Abstract
Border cell migration in the Drosophila ovary has emerged as a genetically tractable model for studying collective cell movement. Over many years border cell migration was exclusively studied in fixed samples due to the inability to culture stage 9 egg chambers in vitro. Although culturing late-stage egg chambers was long feasible, stage 9 egg chambers survived only briefly outside the female body. We identified culture conditions that support stage 9 egg chamber development and sustain complete migration of border cells ex vivo. This protocol enables one to compare the dynamics of egg chamber development in wild-type and mutant egg chambers using time-lapse microscopy and taking advantage of a multiposition microscope with a motorized imaging stage. In addition, this protocol has been successfully used in combination with fluorescence resonance energy transfer biosensors, photo-activatable proteins, and pharmacological agents and can be used with wide-field or confocal microscopes in either an upright or an inverted configuration.
| Original language | English (US) |
|---|---|
| Title of host publication | Drosophila Oogenesis: Methods and Protocols |
| Publisher | Springer New York |
| Pages | 89-97 |
| Number of pages | 9 |
| ISBN (Print) | 9781493928514, 9781493928507 |
| DOIs | |
| State | Published - Aug 31 2015 |
Keywords
- Border cell migration
- Collective cell migration
- Drosophila stage 9 egg chambers
- Organ culture
- Time-lapse live imaging
ASJC Scopus subject areas
- General Biochemistry, Genetics and Molecular Biology
- General Agricultural and Biological Sciences