Preserving functional connectivity is a key goal of conservation management. However, the spatially confined conservation areas may not allow for dispersal and gene flow for the intended long-term persistence of populations in fragmented landscapes. We provide a regional multi-species assessment to quantify functional connectivity for five amphibian species in a human dominated landscape in the Swiss lowlands. A set of resistance maps were derived based on expert opinion and a sensitivity analysis was conducted to compare the effect of each resistance scenario on modelled connectivity. Deriving multi-species corridors is a robust way to identify movement hotspots that provide valuable baseline information to reinforce protective measures and green infrastructure.