Halimeda tuna (J. Ellis & Solander) J.V. Lamouroux is the only Halimeda species found in the Mediterranean Sea, and it is an important habitat former. In the northern Adriatic, H. tuna is among the ten most abundant seaweeds in the upper-infralittoral belt in spring and autumn. The modular thalli consist of serially arranged calcified segments. Calcification is closely related to photosynthesis, which causes alkalinization of the inter-utricular space and triggers aragonite formation. Understanding of the complex patterns of segment shape plasticity in relation to CaCO3content at different depth levels is still incomplete. Geometric morphometrics was used to investigate H. tuna segment shape variation on the depth transect at Cape Madona Nature Monument in the northern Adriatic Sea. The position on the thallus and the CaCO3 content of each studied segment were recorded, allowing slight changes in mineral content to be detected at the microscale of the segments. Our results showed that shape, size, or asymmetry of H. tuna segments were not significantly affected by depth. On the other hand, plants that grew deeper were generally more calcified. The apical and subapical segments contributed to the increase in CaCO3 content at the deeper sites, whereas the basal segments did not. This indicates that reniform or oval segments positioned apically or subapically play a key role in calcification of H. tuna in Mediterranean ecosystems.