Cells require ordered movement of proteins and lipids from one membrane-bound compartment to another. Clathrin-coated vesicles are the most prominent carriers of membrane traffic from cell surface to intracellular organelles (endocytosis), a pathway by which hormones, nutrients, viruses, and their receptors enter cells. They are also important for traffic between organelles. In this presentation, I will discuss (i) technological and analytical advances that I developed to directly visualize clathrin-mediated membrane traffic in three dimensions and in living cells; (ii) data obtained using these advances that defined the mechanism used by cells to counteract membrane tension during clathrin-coated vesicle budding; and (iii) how these advances can be used to study a wide variety of biological processes that occur in living cells and tissues.
