Subcellular localization of diffusible ions in the yeast Saccharomyces Cerevisiae: quantitative microprobe analysis of thin freeze-dried sections

Abstract

In yeast cells, of which the intracellular potassium had been partly replaced by rubidium or caesium, the intracellular ion distribution was studied by means of energy-dispersive X-ray microanalysis. The cells were rapidly frozen and thin sections were cut at low temperature on a cryo-ultramicrotome without the use of a trough liquid. By this dry cryosectioning procedure, complete retention of the diffusible ions in the cells was obtained. Unless the sections had been exposed to moisture, no signs of redistribution were apparent. For quantitative determinations a gelatin standard, containing known amounts of the elements of interest, was prepared in the same way as the cells. The concentrations of potassium, rubidium, caesium and chloride in the nucleus, the cytoplasm and the vacuole could be measured. The intracellular distributions of potassium, rubidium and caesium were very similar. The concentrations of these ions in the cytoplasm were about equal to those in the nucleus and twice those in the vacuole. The total concentration in the cytoplasm was 180–190 mmol/kg fresh weight, in the nucleus 190—200 mmol/kg fresh weight and in the vacuole 75–0 mmol/kg fresh weight. The permeability of the yeast cell for chloride is markedly lower than for the cations.