Abstract The Physiology Core provides a series of graded technologies and expertise to investigators who wish to address fundamental questions about transport processes that occur in the tubular structures of the nephron and other relevant systems. To gain understanding of biological processes that mediate kidney function in health and disease states, in light of the phenotypic diversity and functional complexity of this organ, the use of strategies that allow specific nephron segments/cells/proteins to be studied in isolation under defined conditions is required. The Core, through its three Subcores, offers expertise and state-of-the-art technologies for studying the function and regulation of transport and other membrane resident proteins with progressive degrees of complexity from single molecules to model systems to native epithelia. The Single Tubule and Microperfusion Subcore provides tools and technologies to study the function and expression of transport proteins and associated regulatory pathways in single nephron segments isolated from small mammals and kidney organoids. The Electrophysiology Subcore provides expertise and a toolbox of electrophysiological approaches to study the function and regulation of transport proteins at the molecular and cellular levels in model systems and native epithelia. The Live-Cell Imaging Subcore offers imaging tools and technologies to assess ion transport, changes in membrane potential, cellular volume, intracellular mediators, and signaling molecules in immortalized cells and native preparations using chemical and genetically encoded fluorescent sensors. The Core will apply a multipronged approach and collaborate with the O’Brien Consortium to share specialized resources and technologies throughout the kidney research community, to lower the barrier to entry for early-stage investigators and researchers new to the kidney field, and to advance kidney research and discovery nationwide.