Abstract Species of didelphid marsupials (Didelphimorphia, Didelphidae) differ in their use of the forest strata, but it is not clear whether these differences are in fundamental or realized niches. The fundamental niche of seven species of didelphids (Caluromys philander, Didelphis aurita, Gracilinanus microtarsus, Marmosops incanus, Metachirus nudicaudatus, Micoureus demerarae, and Philander frenatus) was compared using their performance in arboreal walking. The association between performance and vertical use of the forest also was tested accounting for phylogenetic and allometric effects. Tests consisted of making the animal cross five 3 m long horizontal supports of different diameters, 1 m from the ground. The cycle of maximum speed was chosen to measure stride length, frequency and velocity. Arboreal species performed better than the terrestrial ones, but a major part of the variation in stride length (70.95%) and stride frequency (88.10%) was associated with body size. Part of the variation in stride length independent of body size (14.05%) was associated with the degree of vertical use of the forest, after phylogenetic effects were accounted for. Fundamental niches of six of the seven species were discriminated with the performance tests used. Discrepancies between the realized and fundamental niches can be inferred for two of these species, D. aurita and P. frenatus.