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Natal dens and neonatal behavior are a critical but understudied component of reproductive success in cryptic solitary carnivores. Mountain lion (<i>Puma concolor</i>) nursery sites tend to be in remote and difficult-to-observe areas-as a result, there are few in-depth behavioral observations of neonatal mountain lions. In this study, we present detailed opportunistic observations from a week of continuous monitoring of a mountain lion nursery located in the Gilbert-Baker Wildlife Management Area in northwestern Nebraska. Using a non-invasive video camera trap, we recorded 403 videos comprising 76 distinct bouts of activity. We confirmed the presence of four kittens during the first day of monitoring; however, subsequent recordings consistently captured three or fewer individuals-with most videos featuring one (50.7%) or two (34.8%) kittens. This variation demonstrates the inherent limitations of camera traps to reliably detect all individuals, potentially biasing abundance estimates that rely on repeated count data. Play (chasing, pouncing, wrestling, and tree climbing) and vocalizations (high-pitched contact calls by kittens and low growling purrs from the mother) were the most frequent behaviors we observed. We also documented allogrooming and autogrooming, as well as the mother carrying kittens in her mouth. Although no nursing or feeding was directly observed, prey remains found near the nursery on a subsequent visit confirmed maternal foraging and provisioning. Activity patterns were crepuscular overall but differed by mountain lion class and temporal resolution. Specifically, these patterns varied depending on whether we analyzed times from individual videos or the beginning of activity bouts, underscoring the importance of the data resolution used when analyzing camera trap data. Our findings demonstrate the value of camera traps for studying neonatal carnivore behavior and highlight the need for suitable nursery habitats as mountain lions recolonize portions of their former range.