The Isolation and Properties of the Yeast Cell Vacuole

SUMMARY: When glucose and a chelating agent such as EDTA were added to yeast protoplasts suspended in 8.5% (w/v) mannitol solution at pH 6.4 almost all the protoplasts lysed and the main vacuole was released as a discrete structure from about 75% of the protoplasts. The isolated vacuoles were themselves sensitive to osmotic shock and exhibited vital-staining reactions. They are therefore considered to be relatively undamaged. Some of the properties of the isolated vacuoles are described.