Variations in morpho-anatomy, chemical composition, gas exchange and stem water potential (Ψstem) were studied in leaves of Prunus avium L. cultivars with different growth habit: Burlat and Summit (upright, dense canopies), and Van (spreading, open canopy). Light transmittance was highest in Van canopies. Therefore, Van allowed an easier penetration of the visible radiation through the canopy, which improves photosynthetic rates, especially at harvest. Conversely, no differences in Ψstem were found among cultivars. Van showed the highest leaf mass per unit area and variations in this parameter were mainly associated with alterations in leaf thickness than in leaf density. Leaves of Summit and Van were thicker than those of Burlat, mainly due to increased palisade and spongy parenchyma thickness. However, Burlat leaves had the highest palisade/spongy ratio. Total chlorophyll concentration per area and the chlorophyll a/b ratio were always higher in Van leaves than in the other two cultivars. In contrast, Van leaves had the lowest concentrations of soluble carbohydrates and starch. In dense canopies of Summit and Burlat, light reduction caused a notable increase in the relative concentration of total phenols. The above results provide useful information about the biological behaviour of sweet cherry cultivars with open and dense canopies in order to help the agronomical decisions regarding the local environmental conditions and the consumers’ preference. In particular, it is important to manipulate factors such as orchard exposure, row orientation, plant spacing, training system and pruning, which are able to reduce shading in the canopy.