Analyzing gaze behavior with dynamic stimulus material is of growing importance in experimental psychology; however, there is still a lack of efficient analysis tools that are able to handle dynamically changing areas of interest. In this article, we present DynAOI, an open-source tool that allows for the definition of dynamic areas of interest. It works automatically with animations that are based on virtual three-dimensional models. When one is working with videos of real-world scenes, a three-dimensional model of the relevant content needs to be created first. The recorded eye-movement data are matched with the static and dynamic objects in the model underlying the video content, thus creating static and dynamic areas of interest. A validation study asking participants to track particular objects demonstrated that DynAOI is an efficient tool for handling dynamic areas of interest.