Abstract : Designing 3D User Interfaces (UI) requires adequate evaluation tools to ensure good usability and user experience. While many evaluation tools are already available and widely used, existing approaches generally cannot provide continuous and objective measures of usa-bility qualities during interaction without interrupting the user. In this paper, we propose to use brain (with ElectroEncephaloGraphy) and physiological (ElectroCardioGraphy, Galvanic Skin Response) signals to continuously assess the mental effort made by the user to perform 3D object manipulation tasks. We first show how this mental effort (a.k.a., mental workload) can be estimated from such signals, and then measure it on 8 participants during an actual 3D object manipulation task with an input device known as the CubTile. Our results suggest that monitoring workload enables us to continuously assess the 3DUI and/or interaction technique ease-of-use. Overall, this suggests that this new measure could become a useful addition to the repertoire of available evaluation tools, enabling a finer grain assessment of the ergonomic qualities of a given 3D user interface.