Experimental manipulations of streamflow have been used globally in recent decades to mitigate the impacts of dam operations on river systems. Rivers are challenging subjects for experimentation, because they are open systems that cannot be isolated from their social context. We identify principles to address the challenges of conducting effective large-scale flow experiments. Flow experiments have both scientific and social value when they help to resolve specific questions about the ecological action of flow with a clear nexus to water policies and decisions. Water managers must integrate new information into operating policies for large-scale experiments to be effective. Modeling and monitoring can be integrated with experiments to analyze long-term ecological responses. Experimental design should include spatially extensive observations and well-defined, repeated treatments. Large-scale flow manipulations are only a part of dam operations that affect river systems. Scientists can ensure that experimental manipulations continue to be a valuable approach for the scientifically based management of river systems.biological conditions in these systems may not be attributed solely to the level of streamflow during the experiment. Unlike experiments on land, lakes, and small streams in experimental watersheds, flow manipulations involving whole rivers or estuaries can rarely, if ever, be isolated from their social context. Stakeholders have diverse interests in how water is used, and water managers operate facilities and systems to achieve multiple objectives. The overarching issue for scientists involved in large-scale flow experiments, then, is to design scientifically credible and tractable investigations that simultaneously inform water management about policies to achieve long-term objectives.We review the global practice of flow manipulations in rivers as large-scale experiments to guide future efforts in this burgeoning area of interest using examples from over 40 systems (see the supplementary material, available online at http: //dx.doi.org/10.1525/bio.2011.61.12.5). We focus on flow manipulations intended to achieve ecological objectives because of their direct relevance for informing dam operations but recognize that investigations of natural flow events and manipulations not intended for ecological outcomes provide useful information for managing rivers and advancing river ecology. We identify how flow experiments have elucidated and addressed facets of the complexity in river, floodplain, and estuary ecosystems. These examples lead us to a core set of challenges and principles for conducting effective large-scale flow experiments that have both scientific and social value.