A microstructural study of gallbladder stones using scanning electron microscopy

Objective: The objective of this study was to describe the microstructure of different types of gallbladder stones to better understand the basis of gallbladder stone formation. Methods: Gallbladder stones from 387 patients with cholecystolithiasis were first analyzed by Fourier transform Infrared spectroscopy to identify the type of the gallbladder stone, and they were then examined using scanning electron microscopy to define their microstructure. Results: Cholesterol stones were mainly composed of plate‐like or lamellar cholesterol crystals stacked tightly in a radial, cord‐like, or irregular staggered arrangement. A small number of bilirubinate particles were seen occasionally. Pigment stones were mainly composed of loosely arranged bilirubinate particles with different shapes (sphere‐like, clumping‐like, or amorphous). Calcium carbonate stones were composed of calcium carbonate crystals having many shapes (bulbiform, ellipsoid, fagot‐shaped, fusiform, hawthorn‐shaped, cuboid, button‐shaped, lamellar, broken firewood‐shaped, rod‐shaped, acicular, or crushed sugar cane‐shaped). Bulbiform shaped crystals were the most common. Phosphate stones were mainly composed of different sized echin‐sphere‐like or rough bulbiform crystals. Bilirubinate particles were always adherent. Calcium stearate stones usually had a network structure with adherent bilirubinate particles. Protein stones usually had a honeycomb or chrysanthemum petal‐like structure. Cystine stones were composed of hexagonal cystine crystals, some of which had prominent edges. Mixed stones presented different shapes according to their mixed components. Conclusion: Different types of gallbladder stones had characteristic shapes and elements. This study provides an objective basis for further research regarding gallbladder stone formation. Microsc. Res. Tech. 76:443–452, 2013. © 2013 Wiley Periodicals, Inc.     

A comparative study on the self‐assembly of an amyloid‐like peptide at water–solid interfaces and in bulk solutions

In the past years the self‐assembly of amyloid‐like peptides has attracted increasing attentions, because it is highly related to neurodegenerative diseases and has a potential for serving as nanomaterial to fabricate novel and useful nanostructures. In this paper, we focused on the role of interfacial conditions in the self‐assembly of an amyloid‐like peptide, termed Pep11. It was found that, when dissolved in bulk solutions, Pep11 formed into β‐sheet structures and assembled into long filaments in several hours, as revealed by Thioflavin T fluorescence and transmission electron microscopy (TEM) morphology characterization, respectively. When the peptide solution was added onto a mica/HOPG substrate, peptide filaments with three preferred orientations with an angle of 60° to each other were formed immediately, as imaged in situ by atomic force microscopy (AFM). However, the kinetics in filament formation and the morphologies of the formed beta sheet either on HOPG and mica or in bulk solutions were quite different. These results indicate that the interfacial properties dramatically affect the peptide self‐assembly process. Microsc. Res. Tech. 78:375–381, 2015. © 2015 Wiley Periodicals, Inc.     

Quantitative evaluation of bone resorption activity of osteoclast‐like cells by measuring calcium phosphate resorbing area using incubator‐facilitated and video‐enhanced microscopy

Quantitative evaluation of the ability of bone resorption activity in live osteoclast‐like cells (OCLs) has not yet been reported on. In this study, we observed the sequential morphological change of OCLs and measured the resorbing calcium phosphate (CP) area made by OCLs alone and with the addition of elcatonin utilizing incubator facilitated video‐enhanced microscopy. OCLs, which were obtained from a coculture of ddy‐mouse osteoblastic cells and bone marrow cells, were cultured on CP‐coated quartz cover slips. The CP‐free area increased constantly in the OCLs alone, whereas it did not increase after the addition of elcatonin. This study showed that analysis of the resorbed areas under the OCL body using this method enables the sequential quantitative evaluation of the bone resorption activity and the effect of several therapeutic agents on bone resorption in vitro. Microsc. Res. Tech, 2009. © 2008 Wiley‐Liss, Inc.     

A new method of scanning tunneling spectroscopy for study of the energy structure of semiconductors and free electron gas in metals

In this study, anew method for the scanning tunneling spectroscopy (STS) of direct and indirect gap semiconductors and free electron gas in metals is proposed. Band structures of Si, porous Si, and Ge were studied. The tunneling current-voltage characteristics of Si and porous Si surfaces were measured over different voltage intervals from tens of mV to 20 V under incident light from an Xe lamp and those of a Ge surface in the dark. The correlation between the shapes of the I-V curves and band structure of the materials was calculated. It was found that the curves are linear if measured in the voltage range V₀ E_g/(2e) and nonlinear when V₀ α E_g/(2e) (in the measurements the applied voltage was changed from -V₀ to V₀). The method was used for the observation of a new effect of tunneling of free electron gas having thermal energies from a metal tip to a band gap state of the semiconductor. The energy spectrum of free electron gas was measured.     

Angioarchitecture of the terminal vascular system in inflammatory bowel disease—A comparative corrosion-anatomic study

The vascular origin of Crohn's disease (CD) and ulcerative colitis has been discussed but is poorly understood and investigated. A comparison of both diseases on the basis of the angioarchitecture had yet to be performed to investigate vascular morphology further and to evaluate its relevance for the etiology and differential diagnosis. The vascular system of 29 human colonic specimens of patients with CD (n=10), ulcerative colitis (n=11), and colon cancer (n=8), which were taken as controls, was perfused blood-free immediately after resection. Corrosion casts were obtained by injection of Mercox Cl and consecutive digestion with potassium hydroxide. Casts were then cut into sections, sputter coated with gold, and examined in a blinded fashion by scanning electron microscopy. Normal intestine showed a typical honeycomb appearance, the mucosal capillary network was smooth and regular, and the subepithelial vascular plexus was supplied by regularly arranged arterioles. In CD, a partially intact capillary net with alterations, dominant collecting veins, and a characteristic pattern of diameter changes with narrow segments in the capillary bed was seen. In ulcerative colitis a disruption of the colonic vascular architecture was found. The capillarization was rarefied and irregular lumps of subepithelial capillary structures were observed. The morphology of the terminal vascular bed in inflammatory bowel disease in contrast to normal controls is uniquely described. The technique of injecting fresh, human specimens was applied for the first time. The homogeneity of vascular changes within each entity seems to allow a differentiation between CD and ulcerative colitis.     

Three‐dimensional morphology of cerebellar climbing fibers. A study by means of confocal laser scanning microscopy and scanning electron microscopy

The intracortical pathway of cerebellar climbing fibers have been traced by means of scanning electron microscpy (SEM) and confocal laser scanning microscopy (CLSM) to study the degree of lateral collateralization of these fibers in the granular Purkinje cell and molecular layers. Samples of teleost fish were processed for conventional and freeze‐fracture SEM. Samples of hamster cerebellum were examined by means of CLSM using FM4–64 as an intracellular stain. High resolution in lens SEM of primate cerebellar cortex was carried out using chromium coating. At scanning electron and confocal laser microscopy levels, the climbing fibers appeared at the white matter and granular layer as fine fibers with a typical arborescence or crossing‐over branching pattern, whereas the mossy fibers exhibited a characteristic dichotomous bifurcation. At the granular layer, the parent climbing fibers and their tendrils collaterals appeared to be surrounding granule and Golgi cells. At the interface between granule and Purkinje cell layers, the climbing fibers were observed giving off three types of collateral processes: those remaining in the granular layer, others approaching the Purkinje cell bodies, and a third type ascending directly to the molecular layer. At this layer, retrograde collaterals were seen descending to the granular layer. By field emission high‐resolution SEM of primate cerebellar cortex, the climbing fiber terminal collaterals were appreciated ending by means of round synaptic knobs upon the spines of secondary and tertiary Purkinje cell dendrites.     

Effect of in‐office bleaching with 35% hydrogen peroxide with and without addition of calcium on the enamel surface

This study aimed to evaluate effectiveness and effects of bleaching with 35% hydrogen peroxide with and without calcium on color, micromorphology, and the replacement of calcium and phosphate on the enamel surface. Thirty bovine enamel blocks (5.0 × 5.0 mm) were placed into the following groups: G1: artificial saliva (control); G2: 35% hydrogen peroxide gel without calcium (Whiteness HP Maxx–FGM); and G3: 35% hydrogen peroxide gel with calcium (Whiteness HP Blue–FGM). Three color measurements were performed with a spectrophotometer: untreated (baseline), after performing staining, and after application of bleaching agents. Calcium deposition on the enamel was evaluated before and after the application of bleaching agents using energy‐dispersive X‐ray spectrometry. The enamel surface micromorphology was observed under scanning electron microscopy. The pH of each product was measured. The data were subjected to one‐factor analysis of variance (ANOVA), and any differences were analyzed using Tukey's test (P < 0.05). G3 showed greater variation in total color after the experiment than G2 and G1; there was no significant difference in calcium or phosphorus concentration before and after the experimental procedures; morphological changes were observed only in G2 and G3; and the pH values of the Whiteness HP Maxx and Whiteness HP Blue bleaching agents were 5.77 and 7.79, respectively. The 35% hydrogen peroxide with calcium showed greater bleaching potential, but the addition of calcium had no effect in terms of reducing morphological changes or increasing the calcium concentration on the enamel surface. Microsc. Res. Tech. 78:975–981, 2015. © 2015 Wiley Periodicals, Inc.     

Independent trafficking of flavocytochrome b558 and myeloperoxidase to phagosomes during phagocytosis visualised by energy‐filtering and energy‐dispersive spectroscopy‐scanning transmission electron microscopy

When polymorphonuclear leukocytes (PMNs) phagocytose opsonised zymosan particles (OPZ), free radicals and reactive oxygen species (ROS) are formed in the phagosomes. ROS production is mediated by NADPH oxidase (Nox), which transfers electrons in converting oxygen to superoxide (O₂^?). Nox‐generated O₂^? is rapidly converted to other ROS. Free radical‐forming secretory vesicles containing the Nox redox center flavocytochrome b558, a membrane protein, and azurophil granules with packaged myeloperoxidase (MPO) have been described. Presuming the probable fusion of these vesicular and granular organelles with phagosomes, the translation process of the enzymes was investigated using energy‐filtering and energy‐dispersive spectroscopy‐scanning transmission electron microscopy. In this work, the primary method for imaging cerium (Ce) ions demonstrated the localisation of H₂O₂ generated by phagocytosing PMNs. The MPO activity of the same PMNs was continuously monitored using 0.1% 3,3′‐diaminobenzidine‐tetrahydrochloride (DAB) and 0.01% H₂O₂. A detailed view of these vesicular and granular structures was created by overlaying each electron micrograph with pseudocolors: blue for Ce and green for nitrogen (N).     

Tannase‐mediated biotransformation assisted separation and purification of theaflavin and epigallocatechin by high speed counter current chromatography and preparative high performance liquid chromatography: A comparative study

A large scale isolation and purification of theaflavin (TF) and epigallocatechin (EGC) has been successfully developed by tannase‐mediated biotransformation combining high‐speed countercurrent chromatography. After tannase hydrolysis of a commercially available theaflavins extract (TE), the content of TF and EGC in tannase‐mediated biotransformation product (TBP) achieved approximately 3 times enrichment. SEM studies revealed smooth tannase biotransformation and the possibility of recovery of the tannase. A single 1.5 hours' HSCCC separation for TF and EGC employing a two‐phase solvent system could simultaneously produce 180.8 mg of 97.3% purity TF and 87.5 mg of 97.3% purity EGC. However, a preparative HPLC separation of maximum injection volume containing 120 mg TBP prepared 11.2 mg TF of 94.9% purity and 7.7 mg EGC of 89.9% purity. HSCCC separation demonstrated significant advantages over Prep HPLC in terms of sample loading size, separation time, environmental friendly solvent systems, and the production.     

Adverse effects of respiratory disease medicaments and tooth brushing on teeth: A scanning electron microscopy,X‐ray fluorescence and infrared spectroscopy study

The present study aims to evaluate the effect of brushing with fluoride dentifrice on teeth severely affected by erosion due to respiratory medicaments. Enamel (n = 50) and dentin (n = 50) bovine specimens were prepared and treated with artificial saliva (S‐control), acebrofilin hydrochloride (AC), ambroxol hydrochloride (AM), bromhexine hydrochloride (BR), and salbutamol sulfate (SS) and subjected to cycles of demineralization (immersing in 3 mL, 1 min, three times a day at intervals of 1 hr, for 5 days) followed by remineralization (saliva, 37°C, 1 hr). Simulated brushing with fluoridated toothpaste was performed using 810 strokes in a reciprocal‐action brushing simulator. Scanning electron microscopy, micro energy dispersive X‐ray fluorescence (μ‐EDXRF) spectroscopy and attenuated total reflection Fourier transform infrared (ATR FTIR) spectroscopy were then performed. μ‐EDXRF images showed extensive erosion after treatment with all medicaments. SEM images showed enamel erosion in order SS > BR > AC = AM > S after brushing and fluoridation. FTIR results were in agreement. In case of dentin, μ‐EDXRF measurements showed significant difference in mineral content (percent weight of calcium and phosphate) in SS + brushing + fluoridation treated enamel compared to control, while μ‐EDXRF images showed erosive effects in the order SS > AM>BR > AC = S post brushing + fluoridation. SEM images showed erosion in the order SS > AM = BR > AC > S post brushing + fluoridation. Again, FTIR multivariate results were in agreement. Overall, our study shows that proper oral care is critical when taking certain medication. The study also demonstrates the possible use of FTIR for rapid clinical monitoring of tooth erosion in clinics.     

A comparative study of thin coatings of Au/Pd, Pt and Cr produced by magnetron sputtering for FE-SEM

Visualization of structural details of specimens in field emission scanning electron microscopy (FE-SEM) requires optimal conductivity. This paper reports on the differences in conductive layers of Au/Pd, Pt and Cr, with a thickness of 1.5–3.0 nm, deposited by planar magnetron sputtering devices. The coating units were used under standard conditions for source–substrate distance, current, HT and argon pressure. Carbon films, deposited by high-vacuum evaporation on small, freshly cleaved pieces of mica, were used as substrate and mounted on copper grids for TEM and SEM inspection. Au/Pd, Pt and, to a lesser extent, Cr coatings varied in particle density, size and shape. Au/Pd coatings have a slightly more granular appearance than Cr and Pt coatings, but this is strongly dependent on the type of sputtering device employed. In FE-SEM images there is almost no difference in contrast and particle size between the Au/Pd layer and the Pt layers of a similar thickness. The nuclei of Au/Pd are rather small with almost no growth to the sides or in height, making Au/Pd coatings a good alternative to chromium and platinum for FE-SEM of biological tissues because of its higher yield of secondary electrons.     

Chemical and morphological evaluation of enamel and dentin near cavities restored with conventional and zirconia modified glass ionomer subjected to erosion‐abrasion

Microenergy dispersive X‐ray fluorescence (μ‐EDXRF) spectroscopy and scanning electron microscopy (SEM) were used to test the hypothesis that zirconia modified glass ionomer cement (GIC) could improve resistance to erosion‐abrasion to a greater extent than conventional cement. Bovine enamel (n = 40) and dentin (n = 40) samples were prepared with cavities, filled with one of the two restorative materials (GIC: glass‐ionomer cement or ZrGIC: zirconia‐modified GIC). Furthermore, the samples were treated with abrasion‐saliva (AS) or abrasion‐erosion cycles (AE). Erosive cycles (immersion in orange juice, three times/day for a duration of 1 min over a 5 day period) and/or abrasive challenges (electric toothbrush, three times/day for a duration of 1 min over a 5 day period) were performed. Positive mineral variation (MV%) on the enamel after erosion‐abrasion was observed for both materials (p < 0.05), whereas a negative MV% on the dentin was observed for both materials and treatments (p < 0.05). The SEM images showed clear enamel loss after erosion‐abrasion treatment and material degradation was greater in GIC_AE compared to those of the other groups. Toothbrush abrasion showed a synergistic effect with erosion on substance loss of bovine enamel, dentin, GIC, and ZrGIC restorations. Zirconia addition to the GIC powder improved the resistance to abrasive‐erosive processes. The ZrGIC materials may find application as a restorative material due to improved resistance as well as in temporary restorations and fissure sealants.     

Influence of Trigonella foenum graecum seed extract as root canal irrigation agent on root surfaces: A scanning electron microscope/energy dispersive X‐ray analysis study

The purpose of this in vitro study was to investigate the effect of Trigonella foenum graecum seed extract (TFGSE) as a root canal irrigation agent on the surface of instrumented root canals and on changes in the mineral contents of the root dentin. A total of 90 extracted human mature maxillary central incisor teeth with a single root and without root resorption were selected. The specimens were subdivided into five groups according to irrigation protocol groups G0: distilled water (n = 15): G1: Sodium hypochlorite (5.25% NaOCl); G2: 1% TFGSE G3: NaOCl +17% EDTA (3 min); G4: NaOCl+1% TFGSE; G5: 1% TFGSE +17% EDTA. All the roots were sectioned longitudinally into two halves and examined under scanning electron microscope/energy dispersive X‐ray analysis (SEM/EDX) to assess changes in the mineral content of the tooth. Smear layer removal from root canals was evaluated with scanning electron microscopy (SEM). A statistical analysis was performed using the Kruskal–Wallis test. The most effective irrigation protocols in removing smear layer occurred with TFGSE and NaOCl, both with rinsing with 17% EDTA. TFGSE was more effective than NaOCl for removing the smear layer. The results showed the ability of 1% TFGSE to remove the smear layer from the root canals. TFGSE is a promising alternative irrigation agent for root canals.     

A comparison study of detecting gold nanorods in living cells with confocal reflectance microscopy and two‐photon fluorescence microscopy

Two‐photon fluorescence microscopy and confocal reflectance microscopy were compared to detect intracellular gold nanorods in rat basophilic leukaemia cells. The two‐photon photoluminescence images of gold nanorods were acquired by an 800 nm fs laser with the power of milliwatts. The advantages of the obtained two‐photon photoluminescence images are high spatial resolution and reduced background. However, a remarkable photothermal effect on cells was seen after 30 times continuous scanning of the femto‐second laser, potentially affecting the subcellular localization pattern of the nanorods. In the case of confocal reflectance microscopy the images of gold nanorods can be obtained with the power of light source as low as microwatts, thus avoiding the photothermal effect, but the resolution of such images is reduced. We have noted that confocal reflectance images of cellular gold nanorods achieved with 50 μW 800 nm fs have a relatively poor resolution, whereas the 50 μW 488 nm CW laser can acquire reasonably satisfactory 3D reflectance images with improved resolution because of its shorter wavelength. Therefore, confocal reflectance microscopy may also be a suitable means to image intracellular gold nanorods with the advantage of reduced photothermal effect.     

Real color cathodoluminescence scanning electron microscopy—A new effective method for study of SiC materials and devices

Color cathodoluminescence scanning electron microscopy (CCL-SEM) studies prove the correctness of available information concerning space distribution of luminescence centers in SiC layers. Established facts may be applied for investigation of the polytypism, space distribution of impurities, and native defects induced during growth by high-energy particles radiation or by diffusion, as well as by the mechanisms of crystal formation. This paper presents a short survey of diagnostic availability of the CCL-SEM technique for inspection of the SiC materials and devices.     

A comparative study of phenol‐type antioxidants in methyl oleate with quantum calculations and experiments

Chemical software (Gaussian 98w) was used to calculate the highest occupied molecular orbital (HOMO) energy level of three phenol‐type antioxidants to evaluate their oxidation resistance performance. The conclusion drawn from the quantum calculations was that the decreasing order of the oxidation resistance effectiveness was 4,4‐methylene‐di(2,6‐di‐t‐butyl‐phenol) (T511)>2,6‐di‐t‐butyl‐4‐methylphenol (BHT)>phenol. Pressure differential scanning calorimetry (PDSC) and the rotary bomb oxidation test were also used to assess the antioxidants in a methyl oleate base oil. According to the experimental results, the oxidation resistance capability of the three antioxidants decreased in the order BHT > T511 > phenol. The results indicated that the quantum calculation method could be used to screen antioxidants with regard to their oxidation resistance effectiveness.