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  • Articles  (2,506)
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  • 1
    Publication Date: 2013-09-13
    Description: ABSTRACT The objective of this study was to analyze the dentin-resin cements interfacial ultramorphologies using two different methods: scanning (SEM) and transmission electron microscopy (TEM). Four commercial products were evaluated: two conventional cementing system (RelyX ARC/Adper™ Scotchbond™ Multi-Purpose Plus, 3M ESPE and Clearfil Esthetic Cement/DC Bond, Kuraray) and two self-adhesive resin cements (RelyX Unicem, 3M ESPE and Clearfil SA Cement, Kuraray). Prepolymerized resin disks (Sinfony, 3M ESPE) were cemented on oclusal dentin surfaces of 24 third human molars, simulating the indirect restorations. After 24 h, teeth were sectioned into 0.9-mm thick slabs and processed for microscopy analyses (SEM or TEM/ n = 3). Qualitative characterization of dentin-resin cement interface was performed. Hybrid layer formation with long and dense resin tags was observed only for RelyX ARC cementing system. Clearfil Esthetic Cement/DC Bond system revealed few and short resin tags formation, whereas no hybridization and resin tags were detected for self-adhesive resin cements. Some interfacial regions exhibited that the self-adhesive resin cements were not bonded to dentin, presenting bubbles or voids at the interfaces. In conclusion, TEM and SEM bonding interface analyses showed ultramorphological variations among resin cements, which are directly related to dental bonding strategies used for each resin cement tested. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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    Topics: Natural Sciences in General
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  • 2
    Publication Date: 2013-09-13
    Description: ABSTRACT Mimosa pudica has three distinct specialized organs, namely, pulvinus, secondary pulvinus, and pulvinule, which are respectively controlling the movements of petioles, leaflets, and pinna in response to external stimuli. Water flow is a key factor for such movements, but detailed studies on the organization of the vascular system for water transport in these organs have not been published yet. In this study, organizations of the xylem vessels and morphological features of the pulvinus, the secondary pulvinus, and the pulvinule were experimentally investigated by X-ray computed tomography and histological technique. Results showed that the xylem vessels were circularly distributed in the specialized motile organs and reorganized into distinct vascular bundles at the extremities. The number and the total cross-sectional area of the xylem vessels were increased inside the specialized motile organs. Morphological characteristics obtained in this study provided new insight to understand the functions of the vascular networks in the dynamic movements of M. pudica . Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 3
    Publication Date: 2013-09-07
    Description: ABSTRACT Estimates of the radii and solute concentrations of simulated microstructures containing ultrafine spherical precipitates were determined from isoconcentration surfaces and proximity histograms. The accuracy of the estimates of the solute concentrations and the radii of precipitates was found to depend on the size of precipitates. Optimized parameters for analyzing 0.5- to 2-nm-radius precipitates are proposed. The solute content of 0.5-nm-radius precipitates was not estimated correctly by this method. The accuracy of the estimates of the solute concentration and the radius of precipitates were primarily influenced by the solute concentrations of the precipitates. The ranges of error of the solute concentration in the precipitates, which are associated with the analytical limitations of the ultrafine precipitates, were determined, and the results indicated a limitation of the estimates. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 4
    Publication Date: 2013-09-07
    Description: ABSTRACT Ovate mitochondria in cultured human fibroblasts divide by pinching. In the process, as observed by transmission electron microscopy, a deep incisure of the surface membranes separates the organelle into two lobes connected by a slender isthmus. A single element of smooth endoplasmic reticulum (SER) invariably accompanies each incisure, extending deep into the cleft. When the ingrowing membranes meet and fuse with the antipodal membranes, fission occurs. Elongated mitochondria that give no indication of division often are cloaked by a single, continuous cistern of SER. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 5
    Publication Date: 2013-09-12
    Description: ABSTRACT The regeneration of axons after a spinal cord injury or disease is attracting a significant amount of interest among researchers. Being able to assess these axons in terms of morphology, length and origin is essential to our understanding of the regeneration process. Recently, two specific axon tracers have gained much recognition; biotinylated dextran amine (BDA) 10 kDa as an anterograde tracer and cholera toxin-B as a retrograde tracer. However, there are still several complexities when using these tracers, including the volume that should be administered and the best administration site so that a significant amount of axons are labeled in the area of interest. In this article, we describe some simple procedures for injecting the tracers and detecting them. We also quantified the number of axons at different locations of the spinal cord. Our results show axons labeled from motor cortex injections traveled down to the lumbosacral spinal cord in 2 weeks, while BDA injections into the lateral vestibular nucleus and reticular formation took 3 weeks to label axons in the lumbosacral spinal cord. Moreover, this protocol outlines some basic procedures that could be used in any laboratory and gives insight into the number of axons labeled and how procedures could be tailored to meet specific researcher's needs. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 6
    Publication Date: 2013-09-26
    Description: ABSTRACT The usefulness of embedment-free section transmission electron microscopy (TEM) is stressed for present and future morphological analyses, and several examples are demonstrated which are revealed in sections for the first time by this method: en-face views of slit diaphragm of renal glomerulus and fenestrated diaphragm of capillary endothelium, transparency of neural myelin, attenuated endothelium and some basement laminae, labyrinth architecture of vacuoles within lipid droplets, and enhanced 3D effect of ultrastructures, the latter of which is the case in electron tomography. In addition, the biological significance of structured appearance (microtrabecular lattices) of the cytoplasmic matrix, which is disclosed by this method, are briefly reviewed in relation to the sol–gel transition of cytoplasmic heterogenous proteins. Since the ultrastructures of various cells and tissues in this method are confirmed to be well correspondent to those in conventional epoxy section TEM except for isotropic dimensional changes, and because there is no necessity for any special expensive equipments other than those for the conventional TEM, the embedment-free section TEM method with these advantages, deserves much more wide application to the morphological research including electron tomography. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 7
    Publication Date: 2013-10-03
    Description: ABSTRACT Combined microscopy techniques are used to establish the usability of phosphonic acid layers as promoters of hydroxyapatite (HAp) growth. Using spread coating, octadecylphosphonic acid (OPA) self-assembled bilayers are delivered to the thin natural oxide layer of a titanium film surface with no prior treatment. These bilayers aggregate two major advantages of phosphonic moieties to titanium surfaces: nucleation of hydroxyapatite crystals from ionic solution and affinity for both titanium oxide surface and HAp crystals. The functionalized substrates and bare titanium (control) samples are immersed in an aqueous solution containing calcium and phosphorus ions. Over a 4-week immersion time, OPA-functionalized substrates present numerous large agglomerates of inorganic crystals, in contrast to control samples, with no significant amount of deposits. Initial sample characterization was performed with atomic force microscopy (AFM). Compositional and structural characterization of these agglomerates (using TEM, EDS, and electron diffraction), revealed that they are indeed HAp, the main component of the inorganic bone matrix. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 8
    Publication Date: 2013-06-06
    Description: ABSTRACT This study focused on adhesive interface morphologic characterization and nanoleakage expression of resin cements bonded to human dentin pretreated with 1% chlorhexidine (CHX). Thirty-two non-carious human third molars were ground flat to expose superficial dentin. Resin composite blocks were luted to the exposed dentin using one conventional (RelyX ARC) and one self-adhesive resin cement (RelyX U100), with/without CHX pretreatment. Four groups ( n  = 8) were obtained: control groups (ARC and U100); experimental groups (ARC/CHX and U100/CHX) were pretreated with 1% CHX prior to the luting process. After storage in water for 24 h, the bonded teeth were sectioned into 0.9 × 0.9 mm 2 sticks producing a minimum of 12 sticks per tooth. Four sticks from each tooth were prepared for hybrid layer evaluation by scanning electron microscope analysis. The remaining sticks were immersed in silver nitrate for 24 h for either nanoleakage evaluation along the bonded interfaces or after rupture. Nanoleakage samples were carbon coated and examined using backscattered electron mode. Well-established hybrid layers were observed in the groups luted with RelyX ARC. Nanoleakage evaluation revealed increase nanoleakage in groups treated with CHX for both resin cements. Group U100/CHX exhibited the most pronouncing nanoleakage expression along with porous zones adjacent to the CHX pretreated dentin. The results suggest a possible incompatibility between CHX and RelyX U100 that raises the concern that the use of CHX with self-adhesive cements may adversely affect resin-dentin bond. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 9
    Publication Date: 2013-06-09
    Description: ABSTRACT Owing to the highly efficient two-photon fluorescence of gold nanorods and very short fluorescence lifetime compared with the rotational correlation time, the rotation and diffusion of a single gold nanorod can be easily observed by two-photon fluorescence correlation spectroscopy (TP-FCS). This property, along with the previous successful use as a contrast agent in two-photon fluorescence imaging, suggests a potential application in TP-FCS as well. Although the FCS measurement becomes highly efficient with gold nanorods as probes, the amplitude and temporal decay of the measured correlation functions depend critically on excitation power. Here, we investigate various photophysical processes of gold nanorods to determine the cause of such a sensitive power dependency. This understanding provides a basis for choosing appropriate FCS models to recover reasonable physical parameters. Although the correlation function amplitude G (0) is 32 times lower when the excitation power increases from 20 µW to 1.12 mW, the application of a saturation-modified FCS model yields very good fit to each data set and the fitted concentration of 0.64 nM is comparable to the 0.7 nM given by the inductively coupled plasma mass spectrometry measurement. The FCS assay appears to be an efficient method for the quantification of gold nanorods when correctly interpreted. However, even with the saturation considered in the fitting model, the fitted rotational and translational diffusion rates are getting faster as the power increases. This indicates that other effects such as photothermal effects may raise the local temperature, and thus increasing the rotational and translational diffusion rate. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 10
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    Oxford University Press
    Publication Date: 2013-06-12
    Print ISSN: 0022-0744
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    Topics: Electrical Engineering, Measurement and Control Technology , Natural Sciences in General , Physics
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  • 11
    Publication Date: 2013-06-12
    Description: Polysaccharides account for more than 90% of the content of fungal cell walls, but the mechanism underlying the formation of the architecture of the cell walls, which consist of microfibrils embedded in an amorphous wall matrix, remains unknown. We used electron microscopy to investigate ten different fungal cell-wall polysaccharides to determine whether they could self-assemble into the fibrillar or amorphous component of fungal cell walls in a test tube without enzymes. The ultrastructures formed by precipitating β-1,3-glucan and β-1,6-glucan are different depending on the existence of branching in the molecule. Linear β-1,3-glucan and linear β-1,6-glucan precipitate into a fibrillar ultrastructure. Branched β-1,6-glucan, mannan and glycogen precipitates are amorphous. Branched β-1,3-glucan forms a fibrillar plus amorphous ultrastructure. Self-assembly among combinations of different linear and branched cell-wall polysaccharides results in an ultrastructure that resembles that of a yeast cell wall, which suggests that self-assembly of polysaccharides may participate in the development of the three-dimensional architecture of the yeast cell wall.
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  • 12
    Publication Date: 2013-06-12
    Description: When considering drug delivery, the amount of drug that can be carried at a particular time and how the drug is incorporated efficiently into cells are important parameters. Transferrin (Tf)-conjugated nanocarriers have been used for the targeted delivery of drugs to cancer cells due to the availability of receptor-mediated clathrin-dependent endocytosis. In general, however, endocytosis seems to differ according to the size and shape of carriers. Large substances are generally internalized into cells by phagocytosis. We studied the internalization mechanism of Tf-conjugated nanoparticles (diameter, 522 nm). Tf-conjugated polystyrene particles were incorporated into cells by receptor-mediated endocytosis with large clathrin-coated vesicles even though their diameter was 〉500 nm and despite that fact that clathrin-coated vesicles have a diameter of 100 nm. This finding suggests that signals for internalization generated by stimulated Tf receptors (TfRs) activate clathrin-mediated endocytosis preferentially. Whether these larger particles could deliver drugs more efficiently than smaller particles was then examined. The toxicity of larger Tf-conjugated biodegradable nanoparticles (poly(lactic-co-glycolic acid)) encapsulating doxorubicin (diameter, 216 ± 38 nm) was appreciably dependent on the number of Tf molecules conjugated on a particle and the number of TfRs expressed on the cell membrane. Larger Tf-conjugated particles delivered drugs to cancer cells expressing many TfRs more selectively than their smaller counterparts (diameter, 56 ± 9 nm) if they were decorated with an appropriate number of Tf molecules.
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  • 13
    Publication Date: 2013-06-12
    Description: Structural biology relies on good-quality protein crystals in order for structure determination. Many factors affect the growth process of a protein crystal including the way it nucleates and the types of damage and contamination during its growth. Although the nucleation process and quality of a crystal is vital to structure determination, they are both under-studied areas of research. Our research begins to explore ways of measuring the quality of protein crystals, using TEM, thus overcoming the problems associated with viewing wet specimens in a vacuum. Our current understanding of nucleation is that it is a two-step mechanism involving the formation of nuclei from dense liquid clusters; however; it is still unclear whether nuclei first start as amorphous aggregates or as crystalline lattices. Potentially, electron diffraction may be capable of uncovering this process. Using TEM imaging and diffraction of lysozyme as a model protein crystal, we report the internal two-dimensional strain and the density of crystallites in a protein crystal, at a resolution never seen before. The TEM diffraction shows unique features of crystal mosaicity that can be directly correlated to TEM images.
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  • 14
    Publication Date: 2013-06-12
    Description: The cotyledon of legume seeds is a storage organ that provides nutrients for seed germination and seedling growth. The spatial and temporal control of the degradation processes within cotyledons has not been elucidated. Calcium oxalate (CaOx) crystals, a common calcium deposit in plants, have often been reported to be present in legume seeds. In this study, micro-computed tomography (micro-CT) was employed at the SPring-8 facility to examine the three-dimensional distribution of crystals inside cotyledons during seed maturation and germination of Lotus miyakojimae (previously Lotus japonicus accession Miyakojima MG-20). Using this technique, we could detect the outline of the embryo, void spaces in seeds and the cotyledon venation pattern. We found several sites that strongly inhibited X-ray transmission within the cotyledons. Light and polarizing microscopy confirmed that these areas corresponded to CaOx crystals. Three-dimensional observations of dry seeds indicated that the CaOx crystals in the L. miyakojimae cotyledons were distributed along lateral veins; however, their distribution was limited to the abaxial side of the procambium. The CaOx crystals appeared at stage II (seed-filling stage) of seed development, and their number increased in dry seeds. The number of crystals in cotyledons was high during germination, suggesting that CaOx crystals are not degraded for their calcium supply. Evidence for the conservation of CaOx crystals in cotyledons during the L. miyakojimae germination process was also supported by the biochemical measurement of oxalic acid levels.
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  • 15
    Publication Date: 2013-06-12
    Description: The electron irradiation damage of MFI-type zeolite was estimated under various accelerating voltages of 100, 200 and 300 kV from successively captured high-resolution transmission electron microscopy (HRTEM) images. To determine the optimal accelerating voltage for HRTEM imaging of electron-sensitive MFI zeolite, the critical dose was estimated from the disappearance of a specific fast Fourier transform spot calculated from experimental images. Based only on the electron dose, a higher voltage was more advantageous. However, taking into account the minimum dose for imaging with a CCD camera, the optimal accelerating voltage for imaging MFI zeolite was 200 kV. The minimum dose for image detection with a CCD camera was surmised from the output/input signal ratio dependence on the accelerating voltage and the contrast range in simulated HRTEM images.
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  • 16
    Publication Date: 2013-06-12
    Description: The magnetic field generated by a magnetic recording head is evaluated using electron holography. A magnetic recording head, which is connected to an electric current source, is set on the specimen holder of a transmission electron microscope. Reconstructed phase images of the region around the magnetic pole show the change in the magnetic field distribution corresponding to the electric current applied to the coil of the head. A simulation of the magnetic field, which is conducted using the finite element method, reveals good agreement with the experimental observations.
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  • 17
    Publication Date: 2013-06-12
    Description: A new multilayer-coated varied line-spaced grating, JS4000, was fabricated and tested for extending the upper limit of a grating X-ray spectrometer for electron microscopy. This grating was designed for 2–3.8 keV at a grazing incidence angle of 1.35°. It was revealed that this new multilayer structure enables us to take soft-X-ray emission spectra continuously from 1.5 to 4.3 keV at the same optical setting. The full-width at half maximum of Te-L α1,2 (3.8 keV) emission peak was 27 eV. This spectrometer was applied to indium tin oxide particles and clearly resolved Sn-L α (3444 eV) and In-L β1 (3487 eV) peaks, which could not be resolved by a widely used energy-dispersive X-ray spectrometer.
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  • 18
    facet.materialart.
    Unknown
    Oxford University Press
    Publication Date: 2013-04-02
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  • 19
    Publication Date: 2013-04-02
    Description: For development of advanced materials, characterization using a scanning transmission electron microscope (STEM) including analysis via X-ray energy-dispersive spectrometry and electron energy-loss spectrometry is essential. Recent advances in aberration-corrected instruments have offered large-scale data acquisition at a high resolution for limited acquisition times both in imaging and in analysis. Further advanced procedures are required to analyze such large-scale datasets more efficiently including quantification. In addition, more simplified tuning procedures are crucial to the best possible resolution in the latest aberration-corrected instruments. In this review article, several approaches to perform advanced electron microscopy, which the author has been developing with his colleague, are described as ‘Microscopy Hacks’. These are (i) quantification and elemental/chemical-imaging procedures, (ii) advanced statistical approaches to handle large-scale datasets and (iii) instrument characterization and tuning procedures including the latest development of an ad hoc autotuning procedure for aberration-corrected STEM imaging.
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  • 20
    Publication Date: 2013-04-02
    Description: The crystal structure of a new type of molybdenum oxide crystal encapsulated in a single-walled carbon nanotube (CNT) was examined via diffraction and spectroscopic techniques using both X-rays and electron beams. This new type of molybdenum oxide crystal has a chemical bonding state of MoO 3 , as confirmed by X-ray absorption spectroscopy, and the MoO 3 units exhibit axial symmetry, as clarified by electron diffraction from bundled and individual CNTs encapsulating the crystal. To obtain three-dimensional information on the structure, a cross-sectional sample was prepared using a conventional dimple and ion-mill method. High-resolution transmission electron microscopy images exhibit ring-like shapes that originated from the arrangement of the MoO 3 units inside the CNTs, as observed along the tube axis. The units are spaced 0.36 nm from each other in a ring arrangement and the distance between each ring is 0.391 nm.
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  • 21
    Publication Date: 2013-04-02
    Description: This review summarizes the recent advances in three-dimensional (3D) imaging techniques and their application to polymer nanostructures, for example, microphase-separated structures of block copolymers. We place particular emphasis on the method of transmission electron microtomography (electron tomography for short; hereafter abbreviated as ET). As a result of recent developments in ET, truly quantitative 3D images of polymer nanostructures can now be obtained with subnanometer resolution. The introduction of scanning optics in ET has made it possible to obtain large amounts of 3D data from micrometer-thick polymer specimens by using conventional electron microscopes at a relatively low accelerating voltage, 200 kV. Thus, ET covers structures over a wide range of thicknesses, from a few nanometers to several hundred nanometers, which corresponds to quite an important spatial range for hierarchical polymer nanostructures. ET provides clear 3D images and a wide range of new structural information that cannot be obtained using other methods. Information traditionally derived from conventional microscopy or scattering methods can be directly acquired from 3D volume data. ET is a versatile technique that is not restricted to only polymer applications; it can also be used as a powerful characterization tool in energy applications such as fuel cells.
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  • 22
    Publication Date: 2013-04-02
    Description: The performances of a newly developed 80–200 kV cold field emission gun (CFEG) transmission electron microscope (TEM) integrating a spherical aberration corrector for a TEM image-forming lens have been evaluated. To begin, we show that the stability of both emission and probe currents makes use of this new CFEG much friendlier. The energy spread of electrons emitted from the CFEG has been measured as a function of emission current and shows a very last 0.26 eV energy resolution at 200 kV and even 0.23 eV at 80 kV. The combination of the CFEG and the CEOS™ aberration corrector, associated with enhanced mechanical and electrical stabilities of this new microscope, allows reaching an information transfer below 75 pm at 200 and 80 pm at 80 kV. This unseen resolution at 200 kV has allowed us to study the structure of CoPt nanoparticles by observing direct images of their atomic arrangement along the high indexes zone axis. We have evidenced the presence of defects in these nanostructures that are not parallel to the electron beam. The precise stoichiometry of two iron oxides, FeO and Fe 2 O 3 , has been determined from an analysis of iron valence state that was obtained from a direct analysis of EELS fine structures spectrum of the two oxides.
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  • 23
    Publication Date: 2013-04-02
    Description: Data mining from noisy data/images is one of the most important themes in modern science and technology. Statistical image processing is a promising technique for analysing such data. Automation of particle pickup from noisy electron micrographs is essential, especially when improvement of the resolution of single particle analysis requires a huge number of particle images. For such a purpose, reference-based matching using primary three-dimensional (3D) model projections is mainly adopted. In the matching, however, the highest peaks of the correlation may not accurately indicate particles when the image is very noisy. In contrast, the density and the heights of the peaks should reflect the probability distribution of the particles. To statistically determine the particle positions from the peak distributions, we have developed a density-based peak search followed by a peak selection based on average peak height, using multi-reference alignment (MRA). Its extension, using multi-reference multiple alignment (MRMA), was found to enable particle pickup at higher accuracy even from extremely noisy images with a signal-to-noise ratio of 0.001. We refer to these new methods as stochastic pickup with MRA (MRA-StoPICK) or with MRMA (MRMA-StoPICK). MRMA-StoPICK has a higher pickup accuracy and furthermore, is almost independent of parameter settings. They were successfully applied to cryo-electron micrographs of Rice dwarf virus. Because current computational resources and parallel data processing environments allow somewhat CPU-intensive MRA-StoPICK and MRMA-StoPICK to be performed in a short period, these methods are expected to allow high-resolution analysis of the 3D structure of particles.
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  • 24
    Publication Date: 2013-04-02
    Description: The sensory nerve endings of the rat tongue, cheek and palate were studied using immunohistochemical staining and transmission electron microscopy analysis. The specimens were fixed in modified Karnovsky solution and embedded in Spurr resin. Substance P, calcitonin gene-related peptide (CGRP)- and protein gene product 9.5 (PGP b9.5)-containing nerve fibers in the rat tongue, cheek and palate were examined by electronic microscopical analysis and immunohistochemical localization. These fibers run very close to the basal lamina of the epithelium and extend into the filliform and fungiform papillae. Numerous plexiform fibers immunoreactive for substance P, CGRP and PGP 9.5 were found in the connective tissue of mucosa. Electron microscopic observations showed clearly immunostained nerve fibers, which are located very close to the basal lamina of epithelial cells. Some electron-dense granules may be observed in the axoplasms of both substance P and CGRP immunoreactive fibers. Several lamellar corpuscles into the subepithelial connective tissue papillae, Merkel corpuscles and numerous thin unmyelinated and myelinated axons were observed. The terminal axons revealed numerous mitochondria, neurofilaments, microtubules and clear vesicles in the base of axoplasmic protrusions. The lamellar cells showed caveolae and interlamelar spaces filled by amorphous substance. Between the lamellar cells and axoplasmic membrane, and in the adjacent lamellae region, desmosome-type junctions were observed. The quantitative and morphometric analysis showed nerve endings with an average area of 4.83 ± 3.4 μm 2 and 19.4 internal mitochondria in this site and the organized corpuscles with an average area of 79.24 ± 27.24 μm 2 and 24.23 internal mitochondria in this place. All the structures observed are involved in the transmission of pain and mechanoreceptors stimulus of these oral mucosae.
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  • 25
    Publication Date: 2013-04-09
    Description: ABSTRACT Wistar rats (male) were daily administered chlorpyrifos at a dose of 5 mg/kg b wt. and 10 mg/kg b wt. and sacrificed on 1st, 2nd, 4th, 6th, and 8th week. In chlorpyrifos exposed rats hypocalcemia, hypophosphatemia and hypomagnesemia were recorded. At later intervals an increased levels of serum calcium and phosphate were observed. The parathyroid glands and calcitonin cells exhibited increased activity which is evident by increased nuclear volume of these cells. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 26
    Publication Date: 2013-04-09
    Description: ABSTRACT In the dry northern temperate regions of the northern hemisphere, the genus Ephedra comprises a series of native shrub species with a cumulative application history reaching back well over 2,000 years for the treatment of asthma, cold, fever, as well as many respiratory system diseases, especially in China. There are ethnological and philological evidences of Ephedra worship and utilization in many Eurasia Steppe cultures. However, no scientifically verifiable, ancient physical proof has yet been provided for any species in this genus. This study reports the palaeobotanical finding of Ephedra twigs discovered from burials of the Gumugou archaeological site, and ancient community graveyard, dated around 3800 BP, in Lop Nor region of northwestern China. The macro-remains were first examined by scanning electron microscope (SEM) and then by gas chromatography-mass spectrometry (GC-MS) for traits of residual biomarkers under the reference of modern Ephedra samples. The GC-MS result of chemical analysis presents the existence of Ephedra -featured compounds, several of which, including benzaldehyde, tetramethyl-pyrazine, and phenmetrazine, are found in the chromatograph of both the ancient and modern sample. These results confirm that the discovered plant remains are Ephedra twigs. Although there is no direct archaeological evidence for the indication of medicinal use of this Ephedra , the unified burial deposit in which the Ephedra was discovered is a strong indication of the religious and medicinal awareness of the human inhabitants of Gumugou towards this plant. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 27
    Publication Date: 2013-04-09
    Description: ABSTRACT Objective : This in vitro study evaluated the effect of different endodontic auxiliary chemical substances over Enterococcus faecalis (Ef) biofilm through confocal laser scanning microscopy (CLSM). Methods : Forty-five bovine incisors were infected with Ef for 21 days. Teeth were divided into five groups: group 1: 2.5% NaOCl + EDTA, group 2: 2% CHX gel + EDTA, group 3: 2% CHX liquid + EDTA, group 4: 2.5% NaOCl + 2% CHX gel + EDTA, group 5: 2.5% NaOCl + 2% CHX liquid + EDTA and a negative and a positive control group (NCG; PCG). The samples were stained with SYTO9 and propidium iodide and analyzed by CLSM. Bacterial viability was quantitatively analyzed by the proportions of dead and live bacteria in the biofilm remnants. Scores were standardized according to the total bacterial load (TBL)—1: ≤25%, 2: 〉25 ≤50%, 3: 〉50 ≤75%, 4: 〉75% and debris—1: absence of debris; 2: presence of debris. Statistical analysis was carried out through the Kruskal–Wallis and the Fischer exact tests ( P  = 0.05). Results : No statistical differences were observed to CFU, debris and bacterial viability. Conclusion : None of the tested substances could completely eliminate Ef from the root canal space. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 28
    Publication Date: 2013-04-09
    Description: ABSTRACT The aim of the present research was to analyze ultrastructural and immunohistochemical aspects of the alveolar repair after the extraction of molars of alendronate (ALN)-treated rats. Wistar rats received 2.5mg/kg body wt/day of ALN during 14 days previously and 7, 14 and 21 days after the extraction of the second mandibular molar. Specimens were fixed in 2% glutaraldehyde + 2.5% formaldehyde under microwave irradiation, decalcified in 4.13% EDTA and paraffin embedded for TRAP histochemistry and immunohistochemistry for OPN, BSP and endoglin, or embedded in Spurr epoxy resin for TEM analysis. Additional specimens had their soft tissues removed and were processed for scanning electron microscopy. The ALN group presented latent TRAP-positive osteoclasts and nonresorbed alveolar crests with bacterial infection. Mild bone necrosis signs were observed at all time points studied. Ultrastructurally, empty osteocyte lacunae were observed and bone trabeculae surface presented hyalinized aspect. A significant delay in alveolar repair occurred, as well as decreased angiogenesis. ALN treatment provoked mild signs of bone necrosis, despite the high dose employed. The present findings add new information about the ultrastructural aspect of the early repair of rats under ALN treatment and highlight for giving attention when oral surgeries are performed in patients using this drug. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 29
    Publication Date: 2013-04-09
    Description: ABSTRACT In the long run, the widespread use of slide scanners by pathologists requires an adaptation of teaching methods in histology and cytology in order to target these new possibilities of image processing and presentation via the internet. Accordingly, we were looking for a tool with the possibility to teach microscopic anatomy, histology, and cytology of tissue samples which would be able to combine image data from light and electron microscopes independently of microscope suppliers. With the example of a section through the villus of jejunum, we describe here how to process image data from light and electron microscopes in order to get one image-stack which allows a correlation of structures from the microscopic anatomic to the cytological level. With commercially available image-presentation software that we adapted to our needs, we present here a platform which allows for the presentation of this new but also of older material independently of microscope suppliers. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 30
    Publication Date: 2013-04-02
    Description: The F-actin cytoskeleton of Cryptococcus neoformans is known to comprise actin cables, cortical patches and cytokinetic ring. Here, we describe a new F-actin structure in fungi, a perinuclear F-actin collar ring around the cell nucleus, by fluorescent microscopic imaging of rhodamine phalloidin-stained F-actin. Perinuclear F-actin rings form in Cryptococcus neoformans treated with the microtubule inhibitor Nocodazole or with the drug solvent dimethyl sulfoxide (DMSO) or grown in yeast extract peptone dextrose (YEPD) medium, but they are absent in cells treated with Latrunculin A. Perinuclear F-actin rings may function as ‘funicular cabin’ for the cell nucleus, and actin cables as intracellular ‘funicular’ suspending nucleus in the central position in the cell and moving nucleus along the polarity axis along actin cables.
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  • 31
    Publication Date: 2013-04-02
    Description: To evaluate the advantages of combination of two advanced electron microscopic technologies such as serial block-face scanning electron microscopy and double-axis electron beam tomography, we analyzed the three-dimensional morphology of cellular relationships between dendritic and plasma cells in the synovial membrane from patients with rheumatoid arthritis, using the combined approach.
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  • 32
    Publication Date: 2013-04-02
    Description: The low-energy Ar-ion milling method was used to prepare ultrathin specimens for transmission electron microscope observation. The samples were thinned initially by a usual focused ion beam technique or typical Ar-ion milling with a high energy of 2–10 keV and were thinned additionally by an Ar-ion beam with an energy less than 1 keV, typically 500–900 eV. This low-energy ion beam was scanned over the specimen, and secondary electrons induced by the ion beam could be detected to form secondary electron images with a resolution of a few micrometre. Because a desired area can be selected and thinned by the low-energy ion beam, redeposition or cross contamination from irradiation of a metal grid that supports the sample can be prevented. It was shown that the low-energy Ar-ion beam thins a surface amorphous damage layer preferentially and effectively rather than a crystal specimen. Images from ultrathin specimens of two different materials revealed a detailed structure.
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  • 33
    Publication Date: 2013-03-30
    Description: ABSTRACT This article presents a pixellated solid-state photon detector designed specifically to improve certain aspects of the existing Everhart–Thornley detector. The photon detector was constructed and fabricated in an Austriamicrosystems 0.35 µm complementary metal-oxide-semiconductor process technology. This integrated circuit consists of an array of high-responsivity photodiodes coupled to corresponding low-noise transimpedance amplifiers, a selector-combiner circuit and a variable-gain postamplifier. Simulated and experimental results show that the photon detector can achieve a maximum transimpedance gain of 170 dBΩ and minimum bandwidth of 3.6 MHz. It is able to detect signals with optical power as low as 10 nW and produces a minimum signal-to-noise ratio (SNR) of 24 dB regardless of gain configuration. The detector has been proven to be able to effectively select and combine signals from different pixels. The key advantages of this detector are smaller dimensions, higher cost effectiveness, lower voltage and power requirements and better integration. The photon detector supports pixel-selection configurability which may improve overall SNR and also potentially generate images for different analyses. This work has contributed to the future research of system-level integration of a pixellated solid-state detector for secondary electron detection in the scanning electron microscope. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 34
    facet.materialart.
    Unknown
    Oxford University Press
    Publication Date: 2013-04-02
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  • 35
    facet.materialart.
    Unknown
    Oxford University Press
    Publication Date: 2013-04-02
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  • 36
    Publication Date: 2013-09-07
    Description: ABSTRACT Birds occupy a prominent place in the Brazilian economy not only in the poultry industry but also as an animal model in many areas of scientific research. Thus the aim of this study was to provide a description of macro and microscopic aspects of the ectoderm-derived structures in chicken embryos / fetuses poultry ( Gallus gallus domesticus ) from 1st to 19th day of incubation. 40 fertilized eggs, from a strain of domestic chickens, with an incubation period of 2–19 days were subjected to macroscopic description, biometrics, light, and scanning microscopy. All changes observed during the development were described. The nervous system, skin and appendages and organs related to vision and hearing began to be identified, both macro and microscopically, from the second day of incubation. The vesicles from the primitive central nervous system—forebrain, midbrain, and hindbrain—were identified on the third day of incubation. On the sixth day of incubation, there was a clear vascularization of the skin. The optic vesicle was first observed fourth day of development and on the fifth day there was the beginning of the lens formation. Although embryonic development is influenced by animal line as well as external factors such as incubation temperature, this paper provides a chronological description for chicken ( Gallus gallus domesticus ) during its embryonic development. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 37
    Publication Date: 2013-09-07
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  • 38
    Publication Date: 2013-09-13
    Description: ABSTRACT The dog ( canis lupus familiaris ) is the only other species besides humans that develop spontaneous prostatic carcinomas (PCa) at a high frequency. The canine model is primarily utilized for the study of the PCa molecular mechanisms and provides a natural animal model for the study of potential therapies. In humans, the PCa frequently exhibits mutations in the C-MYC and a reduced expression of the E-cadherin and NKX3.1 proteins. This study's objective was to evaluate the NKX3.1, C-MYC, and E-cadherin expression in the canine normal prostate, benign prostatic hyperplasia (BPH), proliferative inflammatory atrophy (PIA) and PCa and to verify differences in expression and subcellular localization of these proteins in the prostatic carcinogenesis. A tissue microarray (TMA) slide was constructed, and immunohistochemistry with antibodies raised against C-MYC, NKX3.1, E-cadherin and p63 was performed using the peroxidase and DAB methods. The C-MYC protein expression was elevated in the cytoplasm and nuclei of the canine PCa and PIA compared with the normal prostate ( P  = 0.004. The NKX3.1 protein expression was reduced in 94.75% of the PCa and 100% of the PIA compared with the normal prostate ( P  = 0.0022). In fact, the expression of E-cadherin trended towards a decrease in carcinomas when compared to normal prostate and PIA. By immunohistochemistry, more p63-positive basal cells were observed in the PCa and PIA when compared with the normal prostate ( P  = 0.0002). This study has demonstrated that the carcinogenesis of canine prostatic tissue may be related to basal cell proliferation, the gain of C-MYC function and the loss of NKX3.1 protein expression. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 39
    Publication Date: 2013-09-23
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  • 40
    Publication Date: 2011-06-16
    Description: Root canal preparation may damage NiTi instruments resulting in wear and deformation. The aim of this study was to make a comparative evaluation of the surface topography of the cervical third of four different rotary systems, before and after being used twelve times, in 1.440 resin blocks with simulated root canals with standardized 45° curvatures, and analyzed by atomic force microscopy – AFM. The blocks were divided into four groups and prepared according to the manufacturers recommendations: Group 1 - K3®; Group 2 - Protaper Universal®; Group 3 - Twisted Files® and Group 4 - Biorace®. After each preparation, the instruments were washed and autoclaved. A total of 240 instruments were selected, being 30 new instruments and 30 after having been used for the 12th time, from each group. These instruments were analyzed by AFM and for quantitative evaluation, the mean RMS (Root mean square) values of the cervical third of the specimens from the four groups were used. The result showed that all the rotary files used for the 12th time suffered wear with change in the topography of the cervical region of the active portion of the file (ANOVA p 〈 0.01). Classifying the specimens in increasing order, from the least to the greatest wear suffered, Group 3 (2.8993 nm) presented the least wear, followed by Group 4 (12.2520 nm), Group 1 (36.0043 nm) and lastly, Group 2 (59.8750 nm) with the largest amount of cervical surface wear. Microsc. Res. Tech., 2011. © 2011 Wiley-Liss, Inc.
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  • 41
    Publication Date: 2011-06-16
    Description: We report the use of superresolution fluorescence microscopy for studying the nanoscale distribution of protein colocalization in living mammalian cells. Nanoscale imaging is attained both by a targeted and a stochastic fluorescence on-off switching superresolution method, namely by stimulated emission depletion (STED) and ground state depletion microscopy followed by individual molecular return (GSDIM), respectively. Analysis of protein colocalization is performed by bimolecular fluorescence complementation (BiFC). Specifically, a nonfluorescent fragment of the yellow fluorescent protein Citrine is fused to tubulin while a counterpart nonfluorescent fragment is fused to the microtubulin-associated protein MAP2 such that fluorescence is reconstituted on contact of the fragment-carrying proteins. Images with resolution down to 65 nm prove a powerful new way for studying protein colocalization in living cells at the nanoscale. Microsc. Res. Tech., 2011. © 2011 Wiley-Liss, Inc.
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  • 42
    Publication Date: 2011-06-16
    Description: Trk neurotrophin receptors are transmembrane tyrosine kinase proteins known as TrkA, TrkB, and TrkC. TrkA is the high affinity receptor for nerve growth factor, TrkB is the one for both brain-derived neurotrophic factor and neurotrophin-4, and TrkC is the preferred receptor for neurotrophin-3. In the adult mammalian brain, neurotrophins are important regulators of neuronal function and plasticity. This study is based on Nothobranchius furzeri, a teleost fish that is becoming an ideal candidate as animal model for aging studies because its life expectancy in captivity is of just 3 months. In adult N. furzeri , all three investigated neurotrophin Trk receptors were immunohistochemically detected in each brain region. TrkA positive neuronal perikarya were localized in the dorsal and ventral areas of the telencephalon and in the cortical nucleus; TrkB immunoreactivity was observed in neuronal perikarya of the dorsal and ventral areas of the telencephalon, the diffuse inferior lobe of the hypothalamus, and Purkinje cells; TrkC positive neuronal perikarya were detected in the most aboral region of the telencephalon, in the magnocellular preoptic nucleus and in few neurons dispersed in the hypothalamus. Numerous positive fibers were widely distributed throughout the brain. Radial glial cells lining the mesencephalic and rhombencephalic ventricles showed immunoreactivity to all three Trks. These findings suggest an involvement of neurotrophins in many aspects of biology of adult N. furzeri . Microsc. Res. Tech., 2011. © 2011 Wiley-Liss, Inc.
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  • 43
    Publication Date: 2011-06-16
    Description: TRPV4 is a nonselective cation channel that belongs to the vanilloid (V) subfamily of transient receptor potential (TRP) ion channels. While TRP channels have been found to be involved in sensing temperature, light, pressure, and chemical stimuli, TPRV4 is believed to be primarily a mechanosensor although it can also respond to warm temperatures, acidic pH, and several chemical compounds. In zebrafish, the expression of trpv4 has been studied during embryonic development, whereas its pattern of TPRV4 expression during the adult life has not been thoroughly analyzed. In this study, the occurrence of TRPV4 was addressed in the zebrafish sensory organs at the mRNA (RT-PCR) and protein (Westernblot) levels. Once the occurrence of TRPV4 was demonstrated, the TRPV4 positive cells were identified by using immunohistochemistry. TPRV4 was detected in mantle and sensory cells of neuromasts, in a subpopulation of hair sensory cells in the macula and in the cristae ampullaris of the inner ear, in sensory cells in the taste buds, in crypt neurons and ciliated sensory neurons of the olfactory epithelium, and in cells of the retina. These results demonstrate the presence of TRPV4 in all sensory organs of adult zebrafish and are consistent with the multiple physiological functions suspected for TRPV4 in mammals (mechanosensation, hearing, and temperature sensing), but furthermore suggest potential roles in olfaction and vision in zebrafish. Microsc. Res. Tech., 2011. © 2011 Wiley-Liss, Inc.
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  • 44
    Publication Date: 2011-06-16
    Description: In recent years, bioinorganic nanohybrids composed of biological macromolecules and functional inorganic nanomaterials have revealed many unique properties that show promise for the future. Transmission electron microscopy (TEM) is a popular and relatively simple tool that can offer a direct visualization of the nanomaterials with high resolutions. When TEM is applied to visualize bioinorganic nanohybrids, a treatment of negative staining is necessary due to the presence of biological molecules in the nanohybrids except for those with densely packed inorganic materials. However, the conventional negative-staining procedure for regular biological samples cannot be directly applied to such bioinorganic nanohybrids. To image a specific bioinorganic nanohybrid, negative-staining factors such as negative stain type, working pH, staining time, and drying method, should be identified. Currently, no detailed studies have been done to investigate how to adjust negative-staining factors based on specific bioinorganic nanohybrids. In this study, bacteriophage-gold nanoparticle hybrids were chosen as a model to systematically study the effects of each factor on the negative staining of the nanohybrids. The best staining conditions for gold nanoparticle-phage nanohybrids were obtained and the effects of each factor on the negative staining of general nanohybrids were discussed. This work indicates that with proper staining it is possible to use TEM to visualize directly both biological and inorganic components without introducing any artifact. Microsc. Res. Tech. 2011. © 2011 Wiley-Liss, Inc.
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  • 45
    Publication Date: 2011-06-23
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  • 46
    Publication Date: 2011-05-12
    Description: The morphological similarities among the species of Solanum are remarkable, and are often very difficult one clear distinction between them. This paper presents a comparative anatomical study of the leaf epidermis of five Brazilian species of Solanum sect. Polytrichum , carried out using light and scanning electron microscopy. The leaf epidermis surfaces were investigated to evaluate their taxonomic significance to be used for separation and delimitation of the species of the section. As results, some micro-morphological characters of the leaf epidermis, such as density, distribution and type of stellate trichomes, and the anticlinal walls of epidermal cells, and also the type and distribution of stomata proved to be the most useful and distinctive characters for the separation and delimitation of the species, and also may contribute as an additional support to the interspecific taxonomy and systematic of Solanum sect. Polytrichum . Microsc. Res. Tech., 2011. © 2011 Wiley-Liss, Inc.
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  • 47
    Publication Date: 2011-05-12
    Description: 3D surface profiling and high resolution imaging were performed to refine the Florin rings and epicuticular wax crystals of Pinus koraiensis needles. Needles were collected from four-year-old seedlings and air-dried for surface observations. Field emission scanning electron microscopy revealed that stomata were found on the abaxial (lower) surface of needles. Measured as ca. 40 μm long, they were largely elliptical or oval-shaped. Epicuticular wax crystals were present in the epistomatal chambers as well as on the surrounding epidermis. Rodlets were prevalently found on the stomatal bands and furrows as well as within the epistomatal chambers. The presence of wax tubules was ascertained by the distinct terminal openings at their ends. The occurrence of wax ridges was evident on the epidermis near the saw-tooth margins (nonstomatal areas). No distinct wax ridges were detected on the dewaxed needles. Raised Florin rings were distinct on the stomata. White light scanning interferometry showed that the diameter and width of stomata were ca. 44.02 ± 3.33 μm and 32.10 ± 3.30 μm, respectively. Measured from the neighboring epidermis to the stomatal aperture, the mean height of the stoma reached ca. 6.23 ± 1.28 μm. Focus variation metrology allowed measuring the mean elevation angle of the stoma, reaching ca. 41.41 ± 11.25°. This is the first report on a novel approach to the establishment of quantitative criteria of Florin ring classification by nontactile 3D surface profiling beyond the previous qualitative descriptions of Florin rings of coniferous species. Microsc. Res. Tech., 2011. © 2011 Wiley-Liss, Inc.
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  • 48
    Publication Date: 2014-11-29
    Description: Off-axis electron holography can be used to measure the inner potential of a specimen from its reconstructed phase image and is thus a powerful technique for materials scientists. However, abrupt reversals of contrast from white to black may sometimes occur in a digitally reconstructed phase image, which results in inaccurate information. Such phase distortion is mainly due to the digital reconstruction process and weak electron wave amplitude in some areas of the specimen. Therefore, digital image processing can be applied to the reconstruction and restoration of phase images. In this paper, fringe reconnection processing is applied to phase image restoration of a crystal structure image. The disconnection and wrong connection of interference fringes in the hologram that directly cause a 2 phase jump imperfection are correctly reconnected. Experimental results show that the phase distortion is significantly reduced after the processing. The quality of the reconstructed phase image was improved by the removal of imperfections in the final phase.
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  • 49
    Publication Date: 2014-11-29
    Description: Diabetes causes oxidative stress, which in turn generates excessive free radicals resulting in cellular damage. Vitamin C is an antioxidant that protects tissues and organs from oxidative stress. The thymus is one of the most important lymphoid organs, which regulates T-lymphocyte proliferation and maturation. The aim of this study is to investigate the protective effects of vitamin C on the thymus of streptozotocin (STZ)-induced diabetic rats. The mitotic activity and cell integrity of thymic lymphocytes were explored. Wistar Albino rats were divided into three groups: control (Group 1), STZ-diabetes (Group 2) and vitamin C-treated STZ-diabetics (Group 3). Rats received a single intraperitoneal injection of 45 mg/kg STZ to induce diabetes. Vitamin C (20 mg/kg) was administered intragastrically. Semithin and ultrathin sections were examined under a light or an electron microscope, respectively. Considerable numbers of mitotic lymphocytes were observed in the thymus of control rats. In the diabetic rats, however, numbers of mitotic lymphocytes decreased to ~57% of controls, and cell division abnormalities were observed. Additionally, diabetic rats showed degeneration in the structure of the thymus including trabecular thickening, accumulation of lipid vacuoles, heterochromatic nuclei and loss of mitochondrial cristae. Degradation of medullar and cortical integrity was also detected. In the vitamin C-treated STZ-diabetic group, the structure of the thymus and mitotic activity of the lymphocytes were similar to the control group. These results suggest that vitamin C protects the thymus against injury caused by diabetes and restores thymocyte mitotic activity.
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  • 50
    facet.materialart.
    Unknown
    Oxford University Press
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  • 51
    Publication Date: 2014-11-29
    Description: We investigated the stability of Pd/ZnO polar interfaces formed by internal oxidation of Pd–Zn alloys by using high-resolution transmission electron microscopy, electron energy-loss spectroscopy and convergent-beam electron diffraction. At 1273 K, a $$(111{)}_{\mathrm{Pd}}/(000\overline{2}{)}_{\mathrm{ZnO}}$$ polar interface defaceted and transformed into a curved interface, while another (111) Pd /(0002) ZnO polar interface retained its flatness. The $$(111{)}_{\mathrm{Pd}}/(000\overline{2}{)}_{\mathrm{ZnO}}$$ polar interface lost some stability over non-polar interfaces at 1273 K, while the (111) Pd /(0002) ZnO polar interface remained stable.
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  • 52
    Publication Date: 2014-11-29
    Description: Transmission electron microscope (TEM) observation of light metal hydrides is complicated by the instability of these materials under electron irradiation. In this study, the electron kinetic energy dependences of the interactions of incident electrons with lithium, sodium and magnesium hydrides, as well as the constituting element effect on the interactions, were theoretically discussed, and electron irradiation damage to these hydrides was examined using in situ TEM. The results indicate that high incident electron kinetic energy helps alleviate the irradiation damage resulting from inelastic or elastic scattering of the incident electrons in the TEM. Therefore, observations and characterizations of these materials would benefit from increased, instead decreased, TEM operating voltage.
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  • 53
    Publication Date: 2014-11-29
    Description: Synaptic plasticity is the process by which long-lasting changes take place at synaptic connections. The phenomenon itself is complex and can involve many levels of organization. Some authors separate forms into adaptations that have positive or negative consequences for the individual. It has been hypothesized that an increase in the number of synapses may represent a structural basis for the enduring expression of synaptic plasticity during some events that involve memory and learning; also, it has been suggested that perforated synapses increase in number after some diseases and experimental situations. The aim of this study was to analyze whether dopamine depletion induces changes in the synaptology of the corpus striatum of rats after the unilateral injection of 6-OHDA. The findings suggest that after the lesion, both contralateral and ipsilateral striata exhibit an increased length of the synaptic ending in ipsilateral (since third day) and contralateral striatum (since Day 20), loss of axospinous synapses in ipsilateral striatum and a significant increment in the number of perforated synapses, suggesting brain plasticity that might be deleterious for the spines, because this type of synaptic contacts are presumably excitatory, and in the absence of the modulatory effects of dopamine, the neuron could die through excitotoxic mechanisms. Thus, we can conclude that the presence of perforated synapses after striatal dopamine depletion might be a form of maladaptive synaptic plasticity.
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  • 54
    Publication Date: 2014-11-29
    Description: Unlike in vitro protein dynamics, intracellular protein dynamics are intricately regulated by protein–protein interactions or interactions between proteins and other cellular components, including nucleic acids, the plasma membrane and the cytoskeleton. Alteration of these dynamics plays a crucial role in physiological phenomena such as gene expression and cell division. Live-cell imaging via microscopy with the inherent properties of fluorescent proteins, i.e. photobleaching and photoconversion, or fluorescence correlation spectroscopy, provides insight into the movement of proteins and their interactions with cellular components. This article reviews techniques based on photo-induced changes in the physicochemical properties of fluorescent proteins to measure protein dynamics inside living cells, and it also discusses the strengths and weaknesses of these techniques.
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  • 55
    facet.materialart.
    Unknown
    Oxford University Press
    Publication Date: 2014-11-29
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  • 56
    Publication Date: 2014-11-29
    Description: We have developed a method of atomic force microscopy (AFM)-assisted scanning tunneling spectroscopy (STS) under ambient conditions. An AFM function is used for rapid access to a selected position prior to performing STS. The AFM feedback is further used to suppress vertical thermal drift of the tip–sample distance during spectroscopy, enabling flexible and stable spectroscopy measurements at room temperature.
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  • 57
    Publication Date: 2014-11-29
    Description: To elucidate high-temperature plastic deformation (creep) mechanism in materials, it is essential to observe dislocation motion under tensile loading. There are many reports on in situ transmission electron microscopy (TEM) observations in the literature; however, the relationship between the dislocation motion and shear stress in 9Cr steel is still not clear. In this study, in order to evaluate this relationship quantitatively, in situ TEM observations were carried out in conjunction with finite element method (FEM) analysis. A tensile test sample was strained at an elevated temperature (903 K) inside a transmission electron microscope, and the stress distribution in the strained sample was analyzed by FEM. The dislocation behavior was clearly found to depend on the shear stress. At a shear stress of 66 MPa, both the dislocation velocity and mobile dislocation density were low. However, a high shear stress level of 95 MPa caused a noticeable increase in the dislocation velocity and mobile dislocation density. Furthermore, in this article, we discuss the dependence of the dislocation behavior on stress. The results presented here also indicate that the relationship between the microstructure and the strength of materials can be revealed by the methods used in this work.
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  • 58
    Publication Date: 2014-11-29
    Description: A dual-axis 360° rotation specimen holder was developed for use in reconstructing the three-dimensional (3D) distribution of a magnetic field using a combination of electron holography and tomography. Pillar-shaped specimens are used to obtain accurate reconstruction without a missing angle. The holder's rotation rod can be turned 〉360°; the pillar is set ±45° to the azimuth for both x - and y -axis rotation. Two rotation series of holograms in individual axes are recorded for vector field tomography. The two vector components of the magnetic field are reconstructed directly from the two series of holograms, and the remaining component is calculated using Maxwell's equation, div B = 0. As a result, all 3D magnetic fields are reconstructed.
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  • 59
    Publication Date: 2014-11-29
    Description: A new type of electrochemical cell was developed for in situ transmission electron microscopy observation that enables the electrode materials to be conveniently changed. The electrochemical cell was used to observe the electrochemical growth or dissolution of copper islands on a gold film with simultaneous cyclic voltammetry measurements. The copper islands could be explained by three-dimensional growing model.
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  • 60
    Publication Date: 2011-10-04
    Description: We present combined epi-coherent anti-Stokes Raman scattering (CARS) and multiphoton imaging with both chemical discrimination and subcellular resolution on human skin invivo. The combination of both image modalities enables label-free imaging of the autofluorescence of endogenous fluorophores by two-photon excited fluorescence, as well as imaging of the distribution of intercellular lipids, topically applied substances and water by CARS. As an example for medical imaging, we investigated healthy and psoriasis-affected human skin with both image modalities in vivo and found indications for different lipid distributions on the cellular level. Microsc. Res. Tech., 2011. © 2011 Wiley-Liss, Inc.
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  • 61
    Publication Date: 2011-10-07
    Description: In this article, we propose a new shape from focus (SFF) method to estimate 3D shape of microscopic objects using surface orientation cue of each object patch. Most of the SFF algorithms compute the focus value of a pixel from the information of neighboring pixels lying on the same image frame based on an assumption that the small object patch corresponding to the small neighborhood of a pixel is a plane parallel to the focal plane. However, this assumption fails in the optics with limited depth of field where the neighboring pixels of an image have different degree of focus. To overcome this problem, we try to search the surface orientation of the small object patch corresponding to each pixel in the image sequence. Searching of the surface orientation is done indirectly by principal component analysis. Then, the focus value of each pixel is computed from the neighboring pixels lying on the surface perpendicular to the corresponding surface orientation. Experimental results on synthetic and real microscopic objects show that the proposed method produces more accurate 3D shape in comparison to the existing techniques. Microsc. Res. Tech. 2011. © 2011 Wiley-Liss, Inc.
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  • 62
    Publication Date: 2011-10-07
    Description: The aim of this study is to show histological and immunofluorescence analysis of renal parenchyma of agoutis affected by gentamicin-induced renal disease after the infusion of bone marrow mononuclear cells (BMMC) stained with Hoechst®. Nine agouti's males were divided into three groups: Test group (TG): renal disease by gentamicin induced ( n = 3), cell therapy group (CTG): renal disease by gentamicin induced and BMMC infusion ( n = 3), and control group (CG): nonrenal disease and BMMC infusion ( n = 3). TG and CTG were submitted to the protocol of renal disease induction using weekly application of gentamicin sulfate for 4 months. CG and CTG received a 1 × 108 BMMC stained with Hoechst and were euthanized for kidney examination 21 days after BMMC injection and samples were collected for histology and immunofluorescence analysis. Histological analysis demonstrated typical interstitial lesions in kidney similarly to human disease, as tubular necrosis, glomerular destruction, atrophy tubular, fibrotic areas, and collagen deposition. We conclude that histological analysis suggest a positive application of agouti's as a model for a gentamicin inducing of kidney disease, beyond the immunofluorescence analysis suggest a significant migration of BMMC to sites of renal injury in CTG. Microsc. Res. Tech., 2011. © 2011 Wiley-Liss, Inc.
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  • 63
    Publication Date: 2011-10-07
    Description: An experimental approach was performed on 20 giant rabbits to establish the possibilities and limitations of μ-CT for routine processing of nondemineralized bone tissue. Hydroxyapatite (HA) or β-tricalciumphosphate (β-TCP) bead implants or a melange of both, microchambered and solid, were implanted into a standardized and precise defect in the patellar groove. The bone-healing phase was chosen for the histology considering 1 or 2 days, and 2, 3, and 6 weeks. Normal X-ray and μ-CT were applied on all specimens; five specimens in the 6-week stage were additionally processed according to the full range of conventional nondemineralized bone processing methods. μ-CT increased the possibilities of nondemineralized histology with respect to bone morphometry and a complete sequence of sections, thus providing a complete analysis of the bone response. μ-CT was limited in differentiating bone quality, cell analyses, and mineralization stages. The investigation based on normal X-rays is limited to defining integration and excluding the fibrous and bony encapsulation of loose implants. μ-CT allows a 3D evaluation of newly formed bone which is clearly marked against the ceramic implant. It does not allow, however, for the differentiation between woven and lamellar bone, the presentation of the canalicular lacunar system, or on the cell level, revealing canaliculi or details of the mineralization process which can be documented by high-resolution microradiography. Titer dynamics of bone formation remains the domain of polychromatic sequential labeling. The complete sequence of μ-CT slices enhances the possibilities for routine histology, tremendously allowing to the focus on detail histology to topographically well-defined cuts, thus providing more precise conclusions which take into consideration the whole implant. Microsc. Res. Tech., 2011. © 2011 Wiley-Liss, Inc.
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  • 64
    Publication Date: 2011-10-15
    Description: This article presents a new approach for extracting high level semantic concepts from digital histopathological images. This strategy provides not only annotation of several biological concepts, but also a coarse location of these concepts. The proposed approach is composed of five main steps: (1) a stain decomposition stage, which separates the contribution of hematoxylin and eosin dyes, (2) a color standardization that corrects color image differences, (3) a part-based representation, which describes the image in terms of the conditional probability of relevant local patches, selected by their stain contributions, (4) a discriminative classification model, which bridges out the found patterns and the biological concepts, (5) a block-based annotation strategy that identifies the multiple biological concepts within an image. A set of 655 skin images, containing 10 biological concepts of skin tissues were used for assessing the proposed approach, obtaining a sensitivity of 84% and a specificity of 67% when annotating images with multiple concepts. Microsc. Res. Tech., 2011. © 2011 Wiley-Liss, Inc.
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  • 65
    Publication Date: 2011-10-15
    Description: The main purpose of this article was to describe the morphology of mesenchymal stem cells (MSCs) differentiated in vitro towards osteogenic and chondrogenic lineages and to focus on the ultrastructural features associated with these processes. Human mononuclear cells (hMNC) were isolated, expanded, and analyzed for the expression of specific cell surface markers to demonstrate their stem cell characteristics. Human mononuclear cells were differentiated in vitro in an osteogenic and in a chondrogenic sense for 7, 14, 21, and 28 days. Subsequently, they were processed using electron microscopic analysis (FEISEM). Alizarin red and alcian blue staining were carried out to demonstrate the deposition of mineral salts and proteoglycans in the extracellular matrix. Undifferentiated MSCs showed a cell surface covered by filopodia and ondulopodia. During differentiation, the MSCs changed their shape from a round to a fibroblastic-like shape. At the end of the differentiation, several filaments with a parallel orientation in the osteogenic samples as well as a network organization in the chondrogenic samples were detected in the extracellular spaces. This study demonstrated that there are morphological features associated with the undifferentiated and differentiated states of the MSCs, which could be utilized as new parameters for identifying and classifying these cells. Microsc. Res. Tech., 2011. © 2011 Wiley-Liss, Inc.
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  • 66
    Publication Date: 2011-10-15
    Description: Starch is the most important carbohydrate in the human diet and contained in many staple foods such as rice, wheat, corn, potatoes and cassava, etc. Currently, microscopic imaging technology is one of the most effective ways to investigate and analyze the structure of starches. Segmentation of starch granules is a necessary step for starch granule structure analysis such as starch granules detection, shape recognition, and size computation. This article investigates a new method based on GVF (gradient vector flow) snake deformable model for starch granules segmentation. The proposed method focuses on full automatic segmentation on granules, especially on separation of adjacent and contacted starch granules, which occur widely in microscopic images. A novel energy function based on position and intensity is introduced into the directional gradient computation, thus the directional gradient is used to obtain the directional GVF snake, which drives the deforming contours to the real contours of multiple granules. To demonstrate the good ability of the proposed method, we segment 30 starch granule images and compare it with the level set method, experimental results show that the new method can separate multiple starch granules successfully; especially it works much better on overlapping objects segmentation than the level set method. Microsc. Res. Tech. 2011. © 2011 Wiley-Liss, Inc.
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  • 67
    Publication Date: 2011-10-15
    Description: Functional restoration for orthopedic, systemic, or dental purposes uses a diverse range of biomaterials. Monitoring for any subsequent failures associated with pathology is vital. To this end, an alternative methodology beyond that of cutting sections by conventional microtome knife-based histomethodologies was pioneered. The method was to saw a block of tissue followed by grinding the unsupported slice to the required thickness. The sawing and grinding of unsupported bones or teeth exposed not only the specimen fragility, but also the delicacy of the associated soft tissues. Although incomplete, the significance of the technique was recognized and improvements were incorporated which persist to the present day addressing the histopathology of challenging specimens. One major improvement for preserving cellular structure as well as reducing fracture incidence was achieved by impregnating tissue samples with cross-linked resins prior to the cutting and grinding processes. Methyl and glycol methacrylate resins superseded all other choices because of the advantages of photo-initiated curing. Further pioneering work led to the formulation of increasingly appropriate resin systems and the subsequent processing protocols evolved, as clinical needs for histology-specific techniques became apparent. Over the decades, several key pioneers entered this scientific arena to develop the now established resin formulation, processing protocols, and allied applications. This has led to the commercialization of the various resin systems. The aim here is to discuss the commercially available glycol and methyl methacrylate resin systems and their practical application to a variety of important techniques used in the histological assessment of complex pathology specimens. Microsc. Res. Tech., 2011. © 2011 Wiley-Liss, Inc.
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  • 68
    Publication Date: 2011-12-07
    Description: Nanofillers are added to dental adhesives to improve mechanical properties of the hybrid layer. Ethanol or water added to the demineralized dentin to improve adhesive infiltration may produce filler aggregation. OBJECTIVE: To assess the effect of 5 vol% water or ethanol addition on nanoparticles distribution in dental adhesives. METHODS: Six available commercial adhesives systems were selected: Clearfil SE Bond (CSE), Clearfil Protect Bond (CPB), FL-Bond (FLB), Clearfil S3 (CS3), Bond Force (BF), One Up Bond F plus (OUB), and an experimental adhesive system without filler (EXP). Polymer films were obtained by adding 0 (control) or 5 vol% water or ethanol into the bonding resins. Preparations were light-cured (40 s). Three specimens were analyzed for each mixture. Three phases and 3D images were taken from each specimen by means of an atomic force microscope in taping mode (TM/AFM). Cluster sizes and surface nanoroughness were assessed. RESULTS: Control specimens from CSE, FLB, OUB, and BF presented clusters. The addition of solvents lead to particles aggregation in tested bonding resins. Ethanol addition produced more aggregates, particularly in adhesives containing fluoraluminosilicate as fillers. CONCLUSIONS: Nanofillers aggregation occurred in all adhesive systems in presence of additional solvents. In general, aggregate sizes were higher after the addition of ethanol. Formed clusters size values are always above the dimensions of the spaces existing between the demineralized collagen fibers. Microsc. Res. Tech. , 2011. © 2011 Wiley Periodicals, Inc.
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  • 69
    Publication Date: 2011-12-07
    Description: This study aimed to evaluate, ex vivo , the nanoleakage in dentinal tubules, the linear infiltration of silver nitrate in the dentin wall/root-end filling material interface, and the presence of gaps in this interface in root-end cavities filled with 4 filling materials. Forty-eight disto-buccal root canals of maxillary molars were instrumented and filled. Retrograde cavities were prepared with ultrasonic points (apical 2 mm). The samples were divided into 2 control groups ( n = 4) and 4 experimental groups ( n = 10): Group I – white mineral trioxide aggregate (MTA); Group II – Super EBA; Group III – Portland cement; and Group IV – Sealer 26. After 1 week, the specimens were subjected to silver nitrate and prepared for SEM (backscattered electrons). In the apical-apical segment, an area with significantly higher leakage was observed for Super EBA, followed by Portland cement, MTA, and Sealer 26 ( P = 0.0054). In the medium and cervical segments, all materials showed the same leakage behavior ( P = 0.1815 and P = 0.1723, respectively). The linear infiltration at the dentin wall/root-end filling material interface was higher with Super EBA than the other groups. No differences in the percentage of gaps along the 3 mm of dentin wall/root-end filling material interface between the 4 materials were evident ( P 〉 0.05). Nanoleakage occurred mainly in the apical segment of the samples, and Super EBA showed the highest values. The area and linear leakage were lower in the middle and coronal segments, regardless of the root-end filling material. No material perfectly sealed the root-end cavities, which allowed for the leakage occurrence. Microsc. Res. Tech., 2011. © 2011 Wiley Periodicals, Inc.
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  • 70
    Publication Date: 2011-12-02
    Description: This article explores the achievable transmission electron microscopy specimen thickness and quality by using three different preparation methods in the case of a high-strength nanocrystalline Cu–Nb powder alloy. Low specimen thickness is essential for spatially resolved analyses of the grains in nanocrystalline materials. We have found that single-sided as well as double-sided low-angle Ar ion milling of the Cu–Nb powders embedded into epoxy resin produced wedge-shaped particles of very low thickness (〈10 nm) near the edge. By means of a modified focused ion beam lift-out technique generating holes in the lamella interior large micrometer-sized electron-transparent regions were obtained. However, this lamella displayed a higher thickness at the rim of ≥30 nm. Limiting factors for the observed thicknesses are discussed including ion damage depths, backscattering, and surface roughness, which depend on ion type, energy, current density, and specimen motion. Finally, sections cut by ultramicrotomy at low stroke rate and low set thickness offered vast, several tens of square micrometers uniformly thin regions of ∼10-nm minimum thickness. As major drawbacks, we have detected a thin coating on the sections consisting of epoxy deployed as the embedding material and considerable nanoscale thickness variations. Microsc. Res. Tech., 2011. © 2011 Wiley Periodicals, Inc.
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  • 71
    Publication Date: 2011-12-02
    Description: Purpose: This work aims to study the erosion on restorative materials and on surrounding dentin. Fifty root dentin samples were obtained from bovine incisors. Methods: Twenty samples were not restored and thirty received cavity preparations. Samples were assigned to five groups: G1, G2: sound dentin (D); G3: composite resin (CR); G4: resin-modified glass-ionomer cement (RMGIC); G5: glass-ionomer cement (GIC). The samples of groups 2–5 were submitted to six cycles (demineralization–remineralization). Samples were analyzed by micro energy-dispersive X-ray fluorescence spectrometry (μ-EDXRF) and by scanning electron microscopy (SEM). Results: Mineral loss was greater in G2 samples than in RMGI 〉 CR 〉 GIC 〉 D (control). SEM images showed pronounced dentin demineralization in groups 2 and 4. The acid erosion has a significant effect on mineral loss (Ca and P) of root dentin without restoration. Conclusions: Composite resin had the best chemical resistance to erosion among all the materials. Fluoride contained in GIC seemed to cause some protection, however, with material degradation. Chemical interaction of tooth-colored dental materials with root dentin could be assessed by μ-EDXRF. Microsc. Res. Tech., 2011. © 2011 Wiley Periodicals, Inc.
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  • 72
    Publication Date: 2012-02-25
    Description: A reliable procedure for measuring parameters connected to surface roughness is needed to compare the gas sensing properties of various thin films or the effect of different fabrication procedures on the surface roughness and the sensing properties. In this article, we propose to investigate how the acquisition parameters specific to atomic force microscopy investigations such as pixel size, scan area and scan speed influence the roughness parameters, namely root mean square and surface area ratio, commonly used for characterizing the gas sensing properties of porphyrins and other materials. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.
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  • 73
    Publication Date: 2012-03-15
    Description: Virus particles and viral inclusions were detected by transmission electron microscopy examination of sections of the seminal vesicles and mucus gland of asymptomatic young drones from colonies of Apis mellifera lightly infested by Varroa mite. In the mucus gland the infection was found in the muscular sheath and epithelium, while in the seminal vesicle in cells of the outer serosa. Isolated viral particles were also observed in the hemolymph occupying the intercellular spaces of the muscular sheath fibers. In the muscle the virus appeared as polygonal crystalloid inclusions, while in the epithelium mainly inside cytoplasmic vesicles. The infected cells apparently are not damaged. The virus particles are present in the hemolymph and forming more mature structures, as crystalloids, in the muscle. This suggests that the virus is liberated in the body fluid and infects the tissues penetrating the cells through endocytosis. The presence of virus in mucus gland epithelial vesicles raise the possibility of its transference to the gland secretion and therefore, to the semen. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.
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  • 74
    Publication Date: 2012-03-15
    Description: The aim of this study was to evaluate the persistence of resin cement residues after dentin surface cleaning with different alcohol-based solutions or an essential oil (eucalyptol). Forty bovine teeth were sectioned in order to expose pulp chamber dentin to be washed with 1.0 mL of 2.5% sodium hypochlorite (NaOCl), followed by 0.1 mL of 17% EDTA application for 3 min, and final irrigation with 2.5% NaOCl. The specimens were air dried and resin-based cement was rubbed onto the dentine surface with a microbrush applicator. After 15 min, the surface was scrubbed with a cotton pellet and moistened with different dentin cleaning solutions, compounding the following groups: G1—95% ethanol, G2—70% ethanol, G3—70% isopropyl alcohol, or G4—eucalyptol. The dentin was scrubbed until the cement residues could not be visually detected. Sections were then processed for SEM and evaluated at ×500 magnification. Scores were attributed to each image according to the area covered by residual sealer, and data were subjected to Kruskal–Wallis at 5% significance. Eucalyptol promoted the most adequate dentin cleaning, although no statistical difference was detected amongst the groups ( P 〉 0.05), except between the eucalyptol and 70% ethanol groups ( P 〈 0.05). All the evaluated dentin cleaning solutions were unable to completely remove the cement residues from the dentin surface. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.
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  • 75
    Publication Date: 2012-03-16
    Description: This study evaluated the effects on human enamel after two bleaching procedures: with a fluoridated bleaching agent and with topical fluoride application postbleaching. It used 43 enamel blocks (3 mm 2 ) that were ground flat (600–2,000 grit) and polished with polishing paste (one and one-fourth). Specimens were randomly divided into three groups according to the bleaching procedure: (1) control group, (2) hydrogen peroxide 35% (HPF) and topical application of fluoride 1.23%, and (3) HP 38% (OP) with fluoride in its composition. Bleaching agents were used according to the manufacturer's instructions. Three methodologies were used: nanoindentation, to observe surface hardness and elastic modulus; atomic force microscopy, to observe surface roughness ( R a − R z ); and scanning electron microscopy, to observe the enamel surface effects. Group OP had a decrease in the elastic modulus after bleaching, which was recovered at 14 days. An increased roughness ( R a ; 32%) was observed on group HPF and had an increased erosion on enamel surface (67%). It was concluded that topical application of fluoride, after using the nonfluoridated whitening agent, increased the roughness values and erosion of enamel. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.
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  • 76
    Publication Date: 2012-03-16
    Description: Feature/edge-preserving noise removal techniques have a strong potential in several application domains including medical image processing. Magnetic resonance (MR) images have a tendency to gain Rician noise during acquisition. In this article, we have presented genetic algorithms based adapted selective non-local means (GASNLM) filter-based scheme for noise suppression of MR images while preserving the image features as much as possible. We have applied GASNLM filter with optimal parameter values for different frequency image regions to remove the noise. Filter parameter values are optimized by genetic algorithm (GA). A change in NLM filter known as selective weight matrix is also proposed to preserve the image features. The results prove soundness of the method. We have compared results with many well known and latest techniques, and the improvements are discussed. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.
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  • 77
    Publication Date: 2012-03-11
    Description: OBJECTIVES: The aim of this in vitro study was to investigate the efficacy of in-office bleaching technique combined with the application of a casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) paste (MI Paste–MI) at different moments and its influence on enamel surface properties. METHODS: Eighty bovine dental crowns were randomly allocated into eight groups ( n = 10), and bleached with either 35% hydrogen peroxide (HP) or 37% carbamide peroxide (CP). Four different protocols of application of MI were considered: without MI, MI applied before bleaching, MI applied after bleaching, and MI applied both before and after bleaching. Bleaching effectiveness was measured by the VITA EasyShade spectrophotometer utilizing the CIEL*a*b* system (Δ E , Δ L *, Δ a *, and Δ b *). Color readings were measured at baseline, 7, 14, and 21 days. Hardness and roughness were measured at baseline (T0) and immediately after bleaching (T14). Data were subjected to the two-way ANOVA for repeated measurements and Tukey's test at 5%. RESULTS: HP groups achieved the greatest color change. The application of a CPP-ACP paste did not reduce the efficacy of bleaching peroxides. Samples bleached with CP showed decreased hardness at T14. Samples bleached with HP that received the application of MI before and before/after bleaching did not present hardness decrease at T14. Samples bleached with peroxides only and received MI after bleaching showed increased roughness at T14. CONCLUSIONS: The use of CPP-ACP was able to prevent negative changes in roughness and hardness of bovine enamel when associated to hydrogen peroxide, and might be applied before/after the bleaching protocol. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.
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  • 78
    Publication Date: 2012-03-11
    Description: Ticks are ectoparasites of great medical and veterinary importance around the world and synthetic chemicals such as permethrin have been used for their control. This study provides a cytochemistry analysis of both degenerative and cell death processes in salivary glands of the brown dog tick Rhipicephalus sanguineus semi-engorged females exposed to 206, 1,031, and 2,062 ppm of permethrin. The results presented herein demonstrate that permethrin is a potent chemical acaricide that would act on the glandular tissue's morphophysiology in this tick species by eliciting severe changes in the acinus shape, intense vacuolation of the acinar cells' cytoplasm, marked glandular tissue disorganization, culminating in an advanced degenerative stage with consequent formation of many apoptotic bodies (cell death). In addition, permethrin induced major changes in the acinar cells' nucleus, such as a change both in its shape and size, chromatin marginalization, nuclear fragmentation, and appearance of picnotic nuclei, especially when the highest concentrations of the product were used. Thus, permethrin induced early degeneration of this tissue characterized by significant changes in the structure of acinar cells and production of enzymes related to the cell death process, in addition to interfering directly in the genetic material of these cells. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.
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  • 79
    Publication Date: 2012-03-15
    Description: The pumping organ of blood-sucking female mosquitoes has a three-dimensional (3D) structure. However, conventional two-dimensional imaging methods are insufficient for visualizing the 3D structure in detail. Furthermore, their 3D imaging tasks are highly time consuming and sample preparation process requires elaborate skill. Among 3D imaging techniques, synchrotron X-ray microscopic computed tomography (SR-μCT) is especially suitable for small insects with opaque cuticles, such as mosquitoes. In this study, the 3D morphological structure of the pump system of a female mosquito was visualized using SR-μCT. Expandable volume capacities of two pump chambers were measured for several mosquito samples of similar size. To verify the cross-sectional images acquired by SR-μCT, complementary paraffin-sectioning data were compared. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.
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  • 80
    Publication Date: 2012-03-16
    Description: Frictional interactions between wires and brackets reduce the efficacy in orthodontic treatments. Self-ligating brackets (SLBs) are now more often used due to lower frictional forces when compared with conventional-ligating brackets. In this study, scanning electron microscopy and atomic force microscopy were used to examine the microstructural effects of stainless steel and ceramic SLBs on the surface roughness of stainless steel and superelastic NiTi wires both after in vivo clinical orthodontic treatment as well as in in vitro three-point bending experiments. A combination of two wires—0.019 in. × 0.025 in. stainless steel wires and 0.016 in. superelastic NiTi wires—and two SLBs—both passive-type stainless steel SLBs and active-type ceramic SLBs—was applied for 4 months (bicuspid-extraction) in an in vivo setting and for 1 month in an in vitro setting (200 g loads). After the SLB treatments, all wires exhibited severe scratches secondary to frictional interactions with the brackets. When used with the stainless steel SLBs (Damon 3MX®), the surfaces of 0.019 in. × 0.025 in. stainless steel ( P 〈 0.0001) and 0.016 in. superelastic NiTi wires ( P 〈 0.05) were significantly smoother than when used with the ceramic SLBs (Clippy-C®). Such results suggest that orthodontic treatments with stainless steel SLBs are more effective than with ceramic SLBs. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.
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  • 81
    Publication Date: 2012-03-16
    Description: To determine whether sodium hypochlorite (NaOCl) with 0.2% chlorhexidine gluconate (CHX) leads to colour change and precipitate formation, and to ultrastructurally analyse the dentine surface after simultaneous irrigation with 0.5% NaOCl and 0.2% CHX. Four tubes in which 5ml of different NaOCl concentrations and 5ml 0,2% CHX were placed, were observed every 15 minutes in the first two hours and after 7 days. Mixture solutions were centrifuged at 800 rpm/4 min. A precipitate is observed under light-microscopy. Thirty-five single-rooted teeth were instrumented using crown-down technique with irrigation: Positive control: distilled water, Negative control: 0,5% NaOCl+15% EDTA, Experimental group: identical to the negative control, then canals were treated with 0.5% NaOCl+0.2% CHX. The longitudinal root sections were observed under scanning-electron-microscopy. The amount of debris was assessed with 5/score-system, and the results were analysed using the Mann-Whitney U test (p 〈 0.001). Change in colour was noticed immediately after the merger and it did not change with time. Mixture solutions showed considerable turbidity, but precipitate was observed only after centrifugation. Statistical analysis showed a significant difference in the dentinal wall purity when comparing the cervical and middle root canal thirds between the experimental and negative control group. The difference was also observed between the cervical and middle thirds compared to the apical third of the root canal within these groups. Interaction between NaOCl and CHX, as well as the creation of precipitates, depends not only on the concentration of NaOCl, but also on the concentration of CHX. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.
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  • 82
    Publication Date: 2012-03-16
    Description: Leafcutter ants of the genus Atta Fabricius are serious agricultural pests. Morphological studies of immature stages within this group are few, and the data provided for species of considerable importance are usually incomplete. In this study, the immatures of Atta sexdens Linnaeus are described and compared using light and scanning electron microscopy. Only specimens from founding stage colonies (i.e., lacking adult workers) were used. The existence of four larval instars was estimated by a frequency plot of maximum head widths, and the larvae of different instars differed from each other mainly by their bodily dimensions. Worker larvae belonged to two distinct morphological castes: (1) gardeners and nurses and (2) within-nest generalists. The worker larvae described in this study differed from a previous description of the same species by the following traits: the existence of a genal lobe, the number of clypeal hairs, the presence of two hairs on the ninth abdominal somite, the presence of hairs on the anterior surface of the labrum, and the shape of the maxillary palpus. This study provides a comparative analysis of immature stages of A. sexdens that may be relevant to future morphological and biological studies of the Attini. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.
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  • 83
    Publication Date: 2012-12-29
    Description: ABSTRACT Microstructure of gelatin particles played a key role in determining the physicochemical properties of gelatin. Ionic strength and pH as systematic manners were considered to affect gelatin particles structure on the micrometer scale. Scanning electron microscopy was used for depicting the morphologies of gelatin particles. Increasing pH to 10.0 or decreasing pH to 4.0, spherical, spindle, and irregular aggregates of gelatin particles at 2, 6, 10, and 14% solution (w/w) were all transformed to spindle aggregates. When NaCl was added to the system, the molecular chains of gelatin possibly rearranged themselves in a stretched state, and the ribbon aggregates was observed. The structural transitions of gelatin aggregates were strongly depended on the electrostatic repulsion. In the gelatin–sodium dodecyl sulfate (SDS) case, the micrometer scale of aggregates was larger and the different degrees of cross-links were induced through hydrophobic interaction and electrostatic repulsion. Microsc. Res. Tech., 2012 . © 2012 Wiley Periodicals, Inc.
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  • 84
    Publication Date: 2012-12-29
    Description: Purpose: Till date there is no exclusive marker for human corneal epithelial stem cells (CESCs). In this study, our strategy is to combine high expression of ABCG2, a putative SC marker with high N/C ratio to develop a specific method for identification of CESCs. Methods Limbal/corneal epithelial cells (LECs/CECs) were isolated from cadaver eyes by enzymatic treatment and the cytospin smears/cryosections were immunostained for ABCG2, p63, or connexin-43 (Cx-43) and counterstained with propidium iodide/DAPI. Analysis was carried out for (1) ABCG2 expression, N/C ratio, (2) p63, ABCG2, N/C ratio, and (3) ABCG2, Cx-43, N/C ratio using confocal/fluorescence microscopy. Results ABCG2 was highly expressed by cells in the basal and suprabasal layers of limbus. The SCs identified on the basis of our earlier method using high levels of p63 expression along with high (〉0.7) N/C ratio were strongly positive for ABCG2 and negative for differentiation marker Cx-43. High expression of ABCG2 was also observed in cells with high p63 expression and low (〈0.7) N/C ratio; however, they were positive for Cx-43. Combining high ABCG2 expression with large N/C ratio in fluorescence microscopy identified 5.3 ± 3.5% LECs as putative SCs and this was confirmed by confocal microscopic analysis. Conclusions We demonstrate the importance of combining ABCG2 expression with N/C ratio for identification and quantification of human CESCs. This method will have application in evaluating the role of niche cells/factors in the maintenance of stemness. Microsc. Res. Tech., 2012 . © 2012 Wiley Periodicals, Inc.
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  • 85
    Publication Date: 2013-01-19
    Description: ABSTRACT Early osteoporosis diagnosis is of important significance for reducing fracture risk. Image analysis provides a new perspective for noninvasive diagnosis in recent years. In this article, we propose a novel method based on machine-learning method performed on micro-CT images todiagnose osteoporosis. The aim of this work is to find a way to more effectively and accurately diagnose osteoporosis on which many methods have been proposed and practiced. In this method, in contrast to the previously proposed methods in which features are analyzed individually, several features are combined to build a classifier for distinguishing osteoporosis group and normal group. Twelve features consisting of two groups are involved in our research, including bone volume/total volume (BV/TV), bone surface/bone volume (BS/BV), trabecular number (Tb.N), obtained from the software of micro-CT, and other four features from volumetric topological analysis (VTA). Support vector machine (SVM) method and k -nearest neighbor ( k NN) method are introduced to create classifiers with these features due to their excellent performances on classification. In the experiment, 200 micro-CT images are used in which half are from osteoporosis patients and the rest are from normal people. The performance of the obtained classifiers is evaluated by precision, recall, and F-measure. The best performance with precision of 100%, recall of 100%, and F-measure of 100% is acquired when all the features are included. The satisfying result demonstrates that SVM and k NN are effective for diagnosing osteoporosis with micro-CT images. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 86
    Publication Date: 2012-11-10
    Description: To well enhance the mineral image and image details obtained by microscopes, an effective mineral image enhancement algorithm through feature extraction using the morphological center operator is proposed in this work. First, mineral image feature extraction based on the morphological center operator is proposed and discussed. Second, the multiscale extension of the mineral image feature extraction is given by using the multiscale structuring elements. Third, the important mineral image features at multiscales of image are extracted and used to construct the final mineral features for mineral image enhancement. Finally, the original mineral image is well enhanced through importing the extracted final mineral image features into the original mineral image. Experimental results on different types of microscopy images of minerals verified the effective performance of the proposed algorithm for microscopy mineral image enhancement. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.
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  • 87
    Publication Date: 2013-03-02
    Description: ABSTRACT This study sought to evaluate the efficacy of passive ultrasonic irrigation (PUI) on removing the smear layer and debris from root dentin using scanning electron microscopy (SEM). Twenty-five bovine incisors were manually prepared and divided into three groups according to the final irrigation protocol: EDTA, final irrigation with 12 mL of 17% EDTA for 3 minutes followed by 5 mL of 2.5% NaOCl; EDTA/PUI, final flush with 4 mL of 17% EDTA and PUI for 30 seconds. These procedures were repeated three times to standardize the volume of the irrigant. Control group, after preparation, the specimens were irrigated only with 17 mL of 2.5% NaOCl. The roots were fractured and analyzed using SEM. The intragroup analysis revealed that the EDTA/PUI protocol removed a higher amount of debris at the cervical third ( P  = 0.03). The intergroup analysis revealed that EDTA/PUI presented the lowest amount of debris at the cervical third ( P  = 0.007). Smear layer scores were higher in the control group compared with the EDTA and EDTA/PUI groups, but only at the cervical third ( P  = 0.02). None of the final irrigant protocols completely removed the smear layer and debris. EDTA/PUI only improved the removal of debris at the cervical third. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 88
    Publication Date: 2013-03-02
    Description: ABSTRACT The bullfrog ( Lithobates catesbeianus ) is an exotic animal that adapted perfectly to the climatic conditions of Brazil after introduction in the country. These climatic conditions favor the reproduction and fattening of bullfrogs. However, the development of the bullfrog is significantly reduced in colder regions because low temperatures affect the secretion of gonadotropins and change the sensitivity of the germinal epithelium to gonadotropic hormones. Thus, the temperature and photoperiod are abiotic factors that influence the life cycle of this species, especially with regard to reproduction. Samples from different regions of the oviducts (the infundibulum, magnum, and isthmus) of 9 bullfrogs were collected in the different seasons of the year. The samples were fixed and processed for analysis with scanning and transmission electron microscopy. A comparison of the morphology of the oviducts collected during different seasonal periods suggests that the frogs have greater reproductive potential in the spring due to the development of the structures related to the oviduct, presence of developed cilia, and electron-dense granules. However, seasonal changes were noted in the animal throughout the year in preparation for reproduction during the spring and summer. Microsc. Res. Tech., 2013 . © 2013 Wiley Periodicals, Inc.
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  • 89
    Publication Date: 2012-09-29
    Description: Analysis of the cellular distributions of coenzymes including NADH may aid in understanding a cells metabolic status. We altered serum concentration (0, 2, and 10%) to induce living myoblast cells to undergo the early stages of differentiation. Through microscopy and phasor-FLIM, we spatially mapped and identified variations in the distribution of free and bound NADH. Undifferentiated cells displayed abundant free NADH within the nucleus along with specific regions of more bound NADH. Complete serum starvation dramatically increased the fraction of bound NADH in the nucleus, indicating heightened requirement for transcriptional processes. In comparison, cells exposed to 2% serum exhibited intermediate free nuclear NADH fraction. Overall our results suggest an order of events in which a cell metabolic status alters significantly during the early stages of serum induced differentiation. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.
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  • 90
    Publication Date: 2012-10-04
    Description: This article presents the SEM-EDX and microFTIR study and the corrosion behavior of new five types of phosphated coatings obtained by coprecipitation in acid aqueous medium of some metal cations, pursuing the influence of the addition of other cations and a moderator of precipitation, on the uniformity and compactness of the layers and on the morphology of dendritic structure of Zn(II) and Fe(II) phosphates. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.
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  • 91
    Publication Date: 2012-10-05
    Description: Because of the limitations of the X-ray hardware systems in mammogram machines, the quality of the breast mammogram images may undergo from poor resolution or low contrast. Quantum noise occurs in the mammogram images during acquisition due to low-count X-ray photons. In this work, an adaptive frost filter has been used to remove quantum noise. Local binary patterns have been extracted to classify breast mammograms into benign and malignant using different classifiers. Results show the superiority of the proposed algorithm in terms of sensitivity, specificity, and accuracy. Mammographic Institute Society Analysis database of mammography has been used for experimentation. Peak signal-to-noise ratio and structural similarity index measure are used to test the validity of adaptive frost filter. Experiment results show that proposed technique produces better results. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.
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  • 92
    Publication Date: 2012-07-14
    Description: The knowledge of the process of egg formation is indispensable for understanding the mechanisms involved in the reproduction of different species. In this context, the objective of this work was to describe the ultrastructure of the oocytes of Urostreptus atrobrunneus (Spirostreptida), a potential plague of urban centers in different locations of São Paulo State. The lack of knowledge about the morphology, physiology, and the reproductive behavior of the species have hindered an effective control of it. The oocytes of U. atrobrunneus presented three development stages: young oocyte or type I; intermediary oocyte or type II; and mature oocyte or type III. During the oocyte development, the cytoplasm become filled with several globules of protein, drops of lipids, and sphaerocrystals, and it was not observed in many organelles in the oocytes with exception of mitochondria, abundant, principally in young oocytes. The vitelline membrane is also deposited in a discontinuous form and the chorion does not present differentiation of layers. The follicular epithelium alters its shape according to the development phase of the oocyte. Part of the vitellus is from exogenous origin and part is endogenous. Before this, only two studies about the ultrastructural analysis of the female germ cells of diplopods were published. Microsc. Res. Tech. © 2012 Wiley Periodicals, Inc.
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  • 93
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    Oxford University Press
    Publication Date: 2012-10-12
    Print ISSN: 0022-0744
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    Topics: Electrical Engineering, Measurement and Control Technology , Natural Sciences in General , Physics
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  • 94
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    Unknown
    Oxford University Press
    Publication Date: 2012-10-12
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  • 95
    Publication Date: 2012-10-12
    Description: The dry weight of tendon tissue is accounted for mainly by collagen fibers. Accordingly, the tendon-healing process primarily involves repair of collagen fibers. During the remodeling phase of tendon healing, newly proliferating collagen fibers are transformed into a mature repaired tendon. Despite the importance of this phenomenon, the details of fibrous rebuilding have not been reported previously. The aim of this study was to visualize the ultrastructural changes and to obtain a clear understanding of the reorganization of the collagen fibers in the tendon repair site, using rat Achilles tendons. We used scanning electron microscopy (SEM) with cell maceration as the main method of analysis. Pretreatment with cell maceration removed the cellular components successfully. This allowed precise visualization of each collagen fiber and the three-dimensional network of the fibers. This study was the first to apply the cell-maceration/SEM method to observe tendon tissue. Seven days after surgery, new collagen fibers grew extensively in the repair site in a random arrangement. Fourteen days after surgery, the collagen fibers began to form an axial arrangement. Near the tendon stump, this change progressed from the outer layer to the core region. On the other hand, in the middle of the repair site, it progressed from the core to the outer layer. Change in the axial arrangement of collagen fibers contributes to the connection between the repair site and the tendon stump and to the separation of the repair site from the paratenon.
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  • 96
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    Oxford University Press
    Publication Date: 2012-10-12
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  • 97
    Publication Date: 2012-10-12
    Description: We have used transient electron-beam-induced current (EBIC) to map minority carrier lifetime distributions in multicrystalline Silicon (mc-Si). In this technique, the electron beam from a scanning transmission electron microscope was on–off modulated while the sample was scanned. The resulting transient EBIC was analyzed to form a lifetime map. An analytical function was introduced as part of the analysis in determining this map. We have verified this approach using numerical simulations and have reproduced a lifetime map for an mc-Si wafer.
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  • 98
    Publication Date: 2012-10-17
    Description: The method to observe the exact morphology of swelled seaweed as an example of biological material by field emission scanning electron microscopy (FE-SEM) with the aid of hydrophilic ionic liquid (IL); 1-butyl-3-methylimidazolium tetrafluoroborate is reported. Seaweed was first swelled in 3.5% NaCl solution and then treated with the IL and water mixture in 1:7 weight ratios and centrifuged to remove the excess IL solution. Thus treated seaweed maintained its morphology even at high magnification and did not show drying in the FE-SEM chamber. This observation technique might be useful for various kinds of biological materials to be observed under FE-SEM. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.
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  • 99
    Publication Date: 2012-10-18
    Description: Purpose: The aim of this in vitro study was to verify the interference of previous application of sodium hypochlorite in the bond strength to dentin irradiated with Er:YAG laser (80 mJ, 2 Hz, 25 J/cm 2 ), using two adhesive systems with different solvents (Adper Single Bond2 – 3M ESPE [SB] and Prime&Bond 2.1 – Dentsply [PB]. Study Design: Ninety standardized dentin disks were treatment with phosphoric acid etching (PA), PA and 10% sodium hypochlorite (SH), Er:YAG laser (ER), ER and 10% SH. After surface preparation, samples received SB or PB followed by composite resin Z250 (3M ESPE)( n = 10). Tensile bond strength test was performed (0.5 mm/min) 24 h after bonding procedure and one treated dentin disk of each group was selected to scanning and transmission electron microscopy (SEM and TEM) observations. Bond strength test data were evaluated by three-way ANOVA and Tukey ( P 〈 0.05) tests. Results: Tested systems had similar bonding behavior when applied with conventional method. SH produced no improvement in bond strength. Phosphoric acid etching either associated with SH, or not, provided higher bond strength values than ER. SEM showed typical ablation pattern on dentin after ER irradiation. SH application after PA promoted greater porosity in intertubular dentin exposing secondary tubules. SH used after ER irradiation showed more evident peritubular dentin. TEM results showed that both PA and ER exposed inter- and peri-tubular dentin collagen fibers. Conclusions: Er:YAG irradiation had a negative influence on SB adhesive, and in both methods, association of SH did not completely remove collagen fiber network, but altered its configuration. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.
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  • 100
    Publication Date: 2012-10-18
    Description: Environmental factors can influence the reproductive rates in bats, and since morphometric information of bats testis is scarce, we aimed to compare the organization and quantification of the intertubular components in the testes of the bat Molossus molossus , collected in different seasons. Testicular histological sections were evaluated using light and electron microscopy. The intertubular compartment occupied an average 10% of the testes, being predominately constituted of Leydig cells (LC). The percentages of the testes occupied by the intertubular compartment and by LC were significantly higher in summer, while the other intertubular components did not vary significantly among the seasons. As suspected under light microscopy, the ultrastructural analysis confirmed the existence of multinucleated LC during winter. The increase in the nuclear percentage of LC in winter seems to have caused the decrease of the cytoplasmatic measurements in that season, as well as in the volume of LC. The highest cytoplasmatic values and volume of LC registered in the spring, summer, and fall can be related to greater activity of this cell in these seasons. The higher investment in intertubular tissue and in LC observed in summer, compared to winter; suggest an increase in the steroidogenic capacity of this bat during summer. The analyses correlating testicular morphometry and abiotic environmental factors in this study confirm the influence of climatic factors on the reproduction of M. molossus . Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.
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