Scanning electron microscopy was used to investigate leaf epicuticular wax of Prunus instititia L. ‘Pixie’ from aseptically cultured plants before and after acclimatization to the greenhouse. Leaves from plants acclimatized for 2 weeks in the greenhouse had more adaxial wax than those from non-acclimatized (culture flask-grown) plants. Acclimatized plants had more adaxial than abaxial wax. No abaxial wax was observed on leaves of non-acclimatized plants. Stomata were present on the abaxial leaf surface only of both acclimatized and non-acclimatized plants. Epicuticular wax layers surrounded guard cells of acclimatized plant leaves but were not present on non-acclimatized plant leaves. Weight changes in non-acclimatized plant leaves coated with silicon rubber on adaxial, abaxial, and both surfaces indicated that excised leaf water loss occurred only through the abaxial surface. Water loss from plants during the acclimatization process thus may be due to abaxial cuticular and stomatal transpiration.