Temporary rivers are characterized by shifting habitats between flowing, isolated pools, and dry phases. Despite the fact that temporary rivers are currently receiving increasing attention by researchers and managers, the isolated pools phase has been largely disregarded. However, isolated pools in temporary rivers are transitional habitats of major ecological relevance as they support aquatic ecosystems during no-flow periods, and can act as refugees for maintaining local and regional freshwater biodiversity. Pool characteristics such as surface water permanence and size, presence of predators, local physicochemical conditions, time since disconnection from the river flow, or distance to other freshwater habitats challenge a comprehensive understanding of the ecology of these habitats, and challenge ecological quality assessments and conservation practices in temporary rivers. In this paper, we aim at providing a characterization of isolated pools from a hydrological, geomorphological, physicochemical, biogeochemical, and biological point of view as a framework to better conceptualize, conserve, and manage these habitats.