title: Cultivation and Live Imaging of Drosophila Ovaries.
abstract: 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.
authors.affiliation: Department of Molecular Genetics and Cell Biology, The University of Chicago, 920 East 58th Street, Chicago, IL, 60637, USA.|Committee on Development, Regeneration and Stem Cell Biology, The University of Chicago, 920 East 58th Street, Chicago, IL, 60637, USA.
authors.forename: Maureen
authors.initials: M
authors.lastname: Cetera
authors.affiliation: Department of Biological Sciences, Butler University, 4600 Sunset Boulevard, Indianapolis, IN, 46208, USA.
authors.forename: Lindsay
authors.initials: L
authors.lastname: Lewellyn
authors.affiliation: Department of Molecular Genetics and Cell Biology, The University of Chicago, 920 East 58th Street, Chicago, IL, 60637, USA. shorne@uchicago.edu.|Committee on Development, Regeneration and Stem Cell Biology, The University of Chicago, 920 East 58th Street, Chicago, IL, 60637, USA. shorne@uchicago.edu.
authors.forename: Sally
authors.initials: S
authors.lastname: Horne-Badovinac
mesh_terms: D000818:Animals; D003470:Culture Media / Q000494:pharmacology; D004331:Drosophila melanogaster / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure; D005260:Female; D007328:Insulin / Q000494:pharmacology; D007814:Larva / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure; D009866:Oogenesis / Q000235:genetics; D061848:Optical Imaging / Q000379:methods*; D010053:Ovary / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure*; D046509:Tissue Culture Techniques; D015053:Zygote / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism / Q000648:ultrastructure*
chemical_list: D003470:Culture Media; D007328:Insulin
keywords: Collective cell migration; Drosophila; Egg chamber; Follicle; Live imaging; Morphogenesis