Electron probe X‐ray microanalysis of intracellular element concentrations in cryosections in the presence of changes in cell volume

Abstract

SUMMARY The interpretation of element concentration data for X‐ray microanalyses of biological tissues, which are subjected to some experimental treatment, can be complicated by changes in cell volume and total cell dry matter induced by the treatment. We have examined the manner in which such changes would affect the values measured in frozen‐dried cryosections of soft tissues, and how they may be taken into account in the interpretation of the results. The element content (mass per unit dry weight) measured by the peak‐to‐continuum or Hall method is independent of changes in cell volume, but is sensitive to a change in the local dry mass. Conversely, intracellular concentrations in terms of mass per unit volume, as determined by the peripheral or internal standard technique, are dependent on volume changes but independent of dry mass. The estimated dry weight fraction is affected by changes in both volume and dry mass. The results obtained from both quantification methods can therefore provide information on the combination of changes in cellular element levels, volume and total dry mass that may occur following the experimental treatment. In a study of the late effect of the drug cisplatin on electrolyte concentrations in kidney proximal tubules, both quantification methods have been used to obtain wet weight and dry weight concentrations. By applying the above considerations, the analytical results have been interpreted as a combination of changes in element levels and a shrinkage of the tubule cells. Cell shrinkage was confirmed by morphometric analysis of tubular cross‐sections.

Effect of Atrial Natriuretic Peptide on Cellular Element Concentrations in Rat Proximal Tubules: Evidence for Inhibition of the Sodium Pump

“…Quantification of the cellular elemental concentration in mmol/ kg wet weight was achieved by a comparison of the element characteristic X-ray signals obtained in the nuclei of cells with those of the albumin standard. The dry weight of the cells was determined by comparing the background radiation of the cellular spectra with that of the standard albumin layer as described by Bostrom et al (1991). The same authors also observed a high degree of correlation between cell volume and total cell dry matter.…”