Objective: The notion of distinguishable processing mechanisms for spatial and spatiotemporal information has largely been neglected in the context of navigation. Only a recent neuropsychological case study has provided initial evidence for the idea that these elements can be differentiated at a functional level. The aim of the current study was therefore to critically verify this double dissociation by adopting a systematic, large-scale approach. Method: Sixty-five chronic stroke patients and 60 matched healthy controls watched a route through a realistic virtual environment. They were assessed on their knowledge of this route in 4 different tasks after the learning phase. Performance on the scene recognition and route continuation tasks was taken as an indication of knowledge of the spatial route aspects. By contrast, spatiotemporal knowledge of the route was assessed in the route order and route progression tasks. Results: Based on single case statistics, 6 patients showed an exceptionally large difference in their performance on the spatial and spatiotemporal tasks. Moreover, 2 patients satisfied formal criteria for a classical dissociation. Conclusions: Our findings showed that spatial and spatiotemporal performance was closely associated in most patients. Nonetheless, the study also provided partial support for the notion of separate space- and time-based processing mechanisms in the context of navigation. This distinction is of particular relevance to the investigation into the cognitive structure underlying navigation behavior.