Microspectrophotometry indicates the presence of at least three cone visual pigments in two Australian marsupials, the fat-tailed dunnart (Sminthopsis crassicaudata) and honey possum (Tarsipes rostratus). Here we have examined the distribution of cone types using antisera, JH455 and JH492, that recognize short-wavelength-sensitive (SWS) and medium-to-long-wavelength-sensitive (M/LWS) cone opsins, respectively. SWS cones were concentrated in dorso-temporal retina in the dunnart with a shallow decreasing gradient extending to the periphery (2300-1500/mm(2)). In the honey possum, SWS cones showed a uniform distribution (2700/mm(2)), except for a slight increase in a narrow peripheral band (3100/mm(2)). In both species, M/LWS cones dominated and their distributions were similar to those of retinal ganglion cells: a horizontal streak in the dunnart (31,000-21,000/mm(2)) and a shallow mid-ventral to peripheral gradient in the honey possum (37,000-26,000/mm(2)). A low number of cones remained unlabeled when the antisera were combined revealing further minority cone population(s). We discuss cone distributions in relation to visual capabilities and requirements of the species.