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Results for “Electron Probe Microanalysis”

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PubMed1983

Intracellular calcium store and transport of elements in acinar cells of the salivary gland determined by electron probe X-ray microanalysis.

Electron probe X-ray microanalysis using freshly frozen hydrated and dried thin sections of dog submandibular gland was performed to determine the distribution of elements and water in the acinar cells of resting and stimulating states. The results obtained are as follows: (a) The secretory granules contained high concentrations of Ca and S while high concentrations of K and P were present in the cytoplasm and/or nucleus of acinar mucous cells of the gland in the resting state. (b) With pilocarpine stimulation, the concentration of Ca increased in the cytoplasm and decreased in the secretory granules, while there was an increase in the concentration of Na and Cl in both the cytoplasm and secretory granules of the cells. (c) The local dry-mass fractions of acinar cells, estimated by comparing the continuum radiation of X-ray spectrum from the frozen hydrated sections with that from the frozen dehydrated sections, were approximately 20 and 33% in the cytoplasm and secretory granules of resting acinar cells, respectively, and each value was not significantly altered under conditions of stimulation having a tendency to decrease slightly. Therefore, the passive Na and Cl influx and the cytoplasmic Ca flowed in from extracellular spaces and released from secretory granules, an intracellular calcium store, by secretory stimulation probably triggers the passive or active Na and Cl extrusion and consequently the osmotic water flux from the basal part of acinar cells to the secretory granules and the lumen, as well as the serial exocytosis of the granules in the luminal side of the acinar cells.

Animals↗
PubMed1991

Three-dimensional spatial relationship between the collagen fibrils and the inorganic calcium phosphate crystals of pickerel (Americanus americanus) and herring (Clupea harengus) bone.

High-voltage (1.0 MV) electron microscopy and stereomicroscopy, electron probe microanalysis, electron diffraction and three-dimensional computer reconstruction, have been used to examine the spatial relationship between the inorganic crystals of calcium phosphate and the collagen fibrils of pickerel and herring bone. High-voltage stereo electron-micrographs were obtained of cross-sections of the cylinder-shaped intramuscular bones in uncalcified regions, in regions where only one or only several crystals had been deposited in some of the fibrils, and in successive sections containing progressively more mineral crystals until the stage of full mineralization was reached. High-resolution electron probe microanalysis confirmed that the electron-dense particles contained calcium and phosphorus. In the earliest stages of mineralization and progressing throughout the mineralization process, the crystals are located only within the collagen fibrils; crystals are not observed free in the extracellular spaces between collagen fibrils. The progressive increase in the mass of mineral deposited in the bone tissue with time occurs, essentially, completely within the collagen fibrils including the stage of full mineralization. At this stage, cross-sectional profiles of collagen fibrils are completely obliterated by mineral. A small number of crystals that are located on or close to the surface of the fibrils appear to extend a very short distance into the spaces between the fibrils. These ultrastructural observations of the very onset of calcification in which nucleation of the calcium phosphate crystals is clearly shown to begin within specific volumes of collagen fibrils, and of the subsequent temporal and spatial sequences of this phenomenon, which shows that calcification continues wholly within the collagen fibrils until maximum calcification is achieved, add important information on the basic physical chemical mechanism of the calcification and the structural elements that are involved. The spatial and temporal independence of the sites where mineralization is initiated establishes that such ultrastructural locations within individual collagen fibrils represent independent, physical chemical nucleation loci. The findings are totally inconsistent with the proposal that crystals must first be deposited in matrix vesicles, or other components such as mitochondria, and subsequently released and propagated in the interfibrillar space, until they eventually reach and impregnate the hole zone regions of the collagen fibrils. Three-dimensional computer reconstruction of serial transverse and longitudinal sections demonstrates periodic swellings along the collagen fibrils, corresponding to the hole zone region of their axial period as mineralization proceeds.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1999

The crystalline pattern of calcium in different topographical regions of the otoconial membrane. An electron probe and spectroscopic diffraction study.

Quantitative electron probe microanalysis and electron spectroscopic diffraction analysis was used to determine the gradient of distribution of calcium and its crystalline pattern at different levels (lower gelatinous membrane, upper gelatinous membrane and otoliths) in the otoconial membrane of adult OF1 mice. Our quantitative electron probe microanalytical data, obtained with scanning-transmission electron microscopy, indicated that there was a gradient in calcium concentration which increased from the vestibular surface towards the otoliths. Differences between the three regions of the otoconial membrane were statistically significant in both the utricle and saccule. Our results with electron spectroscopic diffraction revealed an increasing crystalline development from the lower gelatinous membrane towards the otoliths. Our findings with both techniques suggest that the gelatinous membrane is involved in the maturation and crystallization of the otoliths.

Animals↗
PubMed1990

Visualization of sulfur-containing components associated with proliferating chondrocytes from rat epiphyseal growth plate cartilage: possible proteoglycan and collagen co-migration.

Electron microscopy of epiphyseal growth plate cartilage from normal 4-5-week-old rats has revealed extensive fibrillar aggregates and globules in the pericellular spaces of proliferating chondrocytes. These cells contained small globules and diffusely coiled, fine filaments located within large, membrane-invested vacuoles. All such structures were observed after a variety of different tissue fixation regimes, including glutaraldehyde, osmium tetroxide, and potassium pyroantimonate. The fibrillar aggregates and globules were often overlapping and intermeshed and extended to 0.5 micron in length from their point of origin at cell membranes. Vacuoles were usually found at the periphery of cells, and some, by membrane fusion with the cell envelope, appeared contiguous with extracellular spaces wherein their contents could be discharged. Fine filaments and globules were occasionally observed in the Golgi complex and cisternae of endoplasmic reticulum of the chondrocytes. Further characterization of the cellular and pericellular components by electron microscopic radioautography, electron probe microanalysis, and electron spectroscopic imaging indicated the presence of sulfur, a result suggesting these aggregates, filaments, and globules in part represent proteoglycans in various stages of synthesis, secretion, and assembly. Additional radioautography utilizing 3H-proline implied that filament bundles are also composed of collagen, a result posing the possibility that this protein and the putative proteoglycans may co-migrate both intracellularly and within pericellular matrices. In extracellular matrices adjacent to cell lacunae, the fibrillar aggregates appeared in close association with typical collagen type II fibrils, an observation providing evidence for proteoglycan-collagen network formation in this region of the rat epiphysis. These microscopic and analytical data in situ would support certain studies in vitro of proteoglycan-collagen type II and IX association and are important in describing the interaction of such cartilage components ultimately involved in matrix formation.

Animals↗
PubMed2002

Compositional averaging of continuum intensities in multielement compounds.

We find no physical basis for traditional mass-fraction weighted averaging of X-ray continuum measured in pure elements to estimate continuum production from compounds. Measurements of continuum from pairs of stable isotopes demonstrate that mass alone (the presence of more or fewer neutrons) has no effect on continuum production in electron probe microanalysis. Electron-fraction weighted averaging, which is based on the relative contribution of each constituent element to the electron (or proton) composition of a compound, proved superior to mass-fraction weighted averaging, in predicting the continuum of a compound from the measured continuum of its elements.

Chemical Phenomena↗
PubMed1997

Role of alveolar macrophage lysosomes in metal detoxification.

The intracellular behaviour of different toxic mineral elements inhaled as soluble aerosols or as insoluble particles was studied in the rat by electron microscopy, electron probe microanalysis, and electron microdiffraction. This study showed that, after inhalation, aerosols of soluble elements like cerous chloride, chromic chloride, uranyl nitrate, and aluminium chloride, are concentrated in the lysosomes of alveolar macrophages and are precipitated in the lysosomes in the form of insoluble phosphate, probably due to the activity of acid phosphatase (intralysosomial enzyme). Also, after inhalation of crystalline particles that are insoluble or poorly soluble in water such as the illites (phyllosilicates), ceric oxides (opaline), and industrial uranium oxides (U3O8), the small crystals are captured by the alveolar macrophage lysosomes and transformed over time into an amorphous form. This structural transformation is associated with changes in the chemical nature of particles inhaled in the oxide form. Microanalysis of amorphous deposits observed after inhalation of uranium or ceric oxides has shown that they contain high concentrations of phosphorus associated with the initial elements cerium and uranium. These different processes tend to limit the diffusion of these toxic elements within the organism, whether they are inhaled in soluble form or not.

Aerosols↗
PubMed1977

Crystalloid nature of mineral deposits in Bowman's capsule of the human glomerulus. Study by X-ray microanalysis and electron microdiffraction.

Abnormal crystalline mineral deposits are often observed in the basement membrane of Bowman's capsule in the human glomerulus. The nature of these deposits has been the subject of much discussion. By means of electron probe microanalysis and electron diffraction techniques, we have identified these abnormal deposits as whitlockite (beta-calcium-orthophosphate).

Calcium Phosphates↗
PubMed1993

Pigmenting agents in Martian soils: inferences from spectral, Mossbauer, and magnetic properties of nanophase and other iron oxides in Hawaiian palagonitic soil PN-9.

We have examined a Hawaiian palagonitic tephra sample (PN-9) that has spectroscopic similarities to Martian bright regions using a number of analytical techniques, including Mossbauer and reflectance spectroscopy, X-ray diffraction, instrumental neutron activation analysis, electron probe microanalysis, transmission electron microscopy, and dithionite-citrate-bicarbonate extraction. Chemically, PN-9 has a Hawaiitic composition with alkali (and presumably silica) loss resulting from leaching by meteoric water during palagonitization; no Ce anomaly is present in the REE pattern. Mineralogically, our results show that nanophase ferric oxide (np-Ox) particles (either nanophase hematite (np-Hm) or a mixture of ferrihydrite and np-Hm) are responsible for the distinctive ferric doublet and visible-wavelength ferric absorption edge observed in Mossbauer and reflectivity spectra, respectively, for this and other spectrally similar palagonitic samples. The np-Ox particles appear to be imbedded in a hydrated aluminosilicate matrix material; no evidence was found for phyllosilicates. Other iron-bearing phases observed are titanomagnetite, which accounts for the magnetic nature of the sample; olivine; pyroxene; and glass. By analogy, np-Ox is likely the primary pigmenting agent of the bright soils and dust of Mars.

Evolution, Planetary↗
PubMed1997

In situ compositional analysis of acidocalcisomes in Trypanosoma cruzi.

We measured the elemental content of different compartments in Trypanosoma cruzi epimastigotes using quick freezing, ultracryomicrotomy, and electron probe microanalysis. Vacuoles identified by high electron density contained (in units of mmol/kg dry weight +/- S.E.) large amounts of phosphorus (1390 +/- 13), magnesium (646 +/- 19), calcium (171 +/- 5), sodium (161 +/- 18), and zinc (148 +/- 6). No other compartment had appreciable calcium or zinc content. Iron (128 +/- 16 mmol/kg) was detected only in vacuoles distinct from the electron-dense vacuoles and other organelles. Incubation of cells for 70 min in culture medium in the presence of ionomycin plus nigericin led to a very significant 3- or 2-fold increase in potassium in the electron-dense vacuoles and the iron-rich vacuoles, respectively, with no significant change in the other elements investigated. This indicated the acidic nature of the vacuoles and demonstrated that the electron-dense vacuoles correspond to what were described previously as acidocalcisomes, i.e. acidic compartments rich in Ca2+. The acidocalcisomes were investigated by separation of epimastigote fractions on Percoll gradients in combination with Triton WR-1339 treatment. This detergent caused a rapid vacuolation; these vacuoles were shown by electron microscopy to be largely transparent, with a diffuse matrix. Percoll gradient fractionation demonstrated decreases in the density of various organelle markers in detergent-treated cells compared with controls. Large decreases in the density of the acidocalcisome and the mitochondrion were seen, as well as smaller decreases in the density of the other markers. Conventional electron microscopy of epimastigotes loaded with gold-labeled transferrin indicated that the endosomal system was separate from vacuoles that probably corresponded to the calcium-containing organelles detected by electron probe microanalysis. The combined results provide evidence that acidocalcisomes are organelles different from lysosomes or other organelles previously described in these parasites.

Animals↗
PubMed1996

Effect of a specially formulated remineralizing solution on the remineralizing capacity of saliva.

The present study was carried out to assess whether the remineralization of artificial caries like lesion by artificial saliva can be improved by daily 5 minute treatment in a specially formulated remineralizing solution containing Ca, P, Zn, Sr, F. Extracted human premolars were dipped for 24 hours in artificial saliva and subsequently daily for 5 minutes in the remineralizing solution for 3 weeks. Lesions were assessed using transverse microradiography, electron probe microanalysis (EPMA) and scanning electron microscope (SEM). The results showed that remineralization by specially formulated solutions repair the enamel lesion and enhances the salivary remineralizing potential.

Bicuspid↗
PubMed1980

Quantitation in x ray microanalysis of biological bulk specimens.

Electron probe microanalysis of biological bulk specimens, as compared to thin specimens, is limited by poor X-ray spatial resolution. On the other hand preparation methods for bulk specimens are often much simpler than for thin specimens. With respect to quantitation, however, numerous problems arise, such as poorly known composition of the matrix, local density differences, charging, mass loss, contamination, poorly defined surface topography, etc. The various quantitation methods, commonly used, include the use of ideal and non-ideal standards, the use of P/B-ratios, the calculation of ZAF-correction, and the use of so-called no-standard methods. The accuracy or reliability of the various methods has hardly been systematically investigated. It is shown that approximations from model systems will be necessary. Finally it is expected that the method used for particle analysis will be useful for the investigation of biological bulk specimens.

Biology↗