Ecological regions are areas of similar climate, landform, soil, potential natural vegetation, hydrology, or other ecologically relevant variables. The makeup of aquatic biological assemblages (e.g., fish, macroinvertebrates, algae, riparian birds, etc.) varies dramatically over the landscape, as do the environmental stresses that affect the condition of those assemblages. Ecoregions delineate areas where similar assemblages are likely to occur and, therefore, where similar expectations can be established. For this reason, ecological regions have proven to be an important tool for use in the process of ecological assessment. This article describes four examples of the use of ecological regions in important aspects of environmental monitoring and assessment: (1) design of monitoring networks; (2) estimating expected conditions (criteria development); (3) reporting of results; (4) setting priorities for future monitoring and restoration. By delineating geographic areas with similar characteristics, ecological regions provide a framework for developing relevant indicators, setting expectations through the use of regional reference sites, establishing ecoregion-specific criteria and/or standards, presenting results, focusing models based on relationships between landscape and surface water metrics, and setting regional priorities for management and restoration. The Environmental Protection Agency and many state environmental departments currently use ecoregions to aid the development of environmental criteria, to illustrate current environmental condition, and to guide efforts to maintain and restore physical, chemical and biological integrity in lakes, streams, and rivers.