The ecology and dispersal dynamics of unicolonial ant invaders are poorly understood. In this study, we investigated the genetic structure of a well-documented, invading population of Argentine ants in the Jasper Ridge Biological Preserve in northern California to examine the dispersal distances of reproductives, the direction and mode of population expansion, and changes in the genetic differentiation among nests over time. Using microsatellite data, we measure both traditional FST statistics and multi-locus ge- notype assignment distances to determine the patterns of genetic structure at three spatial scales: population-wide gene flow, population substructure, and mixing between neigh- boring nests. At the population level, there was little viscosity across the Jasper Ridge population, suggesting recent rapid expansion and/or considerable long-distance gene flow, presumably mediated by winged males. The pattern of genetic structure across distance indicates that the scale of queen dispersal was limited to less than 100 m. At the level of population substructure, hierarchical F statistics measures were low across subpopulations, locations within subpopulations, and nests. However, multi-locus genotype assignment tests revealed significant structure between subpopulations and between locations. Genetic dis- tances between nests were lower within locations than between locations, indicating that nests are most closely related to neighboring nests and that the expansion of subpopulations is primarily due to the local budding of new nests from existing nests at the invasion front. At the level of nest connectivity, the genetic differentiation among neighboring nests was associated with the time since invasion. As the invasion proceeds, nearby nests tend to be less closely related, indicating that extensive local mixing does not occur among more established nests. Our results show that the genetic structure of nests was not homogenous across unicolonial populations. Instead, the patterns of genetic structure reflect the limi- tations of and barriers to the dispersal of Argentine ant reproductives, the demographic history of the Jasper Ridge invasion, and changes in the genetic and ecological environment during the course of the invasion.