Abstract
Guard cell protoplasts were prepared from young leaves of pea plants. Under hypertonic conditions they shrink and large numbers of endocytotic («osmocytotic») vacuoles are formed by invagination of the plasma membrane. In thin section these are indistinguishable from other small vacuoles («mini-vacuoles») which are formed by fragmentation of the large central vacuole. However, the two types of vacuole can be individually recognized by labelling the central vacuole with neutral red and by performing the osmotic shrinkage with fluorochromes such as Lucifer Yellow-CH or Cascade Blue present in the extracellular medium. Osmocytotic vacuoles do not fuse with the plasma membrane nor with the mini-vacuoles during a subsequent swelling phase
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