Neighborhood deprivation plays an important role in childhood health and development, but defining the appropriate neighborhood definition presents theoretical as well as practical challenges. Few studies have compared neighborhood definitions outside of highly urbanized settings. The purpose of the current study was to evaluate how various administrative and ego-centric neighborhood definitions may impact measured exposure to deprivation across the urban-rural continuum. We do so using the Family Life Project, a prospective longitudinal population-based sample of families living in North Carolina and Pennsylvania (USA), which also sets the stage for future investigations of neighborhood impacts on childhood health and development. To measure neighborhood deprivation, a standardized index of socioeconomic deprivation was calculated using data from the 2007-2011 American Community Survey. Families’ residential addresses when children were 2 months of age (n=1036) were geocoded and overlaid onto a deprivation index layer created at the census block group level to construct multiple administrative and ego-centric neighborhood definitions. Friedman tests were used to compare distributions of neighborhood deprivation across these neighborhood definitions within urbanized areas, urban clusters, and rural areas. Results indicated differences in urbanized areas (Chisquare= 897.75, P<0.001) and urban clusters (Chi-square=687.83, P<0.001), but not in rural areas (Chi-square=13.52, P=0.332). Findings imply that in urban areas, choice of neighborhood definition impacts measured exposure to neighborhood deprivation. Although exposure to neighborhood deprivation appears to be less sensitive to neighborhood definition in rural areas, researchers should apply theoretical reasoning to choose appropriate definitions of children’s neighborhood.