A protocol for culturing drosophila melanogaster stage 9 egg chambers for live imaging

Mohit Prasad, Anna C.C. Jang, Michelle Starz-Gaiano, Mariana Melani, Denise J. Montell

Research output: Contribution to journalArticlepeer-review

112 Citations (Scopus)

Abstract

This protocol describes a method for the dissection of egg chambers from intact Drosophila females and culture conditions that permit live imaging of them, with a particular emphasis on stage 9. This stage of development is characterized by oocyte growth and patterning, outer follicle cell rearrangement and migration of border cells. Although in vitro culture of egg chambers of later developmental stages has long been possible, until recently stage 9 egg chambers could only be kept alive for short periods, did not develop normally, and border cell migration failed entirely. We have established culture conditions that support overall egg chamber development including border cell migration in vitro. This protocol makes possible direct observation of molecular and cellular dynamics in both wild-type and mutant egg chambers, and opens the door to testing of pharmacological inhibitors and the use of biosensors. The entire protocol takes ˜24 h while the preparation of egg chambers for live imaging requires only 15-20 min.

Original languageEnglish
Pages (from-to)2467-2473
Number of pages7
JournalNature Protocols
Volume2
Issue number10
DOIs
Publication statusPublished - 2007 Oct

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

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