Surveys of benthic invertebrates have revealed patterns attributed to the impacts of acid deposition. Unfortunately, these patterns may be confounded by temporal variation that will affect follow-up studies of the recovery of these communities. Here, we assess spatial and temporal variation in time-limited, kick-and-sweep collections of littoral-zone benthos. Spatial variation comprised five sites representing the predominant nearshore substrates in each of three lakes. Temporal variation spanned a different scale in each lake with five sites sampled: (i) twice on the same day, (ii) once a week for 3 weeks, and (iii) four times through the ice-free season. Variation was quantified using a model II analysis of variance. Spatial differences predominated in same-day samples (60.4% of the variation on average) and those collected over a 3-week period (46.1%). In contrast, samples collected over the ice-free season revealed that spatial and temporal factors accounted for 9.4 and 25.6% of the variation. We conclude that our collections of littoral macrobenthos are highly repeatable if sampling is restricted to short periods (e.g., 3 weeks). Surveys spanning longer periods may incorporate considerable temporal variation from seasonal changes in abundance.