Cultivation and Live Imaging of Drosophila Imaginal Discs

In this chapter, I present a method for the ex vivo cultivation and live imaging of Drosophila imaginal disc explants using low concentrations of the steroid hormone 20-hydroxyecdysone (20E). This method has been optimized for analyzing cellular dynamics during wing disc growth and leverages recent insights from in vivo experiments demonstrating that 20E is required for growth and patterning of the imaginal tissues. Using this protocol, we directly observe wing disc proliferation at a rapid rate for at least 13 h during live imaging. The orientation of tissue growth is also consistent with that inferred from indirect in vivo techniques. Thus, this method provides an improved way of studying dynamic cellular processes and tissue movements during imaginal disc development. I first describe the preparation of the growth medium and the dissection, and then I include a protocol for mounting and live imaging of the explants.