Characterizing macroinvertebrate taxa as either sensitive or tolerant is of critical importance for investigating impacts of anthropogenic stressors in aquatic ecosystems and for inferring causality. However, our understanding of relative sensitivity of aquatic insects to metals in the field and under controlled conditions in the laboratory or mesocosm experiments is limited. In this study, we compared the response of 16 lotic macroinvertebrate families to metals in short-term (10-day) stream mesocosm experiments and in a spatially extensive field study of 154 Colorado streams. Comparisons of field and mesocosm-derived EC20 (effect concentration of 20%) values showed that aquatic insects were generally more sensitive to metals in the field. Although the ranked sensitivity to metals was similar for many families, we observed large differences between field and mesocosm responses for some groups (e.g., Baetidae and Heptageniidae). These differences most likely resulted from the inability of short-term experiments to account for factors such as dietary exposure to metals, rapid recolonization in the field, and effects of metals on sensitive life stages. Understanding mechanisms responsible for differences among field, mesocosm, and laboratory approaches would improve our ability to predict contaminant effects and establish ecologically meaningful water-quality criteria.