubling and period-adding cascades are well established. Other routes into chaos, however, also may play an important role. Time series data of a dynamic AFM experiment indicates a chaotic mode that is related to the intermittency route into chaos. The observed intermittency is characterized as a type III intermittency. Understanding the dynamics of the system will help improve the overall system performance by keeping the operation parameters of dynamic AFM in a range, where chaos can be avoided or at least controlled.     

Climbing the steps of viral atomic force microscopy: visualization of Dengue virus particles

In recent years, the application of atomic force microscopy (AFM) to biological systems has highlighted the potential of this technology. AFM provides insights into studies of biological structures and interactions and can also identify and characterize a large panel of pathogens, including viruses. The Flaviviridae family contains a number of viruses that are important human and animal pathogens. Among them, Dengue virus causes epidemics with fatal outcomes mainly in the tropics. In this study, Dengue virus is visualized for the first time using the in air AFM technique. Images were obtained from a potassium-tartrate gradient-purified virus. This study enhances the application of AFM as a novel tool for the visualization and characterization of virus particles. Because flavivirus members are closely related, studies of the morphologic structure of the Dengue virus can reveal strategies that may be useful to identify and study other important viruses in the family, including the West Nile virus.     

Spectroscopy and atomic force microscopy of biomass

Scanning probe microscopy has emerged as a powerful approach to a broader understanding of the molecular architecture of cell walls, which may shed light on the challenge of efficient cellulosic ethanol production. We have obtained preliminary images of both Populus and switchgrass samples using atomic force microscopy (AFM). The results show distinctive features that are shared by switchgrass and Populus. These features may be attributable to the lignocellulosic cell wall composition, as the collected images exhibit the characteristic macromolecular globule structures attributable to the lignocellulosic systems. Using both AFM and a single case of mode synthesizing atomic force microscopy (MSAFM) to characterize Populus, we obtained images that clearly show the cell wall structure. The results are of importance in providing a better understanding of the characteristic features of both mature cells as well as developing plant cells. In addition, we present spectroscopic investigation of the same samples.     

Atomic force microscopy imaging of polycrystalline CuInSe2 thin films

In this study, atomic force microscopy (AFM) imaging has been used to study the structural properties of polycrystalline CuInSe₂ films, which are widely used as absorber materials in thin film solar cell devices. This technique demonstrated an excellent capability for the reproducible imaging of these rough polycrystalline materials. AFM imaging in combination with statistical analysis revealed distinct differences in the structural properties (i.e. grain width and height distributions, root‐mean‐square (RMS) and peak to valley (R_(p–v)) roughness values) as a function of the specific growth technique used and the bulk composition of the films. In the case of Cu‐rich films, prepared by the H₂Se/Ar treatment of Cu/In/Cu alloys, rough surface structures were in general observed. Statistical analysis revealed two distinct distribution of grains in these samples (1.0–2.5 μm and 3–5.5 μm) with large RMS and R_(p–v) roughness values of 380 nm and 2.6 μm, respectively. In‐rich films were characterized by the presence of much smaller, roughly circular clusters with a significant reduction in both the width and height distributions as well as RMS and R_(p–v) roughness values. The most successful growth techniques, in terms of producing homogeneous and dense films, were in the cases of H₂Se/Ar treated metallic InSe/Cu/InSe alloys and the coevaporation of all materials to form CuInSe₂. Both these techniques produced absorber films with very narrow grain width and height distributions as well as small roughness values. It was possible to establish that high efficiency devices are associated with the use of absorber films with narrow width distributions between 0.5 and 2 μm and small RMS (> 300 nm) roughness values. These values are used as a figure of merit in our laboratories to evaluate the structural properties of our CuInSe₂ thin films.