ABSTRACTThe importance of connectivity for freshwater organisms is widely recognised, yet in‐stream barriers associated with population declines and increased risk of extinction remain globally ubiquitous. Despite their negative consequences, these barriers can protect aquatic communities by limiting the spread of invasive species, leading to conflicting management goals in some regions. Selective fish passage is a solution for the conflicting goals of passing native, desirable species while restricting the spread of invasives. Approaches that can target groups of species sharing similar attributes (i.e. guilds) are likely to be more efficient than those that target species individually, particularly in taxonomically diverse systems. We explored the guild structure of 220 Great Lakes freshwater fishes based on morphological, phenological, physiological and behavioural attributes associated with passage and movement. We identified five distinct guilds as well as the attributes most important for defining these groupings: maximum total length, trophic level, relative eye size, spawning temperature, spawning season, presence/absence of ampullary electroreceptors and the presence/absence of hearing specialisations. The approaches outlined in this work can be generalised to enhance selective fish passage in aquatic ecosystems worldwide.