An increasing number of studies have found tolerance variation in populations consistently exposed to contaminants, but few studies have examined whether these laboratory-derived estimates of tolerance have survival implications in field conditions. We examined four populations of the mayfly Stenacron interpunctatum for variation in tolerance to the common agricultural insecticide clothianidin. Using laboratory bioassays, we found a 2.3× range in 96 h EC50 tolerance values to clothianidin between our four populations. We then conducted a common-garden experiment with nymphs from each population placed into the collection stream most heavily impacted by upstream agricultural activities to assess whether our laboratory tolerance estimates predict survival under field conditions. We monitored survival and growth in situ for three weeks during the spring planting season, when clothianidin is applied to croplands upstream of our study site. While growth was similar across all groups, the most tolerant population, which was native to the impacted stream, had higher survival than the more sensitive populations. This suggests that population-level variation in contaminant tolerance as measured in laboratory bioassays could have real-world survival implications for sensitive aquatic macroinvertebrates in contaminated streams.