Electron probe X-ray microanalysis of human muscle biopsies

Wróblewski, Romuald; Roomans, Godfried M.; Jansson, Eva; Edström, Lars

doi:10.1007/bf00508796

Cited by 74 publications

(27 citation statements)

References 24 publications

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“…This also agrees with the data, which show that influx of Br into the cells is dependent on the concentration gradient, i.e., influx is greater when the external Br Ϫ concentration is higher. The fact that the cells gain Na and lose K is likely due to inhibition of the Na ϩ -K ϩ -ATPase, e.g., due to reduced metabolism, and Na ϩ ions will diffuse into the The size of the analytical volume under the experimental conditions chosen can be estimated at 10 -12 m (Wróblewski et al, 1978), with most of the X-rays coming from near the point of impact rather than from further away. Nevertheless, it is conceivable that some of the intercellular space is sampled.…”

Section: Discussionmentioning

confidence: 99%

“…The frozen pieces of intestine were cut into 16-m thick cryosections in conventional cryostat at -30°C (Wróblewski et al, 1978, as modified by McMillan and Roomans, 1990) and mounted on a carbon specimen holder over a layer of thin Formvar film (Merck, Darmstadt, Germany). The sections were freeze-dried for 72 hours in the cryostat and slowly warmed to room temperature.…”

Section: X-ray Microanalysismentioning

confidence: 99%

“…The specimens were analyzed at 20 kV in a Philips SEM 525 scanning electron microscope (Philips Electron Optics, Eindhoven, The Netherlands) with a LINK AN10000 energy-dispersive detector (Oxford Instruments, Oxford, UK). Under the experimental conditions, the electron beam penetrates maximally 12 m into the sections (Wróblewski et al, 1978). All experiments were carried out in triplicate (unless otherwise stated) and 10 -16 intestinal crypt and villous cells per animal were probed.…”

Section: X-ray Microanalysismentioning

confidence: 99%

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In vitro and in situ experimental model for X‐ray microanalysis of intestinal epithelium

Vanthanouvong

Högman

Roomans

2003

Microscopy Res & Technique

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Intestinal chloride (Cl) transport is disturbed in a number of diseases. X-ray microanalysis can be used to study the distribution of Cl and other ions in intestinal epithelial cells. In this study it was attempted to establish an experimental system that retains the in vivo elemental composition of intestinal epithelial cells. An in vitro system was set up in which a segment of rat intestine was mounted in a bath and perfused with different fluids. The chloride in the bath or in the perfusion fluid could be exchanged for gluconate or bromide to determine the direction of chloride fluxes. An in situ system was set up in which the animal was anesthetized and a segment of the intestine was perfused with different solutions. In the in vitro experiments the concentration of Na and Cl in the epithelial cells increased and that of K decreased. These changes occurred within the first 30 minutes of incubation. Uptake of chloride occurred mainly from the bath, as seen in experiments where bromide was used as a chloride analog. The concentration gradient between bath and tissue determined the extent of chloride uptake. Addition of glucose to the perfusion fluid and bath did not improve the results. In the in situ system, preservation of the intracellular ion composition was better. Acceptable results were obtained with perfusion with Krebs-Ringer's buffer without glucose for 30 minutes. In this case, the elemental content of the cell did not change appreciably during incubation. If glucose was added, the Na concentration increased in comparison to the control, both in crypt and villus cells. It is concluded that the intestinal epithelium is a sensitive system, very prone to disturbance of its homeostasis. However, the in situ system can be used in studies of agonist-induced ion transport.

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Section: Discussionmentioning

confidence: 99%

Section: X-ray Microanalysismentioning

confidence: 99%

Section: X-ray Microanalysismentioning

confidence: 99%

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In vitro and in situ experimental model for X‐ray microanalysis of intestinal epithelium

Vanthanouvong

Högman

Roomans

2003

Microscopy Res & Technique

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“…The sections were coated with a thin conductive carbon layer to prevent charging in the electron microscope. 25,26 The first 5 to 10 sections were cut from the dissected edge of the tissue block and discarded because the edge might have been dam- aged. The sections were analyzed in a Philips 525 scanning electron microscope (Philips Electron Optics, Eindhoven, The Netherlands) at 20 kV with a LINK AN10000 energydispersive spectrometer system (Oxford Instruments, Oxford, UK).…”

Section: X-ray Microanalysismentioning

confidence: 99%

Preservation of mouse liver tissue during cold storage in experimental solutions assessed by x-ray microanalysis

Kozlova

Khalid

Roomans

2003

Liver Transplantation

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The increasing use of organs for transplantation necessitates the development of optimal preservation techniques. The goal of this study was to investigate changes in elemental content in mouse liver cells during cold storage by x-ray microanalysis in parallel with morphologic studies. Tissue was stored at 4°C for 4 to 12 hours in normal Krebs-Ringer solution (high sodium/potassium ratio), modified Krebs-Ringer solution (low Na ؉ /K ؉ ratio), Euro-Collins solution, University of Wisconsin (UW) solution, or seven modified versions of the UW solution. Incubation of liver in normal Krebs-Ringer solution caused a significant increase in sodium and decrease in potassium concentrations in contrast to incubation in other solutions. The concentration of sodium, potassium, and chlorine in the cells closely followed the concentration in the storage solution, indicating that the intracellular concentration of these ions during storage is entirely dependent on diffusion processes. The calcium concentration was independent of the storage solution used. Studies by light and transmission electron microscopy showed good preservation of hepatocytes after storage for 8 and 12 hours in UW solution and its variants, modified KrebsRinger solution and Euro-Collins solution, but showed moderate damage to mitochondria and swelling of the endoplasmic reticulum in normal Krebs-Ringer solution. In addition, damage to the sinusoidal endothelial cells was observed after 4 hours in normal Krebs-Ringer solution and after 8 to 12 hours in the other solutions. In conclusion, the only factor determining the intracellular concentration of diffusible ions after cold tissue storage is the ionic composition of the extracellular medium. X-ray microanalysis provides an objective method for assessing whether the intracellular ionic composition of tissue is maintained during storage. (Liver Transpl 2003;9: 268-278.)A persistent problem in organ transplantation is the tissue damage that occurs during organ preservation. University of Wisconsin (UW) solution is considered one of the most effective cold storage solutions and allows successful preservation of the liver. 1-3 UW solution was first developed experimentally for pancreas transplantation 4,5 and only later applied to the preservation of liver, heart, 2,6 and lung. 7 Liver grafts have been preserved for as long as 24 hours, still offering a thriving transplantation and satisfactory bile production. 8 Liver transplantation has been made more effective with UW solution, and the time of preservation of the liver and pancreas has been significantly increased compared with earlier solutions used for simple cold storage. 9,10 Still, it has been suggested that a cold ischemia for more than 12 hours is a great hazard for the recipient. 11,12 It has been suggested that further developments of graft preservation solutions should concentrate on specific organ requirements and not on grafts in general. 2 ␤-Galactosidase has been proposed as a marker of ischemia-reperfusion injury in the liver because Kupffer ...

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“…For EMP, 16 ~tm cryosections were mounted on carbon specimen holders and freeze-dried (Wroblewski et al 1978). The analysis was carried out in a Jeol 100C electron microscope with Asid-4B scanning attachment, in the secondary mode at an accelerating voltage of 20 kV.…”

Section: Methodsmentioning

confidence: 99%

Quantitative correlative proton and electron microprobe analysis of biological specimens

Forslind

Kunst

Malmqvist³

et al. 1985

Histochemistry

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To investigate the possibility of quantitative correlative proton microprobe (PMP) and electron microprobe (EMP) analysis of biological soft tissue, model specimens were analyzed by both techniques. The specimens consisted of freeze-dried sections of gelatin containing known concentrations of nickel chloride. Both for PMP and for EMP, the signal was expressed as the ratio of the characteristic intensity and the continuum intensity in a peak-free region of the spectrum. With both techniques, calibration curves (signal versus known concentration) obtained, showed a deviation from linearity at high nickel concentrations. However, a linear relation (correlation coefficient 0.996) was obtained in a plot of EMP signal versus PMP signal. This indicates that quantitative correlative PMP and EMP analysis can be carried out by using the same standard for both analytical techniques.

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Electron probe X-ray microanalysis of human muscle biopsies

Cited by 74 publications

References 24 publications

In vitro and in situ experimental model for X‐ray microanalysis of intestinal epithelium

In vitro and in situ experimental model for X‐ray microanalysis of intestinal epithelium

Preservation of mouse liver tissue during cold storage in experimental solutions assessed by x-ray microanalysis

Quantitative correlative proton and electron microprobe analysis of biological specimens

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