In eukaryotes, lipids are not only an important constituent of the plasma membrane but are also used to generate specialized membrane-bound organelles, including temporary compartments with critical functions. As such, lipids play a key role in intracellular homeostasis—the ability of a cell to maintain stable internal conditions upon changes in its extracellular environment. Autophagy, one of the cellular processes through which eukaryotic cells strive for survival under stress, is heavily dependent on lipid and membrane trafficking through the de novo formation of autophagosomes—temporary, large, and double-bilayered organelles in which materials are encapsulated for recycling. This chapter discusses what we know about lipid homeostasis and trafficking during autophagy and autophagosome formation, and comments on future directions of the field.