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Utilization of electron probe microanalysis in gadolinium-treated mice.

Gadolinium is used as a contrast media for magnetic resonance imaging and, experimentally, to block Kupffer cell phagocytosis. In this study, we utilize electron probe microanalysis to determine the subcellular localization of gadolinium chloride (GdCl3) administered to mice in a short-term toxicology study. Male CD-1 mice were administered 0.0, 2.5, or 8.0 mg/kg GdCl3 iv for 14 consecutive weekdays. Liver-associated enzymes were significantly elevated in high-dose animals only and correlated histologically with multifocal, hepatocellular degeneration associated with a neutrophilic infiltrate. Morphological investigations were performed on high-dose animals. Hepatocytes and Kupffer cells had morphologic features of cellular injury consisting of swollen mitochondria and vesiculated profiles of endoplasmic reticulum. Hepatocytes, Kupffer cells, bile canaliculi, and neutrophils in the liver contained discrete aggregates of electron-dense granular material, as did pulmonary interstitial macrophages, splenic macrophages, and mesangial cells of the renal glomerulus. The intracellular granular material in the liver, lung, spleen, and kidney was confirmed as gadolinium by qualitative electron probe microanalysis. These results document both hepatic and extra-hepatic accumulation of gadolinium in cells of the mononuclear phagocytic system and highlight the importance of electron probe microanalysis in toxicologic assessment.

Animals↗

[Studies on gallium alloys for dental restorations. 2. Electron probe microanalysis for hardened gallium alloys].

Hardened gallium alloys were analyzed by scanning electron microscopy (SEM) and electron probe microanalysis (EPMA). The cores, matrix and white irregular shape phases were visible with SEM. The round or irregular shape cores contained Ag, Pd, Cu and Zn but did not contain Ga and In. The dark irregular shape matrix, which surrounded the core, consisted of Ga-Ag, Ga-Pd, Ga-Cu and Ga-Sn phases. However, the amount of Ga-Sn phase was very low. The white phase in the matrix was Ag-In phase.

Dental Alloys↗

A combined scanning and transmission electron microscopic study and electron probe microanalysis of human pineal acervuli.

Untreated, decalcified and trypsinized acervuli from human pineal bodies were studied with the scanning and transmission electron microscope as well as by electron probe microanalysis. The mulberry-like acervuli are composed of a various number of spherical lobes (135-800 mum) between which clustered groups of globuli (4--14 mum in diameter) are observed. The acervular lobes are very probably formed by an aggregation of these globuli. Small round particles 125--500 A in diameter are observed on the surface of the pineal concretions. These are not influenced by either decalcification or trypsin treatment. The acervular mineral corresponds morphologically to hydroxyapatite. The electron probe microanalysis reveals the existence of calcium and phosphorus as main components of the acervuli. Small quantities of magnesium and strontium were also detected.

Aged↗

Scanning electron microscopy and electron probe microanalysis of calcified alpha-elastin coacervates.

The uptake of calcium and phosphorus from a serum calcification medium by coacervated alpha-elastin was studied by electron probe microanalysis and scanning electron microscopy. Calcium and phosphorus were bound by the coacervate in ratios similar to that of hydroxyapatite. A significant difference was observed in the secondary electron image of the coacercules from the serum during calcification of the coacervate.

Animals↗

Quantitative biological electron probe microanalysis with a wavelength dispersive spectrometer.

This paper describes the details of quantitative electron probe microanalysis (EPMA) performed with a wavelength dispersive spectrometer (WDS). EPMA was carried out on the giant neuron of a fresh frozen ganglion from the snail Lymnaea stagnalis. The freeze-dried cryosections were compared with sections of freeze-dried, embedded tissue. It was found, that in the ganglion there are two kinds of neurons with a different chlorine concentration of 11 mmole/liter and 32 mmole/liter. Isolated neurons in culture were shown to differ in elemental composition from those in the ganglion tissue.

Animals↗

Measurement of element content in isolated papillary collecting duct cells by electron probe microanalysis.

A method was developed to measure the element content of freshly isolated papillary collecting duct (PCD) cells by electron probe microanalysis in a scanning electron microscope. After isolation, the cells were transferred onto a Thermanox support by centrifugation and the extracellular medium was removed by brief exposure to buffered ammonium acetate; cryofixation, freeze-drying, and coating with carbon followed. Under visual control in the scanning electron microscope the Na, Cl, K and P content of cell clusters (about 30 cells/cluster) was then measured by X-ray microanalysis. Cells incubated in control medium showed potassium:sodium ratios identical to those determined previously in cryosections of the same cells. In ouabain-treated cells sodium influx and potassium efflux was demonstrated. Potassium left the cells with a t1/2 of 21.7 min. The t1/2 of Na influx was 12.6 min for the first 15 min of incubation, whereafter further influx was markedly slower. Ouabain-induced sodium influx was inhibited 40% by amiloride. These results indicate that X-ray microanalysis can be applied to analyze the ion content of isolated cell clusters derived from the papillary collecting duct. Using ouabain and amiloride as inhibitors the suitability of the method to identify transport systems is demonstrated.

Amiloride↗

Scanning electron microscopy and electron probe microanalysis studies of human pineal concretions.

The calcareous concretions of human pineal bodies were investigated with scanning electron microscopy and electron probe microanalysis. The initial concretions measuring 5-7 microns in diameter may have started at the calcified pinealocytes. They grew appositionally forming concentric laminations, and then the simple calcospherulites over 20 microns occasionally aggregated with each other. Some of them became numerous spherulite-aggregated concretions. Others individually grew with scallop-shaped concentric laminations at intervals of 0.05-1 microns and became lobated calcospherulites up to 0.5 mm. The concretions over 0.5 mm were formed by their attachments. The major elements were Ca and P, while traces of S, Mg, and Na were detected. In the calcification and crystallization values, the center of the concretions over 50 microns was significantly higher than the periphery, while there were no differences among the centers and also among the peripheries. The Ca and P amounts in the center were 30.8% and 14.2% by weight and the Ca/P molar ratio was 1.68; thereby the sand-grain-shaped crystals may be nearly hydroxyapatite, as reported previously.

Aged↗

Quantitative electron probe microanalysis of biological thin sections: the use of stem for measurement of local mass thickness.

A method for performing quantitative electron probe microanalysis on ultrathin (less than 30 microgram/cm2) biological samples is described and evaluated. The technique is based on a measurement of the characteristic peak count rate and the degree of beam attenuation as the primary electron beam passes through the sample. Using this method it is possible to measure the concentration of a given element such as sodium in sections ranging in mass-thickness from several microgram/cm2 up to 30 microgram/cm2 with an accuracy of better than 10%. For sections having a mass thickness of approximately 11 microgram/cm2 the minimum detectable concentration for sodium was found to be 20 mmolar or 4 X 10(-2) wt.%. The interaction of the electron beam with the sample is also discussed with emphasis on characterizing the variation in sample mass with radiation dose.

Electron Probe Microanalysis↗

[Electron probe microanalysis of potassium and phosphorus in mouse oocytes and zygotes].

Intracellular concentrations of potassium and phosphorus were determined by Electron Probe Microanalysis in mouse mature oocytes and zygotes. The oocytes were characterized by insignificant variations in the concentrations of these elements in the cytoplasm: 60 +/- 4 and 103 +/- 6 mM, respectively. In zygotes, on the contrary, significant variations were observed: 64 +/- 16 and 84 +/- 14 mM, respectively. Changes in the potassium homeostasis during the first cell cycle have been discussed.

Animals↗

Measurement of subcellular Ca2+ redistribution in cardiac muscle in situ: time resolved rapid freezing and electron probe microanalysis.

To directly assess the physiological roles of sarcoplasmic reticulum (SR) and mitochondria (MT), we have utilized energy dispersive electron probe microanalysis (EPMA) on ultrathin freeze-dried cryosections from isolated papillary muscles, rapidly frozen at precise time points of the contractile cycle. Using this approach, we can detect redistribution of subcellular Ca2+ during the cardiac contractile cycle. Changes in Ca2+ of less than 1.0 mmol/kg dry wt can be detected. By determining the variability of the Ca2+ measurements in preliminary experiments, we have also demonstrated that it is possible to optimize experimental design, i.e., to predict the number of animals per treatment group and the number of X-ray spectra per animal that are required in order to detect a specified Ca2+ difference. Quantitative EPMA of rapidly frozen contracting papillary muscle has also allowed us to correlate the Ca2+ content of SR and MT with the contractile state of the muscle. Our results show a decrease of 40% in the amount of Ca2+ stored in the junctional SR during a cardiac muscle twitch, thus providing direct evidence for a role of the SR as a primary site of Ca2+ release. In addition, we have demonstrated dissociation between MT Ca2+ uptake and activation of regulatory enzymes, such as pyruvate dehydrogenase, indicating that MT Ca2+ uptake is not required for activation of MT metabolism.

Animals↗

Determination of iodine concentration and distribution in rat thyroid follicles by electron-probe microanalysis.

The concentration and the distribution of iodine in various sized follicles of rat thyroid glands have been analyzed by electron-probe microanalysis. The results of the iodine analysis were grouped according to uncorrected lumen diameter size. No significant differences in iodine concentration were observed among the various size categories. When the results for all follicles from a given sample were pooled, the standard error of the mean was approximately 4%. Usually 40-50 follicles per animal were analyzed. The concentration of iodine ranged from 0.9 to 2.1% by weight among individual animals. Scanning pictures and step-scan analysis showed the iodine distribution to be quite uniform across the colloid area. Several techniques of sample preparation were used; they produced no significant differences in the iodine concentrations observed. Sodium concentration, also determined in all samples, was found to vary from 2 to 9% by weight. Because of the mobility of the sodium ion, its distribution was greatly affected by the method of sample preparation. The technique that best preserved the natural chemistry of the sample was that of freezing the tissue, sectioning, and then freeze-drying.

Animals↗

Electron probe microanalysis of human teeth for the determination of individual age.

The calcium and phosphorus weight concentrations as well as the calcium/phosphorus weight ratio were determined by electron probe microanalysis both peritubularly and intertubularly in the dentine of 25 adults of varying age and sex, originating from our autopsy material. The studies were performed with a TESLA BS-300 scanning electron microscope coupled with an EDAX-149-10 X-ray detector and a BMG-38100 X-ray analyser with an accelerating potential of 15 kV, a sample current of 100 pA and a measurement time of 20 s. Peritubularly the Ca/P weight ratio of the hypermineralized part of the dentine decreases with age and is in close correlation with it (r = -0.9052) and this tendency is significant even at the level of 95% (P less than 0.05). The equation of the regression line is: Y = 37.17 x +139.37 where x is the age and y is the Ca/P weight ratio. The weight concentration of phosphorus is in an even closer correlation with age (r = 0.9712) and this correlation is also significant (P less than 0.01). The equation of the regression line is the following: y = 625.0 x -42.06 The close correlation of both the Ca/P weight ratio of the teeth and of the phosphorus weight concentration with the age allows individual age determination to be performed in the course of forensic medical and palaeoanthropological studies.

Age Determination by Teeth↗

Factitial cutaneous ulcers and nodules: the use of electron-probe microanalysis in diagnosis.

Two cases are reported in which persistent nodular or ulcerative skin lesions with granulomatous histology suggested possible injection of foreign material. Electron-probe microanalysis was used to determine the presence and nature of foreign materials present in the skin. This technic identified specific agents used as fillers in tablets, indicating the injection of pulverized tablet material. The patients were counseled regarding this evidence, and psychotherapy and drug rehabilitation were recommended in each instance. This technic provides a valuable tool for accurate identification of foreign substances in the skin.

Adult↗