Cultivation and live imaging of drosophila ovaries

Drosophila egg chamber development depends on a number of dynamic cellular processes that contribute to the final shape and function of the egg. We can gain insight into the mechanisms underlying these events by combining the power of Drosophila genetics and ex vivo live imaging. During developmental stages 1–8, egg chambers rotate around their anterior-posterior axes due to collective migration of the follicular epithelium. This motion is required for the proper elongation of the egg chamber. Here, we describe how to prepare stage 1–8 egg chambers for live imaging. We provide alternate protocols for the use of inverted or upright microscopes and describe ways to stabilize egg chambers to reduce drift during imaging. We discuss the advantages and limitations of these methods to assist the researcher in choosing an appropriate method based on experimental need and available resources.