The first direct observation, by atom probe tomography, of a deuterium dead layer is reported for Fe/V multilayered film loaded with D solute atoms. The thickness of the dead layers was measured to be 0.4–0.5 nm. The dead layers could be distinguished from chemically intermixed layers. The results suggest that the dead layer effect occurs even near the interface of the mixing layers, supporting an interpretation that the dead layer effect cannot be explained solely by electronic charge transfer but also involves a modulation of rigidity.