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At least 163 records · Page 9Linked to original sources

Ultra Flat Gold Surfaces for Use in Chemical Force Microscopy: Scanning Probe Microscopy Studies of the Effect of Preparation Regime on Surface Morphology.

The preparation of ultra flat gold surfaces for use in chemical force microscopy (CFM) has been studied. The surfaces were studied in terms of substrate effects by comparing mica, Si (110) wafer and glass slides. The effect of different annealing regimes was also investigated. Measurements on these surfaces were made by both atomic force microscopy (AFM) (in contact and tapping mode) and by scanning tunneling microscopy (STM). The films contain different morphologies with respect to grain size and topography. Calculations of surface roughness present values less than 2.5 nm for all surfaces studied, making the choice of the "flattest" surface difficult if based on criteria of surface roughness alone. Additionally, it is shown that different acquisition parameters can produce dissimilar images that have stability and reproducibility.

Journal Article↗

Scanning acoustic Doppler microscopy and scanning acoustic correlation microscopy.

Acoustic microscopy with vector contrast at 100 MHz in a fluid with immersed particles is used to detect the flow profile in front of a microscopic orifice. The velocity profile concerning the component in axial direction of the focused beam is derived from the phase contrast. Possibilities to resolve the flow profile also for the components in normal direction with respect to the axis are demonstrated. The methods concerning measurement techniques and data evaluation for scanning acoustic Doppler microscopy are presented. For scanning acoustic correlation microscopy the time dependent phase and amplitude signals resulting from sound waves scattered by the immersed particles (aluminium flakes with a typical diameter of 10 microm) have been analysed by correlation procedures. From the obtained autocorrelation functions the velocity distribution can be derived. Both methods can be applied simultaneously. Data analysis is based on the information contained in the originally obtained images in vector contrast derived from temporal and spatial resolved analogue and digital processing of the acoustic signals.

Journal Article↗

Multi-photon Excitation Microscopy and Confocal Microscopy Imaging of In Vivo Human Skin: A Comparison.

: We compare here multi-photon excitation microscopy and tandem scanning reflected light confocal microscopy for the microscopic observation of human skin in vivo. Multi-photon excitation is induced by a 80-MHz pulse train of femtosecond laser pulses at 780 nm wavelength. This nonlinear microscopic technique is inherently suitable for tissue fluorescence imaging because of its deeper penetration depth and lower specimen photodamage. This technique has noninvasively obtained tissue structural information in human epidermis and dermis. Alternatively, tandem scanning confocal light microscopy based on a white light source can provide video-rate image acquisition with high resolution and high contrast. Reflected light confocal methods have been used to obtain images from the skin surface to the epidermal-dermal junction. The relative merits of these two techniques can be identified by comparing three-dimensionally resolved images obtained from the forearm skin of the same volunteer.

Journal Article↗

Compositional and morphological imaging of CO2 laser irradiated human teeth by low vacuum SEM, confocal laser scanning microscopy and atomic force microscopy.

Enamel and dentin of human teeth irradiated by CO2 laser were investigated by confocal laser scanning microscopy (CLSM), low vacuum scanning electron microscopy (WET-SEM) and atomic force microscopy (AFM). Optical tomographic imaging by CLSM, compositional imaging based on atomic number effect of reflected electrons by WET-SEM, high resolution observation of surface morphology by AFM were done for both the irradiated and non-irradiated area of the same specimen throughout. The crystals of about 50 microm length and the bright spots were observed by CLSM at the bottom of the cavity induced by laser irradiation. They turned out from the observation by WET-SEM as the acicular crystals with the cross section of an irregularly hexagonal shape situated parallel and perpendicular, respectively, to the inner surface of the cavity. The thickness of the thermally deteriorated zone of the cavity was about 25 microm. The crystals unidirectionally grown up to the size of several hundreds nm were also observed by AFM, while the apatite crystallites of 50-150 nm were recognized all over in non-irradiated area. All the results suggest that after instantaneous melting at the surface of teeth by CO2 laser shot the crystals of calcium phosphate were recrystalized and grown to a large size. The compositional imaging in addition to morphological observation was useful to obtain the information of the change in materials induced by laser irradiation.

Journal Article↗

Comparative morphology of granulocytes collected by three methods of leukapheresis. A light microscopy and transmission electron microscopy study.

The morphology of granulocytes collected by continuous-flow centrifugation (CFC), discontinuous-flow centrifugation (DFC), and continuous-flow filtration (CFF) was investigated in 18 healthy donors by means of light microscopy and transmission electron microscopy. Light microscopy study of semithin sections of granulocytes collected by CFC and DFC showed minimal morphologic abnormalities, compared to granulocytes procured by CFF. Ultrastructural study of granulocytes procured by CFF showed more conspicuous qualitative and quantitative abnormalities (the most prominent being "microvilli," degranulation, and bazarre chromatin) than in granulocytes obtained by the other two methods. Controls showed that the bulk of CFF-cell abnormalities was due to the "tapping" of the filters. Factors such as the mechanical compression (plasma extractor) used in DFC method, donor pretreatment with anticoagulants and steroids, hydroxyethyl starch, and duration of leukapheresis scarcely influenced granulocyte morphology.

Adult↗

Scanning electron microscopy and transmission electron microscopy study of hot-deformed gamma-TiAl-based alloy microstructure.

The aim of this work was to assess the changes in the microstructure of hot-deformed specimens made of alloys containing 46-50 at.% Al, 2 at.% Cr and 2 at.% Nb (and alloying additions such as carbon and boron) with the aid of scanning electron microscopy and transmission electron microscopy techniques. After homogenization and heat treatment performed in order to make diverse lamellae thickness, the specimens were compressed at 1000 degrees C. Transmission electron microscopy examinations of specimens after the compression test revealed the presence of heavily deformed areas with a high density of dislocation. Deformation twins were also observed. Dynamically recrystallized grains were revealed. For alloys no. 2 and no. 3, the recovery and recrystallization processes were more extensive than for alloy no. 1.

Journal Article↗

Minocycline-related cutaneous hyperpigmentation as demonstrated by light microscopy, electron microscopy and X-ray energy spectroscopy.

A 70-year-old patient with a chronic cutaneous ulcer treated by minocycline hydrochloride developed hyperpigmentation of the forearms. Biopsy material was studied by light microscopy, electron microscopy and X-ray energy spectroscopy. Granular gold-brown pigment was found in dermal histiocytes and eccrine myoepithelial cells, which gave positive reaction with Prussian blue and Fontana-Masson stains. Electron microscopy revealed intracytoplasmic granules of dark, homogeneous material and small fine particles. X-ray energy spectroscopy showed iron and other elements in smaller amounts. The different types of minocycline-related hyperpigmentation and the possible pathomechanism are discussed with special regard to the importance of the diagnostic methods.

Aged↗

Characterization by atomic force microscopy and cryoelectron microscopy of tau polymers assembled in Alzheimer's disease.

The structure of the Paired Helical filaments (PHF), a polymer of the microtubule associated protein tau, has been studied by Atomic Force Microscopy (AFM) and by cryoelectron microscopy. Mica and graphite were used as substrates in the AFM analysis with no differences in the results. A banding pattern of 8-12 nm width within the helical structure is found when detailed analysis of the data is performed. High AFM resolution images obtained by using an ultra sharp tip confirm the previous results and suggest that the structures observed are compatible with a helical ribbon made up of two parallel strands. These results were confirmed by cryoelectron microscopy experiments.

Journal Article↗

The immotile cilia syndrome: phase contrast light microscopy, scanning and transmission electron microscopy.

In the immotile cilia syndrome, transmission electron microscopy of the cilia shows abnormalities in the arrangement of the central pairs of tubules and in the dynein arms of the peripheral tubules, or in the radial spokes, We studied four nonrelated children, 9/12, 5, 6, and 6 years old, with situs inversus and a history of chronic sinusitis and bronchitis (Kartagener's syndrome) and four children in the same age group and with the same history, but without situs inversus. Under the phase contrast microscope no motile cilia were seen in the four patients with Kartagener's syndrome and in two of the four other children. Transmission electron microscopy showed aberrations in the cilia (absence of dyneim arms, random orientation of central tubules) in the patients with Kartagener's syndrome. Scanning electron microscopy revealed differences in morphology and arrangement of cilia between patients and controls. In the patients much more mucus was present on the mucosal surface. Furthermore, the cilia were in a state of disorder, with a multidirectional orientation instead of the parallel orientation seen in controls.

Child↗

Characterization of the annealed (0001) surface of sapphire (alpha-Al2O3) and interaction with silver by reflection electron microscopy and scanning reflection electron microscopy.

Annealed (0001) surfaces of single-crystal sapphire (alpha-Al2O3) rod have been studied in the electron microscope using reflection electron microscopy (REM), scanning reflection electron microscopy (SREM), and reflection high energy electron diffraction (RHEED). Annealed surfaces of (0001) sapphire are vicinal and characterized by close-packed (0001)-oriented terraces separated by faceted multiple-height steps, with edges parallel to energetically preferred low-index directions (less than 1010 greater than and less than 1120 greater than). These structural features are not seen on cleaved surfaces or polished surfaces treated at temperatures less than 1,250 degrees C. Oxygen-annealing produces clean surfaces which prove useful for investigating the interaction of deposited metals with the (0001) sapphire. Both REM and SREM (with microdiffraction spots) techniques have been used to observe fine structure of flat Ag islands on the scale of 1-100 nm on the (0001)-oriented terraces as well as aggregations at the steps. A preliminary result on interaction with Cu is also included.

Aluminum Oxide↗

3-D morphological characterization of the liver parenchyma by atomic force microscopy and by scanning electron microscopy.

A comparative study of atomic force microscopy (AFM) and scanning electron microscopy (SEM) imaging of the healthy human liver parenchyma was carried out to determine the similarities and the differences. In this study, we compared the fine hepatic structures as observed by SEM and AFM. Although AFM revealed such typical hepatic structures as bile canaliculi and hepatocytes, it also showed the location of the nucleus and chromatin granules in rough relief structure, which was not visible by SEM. By contrast, SEM visualized other structures, such as microvilli, the central vein, and collagenous fibers, none of which was visualized by AFM. For better orientation and confirmation of most of the structures imaged by SEM and AFM, Congo Red-stained specimens were also examined. Amyloid deposits in the Disse's spaces were shown especially clearly in these images. The differences between the SEM and AFM images reflected the characteristics of the detection systems and methods used for sample preparation. Our results reveal that more detailed information on hepatic morphology is obtained by exploiting the advantages of both SEM and AFM.

Adult↗

A study of phase separation in peptide-loaded HPMC films using T(zero)-modulated temperature DSC, atomic force microscopy, and scanning electron microscopy.

Despite the widespread use of drug-loaded polymeric systems, there is still considerable uncertainty with regard to the nature of the distribution of the drug within the polymer matrix. The aim of this investigation was to develop thermal and microscopic techniques whereby the miscibility and spatial distribution of a model peptide, cyclosporin A (CyA), in hydroxypropyl methylcellulose (HPMC) films may be studied. The new technique of T(zero)-modulated temperature differential scanning calorimetry (T(zero) MTDSC), scanning electron microscopy (SEM), and pulse force mode atomic force microscopy (PFM-AFM) were used in conjunction to study films prepared using a solvent evaporation process, with a solvent extraction study performed to elucidate the nature of the observed phases. T(zero) MTDSC studies showed glass transitions for both the HPMC and CycA, with the T(g) for the HPMC and CycA seen for the mixed systems. SEM showed two spherical phases of differing electron density. PFM-AFM also showed spheres of differing adhesion that increased in size on addition of drug. Pixel intensity analysis indicated that the smaller spheres corresponded to CycA. Exposure of the films to dichloromethane, in which CycA is soluble but HPMC is not, resulted in the presence of voids that corresponded well to the spheres suggested to correspond to the drug. It was concluded that the system had undergone extensive or complete phase separation, and that the thermal and microscopic techniques outlined above are an effective means by which this issue may be studied.

Calorimetry, Differential Scanning↗

A study of sulfamerazine single crystals using atomic force microscopy, transmission light microscopy, and Raman spectroscopy.

Sulfamerazine (SMZ) Form I and II single crystals were prepared from aqueous dispersions of SMZ bulk samples and studied using several microscopic and spectroscopic techniques. Transmission light microscopy and Raman spectroscopy were used to observe and identify single crystals. The results indicated that Form I single crystals tended to be rectangular laths while Form II ones tended to be hexagonal laths. Surface morphology of individual single crystals was further investigated by atomic force microscopy (AFM). AFM images revealed a smooth top surface, a uniform height, and sharp edges for both forms of single crystals. Both height and phase images showed crystalline terraces with different step heights for the top surface of Form I. Surface properties of single crystals were evaluated using AFM force measurements. Experimental results indicated that the top surface of Form I single crystals was more hydrophilic than that of Form II. Theoretical calculations predicted a dominant crystal face of (020) for the Form I single crystals and (002) for the Form II ones. The correlations between calculation predictions and experimental results were discussed.

Anti-Infective Agents↗

Transmission electron microscopy and scanning force microscopy of poly r(A-U) and poly r(A-U)-ethidium bromide.

Transmission electron microscopy and scanning force microscopy of negative-stained, carbon-coated replica and mica-adsorbed preparations of 200 microM poly r(A-U) and 50 microM ethidium bromide/200 microM poly r(A-U) have been employed to evaluate ethidium-induced changes in poly r(A-U) topology. Poly r(A-U) alone exhibits elongated conformations 85-115 nm in length that possess a number of hairpin loops as well as single-stranded domains. While the double-stranded domains are found predominately at the base of the hairpin loops (diameter = 5-30 nm), other rod-like (presumably double-stranded) regions ranging from 25-80 nm in length are present in other portions of the poly r(A-U). In contrast with the poly r(A-U) alone, the EB/poly r(A-U) combination appears as a heterogeneous population of condensed structures whose lengths and widths vary from 12-88 nm and 15-45 nm, respectively. These conformational changes are due to a number of factors, including the displacement of ordered water surrounding the poly r(A-U) and charge shielding of the phosphate groups of the poly r(A-U) upon the binding of the ethidium.

Adenine↗

Scanning electron microscopy studies of protein-functionalized atomic force microscopy cantilever tips.

Protein-functionalized atomic force microscopy (AFM) tips have been used to investigate the interaction of individual ligand-receptor complexes. Herein we present results from scanning electron microscopy (SEM) studies of protein-functionalized AFM cantilever tips. The goals of this study were (1) to examine the surface morphology of protein-coated AFM tips and (2) to determine the stability of the coated tips. Based on SEM images, we found that bovine serum albumin (BSA) in solution spontaneously adsorbed onto the surface of silicon nitride cantilevers, forming a uniform protein layer over the surface. Additional protein layers deposited over the initial BSA-coated surface did not significantly alter the surface morphology. However, we found that avidin-functionalized tips were contaminated with debris after a series of force measurements with biotinylated agarose beads. The bound debris presumably originated from the transfer of material from the agarose bead. This observation is consistent with the observed deterioration of functional activity as measured in ligand-receptor binding force experiments.

Adsorption↗

Validation of three-dimensional surface characterising methods: scanning electron microscopy and confocal laser scanning microscopy.

Surface characteristics of enosseous titanium implants have been known to influence the quality of osseointegration. Parameters recommended for metrical analysis should be supplemented by a topographical description. In this study, Ra values obtained by established tactile and optical profilometric methods are correlated with those obtained by stereo scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). As test specimens, standardised CVD diamond-coated titanium alloys with different microwave coating power ranging from 2,000 to 3,000 W were used. A non-CVD-coated corund grit-blasted specimen (NC) was used as reference. After coating with a CVD, power of 2,000 W Ra ranges from 4.33 to 5.69 microm depending on the method used. With increasing power of the coating process, the amplitude of the surface roughness is significantly increased to 4.53 to 6.89 microm. Ra values of the same sample obtained by different methods are also significantly different (p = 0.001). Compared with the established methods of tactile and optical profilometry, SEM and CLSM offer valid data on the surface roughness accompanied by a topographical imaging. In future studies, the underlying method should be specified to interpret roughness values correctly, as not every method is suitable for each specimen and values obtained by different methods vary extremely.

Diamond↗

Quantitative intercomparison of transmission electron microscopy, flow cytometry, and epifluorescence microscopy for nanometric particle analysis.

Nanometric biological particles such as viruses have received increased attention in a wide range of scientific fields. Evaluation of viral contributions to environmental processes and the use of viruses in medical applications such as gene therapy require viruses to be routinely and accurately enumerated. There are a variety of existing techniques for counting viruses, namely, plaque assays, transmission electron microscopy (TEM), epifluorescence microscopy (EFM), and flow cytometry (FCM); each has advantages and disadvantages. While there have been attempts to intercompare some of these techniques to determine the most effective means to count viruses, no previous study used a technique-independent standard for quantitative comparison of collection efficiency, accuracy, and precision. In this work, polystyrene nanospheres were used as standards for the intercomparison of performance characteristics for TEM, EFM, FCM, as well as a custom-built flow cytometer (the Single Nanometric Particle Enumerator, SNaPE). EFM and SNaPE exhibited the highest degree of accuracy and precision, with particle concentrations deviating < or =5% from true and relative errors less than half that of TEM, EFM and SNaPE are also significantly more time and cost efficient than TEM.

Calibration↗

Immunogold labelling of fibroblast focal adhesion sites visualised in fixed material using scanning electron microscopy, and living, using internal reflection microscopy.

A new immunogold labelling method for the visualisation of vinculin, an integral protein in focal adhesions of cells, is reported. Quantification of vinculin is indicative of substrate cytocompatibility (cytocompatibility is one aspect of biocompatibility; it is the cellular response to a biomaterial). For efficient labelling, most of the cell body above the cell-substrate interface was removed with detergent. The antigen blocking procedure, size of label (5 nm) and duration of silver-enhancement (6 min), for visualisation of the labelled sites on the whole cell by scanning electron microscopy (SEM), were determined. Imaging living cells with interference reflection light microscopy, followed by backscattered electron (BSE) imaging of the same fixed and immunolabelled cells confirmed the results. Collecting low voltage BSE images of embedded cells after the substrate had been removed provided 'sectional' views through the cell. This enabled visualisation of vinculin exclusively within the cell-substrate contact zone; the focal adhesions. The method could be of general use in the imaging of protein distribution at biological tissue/substrate interfaces.

Acrylic Resins↗