Observations on calcareous corpuscles using a scanning electron microscope.
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X-ray microanalysis of non-biological and biological specimens was carried out in an environmental scanning electron microscope (ESEM) over a range of atmospheric conditions. Introduction of water vapour into the specimen chamber lead to direct X-ray contribution from oxygen atoms, an increase in extraneous background (causing reduced P/B ratios of other elements), X-ray absorption (also reducing P/B ratios) and broadening (skirting) of the electron beam. Similar results were obtained after introduction of an argon atmosphere. These effects were reduced under conditions of minimal chamber atmospheric pressure and maximal accelerating voltage. Because of beam skirting, quantitative X-ray microanalysis of biological specimens in a water vapour atmosphere was only valid where the sample was spread over a wide area (leading to mean elemental values for the whole preparation). Unless appropriate correction factors or changes in instrumentation can be implemented, quantitative analysis of wet specimens in ESEM cannot be applied to discrete specimens or to limited areas within a mixed sample.
OBJECTIVE: To elucidate the ultrastructural details of calcium oxalate-containing urinary calculi from dogs. Sample Population-38 specimens selected from a collection of 8,297 oxalate-containing urinary calculi from dogs: 22 specimens composed of calcium oxalate (calcium oxalate monohydrate [COM], calcium oxalate dihydrate [COD], or COM and COD) and 16 specimens composed of calcium oxalate with amorphous calcium phosphate. PROCEDURE: Analyses of specimens included use of plain, reflected, and polarized light microscopy, X-ray diffractometry, scanning electron microscopy (SEM) with backscattered electron (BSE) imagery, and electron microprobe analysis. RESULTS: Four texture types were observed in calcium oxalate calculi; 4 texture types of calcium oxalate-calcium phosphate-mixed calculi were recognized. Texture types were delineated through differences in calcium oxalate crystal sizes, which were affected by urine supersaturation and abundance of crystal nucleation sites. Segregation of calcium oxalate from calcium phosphate indicated they do not precipitate under the same conditions. Deposition of calcium phosphate between calcium oxalate crystals decreased the volume of pore spaces within calculi. Porosity was observed along boundaries between COM and COD. Minute pores increased the surface area of calculi exposed to urine, and this increase in liquid-solid interface promotes interaction of crystals with the surrounding urine. CONCLUSIONS AND CLINICAL RELEVANCE: Calcium oxalate urolithiasis is of major concern, because it is often a recurrent disease among dogs, principally treated by surgical removal of calculi, with few effective dissolution strategies. Understanding the ultrastructure and mineralogic content of calcium oxalate and its association with amorphous calcium phosphate is a step toward the solution of this increasingly important medical problem.
Dune reed, as an ecotype of reed plant (Phragmites communis Trin.), is an ideal material for studies on the adaptations of plant to environmental conditions. Scanning electron microscope, energy-dispersive X-ray analysis, and plant tissue culture techniques were used to investigate the effect of extreme temperature, salt, and polyethylene glycol-induced osmotic stress on the intracellular elements K, Na, Ca, and Cl in cultured cells from dune reed and swamp reed (as control). The results indicated that the percentages of the studied elements in dune reed cells exposed to various stresses increased or decreased obviously compared to the swamp reed cells. It has been found that a pattern of absorbing K and discharging Na exists in dune reed cells, which did not exist in swamp reed cells. The pattern is thought to be a significant physiological mechanism of the dune reed response to adverse environmental factors. In addition, the percentages of Ca and Cl in dune reed cells were also shown to increase at high temperature. The growth of cells along with their surface features under different stress conditions were observed and the results are discussed.
A layer of liquid lines the airways in the lung. Previous microscopic studies have suggested that it is in two phases, with a mucous gel lying above a periciliary sol. However, shrinkage artifacts due to chemical fixation, dehydration, and drying have prevented reliable estimates of the depth of these layers. To avoid such problems, we have studied the surface liquid of bovine trachea by low-temperature scanning electron microscopy (LTSEM). A polished copper probe cooled to liquid nitrogen temperature was applied to the mucosal surface of sheets of excised tracheal epithelium to effect rapid freezing of surface liquid. Tissue sheets were then mounted in an LTSEM (AMRay 1000A with Biochamber) which maintains samples at -180 degrees C with a Joule-Thompson refrigerator built into the stage. Tissues were fractured at right angles to the epithelial surface, coated with gold, and viewed, all at 10(-5) to 10(-6) torr without transfer through air. The sample was stable under the electron beam at accelerating voltages up to 20 kV. Epithelial features (nuclei, cilia, microvilli, mucous granules) were well preserved. The mucosal surface of the cells was covered with material on the order of 8 microns in depth. The mucous gel and periciliary sol could be seen as distinct layers and could be distinguished by the size and pattern of ice crystal voids generated by radiant-etching of the fractured surface of the sample.
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Scanning electron microscopy (SEM) analysis and qualitative energy dispersive x-ray microanalysis (EDS) of investment materials for dental titanium casting were performed. Two commercial investment materials, Ohara and Castmatic, and an experimental zirconia material were analyzed. The microstructural constituents and the unfired and fired structures were included. Larger refractory particles and matrix embedding smaller particles were observed with each material. Detection of aluminum, silicon, magnesium, zirconium and oxygen provided a basis to reason the presence of alumina (Al2O3), silica (SiO2), magnesia (MgO), and zirconia (ZrO2). Hence, Ohara contained quartz and an alumino-silicate, Castmatic contained magnesia and quartz and experimental zirconia contained zirconia and an alumino-silicate, taken to be kyanite, as components providing refractoriness and expansion. Even though unequivocal detection of phosphorous in the spectra for Ohara was not obtained (P K alpha = 2.013 keV; Zr L alpha = 2.042 keV), an emission peak at 2.0 keV was taken to be due to P and related, along with MgO, to bonding by magnesium phosphate. For Castmatic, unfired strength was thought to be due to calcium chloride and calcium silicate and fired strength to forsterite, (2MgO.SiO2). Detection of calcium and chlorine also suggested bonding of experimental zirconia via calcium chloride. Extensive microcracking occurred around refractory particles and through matrix in experimental zirconia which is likely to have resulted from the firing of kyanite to 1400 degrees C, to the monoclinic to tetragonal transformation of any unstabilized ZrO2, or to the thermal expansion mismatch between kyanite and matrix.
Environmental scanning electron microscope (ESEM) fitted with an energy dispersive X-ray microanalyzer (EDX) was used to investigate the surface micromorphology and arsenic (As) micro-distribution in Chinese brake (Pteris vittata L.). It was found that amounts of trichome, which possessed multicellular structure with the average length of 160 microm and with an average diameter of 28 microm, existed in the frond of P. vittata, and the density of trichome on the pinnate axial surface was higher than that on the petiole. Visible X-ray peak of As was recorded in the epidermal cell and trichome. The relative weight of As in the pinnate trichome, which contained the highest concentration of As among all tissues of the plant, was 2.4 and 3.9 times as much as that in the epidermal and mesophyllous cells, respectively. The As concentrations in the basal and stalk cells of the same trichome were higher than that in its cap cell. This is the first time to report that the trichome of P. vittata plays an important role in arsenic hyperaccumulation. The finding from the present study implies that much attention should be paid to the role of the trichome in understanding the hyperaccumulation and detoxicity of As in the hyperaccumulator and improving the ability of As accumulation.
Quantitation in biological X-ray microanalysis usually depends on the reference to one or more element standards. This paper deals with the general characteristics of such standards and gives a survey of the various methods of standard preparation reported in literature. Their advantages and disadvantages are briefly discussed. Practical experiments were carried out to prepare a potassium, calcium, and sulfur standard for X-ray microanalysis in the scanning transmission electron microscope.
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The skin is a tissue containing a large number of collagen types. Several collagens are restricted at the dermo-epidermal junction, contrarily to others present throughout the dermis. However, the distribution of the dermal collagen varies during embryonic development. In this contribution, we have been interested in the collagen types associated with the major collagenous components of the dermis, which are the collagen types I and III. Type V collagen, which is mixed with collagen types I and III to form heterotypic fibrils, has been studied during mouse embryo development. Transcripts of the alpha 1 (V) gene have been localized by in situ hybridization, on flattened cells of the stratum germinativum first, and then only on dermal cells. The expression of the gene decreases at birth, while the expression of the alpha 1(I) gene remains constant, with, however, a ring of high intensity around hair follicles. Other collagen types (VI, and the fibril-associated collagens XII and XIV) have been studied during calf embryonic development by immunofluorescence and ultrastructural immunogold detection. Type VI collagen appears homogeneously distributed throughout the dermis. Type XII collagen is first widely distributed and becomes restricted in the upper, papillary dermis after 6 months of gestation. Type XIV collagen, on the contrary, is first located as a delicate framework around hair follicles (at 19 weeks of gestation), and progressively invades the whole dermis where it appears abundant just before birth. The different functions of all these collagens are discussed in terms of dermis architecture, mechanical properties and physiology.
BACKGROUND: Extensive studies on the regulation of the volume and composition of amniotic and allantoic fluid in the sheep have suggested that the amniotic and allantoic membranes must play an active role in these processes. Little is known of the functional morphology of the sheep amnion and allantois beyond the presence of an epithelium overlying connective tissue. METHODS: The ovine amnion and allantois were characterized at a range of gestational ages (27-140 days of gestation, where term is 145-150 days) by electron microscopy (SEM and TEM) and the presence of transporting ATPases examined by use of immunohistochemistry (Ca++-ATPase) and in situ hybridization (Na,K-ATPase). RESULTS: With increasing gestational age, the cell height of epithelium of the membranes increased, as did the number of apical microvilli and the length of zonulae occludentes. Epithelial cell cytoplasm increased in complexity, and cell shape changed from flattened to cuboidal. Proliferation of cells occurred until close to term. Immunoreactivity to Ca++-ATPase was present in the basolateral membranes at all stages of gestation examined, but hybridization with the alpha and beta subunits of Na,K-ATPase was present only at or after 100 days of gestation. CONCLUSIONS: The epithelia of the sheep amnion and allantois display characteristics typical of transporting epithelia. As the epithelia mature, changes related to increased capacity for solute and fluid transport regulation occur.
Cobalt-chromium alloy porous coatings have become increasingly popular as a means of achieving a stable, longer lasting fixation on orthopedic implants. However, sintering heat treatments cause changes in the microstructure that result in changes in the corrosion behavior of the porous coatings. Experiments were conducted to examine the effects of microstructure on the corrosion of CoCr porous coatings. Four distinct microstructures were characterized. Light microscopy revealed microstructures A and B had relatively fewer carbides and a large-grain structure, microstructures C and D displayed a finer grain size with significant carbide formation, predominantly within grains and fusion zones. Accelerated anodic corrosion experiments were conducted to study the localized attack of each microstructure. Experiments were conducted using metallographically polished porous-coated disks for durations of 1, 4, 16, 24, and 72 h. The disks were examined with SEM and EDS to observe preferential attack and element depletion. Results showed a progressive dissolution of the matrix, with preferential attack of the grain boundaries and regions adjacent to the carbides due to sensitization. A precipitation layer was found to be Cr-rich and possibly composed of chromium hydroxide or a chromium orthophosphate. In addition, the solution became yellow in color with longer exposure times, possibly due to the presence of chromate ions, as indicated by an increased chromium level detected by AAS.
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Dense and porous Ti, Ti-alloy, and stainless steel specimens were implanted in canine trabecular bone. After 6 and 12 months the specimens were retrieved together with bone tissue immediately adjacent. The trace metal content in the tissue samples was determined using neutron activation analysis, differential pulse polarography, graphite furnace atomic-absorption spectrophotometry, electron microprobe analysis, and laser microprobe analysis. The results are discussed in view of (i) the release of Ti ions, which is larger for porous than for bulk specimens, (ii) the various artifacts arising in electron microprobe (EMP) and laser microprobe mass analysis (LAMMA) determination of compositional gradients of trace metal-ion content in bone tissue, (iii) the absence of measurable quantities of V in bone tissue, and (iv) the difference in local tissue accumulation between Ti, released from Ti specimens, and Ni, released from stainless steel specimens.
We have produced three kinds of apatite-containing glass ceramics of the same chemical composition: MgO (4.6), CaO (44.9), SiO2 (34.2), P2O5 (16.3), CaF2 (0.5) (in weight ratio). They contain different crystal combinations and have different mechanical properties. The first glass ceramic (A-GC) was prepared by heating a glass plate to 870 degrees C. It contains only oxy- and fluoroapatite (35 wt%). The second glass ceramic (A-W-GC), and the third (A-W-CP-GC), were prepared by heating glass powder compacts to 1050 degrees C and 1200 degrees C, respectively. A-W-GC contains oxyapatite and fluoroapatite (Ca10(PO4)6(O,F2] (35 wt%) and beta-wollastonite (40 wt%). A-W-CP-GC contains oxyapatite and fluoroapatite (20 wt%), beta-wollastonite (CaO X SiO2) (55 wt%), and beta-whitlockite (3CaO X P2O5) (15 wt%). The bending strengths of A-GC, A-W-GC, and A-W-CP-GC were 88MPa, 178MPa, and 213MPa, respectively, in air. Rectangular ceramic plates (15mm X 10mm X 2mm) were implanted into a rabbit tibia. Ten and 25 weeks after implantation, the segment of tibia with implant was excised for examination. The segment was held by a special jig and the traction breaking load (failure load) was measured by an autograph. A-GC showed a lower load than A-W-GC and A-W-CP-GC. The loads for A-W-GC and A-W-CP-GC were almost equal. The failure loads did not change significantly between 10 and 25 weeks for any of the materials. The interface was examined by Giemsa surface staining, contact micro-radiography, and SEM-EPMA. Giemsa surface staining and CMR revealed direct bonding between the materials and the bone for all the three materials. SEM-EPMA showed that Si and Mg content decreased, Ca content did not change, and P content increased at the reaction zone between all three glass ceramics and bone. This was observed at 10 weeks, as well as at 25 weeks, after implantation. The reaction zone was narrowest with A-GC, wider with A-W-GC, and widest with A-W-CP-GC.