Electron microprobe study of the effect of long‐term vacuum storage of freeze‐dried cryosections on distribution of elements in cells

Abstract

The distribution of elements in yeast cells was measured on freshly prepared, freeze-dried cryosections and compared with the distribution obtained on the same sections after storage for 20 months in a vacuum below 2.6 kPa. The average concentration of phosphorus remained unchanged but was equalized throughout the cells, i.e. it migrated from vacuoles into the cytoplasm and mitochondria. Zinc remained preferentially localized in the vacuoles, but the ratio between vacuolar and cytoplasmic zinc concentrations decreased about three-fold. Cytoplasmic and mitochondrial concentrations of potassium remained unchanged while partial release from the vacuoles and subsequently from the cells was observed. This resulted in a homogeneous distribution of potassium in the cells after 20 months and some of the vacuolar potassium appeared in spectra of formvar film measured several micrometres from the cells. A large increase in the sodium (from 160 to 360% more than in fresh sections), magnesium (from 110 to 200% more) and sulphur (from 70 to 350% more) contents was observed in all cellular compartments (except for vacuoles, where only a 20% increase in the magnesium content was observed), while chlorine was almost completely released from the cells. The limitations of the use of long-term vacuum-stored cryosections for electron microprobe analysis of cells are discussed.