Fabrication of hierarchical micro/nanostructures via scanning probe lithography and wet chemical etching

In this study, we propose a simple and effective method for fabricating hierarchical silicon structures via the combination of scanning probe lithography (SPL) and wet chemical etching. Here, silicon oxide structures were protruded from a 100-oriented silicon surface, followed by the passivation of silicon nitride by AFM tip-induced local oxidation. Based on the two-dimensional (2D) silicon oxide patterns, three-dimensional (3D) microstructures with high aspect ratios were formed by wet etching with HF and KOH. A variety of combinations of SPL and the etching process allowed us to fabricate diverse silicon-based structures such as deep-etched microstructures and multi-terraced nanostructures.     

The effect of deformation on the lateral resolution of atomic force microscopy

A computer model based on the elastic properties of rubber is introduced for the evaluation of the lateral resolution in atomic force microscopy of deformable specimens. The computational results show that, if the full width at half-height can be defined as the lateral resolution, it is continuously improved at greater probe forces, at the expense of a reduced molecular height. In fact, even for a probe that is bigger than the molecule, the real size of the molecule can be 'recovered' at about 25% compression. This result demonstrates that for a better lateral resolution, a greater probe force can be beneficial, provided that the molecule is not moved or damaged and the response remains elastic. Measurements on isolated low-density lipoproteins (LDL) show that with 26% vertical compression, the lateral size measured in atomic force microscopy is only about 72% of the value predicted by a simple convolution, and is only slightly larger (≈ 13%) than the known size of LDL. Therefore, the results on LDL provide a direct support for the conclusions of the computational model.     

Quantitative measurement of mRNA at different loci within an individual living cell

Asymmetric localizations of cellular proteins and mRNAs are important for cell functions such as division, differentiation and development. The localization of specific mRNA generates cell polarity by controlling the translation sites of specific proteins and thereby restricting their locations to appropriate cellular regions. We have previously reported a novel method based on atomic force microscopy (AFM) for examining gene expression in a single living cell without killing or destroying it. An AFM tip was inserted into a living cell to extract mRNAs, which were analyzed after multiplication by RT-PCR and quantitative PCR. By applying this method, in this study we performed quantitative measurement of mRNA at different loci within individual living cells.     

Study of the nanoscopic deformation of an annealed nafion film by using atomic force microscopy and a patterned substrate

We demonstrated the repetitive imaging of the same area of a nafion film before and after annealing by using atomic force microscopy (AFM). In order to find the exact same area of the same sample after changing the cantilever and reattaching the sample, a micropatterned substrate was developed. A micropattern with a 250–500 μm pitch was prepared on the backside of a transparent glass substrate. This pattern includes various signs such as colored letters and numbers at the center of each lattice of the pattern. The nanostructures fabricated by AFM nanolithography on a nafion film using this new method were successfully characterized before and after annealing (over 100 °C). The AFM images clearly showed that the nanostructures on a nafion film were dramatically changed by annealing. The data indicated an evidence to understand why the nafion fuel cell does not work well at high temperatures. Our method is probably effective for the study of nanoscopic dynamics in various surface structures.     

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.     

Real-time observations of mechanical stimulus-induced enhancements of mechanical properties in osteoblast cells

Osteoblast, playing a key role in the pathophysiology of osteoporosis, is one of the mechanical stress sensitive cells. The effects of mechanical load-induced changes of mechanical properties in osteoblast cells were studied at real-time. Osteoblasts obtained from young Wister rats were exposed to mechanical loads in different frequencies and resting intervals generated by atomic force microscopy (AFM) probe tip and simultaneously measured the changes of the mechanical properties by AFM. The enhancement of the mechanical properties was observed and quantified by the increment of the apparent Young's modulus, E(*). The observed mechanical property depended on the frequency of applied tapping loads. For the resting interval is 50s, the mechanical load-induced enhancement of E(*)-values disappears. It seems that the enhanced mechanical property was recover able under no additional mechanical stimulus.     

High-resolution constant-height imaging with apertured silicon cantilever probes

We present high-resolution aperture probes based on non-contact silicon atomic force microscopy (AFM) cantilevers for simultaneous AFM and near-infrared scanning near-field optical microscopy (SNOM). For use in near-field optical microscopy, conventional AFM cantilevers are modified by covering their tip side with an opaque aluminium layer. To fabricate an aperture, this metal layer is opened at the end of the polyhedral probe using focused ion beams (FIB). Here we show that apertures of less than 50 nm can be obtained using this technique, which actually yield a resolution of about 50 nm, corresponding to λ/20 at the wavelength used. To exclude artefacts induced by distance control, we work in constant-height mode. Our attention is particularly focused on the distance dependence of resolution and to the influence of slight cantilever bending on the optical images when scanning at such low scan heights, where first small attractive forces exerted on the cantilever become detectable.     

In situ wear studies of surface micromachined interfaces subject to controlled loading

Friction and wear are major limiting factors for the development and commercial implementation of devices fabricated by surface micromachining techniques. These tribological properties are studied using a polycrystalline silicon nanotractor device, which provides abundant, quantitative information about friction and wear at an actual microelectromechanical system (MEMS) interface. This in situ approach to measuring tribological properties of MEMS, combined with high-resolution atomic force microscope (AFM) images of wear tracks, provides insight into the effects of different MEMS surface processing on wear. In particular, monolayer coatings have a significant positive effect, while surface texturing does not strongly affect performance.     

Imaging the native structure of the chaperone protein GroEL without fixation using atomic force microscopy

Most sample preparation methods for scanning probe or electron microscopy require that biomolecules, such as proteins, be fixed. Fixation destroys the molecular functionality and can possibly affect the true molecular structure. Here we report sample preparation conditions that allow the imaging of an unfixed protein, GroEL, under in-vivo conditions, by atomic force microscopy. Under these conditions, the protein should maintain its native structure and biological activity. The typical toroidal shape with pore of the GroEL complex was easily visible in the images. Images of a single complex show dimensions that agree well with crystallographic data. Under in-vivo conditions, it should be possible to study the biological activity and function of proteins.     

Characterization of nanoscale recording mark on Ge2Sb2Te5 film

We have fabricated nanoscale recording marks on Ge₂Sb₂Te₅ (GST) films with conductive atomic force microscope (AFM). GST films were deposited on glass or polyimide film with thickness of 150–200 nm by the rf–sputtering method. Through current–voltage (I–V) spectroscopy, good cantilevers for fabrication and characterization of nanoscale marks on GST were selected. A fresh and highly conductive tip showed voltage-switching characteristic in the I–V curve, where the threshold voltage was 1.6 V. Nanoscale dot and wire arrays of crystalline phases were successfully obtained by varying sample bias voltage from −10 to 10 V. With highly conductive tips, nanowires having full-width at half-maximum of 20 nm could be fabricated, whereas nanowires could not be fabricated with bias voltage below −2 V. The width of the nanoscale mark was increased by repetition of AFM lithography even with same applied voltage and lithography speed. For a thicker nanowire, the width measured in current-image (C-image) was observed to be 2 times of that measured in topography-image (T-image). This result supports that current sensing provides an image of phase-changed GST area with higher resolution than topography sensing.     

Morphological, nanomechanical and cellular structural characterization of human hair and conditioner distribution using torsional resonance mode with an atomic force microscope

Characterization of the cellular structure and chemical and physical properties of hair are essential to develop better cosmetic products and advance the biological and cosmetic sciences. Although the morphology of the fine cellular structure of human hair has traditionally been investigated using scanning electron microscopy and transmission electron microscopy, atomic force microscopy can be used for characterization in ambient conditions without requiring specific sample preparations and surface treatment. In this study, the tapping and torsional resonance modes in an atomic force microscope are compared for measurements of stiffness and viscoelastic properties. The materials were mapped using amplitude and phase angle imaging. The torsional resonance mode showed advantages in resolving the in-plane (lateral) heterogeneity of materials. This mode was used for investigating and characterizing the fine cellular structure of human hair. Various cellular structures (such as the cortex and the cuticle) of human hair and fine sublamellar structures of the cuticle, such as the A-layer, the exocuticle, the endocuticle and the cell membrane complex were easily identified. The distribution and thickness of conditioner on the treated hair surface affects the tribological properties of hair. The thickness of the conditioner was estimated using force distance measurements with an atomic force microscope.     

Effect of interlayer on structure and performance of anode-supported SOFC single cells

To lower the operating temperatures in solid oxide fuel cell (SOFC) operations, anode-supported SOFC single cells with a single dip-coated interlayer were fabricated and the effect of the interlayer on the electrolyte structure and the electrical performance was investigated. For the preparation of SOFC single cells, yttria-stabilized zirconia (YSZ) electrolyte, NiO-YSZ anode, and 50% YSZ-50% strontium-doped lanthanum manganite (LSM) cathode were used. In order to characterize the cells, scanning electron microscopy (SEM) and atomic force microscopy (AFM) were utilized and the gas (air) permeability measurements were conducted for gas tightness estimation. When the interlayer was inserted onto NiO-YSZ anode, the surface roughness of anode was diminished by about 40% and dense crack-free electrolytes were obtained. The electrical performance was enhanced remarkably and the maximum power density was 0.57W/cm(2) at 800 degrees C and 0.44W/cm(2) at 700 degrees C. On the other hand, the effect of interlayer on the gas tightness was negligible. The characterization study revealed that the enhancement in the electrical performance was mainly attributed to the increase of ion transmission area of anode/electrolyte interface and the increase of ionic conductivity of dense crack-free electrolyte layer.     

Atomic force microscopy of histological sections using a new electron beam etching method

In order to examine histological sections of the rat vomeronasal epithelium with the atomic force microscope (AFM), we developed an electron beam etching method that improves the resolution of AFM images. This method results in AFM images comparable to those obtained with the transmission electron microscope (TEM). Ultrathin tissue sections embedded in epoxy resin were observed before and after the treatment with electron beam radiation. Before electron beam treatment, epithelial structures such as the microvilli surface, dendritic processes, the supporting cell layers and the neuronal cell layers were all visible using the AFM. However, only a few subcellular structures could also be resolved. The AFM images were not as clear as those obtained with the TEM. After electron beam treatment, however, the resolution of AFM images was greatly improved. Most of the subcellular structures observed in TEM images, including the inner membrane of mitochondria, ciliary-structure precursor body, junctional complexes between the neurons and supporting cells, and individual microvilli were now visible in the AFM images. The electron beam treatment appeared to melt the embedding resin, bringing subcellular structures into high relief. The result of this study suggests that electron beam etching of histological samples may provide a new method for the study of subcellular structure using the AFM.     

Analysis of force curves obtained on the live cell membrane using chemically modified AFM probes

Force curves were obtained on the live cell surface using an atomic force microscope mounted with a modified tip with the bifunctional covalent crosslinker, disuccinimidyl suberate, which forms a covalent bond with amino-bearing molecules on the cell surface. A ramp delay time of 1.0 s was introduced before the start of the retraction regime of the force curve to increase the stationary reaction time between the crosslinkers on the tip and the amino groups on the cell surface. While live cell surface responses to forced contact with a non-functionalized tip rarely showed evidence of tip–cell interaction, those obtained with modified tips gave clear indication of prolonged adhesion which was terminated by a single step release of the tip to its neutral position. Under the given experimental conditions of this work, 58% of a total of 198 force curves gave only one jump and 70% of those with one jump gave the final rupture force of 4.5±0.22 nN. The result emphasized the uniqueness of the observed mechanical response of the cell surface when probed with chemically modified tips.     

退火温度对二氧化钛薄膜的性能影响

为了获得性能优良的二氧化钛薄膜,采用电子束蒸发沉积方法制备二氧化钛薄膜,并分别在300、600、900℃空气中对样品进行退火处理以改善所制备二氧化钛薄膜的性能。分别采用X射线衍射(XRD)、原子力显微镜(AFM)以及分光光度计研究了退火温度对二氧化钛薄膜结构和光学性能的影响。结果表明,退火处理可以使二氧化钛薄膜由非晶态薄膜转换为金红石型薄膜,且金红石晶型成分随退火温度的加大而增大,同时退火处理可以改善二氧化钛薄膜在300~1 200nm光谱范围的总吸光率以及增大二氧化钛薄膜的应用范围。     

Implementation of a piezoresistive MEMS cantilever for nanoscale force measurement in micro/nano robotic applications

The nanoscale sensing and manipulation have become a challenging issue in micro/nanorobotic applications. In particular, a feedback sensor-based manipulation is necessary for realizing an efficient and reliable handling of particles under uncertain environment in a micro/ nano scale. This paper presents a piezoresistive MEMS cantilever for nanoscale force measurement in microrobotics. A piezoresistive MEMS cantilever enables sensing of gripping and contact forces in nanonewton resolution by measuring changes in the stress-induced electrical resistances. The calibration of a piezoresistive MEMS cantilever is experimentally carried out. In addition, as part of the work on nanomanipulation with a piezoresistive MEMS cantilever, the analysis on the interaction forces between a tip and a material, and the associated manipulation strategies are investigated. Experiments and simulations show that a piezoresistive MEMS cantilever integrated into a microrobotic system can be effectively used in nanoscale force measurements and a sensor-based manipulation.     

Immobilization of DNA on 11-mercaptoundecanoic acid-modified gold (111) surface for atomic force microscopy imaging

Immobilized DNA on preformed 11-mercaptoundecanoic acids (MUDA) self-assembled monolayers (SAMs) on a gold (111) surface was bound by a divalent cation bridges was imaged by atomic force microscopy (AFM). The DNA immobilization was attributed to the formation of ionic bridges between the carboxylate groups of MUDA and the phosphate groups of DNA. AFM images revealed that DNA molecules could be immobilized strongly enough to permit stable and reproducible imaging. The effect of different bridge cations, such as Mg(2+), Zn(2+) and Cu(2+), and the pH of DNA assembled solution on immobilization and conformation of DNA was studied. Plasmid DNA pBR 322/Pst I molecules were straightened by using a molecular combing technique on the MUDA surface.     

Effect on crystalline structure of AISI M2 steel using tungsten–thorium electrode through MRR,EWR, and surface finish

To investigate on the crystalline structure of AISI M2 steel by using tungsten–thorium electrode in electrical discharge machining (EDM) process was studied. Furthermore, the investigation were carried out for finding the value of material removal rate (MRR), electrode wear rate (EWR) and surface roughness (SR) of tool steel material depending upon three variable input process parameters. On the basis of weight loss, the value of MRR and EWR were calculated at optimized process parameter. Subsequently, surface topography of the processed material were examined through different characterization techniques like scanning electron microscopy (SEM), Optical surface profiler (OSP) and Atomic force microscopy (AFM), respectively. In XRD study, broadening of the peak was observed which confirmed the change in material properties due to the homogeneous dispersion of the particles inside the matrix. Lowest surface roughness and MRR of 0.001208 mg/min was obtained. Minimum surface roughness was obtained 1.12 μm and 2.18427 nm by OSP and AFM study, respectively. Also, minimum EWR was found as 0.013986 mg/min.