ALBERT

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

Description: Insects rely heavily on olfaction to locate habitat, mates, and oviposition sites, while odorant molecules and the antennal olfactory sensory cells of insects are two indispensable components of olfactory response. Our previous work identified the sex pheromones and volatile compounds derived from host plant of Dendrolimus tabulaeformis Tsai et Liu, a serious economic pest of pines in northern China. However, little is known about the olfactory system of D. tabulaeformis , especially in females. To make a better understanding of the D. tabulaeformis olfactory response, we investigated the structure, innervation, numbers, and distribution of sensilla on the antennae of male and female moth, based on scanning and transmission electron microscopy. The pinniform antennae of this moth bear five types of sensilla: trichoid (TS), basiconica (BS), styloconic (StS), and chaetica (ChS) sensilla, and belt-type structures (BTSs). The BTSs are hollow and are not chemical sensilla. The ChSs occur on the scape. StS contain dendrite sheaths and four dendrites containing microtubules in their cores. The BS contents are continuous with that in the subsegments. The TSs were sexually dimorphic and could be divided into 12 subtypes based on dendrite number and form. TS1-4 contained two, three, or four dendrites and were similar in both sexes. TS 5-8 and TS 9-12 occurred only on male and female antenna, respectively. We discussed the possible functions of these sensilla. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: PURPOSE: To investigate the changes in the ultrastructural architecture of hydrated corneoscleral button (HCSB) and hydrated corneal button (HCB). METHOD: Corneoscleral buttons ( n = 4) and corneal buttons ( n = 4) were hydrated in deionized water for 24 and 48 h and were fixed in 2.5% glutaraldehyde containing cuprolinic blue in sodium acetate buffer and processed for electron microscopy. The tissue was dehydrated and embedded in TAAB 031 resin. Normal CSB and CB were also processed using the same method. Ultrathin sections were stained with uranyl acetate and lead citrate. The sections were observed under JEOL 1400 transmission electron microscope. RESULTS: In both HCSB and HCB, the lamellae were disorganized and proteoglycans (PGs) were degenerated and detached from the collagen fibrils (CFs). In the HCSB samples, interfibrillar spacing increased but the diameter of the CF had not changed compared with those in the normal tissue. Within the HCSB, the interfibrillar spacing in the posterior stroma was significantly higher compared with those in the anterior and middle stroma. In the HCB, both the interfibrillar spacing and CF diameter had significantly increased compared to those observed in both the HCSB and normal tissue. The CFs were severely damaged exhibiting very thin microfilaments within CFs. Remnants of the CFs were present in the stromal matrix. CONCLUSION: Hydration of the cornea in both the HCSB and HCB increases the interfibrillar spacing in the anterior, middle, and posterior stroma. The CF in the HCB was enlarged and had degenerated. This could be due to penetration of water into the intermolecular spacing within the CF. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: This research aimed to examine the morphological features of the tongue and laryngeal entrance of long-legged buzzard by macroscopic and scanning electron microscopic methods. Two adult buzzards were used as material. The tongue was fairly elongated and terminated in oval shovel-like apex. Scale-like projections were localized on the apex and body of tongue. Both lateral sides of lingual body were contained considerably long thread-like projections. Many orifices of lingual posterior salivary glands were discerned among scale-like projections in median sulcus of the lingual body. Papillary crest of sharp conical papilla were observed on the between the body and root of the tongue. No conical papillae or other projections were existent on the root of the tongue, but numerous orifices of posterior salivary gland ducts were detected. In addition, orifices of anterior salivary gland ducts were present on the dorsal and lateral surfaces of the lingual body. Numerous conical papillae were observed on the caudal region of glottis and no conical papillae or any similar projection which were bordered the glottic fissure was noted. Anatomy of these organs in the long-legged buzzard and white tailed eagle which are the member of the same family, Accipitridae, revealed very high similarity. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: This study aimed to evaluate the effect of four chemomechanical surface treatments on the surface average microroughness and profile of laser-sintered and vacuum-cast dental prosthetic structures. Square-shaped blocks (10 mm × 10 mm × 1.5 mm) were prepared as follows: (1) laser-sintered Co Cr (L) (ST2724G); (2) cast Co Cr (C) (Gemium-cn); and (3) cast Ni Cr Ti (T) (Tilite). Specimens of each alloy group were randomly divided into five subgroups ( n = 10 each), depending on the conditioning method used: (1) no treatment (control); (2) sandblasting (125 μm Al 2 O 3 -particles); (3) silica coating (50 μm silica-modified Al 2 O 3 -particles); (4) oxidation; and (5) oxidation plus opacification. Subgroups 2 and 3 represent “inner” pretreatments proposed for ceramometal restorations to improve the metal surface area available for luting cements. Subgroups 4 and 5 are the “outer” pretreatments required for bonding the aesthetic veneering ceramics to the underlying metal frameworks. Average surface roughness (Ra/μm) was determined using a surface profilometer. Data were analyzed by two-way ANOVA and Student–Newman–Keuls tests (α = 0.05). Metal surface topography was SEM-analyzed. Despite the inner pretreatment applied, L samples resulted in the highest microroughness ( P 〈 0.001), whereas sandblasting produced a surface-smoothing effect in cast specimens. After oxidation, a significant increase in surface roughness occurred in all groups compared with controls, L specimens being the roughest ( P 〈 0.001). Opacification caused a flattening effect of all oxidized structures; all opacified groups resulting in similar microroughness. Laser sintering of Co Cr enhances the roughness of metal structures, which may improve the frameworks' microretention of the cements, and of the opaquer before the copings are veneered with the aesthetic ceramics. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: There is an increasing variety of endocrine disrupting chemicals (EDCs) either with (anti)estrogenic or (anti)androgenic potential widely present in the environment. These xenosteroids may mimic endogenous steroid hormones disrupting the homeostasis of physiological pathways and leading to several disturbances, especially in tissues highly dependent on steroid hormones such as the prostate. Taking this into account, this comparative study aimed to verify the potential of ethinylestradiol (EE) and testosterone acting as ECDs on the prostate of both male and female adult gerbils exposed to these agents during the embryonic phase. Consequently, pregnant gerbils were treated either with 10 μg/kg/day of EE or with a single dose of 1 mg of testosterone cypionate. The pups that were born 6–8 days after testosterone exposure and the pups that were born after 3 days of EE exposure were allowed to grow but were sacrificed within 4 months. Serological, morphological, stereological, and immunohistochemical analyses were used. Overall, the results showed that both sexes exposed to testosterone and EE during gestation had a prostatic gland with an increased stromal and epithelial and a reduced luminal compartment. Moreover, we observed that glands affected with prostatic intraepithelial neoplasia showed intense stromal reshuffling. In conclusion, although these alterations were observed in both sexes, more relevant to this study was the differential responsiveness of males and females exposed to these different drugs. Whereas the EE affected males more, the testosterone was more harmful to the females. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.

Description: Electron microscopy has been used to visualize chromosome since it has high resolution and magnification. However, biological samples need to be dehydrated and coated with metal or carbon before observation. Ionic liquid is a class of ionic solvent that possesses advantageous properties of current interest in a variety of interdisciplinary areas of science. By using ionic liquid, biological samples need not be dehydrated or metal-coated, because ionic liquid behaves as the electronically conducting material for electron microscopy. The authors have investigated chromosome using ionic liquid in conjunction with electron microscopy and evaluated the factors that affect chromosome visualization. Experimental conditions used in the previous studies were further optimized. As a result, prewarmed, well-mixed, and low concentration (0.5∼1.0%) ionic liquid provides well-contrasted images, especially when the more hydrophilic and the higher purity ionic liquid is used. Image contrast and resolution are enhanced by the combination of ionic liquid and platinum blue staining, the use of an indium tin oxide membrane, osmium tetroxide-coated coverslip, or aluminum foil as substrate, and the adjustment of electron acceleration voltage. The authors conclude that the ionic-liquid method is useful for the visualization of chromosome by scanning electron microscopy without dehydration or metal coating. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: The glycoconjugate pattern of developing ovarian follicles in wild and cultured Senegalese sole Solea senegalensis was investigated by means of lectin histochemistry. Ovaries from cultured fish contained oocytes up to the late vitellogenic stage, whereas they reached the hydration stage in wild specimens. The follicular cells bound MAL II, SBA, HPA, DBA, Con A, KOH-sialidase (K-s)-WGA, GSA I-B 4 in the late vitellogenic stage, and in wild fish also SNA and K-s-PNA, whereas in the hydration stage SBA, HPA, DBA, and GSA I-B 4 only. The zona radiata reacted with SBA, HPA, DBA, Con A, and GSA I B 4 in the late vitellogenic stage and in cultured fish also with UEA I, whereas in the hydration stage it stained with SBA only. The cortical alveoli bound SBA, HPA, RCA 120 during the late vitellogenic stage, also SNA, PNA, K-s-PNA, GSA I-B 4 in cultured fish, DBA, and K-s-WGA in wild ones which stained with SBA, HPA, and GSA I-B 4 in the hydrated stage. The yolk reacted with Con A in the late vitellogenic oocytes, and also with MAL II, SNA, K-s-PNA, SBA, HPA, K-s-WGA, GSA I-B 4 , UEA I in the hydrated ones. From perinucleolus to late vitellogenic stages, the oocyte nucleoplasm bound Con A, GSA I-B 4 , GSA II, UEA I, and in wild fish also MAL II, SNA, LTA but only GSA I-B 4 reactivity in the early maturation stage. These findings demonstrate that the glycan pattern of fish ovarian follicles changes during the maturative stages and that it is affected by culture-rearing conditions. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.

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.

Description: Here we report a new sample preparation method for three-dimensional electron tomography. The method uses the standard film deposition and focused ion beam (FIB) methods to significantly reduce the problems arising from the projected sample thickness at high tilt angles. The method can be used to prepare tomography samples that can be imaged up to a ±75° tilt range which is sufficient for many practical applications. The method can minimize the problem of Ga + contamination, as compared to the case of FIB preparation of rod-shaped samples, and provides extended thin regions for standard 2D projection analyses. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: The liver has a remarkable ability to regenerate after partial hepatectomy (PH), although the factors governing such ability are still poorly understood. During the prereplicative phase of the regeneration, ultrastructural alterations of periportal hepatocytes were seen, including mitochondrial swelling, abnormal accumulation of lipids, and myelin figures which could lead to the formation of lipid droplets. As it has been hypothesized that caveolin-1 is involved in lipidogenesis and in mitochondrial homeostasis, we aimed to study the subcellular distribution of caveolin-1 in hepatocytes at an early stage following PH. Liver samples were processed for light and electron microscopy at 0 h, 24 h, and 96 h after PH. The expression and subcellular distribution of caveolin-1 was assessed by immunohistochemical and immunocytochemical techniques. Following PH, at 24 h, membranes of altered mitochondria of periportal hepatocytes exhibited significant decrease of caveolin-1 expression compared with control. Myelin figures showing high expression of caveolin-1 were also seen. At 96 h, hepatocytes became ultrastructurally similar to the control liver, and the expression of caveolin-1 on mitochondria showed a moderate increase compared with 24 h after PH. Decrease of expression of caveolin-1 in the altered liver mitochondrial membranes at 24 h following PH, and the high expression of caveolin-1 observed on myelin figures, suggests involvement of caveolin-1 is in both mitochondrial homeostasis and lipidogenesis. Addressing the role played by caveolin-1 during liver regeneration might disclose additional features of mitochondrial homeostasis and lipidogenesis during frequent metabolic liver diseases. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

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.

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.

Description: Apatura ilia (Denis and Schiffermüller, 1775) and A. iris (Linnaeus, 1758) are fascinating butterflies found in the Palaearctic ecozone (excepting the north of Africa). The wings of these insects are covered with a great number of two types of scales positioned like roof tiles. Type I scales are on the surface, while type II scales are situated below them. The structural color of the type I scales is recognized only on the dorsal side of both the fore and hind wings of the males of the aforementioned species. Both types of scales are responsible for pigment color of the wings, but iridescence is observed only in the type I scales. The brilliant structural color is due to a multilayer structure. The features of the scales, their dimensions and fine structure were obtained using scanning electron microscopy. Cross sections of the scales were then analyzed by transmission electron microscopy. The scales of the “normal” and clytie forms of A. ilia have a different nanostructure, but are of the same type. A similar type of structure, but with a different morphology, was also noticed in A. iris . The scales of the analyzed species resemble the scales of tropical Morpho butterflies. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: Titanium surface texture and chemistry modification successfully improves the host response and consequently the bone-to-implant contact surrounding dental implants. The aim of the present study was to investigate, using histomorphometrical-analysis, the effects of titanium surface modification by laser-ablation (Nd:YAG) followed by thin chemical deposition of HA. Forty-eight rabbits received one implant by tibiae of AS-machined (MS), laser-modified (LMS), or biomimetic hydroxyapatite-coated (HA) surface. Bone-to-implant contact (BIC) and bone area (BBT) were evaluated after 4, 8, and 12 weeks, at cortical and cancellous regions. Average BIC in the cortical region was higher ( P 〈 0.001) on the LMS and HA implants for all periods, with no differences between LMS and HA. For the cancellous area, the LMS and HA implants showed higher ( P 〈 0.01) BIC than MS at the initial periods. The LMS and HA showed similar values in the cortical region, but a tendency of higher values for HA in the cancellous region was observed in all periods. For the BBT, the differences were found only between HA and MS after 4 weeks in the cortical region ( P 〈 0.05), and after 12 weeks in the cancellous area ( P 〈 0.05). Our results showed that HA biomimetic coating preceded by laser treatment induced the contact osteogenesis and allowed the formation of a more stable bone–implant interface, even in earlier periods. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: Chrysomya megacephala is a forensic important fly, and its immature forms also cause myiasis. The adults are the first insects to reach a carcass and can oviposit just a few hours after arrival. Therefore, the knowledge of immature stages of flies is essential for correct identification of the species found on corpses. Scanning electron microscopy (SEM) gives detailed information on the morphological characters that can help identify the immature forms of flies. The aim of this study is to identify larvae and puparia of C. megacephala using SEM. The larval instar body of C. megacephala is similar at all instars. The integument is smooth with small spines located at the limit of all segments. The cephalic region has a group of robust spines with one or two tips. The puparia are very similar to third instar larvae, except for the cephalic structures that are retracted. The integument shows the wrinkles from the third instar larvae and posterior spiracle disc with three spiracular openings localized on the top of an elevation. In conclusion, SEM provides some characteristics to distinguish among Chrysomya species that could help entomologists to identify immature found on corpses. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.

Description: Egg morphology of nine species of the cluster fly genus Pollenia Robineau-Desvoidy was studied with scanning electron microscopy. Funnel-shaped micropylar area, chorion covered with centrally depressed hexagons, well developed hatching pleats on the dorsal surface encompassing the median area were found in eggs of all species: P. amentaria (Scopoli), P. angustigena Wainwright, P. atramentaria (Meigen), P. labialis Robineau-Desvoidy, P. mayeri Jacentkovsky, P. pediculata Macquart, P. rudis (Fabricius), P. similis (Jacentkovsky), and P. vagabunda (Meigen). Untypical for Calliphoridae, a dual morphological structure of plastron-bearing area is described here for the first time. On the basis of our observations, all species studied share the presence of a plastron respiratory function on the entire surface between the hatching lines. Differences between species were found in the shape of the longitudinal hatching pleats and the proportion between ‘island pattern’ and ‘hexagonal pattern’ of their chorionic surface, and in the intensity of perforation of hexagons of the median area. Comparisons of Pollenia egg morphology with that of other representatives of Calliphoridae revealed its unique structure, allowing easy differentiation from other representatives of the family. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.

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.

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.

Description: Background : Diabetes mellitus results in many complications, also compromising the salivary glands. The current treatment for this condition should be a substituting method to exogenous insulin. In this aspect, the immunotherapy has been tested, but, it can be inefficient as an agent for the control of damage caused by diabetes. Thus, the aim of this study was to evaluate the anti-CD3 monoclonal antibody as alternative immunotherapy in the recovery of salivary glands of spontaneously diabetic NOD (nonobese diabetic) mice. Methods : NOD mice were divided into two groups of 10 animals: group I (untreated diabetic mice) and group II (anti-CD3-treated diabetic mice). After treatment, the samples of salivary glands were collected for histological examination under both transmitted and polarized light microscopy. Results : Alterations in tissue architecture; increase in extracellular matrix and presence of inflammatory process were observed in untreated animals. Recovery of the salivary acinar cells occurred in treated animals. The parotid glands demonstrated a smaller amount of collagen fibers and were not observed severe inflammatory processes. Conclusion : These results indicate that immunotherapy contributed to reestablishment of tissue damaged by the hyperglycemic condition, demonstrating that the immunomodulation plays an important role in the recovery of salivary glands. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: Despite the large number of species, their wide distribution, and unique reproductive characteristics, Neotropical bats have been poorly studied, and important aspects of the reproduction of these animals have not been elucidated. We made an ultrastructural analysis of spermatogenesis in Molossus molossus (Molossidae). The process of spermatogonial differentiation is similar to that found in other bats and is also relatively similar to that of Primates, with three main spermatogonia types: A d , A p , and B. Meiotic divisions proceed similarly to those of most mammals, and spermiogenesis is clearly divided into 12 steps, in the middle of the range known for bats (9–16 steps). Formation of the acrosome is similar to that known from other mammals; however, the ultrastructure of spermatozoa was found to have unique characteristics, including many wavy acrosomal projections on its surface, which seems to be specific for the family Molossidae. Comparing the ultrastructure of the spermatozoon of M. molossus with other bats already study, we observed that three characters vary: morphology of the outer dense fibers, of the perforatorium, and of the spermatozoon head. The great similarity of morphological characters between M. molossus and Platyrrhinus lineatus suggests that M. molossus is more closely related to the Phyllostomidae than to the Rhinolophidae and the Vespertilionidae. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.

Description: Although the Geographical Information System (GIS), which integrates computerized drawing computer assisted design (CAD) and relational databases (data base management system (DBMS)), is best known for applications in geographical and planning cartography, it can also use many kinds of information concerning the territory. A multidisciplinary project was initiated since 5 years a multidisciplinary study was initiated to use GIS to integrate environmental and ecological data with findings on animal health, ethology, and anatomy. This study is chiefly aimed at comparing two different methods for measuring the absorptive surface of rumen papillae. To this scope, 21 female sheep ( Ovis aries ) on different alimentary regimes (e.g., milk and forage mixed diet, early herbaceous diet, dry hay diet, and fresh hay diet at the maximum of pasture flowering and at the maximum of pasture dryness) were used; after slaughtering, 20 papillae were randomly removed from each sample collected from four indicator regions of rumen wall, placed near a metric reference and digitally photographed. The images were developed with the ArcGIS™ software to calculate the area of rumen papillae by means of GIS and to measure their mid-level width and length to calculate the papillae area as previously performed with a different method. Spatial measurements were analyzed using univariate and multivariate methods. This work demonstrates that the GIS methodology can be efficiently used for measuring the absorptive surface of rumen papillae. In addition, GIS demonstrated to be a rapid, precise, and objective tool when compared with previously used method. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.

Description: To help understand the mechanism of host discrimination during oviposition of a parasitoid wasp, Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae), the ultrastructure and morphology of the sense organs on the dorsal and ventral stylets of the ovipositor were examined using scanning and transmission electron microscopy. Ovipositor stylets were scanned for sense organs, because only unsheathed ovipositors are used for probing, stabbing, and host discrimination. Five types of sensilla and secretary pores were identified. Based on their pattern of innervation, the surface-dome and pitted-dome sensilla were thought to have a gustatory function and, thus, are the best candidates for host-discrimination. The arrangements of dendrites of type I and type II campaniform sensilla suggested that these acted as pressure- and mechano-receptors, respectively. Coeloconic sensilla might act as thermo-hygroreceptors, whilst secretary pores might provide lubrication during the process of stabbing for oviposition. Surface-dome sensilla were the most numerous type found followed by secretary pores, pitted-dome sensilla, and coeloconic sensilla. Almost all the sensilla and secretary pores were more numerous on the dorsal than on the ventral stylets, and the maximum number of these sensilla and secretary pores occurred on the distal end of the ovipositor stylets and decreased toward the proximal end. The surface-dome sensilla and secretary pores occurred in alternate rows while the pitted-dome sensilla were scattered all over the surface of the ovipositor stylets, especially on the sides of the stylets. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.

Description: The speed and accuracy of Förster resonance energy transfer (FRET) measurements can be improved by rapidly alternating excitation wavelengths between the donor and acceptor fluorophore. We demonstrate FRET efficiency measurements based on a fiber laser and photonic crystal fiber as the source for two-photon excitation (TPE). This system offers the potential for rapid wavelength switching with the benefits of axial optical sectioning and improved penetration depth provided by TPE. Correction of FRET signals for cross excitation and cross emission was achieved by switching the excitation wavelength with an electrically controlled modulator. Measurement speed was primarily limited by integration times required to measure fluorescence. Using this system, we measured the FRET efficiency of calmodulin labeled with Alexa Fluor 488 and Texas Red dyes. In addition, we measured two-photon induced FRET in an E 0 GFP-mCherry protein construct. Results from one-photon and two-photon excitation are compared to validate the rapid wavelength switched two-photon measurements. Microsc. Res. Tech., 2011. © 2011 Wiley Periodicals, Inc.

Description: The purpose of this study was to compare the inorganic content and morphology of one nanofilled and one nanohybrid composite with one universal microhybrid composite. The Vickers hardness, degree of conversion and scanning electron microscope of the materials light-cured using LED unit were also investigated. One nanofilled (Filtek™ Supreme XT), one nanohybrid (TPH® 3 ) and one universal microhybrid (Filtek™ Z-250) composite resins at color A 2 were used in this study. The samples were made in a metallic mould (4 mm in diameter and 2 mm in thickness). Their filler weight content was measured by thermogravimetric analysis (TG). The morphology of the filler particles was determined using scanning electron microscope equipped with a field emission gun (SEM-FEG). Vickers hardness and degree of conversion using FT-IR spectroscopy were measured. Filtek™ Z-250 (microhybrid) composite resin shows higher degree of conversion and hardness than those of Filtek™ Supreme XT (nanofilled) and TPH® 3 (nanohybrid) composites, respectively. The TPH 3 ® (nanohybrid) composite exhibits by far the lowest mechanical property. Nanofilled composite resins show mechanical properties at least as good as those of universal hybrids and could thus be used for the same clinical indications as well as for anterior restorations due to their high aesthetic properties. Microsc. Res. Tech., 2011. © 2011 Wiley Periodicals, Inc

Description: OBJECTIVE: Signal pattern enumeration of Urovysion Fluorescence in Situ Hybridization test is tedious and requires great experience. Our aim was to eliminate human interaction by automating the process, using an adoptable, automated image acquisition, and analysis system. METHODS: For extensive analytical analysis control, cell populations were used, while preliminary clinical study was performed on 21 patients with clinical suspicion for bladder cancer. All investigations were carried out using an automated user-trainable workstation (Metafer4-Metacyte). RESULTS: The system identified nuclei with a specificity and sensitivity of 92.7 and 96.6%, respectively, while signal detection accuracy was 81.1% on average. Both analytical and diagnostic accuracy of automated analysis was comparable to manual approach (94.8 and 71% vs. 97.9 and 76%, respectively), but classification accuracy increased with degree of polysomy, thus diagnostic sensitivity in low grade, low stage cases was poor. CONCLUSION: It is possible to automate signal enumeration of Urovysion using a user-trainable system, and achieve efficiency comparable to manual analysis. Previously introduced automated immunophenotypic targeting should further increase diagnostic sensitivity, while resulting in a comprehensively automated method. However, the problem of reduced detection accuracy in cases featured with low polysomy is likely to remain a great challenge of automated signal enumeration. Microsc. Res. Tech. 2011. © 2011 Wiley Periodicals, Inc.

Description: This study aimed to evaluate the effect of four chemomechanical surface treatments on the surface average microroughness and profile of laser-sintered and vacuum-cast dental prosthetic structures. Square-shaped blocks (10 mm × 10 mm × 1.5 mm) were prepared as follows: (1) laser-sintered Co Cr (L) (ST2724G); (2) cast Co Cr (C) (Gemium-cn); and (3) cast Ni Cr Ti (T) (Tilite). Specimens of each alloy group were randomly divided into five subgroups ( n = 10 each), depending on the conditioning method used: (1) no treatment (control); (2) sandblasting (125 μm Al 2 O 3 -particles); (3) silica coating (50 μm silica-modified Al 2 O 3 -particles); (4) oxidation; and (5) oxidation plus opacification. Subgroups 2 and 3 represent “inner” pretreatments proposed for ceramometal restorations to improve the metal surface area available for luting cements. Subgroups 4 and 5 are the “outer” pretreatments required for bonding the aesthetic veneering ceramics to the underlying metal frameworks. Average surface roughness (Ra/μm) was determined using a surface profilometer. Data were analyzed by two-way ANOVA and Student–Newman–Keuls tests (α = 0.05). Metal surface topography was SEM-analyzed. Despite the inner pretreatment applied, L samples resulted in the highest microroughness ( P 〈 0.001), whereas sandblasting produced a surface-smoothing effect in cast specimens. After oxidation, a significant increase in surface roughness occurred in all groups compared with controls, L specimens being the roughest ( P 〈 0.001). Opacification caused a flattening effect of all oxidized structures; all opacified groups resulting in similar microroughness. Laser sintering of Co Cr enhances the roughness of metal structures, which may improve the frameworks' microretention of the cements, and of the opaquer before the copings are veneered with the aesthetic ceramics. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: We have conducted a morphological study of the ampullae of Lorenzini on two shark species from Squatina Genus. In both species, S. guggenheim and S. occulta , the ampullae were observed like small pores scattered in the head region similar to other species of the Chondrichthyes Class. However, differently of the other species a greatest density of ampullae of Lorenzini was observed along of the body surface. After fixation using 10% formaldehyde, the ampullae were removed and processed for light and scanning electron microscopy. Macroscopically, the two shark species differed by the presence of dorsal spines that appeared from the head to the first dorsal fin in S. guggenheim and were absent in S. occulta . Microscopically, there were no differences between the ampullae of Lorenzini channels in these two species. The wall of the ampulla was formed by a simple squamous epithelium. Bands of connective tissue, hyaline cartilage and collagen fibers were found between the ampulla and the skeletal striated muscle layer. Nerve branches responsible for conducting signal pulses to the central nervous system were visible between the muscle and connective tissue layers. Using scanning electron microscopy and histological analysis, we found that the channels were twisted and positioned parallel to the skin. The inside of the channels contained a large amount of a gelatinous secretion composed by polysaccharides. Therefore, we conclude that the morphological combination of extended distribution of the ampullae of Lorenzini and the body shape may represent an adaptation of these species to their way of life. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: The low-shrink Silorane-based composite could bond effectively to bone and showed the potential be used as a bone cement. Bone organ culture maintains the anatomical order, natural cell-to-cell and cell-to-matrix relationship. The purpose of this study was to evaluate the responses of bone cells to a Silorane-based composite which was compared with a representative polymethyl methacrylate (PMMA) bone cement. The critical size defects were created through the parietal bones from one litter of mice. The paired bones were divided into two groups: Silorane-based composite group and PMMA group. The prepared two groups of disks were put into the defects. The cultures were grown in vitro for 38 days and analyzed with microcomputed-tomography, dissecting-microscope, phase- contrast-microscope, scanning-electron-microscopy, and energy- dispersive-X-ray. At the 10th day, the Silorane disk was almost fully covered by a sheet of cells but the cells hardly attached to the disk surface. The edge of the PMMA disk was covered by a sheet of cells and the migrated individual cells attached to the whole surface of the disk. At the 38th day, some cells attached to the exposed disk area of the Silorane disk while the formed tissues covered the whole surface of the PMMA disk. The collagen fibers, globular deposits and bone formation were visible in both groups. The Silorane-based composite showed promise as a potential bone cement when compared with PMMA which is used in clinical orthopedics. However, the cell attachment to PMMA was evidently better than to Silorane-based composite. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: Stroke is associated with the intracerebral formation of fibrin clots which may lead to irreversible brain damage. Thrombolytic therapies employ a variety of natural and/or recombinant plasminogen activators to initiate fibrinolytic degradation of cerebral thrombi. However, such therapies when installed beyond 4- to 6-h window, may fail to achieve the expected outcome. This is due to the hydroxyl radical-induced modification of fibrin(ogen) molecules rendering them refractory to fibrinolytic degradation, but no cause of the increased free radical generation in stroke was offered. Here, we show by means of electron microscopy that iron ions added to human blood dramatically enhances fibrin fibers formation with thrombin, and significantly delays fibrinolysis during spontaneous clotting of native blood. Iron ions caused the appearance dense matted fibrin deposits, similar, if not identical, to those observed in plasma of patients with stroke. These results may explain a known relationship between thrombotic diseases and the increased body concentrations of free iron and/or hemoglobin derivatives. We conclude that any action resulting in the inhibition of hemostatic abnormalities, as well as in the reduction of body free iron and scavenging of hydroxyl radicals (e.g., by polyphenols) can potentially prevent pathological reactions associated with consequences of stroke. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.

Description: Anastatus japonicus Ashmead (Hymenoptera: Eupelmidae) is an important egg parasitoid of several major insect pests. To better understand its host finding mechanisms, the antennal sensilla of female wasp were investigated by scanning electron microscopy. Sensilla chaetica were found mainly on radicle and pedicel segments of the antennae. i-Type sensilla, s. campaniformia , and corneous sensilla were detected on the leeward side, while s. coeloconica and lance sensilla were presented on the windward side of the antennae. S. trichodea and s. basiconica were more abundant on the leeward side than on the windward side of the antennae. More s. placodea were found on the windward side than on the leeward side of the right antenna, while the opposite results were observed on the left antenna. Overall, more s. placodea were found on the right antenna than that on the left antenna. The numbers of s. trichodea and s. basiconica on the clava or the third flagellum antennomere of the right antenna were more than those of the left antenna, whereas their distribution patterns on the other corresponding antennomeres were reverse. Our results showed that there is a strong asymmetrical antennal sensilla distribution quantitatively and spatially between the left and right antennae. Placoid sensilla are present more on the right antenna than on the left antenna. S. campaniformia , corneous sensilla, and i-type sensilla were found only on the leeward side of the antennal clava, while their external morphology and potential functions were described and discussed in detail for the first time. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.

Description: This research aimed to examine the morphological features of the tongue and laryngeal entrance of long-legged buzzard by macroscopic and scanning electron microscopic methods. Two adult buzzards were used as material. The tongue was fairly elongated and terminated in oval shovel-like apex. Scale-like projections were localized on the apex and body of tongue. Both lateral sides of lingual body were contained considerably long thread-like projections. Many orifices of lingual posterior salivary glands were discerned among scale-like projections in median sulcus of the lingual body. Papillary crest of sharp conical papilla were observed on the between the body and root of the tongue. No conical papillae or other projections were existent on the root of the tongue, but numerous orifices of posterior salivary gland ducts were detected. In addition, orifices of anterior salivary gland ducts were present on the dorsal and lateral surfaces of the lingual body. Numerous conical papillae were observed on the caudal region of glottis and no conical papillae or any similar projection which were bordered the glottic fissure was noted. Anatomy of these organs in the long-legged buzzard and white tailed eagle which are the member of the same family, Accipitridae, revealed very high similarity. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: Anastatus japonicus Ashmead (Hymenoptera: Eupelmidae) is an important egg parasitoid of several major insect pests. To better understand its host finding mechanisms, the antennal sensilla of female wasp were investigated by scanning electron microscopy. Sensilla chaetica were found mainly on radicle and pedicel segments of the antennae. i-Type sensilla, s. campaniformia , and corneous sensilla were detected on the leeward side, while s. coeloconica and lance sensilla were presented on the windward side of the antennae. S. trichodea and s. basiconica were more abundant on the leeward side than on the windward side of the antennae. More s. placodea were found on the windward side than on the leeward side of the right antenna, while the opposite results were observed on the left antenna. Overall, more s. placodea were found on the right antenna than that on the left antenna. The numbers of s. trichodea and s. basiconica on the clava or the third flagellum antennomere of the right antenna were more than those of the left antenna, whereas their distribution patterns on the other corresponding antennomeres were reverse. Our results showed that there is a strong asymmetrical antennal sensilla distribution quantitatively and spatially between the left and right antennae. Placoid sensilla are present more on the right antenna than on the left antenna. S. campaniformia , corneous sensilla, and i-type sensilla were found only on the leeward side of the antennal clava, while their external morphology and potential functions were described and discussed in detail for the first time. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.

Description: We have explored the effects of atmospheric environment on Kelvin force microscopy (KFM) measurements of potential difference between different regions of test polycrystalline diamond surfaces. The diamond films were deposited by microwave plasma-assisted chemical vapor deposition, which naturally produces hydrogen terminations on the surface of the films formed. Selected regions were patterned by electron-beam lithography and chemical terminations of oxygen or fluorine were created by exposure to an oxygen or fluorine plasma source. For KFM imaging, the samples were mounted in a hood with a constant flow of helium gas. Successive images were taken over a 5-h period showing the effect of the environment on KFM imaging. We conclude that the helium flow removes water molecules adsorbed on the surface of the samples, resulting in differences in surface potential between adjacent regions. The degree of water removal is different for surfaces with different terminations. The results highlight the importance of taking into account the atmospheric environment when carrying out KFM analysis. © 2012 Wiley Periodicals, Inc.

Description: To evaluate the efficacy of ProTaper Universal rotary retreatment system and the influence of sealer type on the presence of filling debris in the reinstrumented canals viewed in an operative clinical microscope. Forty-five palatal root canals of first molars were filled with gutta-percha and one of the following sealers: G1, EndoFill; G2, AH Plus; G3, Sealapex. The canals were then reinstrumented with ProTaper Universal rotary system. Roots were longitudinally sectioned and examined under an operative clinical microscope (10×), and the amount of filling debris on canal walls was analyzed using the AutoCAD 2004 software. A single operator used a specific software tool to outline the canal area and the filling debris area in each third (cervical, middle, and apical), as well as the total canal area. Data were analyzed by Kruskal-Wallis test and Tukey test at P 〈 0.05. Sealapex demonstrated significant differences in the average of filling debris area/canal among the 3 thirds. This group revealed that apical third showed more debris than the both cervical and middle third ( P 〈 0.0001). Endofill presented significantly more filling debris than Sealapex in the cervical third ( P 〈 0.05). In the middle ( P = 0.12) and apical third ( P = 0.10), there were no differences amongst groups. Debris was left in all canal thirds, regardless of the retreatment technique. The greatest differences between techniques and sealers were found in the cervical third. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.

Description: We have conducted a morphological study of the ampullae of Lorenzini on two shark species from Squatina Genus. In both species, S. guggenheim and S. occulta , the ampullae were observed like small pores scattered in the head region similar to other species of the Chondrichthyes Class. However, differently of the other species a greatest density of ampullae of Lorenzini was observed along of the body surface. After fixation using 10% formaldehyde, the ampullae were removed and processed for light and scanning electron microscopy. Macroscopically, the two shark species differed by the presence of dorsal spines that appeared from the head to the first dorsal fin in S. guggenheim and were absent in S. occulta . Microscopically, there were no differences between the ampullae of Lorenzini channels in these two species. The wall of the ampulla was formed by a simple squamous epithelium. Bands of connective tissue, hyaline cartilage and collagen fibers were found between the ampulla and the skeletal striated muscle layer. Nerve branches responsible for conducting signal pulses to the central nervous system were visible between the muscle and connective tissue layers. Using scanning electron microscopy and histological analysis, we found that the channels were twisted and positioned parallel to the skin. The inside of the channels contained a large amount of a gelatinous secretion composed by polysaccharides. Therefore, we conclude that the morphological combination of extended distribution of the ampullae of Lorenzini and the body shape may represent an adaptation of these species to their way of life. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: The low-shrink Silorane-based composite could bond effectively to bone and showed the potential be used as a bone cement. Bone organ culture maintains the anatomical order, natural cell-to-cell and cell-to-matrix relationship. The purpose of this study was to evaluate the responses of bone cells to a Silorane-based composite which was compared with a representative polymethyl methacrylate (PMMA) bone cement. The critical size defects were created through the parietal bones from one litter of mice. The paired bones were divided into two groups: Silorane-based composite group and PMMA group. The prepared two groups of disks were put into the defects. The cultures were grown in vitro for 38 days and analyzed with microcomputed-tomography, dissecting-microscope, phase- contrast-microscope, scanning-electron-microscopy, and energy- dispersive-X-ray. At the 10th day, the Silorane disk was almost fully covered by a sheet of cells but the cells hardly attached to the disk surface. The edge of the PMMA disk was covered by a sheet of cells and the migrated individual cells attached to the whole surface of the disk. At the 38th day, some cells attached to the exposed disk area of the Silorane disk while the formed tissues covered the whole surface of the PMMA disk. The collagen fibers, globular deposits and bone formation were visible in both groups. The Silorane-based composite showed promise as a potential bone cement when compared with PMMA which is used in clinical orthopedics. However, the cell attachment to PMMA was evidently better than to Silorane-based composite. Microsc. Res. Tech., 2012. © 2012 Wiley Periodicals, Inc.

Description: The aim of this study was to compare the percentage of gutta-percha (PGP) in mesial root canals of mandibular molars obturated with LC (Lateral Compaction) or SC (Single Cone) ProTaper Universal System techniques at different levels of the root. Mesial root canals of 20 human permanent molars with similar anatomical characteristics were instrumented using the ProTaper Universal rotary system technique until the F2 instrument, with 20 canals filled by SC ProTaper Universal technique and 20 canals by the LC technique. The mesial roots were sectioned transversely to 3, 5, and 7 mm from the root apex. Digital images of specimens were obtained at MIC-D digital microscope in increases of 30 to 35X. The gutta-percha area was measured using ImageTool software. Data were analyzed using two-way ANOVA and Bonferroni test (α = 0.05). The SC technique provided greater PGP than the LC technique in the apical third (3 mm) ( P 〈 0.001). In the other thirds (5 and 7 mm) there was no statistical difference between the two techniques regarding the PGP ( P 〉 0.05). There was no statistically significant difference between thirds of the root canal for both techniques ( P 〉 0.05). It was concluded that SC technique provided greater PGP than the LC technique in the apical third of mesial root canals of mandibular molars. There was no difference between the two techniques regarding the PGP in the cervical and middle thirds. Microsc. Res. Tech. 2012. © 2012 Wiley Periodicals, Inc.