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.
Key wordsDrosophila Egg chamber Follicle Live imaging Collective cell migration Morphogenesis
We thank members of the Horne-Badovinac lab for input, Guillermina Ramirez-San Juan for the dissection video, and Claire Stevenson for the images in Fig. 1c. M.C. was supported by NIH T32 GM007183 and work in the Horne-Badovinac lab is supported by NIH R01 GM094276.
Drosophila ovary dissection. Video showing dissection of Drosophila ovaries using a stereomicroscope. Alternate dissection methods are shown for acquiring stage 6-8 or stage 1-5 egg chambers. After dissection, healthy ovarioles are sorted and older egg chambers are trimmed away. Please see Fig. 2 for stills of this video and a detailed procedural description.
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