Information about reproductive habitat and migration pathways is of paramount importance to restore migratory fish species. This study assesses the availability of spawning and nursery habitats for the European sturgeon (Acipenser sturio) in the delta and lower Rhine (covering over 350 river kilometres) as part of a larger feasibility assessment for a future restoration of this critically endangered species. The general approach has three steps: (1) the identification of the species' specific habitat requirements, based on a systematic literature review; (2) the collection and preprocessing of data from two countries, including the 1D and 2D modelling of water depths and flow velocities; and (3) GIS-based mapping of spawning and nursery habitat. Based on a HSI score of 1, we identify a total of 0.75 km2 as minimal spawning habitat, potentially suitable for approximately 2500 female European sturgeons (one spawning site would use ~300 m2). This is sufficient, as currently, only an estimated maximum number of 750 adults exist. Suitable spawning habitat is mainly located in the German state of North Rhine-Westphalia, whereas suitable nursery habitat is mainly located in the Netherlands. The availability is, however, significantly reduced by coastal infrastructure (damming) and inland navigation. The insights gained can be used to assess the current suitability of the river Rhine for the species' reintroduction and to identify opportunities for habitat restoration and protection for various life stages. The outcomes thus play an essential role in the conservation of the species. In addition, the modelling approach developed could be applied to other northwestern European rivers. This broader application would allow intercomparison and support decisions about which rivers are best suited for future reintroduction of the critically endangered European sturgeon.