Atomic force microscopy of Bacillus spore surface morphology

Bacillus spore surface morphology was imaged with atomic force microscopy (AFM) to determine if characteristic surface features could be used to distinguish between four closely related species; Bacillus anthracis Sterne strain, Bacillus thuringiensis var. kurstaki, Bacillus cereus strain 569, and Bacillus globigii var. niger. AFM surface height images showed an irregular topography across the curved upper surface of the spores. Phase images showed a superficial grain structure with different levels of phase contrast and significant differences in average surface morphologies among the four species. Although spores of the same species showed similarities, there was significant variability within each species. Overall, AFM revealed that spore surface morphology is rich with information, which can be used to distinguish a sample of about 20 spores from a similar number of spores of closely related species. Statistical analysis of spore morphology from a combination of amplitude and phase images for a small sample allows differentiation between, B. anthracis and its close relatives.     

Immobilization and AFM of single 4 x 6-mer tarantula hemocyanin molecules

The high molecular mass respiratory protein of the tarantula Eurypelma californicum, a 4 × 6-mer hemocyanin, was investigated by atomic force microscopy (AFM). Various substrates and methods were evaluated for immobilization of individual hemocyanin molecules on a solid surface. Samples were imaged after physisorption on mica and self-assembled monolayers, and after chemisorption on Au(111) and N-hydroxy-succinimide (NHS) functionalized surfaces. AFM measurements were carried out preferable in solution and contact mode, but also in Tapping mode and on air-dried samples. Adsorption of the protein on mica followed by drying and carrying out the measurements in Tapping mode gave the best results. In the AFM images the four hexamers of the native 4 × 6-mer hemocyanin have been defined. The results were compared with independent available structural data and represent a validation case for this technique applied for the first time on such giant and complex molecules. As observable in images taken by transmission electron microscopy and also proposed from SAXS data, 4 × 6-mers could be found where the half-molecules are tilted against each other. This study is a step in resolving conformational heterogeneities, involved in oxygen binding of hemocyanins, at the single-molecule level by AFM.     

Atomic force microscopy and UV-visible absorption-spectroscopy studies of ZnO nanometer colloidal particles supported on graphite

ZnO nanometer particles were synthesized by depositing ZnO colloidal suspensions onto the substrate of Highly Oriented Pyrolytic Graphite (HOPG). The isolated particles and their aggregation phase were directly imaged by Atomic Force Microscopy (AFM). High-resolution AFM images have been used to measure the diameter of individual particles, and large area images revealed that these ZnO particles on graphite surface form netlike aggregation. Experimental results also indicated that dispersive particles on a flat area are very mobile and easy to be pushed around during scanning due to the their weak adsorption on the substrate and tip-particle interactions.     

微焦点X射线源类同轴相衬成像

微焦点X射线源相衬成像是利用微焦点X射线源透过样品后携带的相位信息对样品内部结构成像. 通过获取相面上的强度信息利用相位复原方法可以得到物体内部的相位信息. 获取了不同放大倍数下的蚂蚁图像, 通过强度传播方程(Transport of Intensity Equation, TIE)方法得到了蚂蚁的相衬图像, 采用 边缘可见度(Edge Visibility)方法和熵法比较了不同放大倍数下的成像效果以及复原前后图像质量.     

Remnant grooves on alumina surfaces

Thermal grooving of grain boundaries in alumina has been studied using a combination of visible-light microscopy (VLM) and atomic-force microscopy (AFM). The partial angles of grooves that develop during heat treatments at 1650 °C were estimated by AFM. The observation of remnant grooves is key to determining the magnitude of the movement of a grain boundary. Imaging using VLM and AFM enabled the smoothing of remnant grooves to be observed through a progression of heat treatments, which enables the anisotropic nature of the surface diffusivity of alumina to be monitored.     

Effect of ultrasonication on the flotation of fine graphite particles: Nanobubbles or not?

It has been reported that nanobubbles can be produced by ultrasonication. However, it remains unclear whether part of the contribution of ultrasonication on flotation performance can be attributed to the generation of nanobubbles. In this work, we systematically studied this point of ultrasonication by combining a series of techniques including flotation testing, AFM (atomic force microscope) measurement, and settling testing. AFM imaging showed that no surface nanobubbles were found at the HOPG-water interface before and after subjection to ultrasonication. Further, surface nanobubbles were generated with solution exchange before ultrasonciation and then they were subjected to ultrasonication. It was found that ultrasonication did not destroy the pre-existing surface nanobubbles at the HOPG (highly oriented pyrolytic graphite) -water interface. Settling tests and flotation tests indicate that ultrasonication has a negligible influence on the interaction between graphite particles and thus flotation performance. Nanobubbles were not one of the outcomes for ultrasonication.     

25 nm resolution single molecular fluorescence imaging by scanning near-field optical/atomic force microscopy

High-resolution single molecular near-field fluorescence images were observed by scanning near-field optical/atomic force microscopy (SNOM/AFM). We modified the SNOM/AFM for both high-resolution fluorescence imaging and high-resolution topographic imaging. The imaged fluorophore, Alexa 532, is prepared with a poly-methyl-methacrylate (PMMA) film coating. A fluorescence resolution of 25 nm was obtained with a simultaneous topographic image of a flat surface. A sample prepared with a lower PMMA concentration exhibited a rough surface in the micro area. The results for the flat surface indicated that the fluorescence resolution is worst in the rough surface sample, that the maximum fluorescence intensities for the individual fluorophore are similar, and that the decay rate is faster. Thus, we concluded that the morphological effect is an important factor in fluorescence image resolution and the apparent lifetimes of the fluorescence molecules.     

A novel atomic force microscope operating in liquid with open probe unit and optimized laser tracking system

A novel atomic force microscope （AFM） for large samples to be measured in liquid is developed. An innovative laser beam tracking system is proposed to eliminate the tracking and feedback errors. The open probe design of the AFM makes the operation in liquid convenient and easy. A standard 1200-lines/mm grating and a sheet of filter paper axe imaged respectively in air and liquid to confirm its performance. The corrosion behavior of aluminum surface in 1-mol/L NaOH solution is further investigated by the AFM. Experimental results show that the system can realize wide range （20 × 20 （μm）） scanning for large samples both in air and liquid, while keeping nanometer order resolution in liquid by eliminating the tracking and feedback error.     

Imaging and manipulation of biological structures with the AFM

Many biologists have dreamt of physically touching and manipulating the biomolecules they were investigating. With the invention of the atomic force microscope (AFM), this dream has come true. Here, recent applications of the AFM to image and to manipulate biological systems at the nanometer scale are reviewed. Macromolecular biological assemblies as well as individual biomolecules can be subjected to controlled nanomanipulation. Examples of AFM application in imaging and nanomanipulation include the extraction of chromosomal DNA for genetic analysis, the disruption of antibody--antigen bonds, the dissection of biological membranes, the nanodissection of protein complexes, and the controlled modulation of protein conformations. Also reviewed is the novel combination of single molecule imaging and force spectroscopy which allows biomolecules to be imaged, and inter- and intramolecular forces to be measured. Future application of these nanotechniques will reveal new information on the structure, function and assembly of biomolecules.     

Surfactants in Mechanical Alloying/Milling: A Catch-22 Situation

Mechanical alloying/milling technique is characterized by the repeated welding and fracturing of powder particles in a high-energy ball mill, which often results in excessive cold welding and agglomeration of ductile particles. To achieve the critical balance between cold welding and fracturing, the surface of the deforming particles is modified by introducing a suitable organic material, called surfactant or process control agent (PCA). However, the use of surfactants is self-contradictory by nature and requires further consideration of the milling variables and type/amount of surfactant. The current article provides a practical approach to the promises and challenges associated with surfactants in mechanical alloying/milling. An attempt has been made to address the most crucial aspects correlated with surfactants, including contamination, the morphology and size of powder particles, formation of alloy and microstructural evolution, and powder yield, as well as the physico-mechanical properties, such as magnetism, density, hardness, and compressive strength. An overview is also given on the adsorption mechanism of surfactants onto the surface of powder particles, with a special emphasis on type, amount, and the addition time of surfactants in the mechanical alloying process.     

Localization and attempted quantification of various functional groups on pulpwood fibres

The distribution of different free chemical functional groups on wood and pulp fibres has been determined by means of atomic force microscopy (AFM) with chemically modified tips. Because these functional groups show a higher affinity to similar groups on the substrate surface during scanning, AFM images determined with an additional digital pulsed-force mode (DPFM) controller allow the distribution of the chemical components to be imaged and to a degree also to be quantified. The investigated tip coatings showed a different sensitivity towards the major chemical components present in wood fibres, determined on spin-coated films and on wood fibres. A clear distinction between cellulose and lignin was possible in both cases. This technique could therefore be used to differentiate between cellulose and lignin present on pulp fibre surfaces and confirm the successful removal of lignin by pulping.     

Detection of impurity segregation in zone-confined, polycrystalline tin-doped indium oxide thin films by STM and AFM

This paper discusses the possibility of using STM and AFM to image the dopant material in a segregated state. Samples of tin-doped indium oxide were prepared using a zone-confining process. The resultant material has dopant species segregated over certain grain boundaries at desired positions while the others remain dopant free. Samples were then imaged using STM, AFM and STEM. Enhanced contrast from the dopant rich grain boundaries and a larger grain size are observed at the surface making an interface with the substrate as compared to the free surface of the sample, while secondary electron STEM images show these grains to be smaller in size and the boundaries to be almost physically flat. This is interpreted to be a consequence of the zone-confining effect.     

Milling of Organic Solids in a Jet Mill. Part 1: Determination of the Selection Function and Related Mechanical Material Properties

The particle size distribution of pharmaceutically active materials and other fine chemicals determines the performance of the final product to a large extent. Often milling of these particles is necessary. It is not possible to determine the milling conditions solely on the basis of the particle size distribution of the starting material, because the (mechanical) properties of the material also determine the desired milling conditions. It is often not possible to optimize milling conditions experimentally because the amount of material available is frequently highly limited. A theoretical approach towards predicting the best milling conditions is needed. The purpose of this study was to develop a method to predict the desired milling conditions given a specific (organic) solid material. The selection function and the breakage distribution function are usually the starting points in modeling the milling process. The selection function is the parameter that includes the material and mill properties. Dimensional analysis made it possible to correlate the selection function with material properties. A set of theories available in the literature enable prediction of the material properties. For different compounds (lactose, paracetamol, a steroidal compound, and two heterocyclic compounds) the selection functions were calculated. The calculations predict differences: lactose reduces slowly in size, while one of the heterocyclic compounds shows the most intense fracture.     

Chemical surface composition of the polyethylene implanted by Ag ions studied by phase imaging atomic force microscopy

High density polyethylene (HDPE) has been modified by Ag⁺ ion implantation with the energy of 60 keV. The total amount of implanted silver ions was 1, 5 and 12 × 10¹⁵ ions/cm². The surface topography was observed by atomic force microscopy (AFM), while the surface composition changes were detected using phase imaging AFM. Surface topography changes were studied in detail using 3D surface parameters analyses. The average roughness decreased for the implanted HDPE indicating the flattening of the surface. Phase AFM images indicated the homogenization of the polyethylene during ion implantation, while histogram analyses confirmed the change in surface composition.     

Surface roughness analysis and magnetic property studies of nickel thin films electrodeposited onto rotating disc electrodes

Ni films were electrodeposited onto polycrystalline gold substrates mounted on a rotating disc electrode. The effects of rotation speed, film thickness and current density on the kinetic roughening and magnetic properties of the films were investigated. The film surface roughness was imaged using an atomic force microscope (AFM). The results indicate that the film roughness increases as the film thickness or deposition current density increases. We found that the electrodeposited Ni films exhibit anomalous scaling since both local and large-scale roughnesses show a power-law dependence on the film thickness. The effect of electrode rotation speed on the film surface roughness was also investigated. Scanning electron microscopy studies (SEM) had a good agreement with the AFM results. The average crystalline size of the film surfaces is also calculated from X-ray line broadening using (220) peak and Debye–Scherrer formula. The obtained results agree with that of AFM and SEM. The Ni thin films which are grown at different deposition current densities and rotation speeds exhibit in-plane magnetization with coercivities less than 110 Oe.     

Lattice-resolution imaging of the sapphire (0 0 0 1) surface in air by AFM

Lattice-resolution images of single-crystal α-alumina (sapphire) (0 0 0 1) surfaces have been obtained using contact-mode AFM under ambient conditions. It was found that the hexagonal surface lattice has a periodicity of 0.47 ± 0.11 nm, which is identical to that reported previously when the same surface was imaged in water. Large lattice corrugations (as high as 1 nm) were observed, but were concluded to be imaging artifacts because of the strong friction which causes additional deflection of the cantilever. The additional deflection of the cantilever is registered by the detector of the optical beam-deflection AFM resulting in an overestimation of the height at each lattice point. Abrupt changes were also resolved in the topography including honeycomb patterns and a transition from 2D lattices to 1D parallel stripes, with scanning direction. These phenomena can be explained by the commensurate sliding between the tip and sapphire surface due to the strong contact force.     

Material anisotropy revealed by phase contrast in intermittent contact atomic force microscopy

Phase contrast in intermittent-contact atomic force microscopy (AFM) reveals in-plane structural and mechanical properties of polymer monolayers. This is surprising, because measurements of nanoscale in-plane properties typically require contact mode microscopies. Our measurements are possible because the tip oscillates not just perpendicular but also parallel to the sample surface along the long axis of the cantilever. This lateral tip displacement is virtually universal in AFM, implying that any oscillating-tip AFM technique is sensitive to in-plane material properties.     

Dislocation generation and motion at the incipient stages of surface plasticity in barite

Force–penetration curves on nanoindented barite (001) surfaces have been recorded and microstructure imaged with atomic force microscopy (AFM). Pop-ins in the penetration curves have been correlated to the appearance of permanent concave traces in the surface and to the emission of specific dislocations, some of which can cross-slip resulting in terraces winding around the nanoindentation contact point. The dislocations identified in barite can be related to special deformation mechanisms previously reported in metals. The effective shear stress, which results in permanent traces associated with plasticity, as obtained from the AFM data, is compared with the theoretical values estimated for a perfect crystal.     

Structural investigation of n-hexadecanoic acid multilayers on mica surface: Atomic force microscopy study

The structure of n-hexadecanoic acid (HA) multilayers formed by spreading an ethanol solution containing this molecule onto a freshly cleaved mica surface has been studied by atomic force microscopy (AFM). AFM images of multilayers obtained with different coating time showed that HA molecules first formed some sporadic domains on mica surface. With the proceeding of the coating process, these domains gradually enlarged and coalesced, until formed a continuous film finally. It was observed that HA molecules were always adsorbed on mica surface with tilted even-numbered layers structure. The height of the repeated tilted bilayer film was measured to be approximately 3.8 ± 0.2 nm, which implied a ∼60° tilt molecular conformation of the HA bilayers on mica surface. Phase image confirmed that the HA multilayers terminated with the hydrophilic carboxylic acid groups. The formation mechanism of the HA multilayers was discussed in detail. Thus, resulted hydrophilic surfaces are of special interest for further study in biological or man-made member systems.     

Thermally-induced morphological transition in WOx deposited on a ZrO2(1 0 0) substrate

A combined atomic force microscopy (AFM) and X-ray photoelectron spectroscopy (XPS) study of tungsten oxide model catalysts is presented. The model catalysts were prepared by applying the real preparation method to a ZrO₂(1 0 0) single crystal support. AFM imaged several granular structures of scattered dimensions on the surface of ZrO₂(1 0 0) in the as prepared samples. After heating, at low loading the tungsten species rearranged into small WO_x particles strongly interacting with the substrate. At high tungsten content large WO₃ aggregates also formed. XPS analysis confirmed these changes. The estimated surface density of the interacting W-containing species closely matched that of real catalysts.