The use of thin specimens for X‐ray microanalysis in biology

Hall, T. A.; Anderson, H. Clarke; Appleton, T. C.

doi:10.1111/j.1365-2818.1973.tb04670.x

Cited by 198 publications

(71 citation statements)

References 5 publications

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“…The peak list was printed together with the integral of counts between 4 and 6 keV energies where no significant peaks occur in the biological specimens. The mass fraction method of Hall et al (14) was used for quantitative analysis, extended for biological bulk specimens by applying a special selection of standards and avoiding the necessity of ZAF correction (28,29) . Calculation of peak to background ratios, preparation of histograms, etc.…”

Section: Methodsmentioning

confidence: 99%

Intracellular Na+:K+ ratios in human cancer cells as revealed by energy dispersive x-ray microanalysis.

Zs.‐Nagy¹,

Lustyik²,

Zs.-Nagy³

et al. 1981

The Journal of Cell Biology

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Intranuclear sodium, potassium, and chloride contents were measured by energydispersive x-ray microanalysis in freeze-fractured, freeze-dried, bulk-tumor samples taken from 10 patients suffering from invasive urogenital cancers. Human biopsies were carried out during the first diagnostic interventions before any cytostatic treatment had been applied. Pathohistological diagnosis established the malignancy in each case . The cancers were classified in three types: keratinizing, transitional cell, and hypernephroid carcinoma. More than 250 cell nuclei were measured from each type of cancer . The results were compared with those obtained in intact human urothelium taken from patients having no malignant processes. Proximal and distal tubular epithelial cell nuclei representing the origin of human hypernephroid cancer were also measured in rat kidney because corresponding healthy human material cannot be obtained . The analyses revealed, in all three types of cancer cells, that the average intranuclear sodium content increased more than three-fold, the potassium content decreased 32, 16, and 13%, respectively; meanwhile the chloride content increased, but to a lesser extent than did the sodium . The intranuclear Na + :K + ratios were more than five-fold higher in the cancer cells on the average, and their distribution histograms were much broader than in the normal human urothelium and in the tubular cell nuclei of the rat kidney . The results obtained fit well with the theory of Cone, C. D., Jr . 1971 . J. Theor. Biol. 30 : 151-181 according to which the sustained depolarization of the cell membrane may be of mitogenic effect .

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Section: Methodsmentioning

confidence: 99%

Intracellular Na+:K+ ratios in human cancer cells as revealed by energy dispersive x-ray microanalysis.

Zs.‐Nagy¹,

Lustyik²,

Zs.-Nagy³

et al. 1981

The Journal of Cell Biology

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“…The spectra were analysed with a dedicated PDP-I 1 microcomputer using a digital filtration and least-squares fitting program (Shuman, Somlyo & Somlyo, 1976). The analysis uses the ratio of peak counts to continuum (1-00-3 90 keV in our case) count to calculate elemental mass fractions as described by Hall et al (1973). Binary salt standards were used to establish relative counting efficiencies, and S in an albumin sample was used as an absolute standard.…”

Section: Elemental Analy8i8mentioning

confidence: 99%

“…Hall (1971) formulated the theory ofcontinuum normalization for the quantification of X-ray data from biological specimens, and Hall, Anderson & Appleton (1973) for thin sections of soft tissues. Gupta, Hall & Moreton (1977) provided a detailed exposition of the procedures and history as well as fully quantitative data from many tissues.…”

Section: Introductionmentioning

confidence: 99%

Changes in elemental concentrations of rat parotid acinar cells following pilocarpine stimulation.

Izutsu¹,

Johnson²

1986

The Journal of Physiology

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SUMMARY1. Quantitative electron microprobe analysis was used to measure elemental dry weight concentrations in cytoplasm, secretary granules and nuclei of resting and pilocarpine-stimulated rat parotid gland acinar cells.2. Secretory granules in resting cells had lower concentrations of Na, Mg, P, Cl and K, and higher concentrations of S and Ca than cytoplasm or nuclei. Nuclei in resting cells had lower S and higher K concentrations than cytoplasm.3. Three major pilocarpine-related changes were found: (i) cytoplasmic dry weight concentrations of Na and Ca increased and the concentration of K decreased, (ii) the nuclear concentration of Na increased while that of K decreased and (iii) the concentrations of Na and Cl increased in secretary granules.4. These results indicate that the nuclear, and cytoplasmic compartments have different mechanisms for regulating their elemental concentrations relative to the secretary granules.5. The present results are largely consistent with X-ray microanalysis results from the pilocarpine-stimulated dog submandibular gland.

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“…Peak-to-continuum ratios were converted to elemental concentrations using weighting factors derived from standard curves obtained from freeze-dried sections ofgelatin-glycerol preparations containing different concentrations of various elements (25). Values obtained were for total electrolyte concentrations, including bound and free, ionized components (24)(25)(26)(27). The data were expressed as millimoles per kilogram dry weight, with the denominator based on the local mass density derived from the continuum ofthe x-ray spectra (24)(25)(26)(27).…”

Section: Methodsmentioning

confidence: 99%

“…Elemental peak intensities were obtained by deconvolution of the spectra using a multiple least-squares fitting technique (25). A computerized method was used for quantitative analysis of spectra, based on the direct relationship between peak-to-continuum ratio and elemental concentration (26,27). Spectral analysis included a computer fitting routine using the first and second derivatives ofthe major (K alpha) potassium peak (27).…”

Section: Methodsmentioning

confidence: 99%

Relationship between calcium loading and impaired energy metabolism during Na+, K+ pump inhibition and metabolic inhibition in cultured neonatal rat cardiac myocytes.

Morris¹,

Hagler²,

Willerson³

et al. 1989

J. Clin. Invest.

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This study tested the hypothesis that the initiating mechanism is a major determinant of the response to calcium (Ca) accumulation in myocardium. Cultured neonatal rat ventriculocytes were exposed to Na', K+ pump inhibition with 1 mM ouabain and metabolic inhibition with 20 mM 2-deoxy--glucose and 1 mM cyanide (DOG-CN) for up to 2 h. Microspectrofluorometry of myocytes loaded with fura-2 showed that ouabain resulted in a relatively rapid increase in [Ca2]i, up to 2-3 tM (two to threefold above peak systolic level) and that DOG-CN produced an initial decrease and then a relatively slow increase in ICa2+li up to peak systolic level. Electron probe x-ray microanalysis (EPMA) showed prominent increases in Na and Ca and decreases in K and Mg in cytoplasm and mitoshondria with both interventions, although the increases in Ca were greater with ouabain than DOG-CN. ATP was reduced by 58% after 1 and 2 h of ouabain and by 70 and 90% after 1 and 2 h of DOG-CN, respectively. Thus, ouabain produced greater calcium accumulation and less ATP reduction than DOG-CN.Upon return to normal medium for 30 min, myocytes showed recovery of most electrolyte alterations and resumption of normal Ca2+ transients after 1 h exposure to either ouabain or DOG-CN; however, recovery was less after 2 h of either treatment, with elevated [Ca2+1, maintained in many myocytes. We conclude that the severity of myocyte injury is influenced by the magnitude and duration of both ATP reduction and calcium accumulation. IntroductionMyocardial ischemia leads to impaired energy metabolism, membrane dysfunction, and altered calcium (Ca) homeostasis (1-10). These metabolic and pathophysiological alterations are thought to lead to the progression of irreversible injury; how-

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The use of thin specimens for X‐ray microanalysis in biology

Cited by 198 publications

References 5 publications

Intracellular Na+:K+ ratios in human cancer cells as revealed by energy dispersive x-ray microanalysis.

Intracellular Na+:K+ ratios in human cancer cells as revealed by energy dispersive x-ray microanalysis.

Changes in elemental concentrations of rat parotid acinar cells following pilocarpine stimulation.

Relationship between calcium loading and impaired energy metabolism during Na+, K+ pump inhibition and metabolic inhibition in cultured neonatal rat cardiac myocytes.

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