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 briefl y outside the female body. We identifi ed 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) |
|---|---|
| Pages (from-to) | 89-97 |
| Number of pages | 9 |
| Journal | Methods in Molecular Biology |
| Volume | 1328 |
| DOIs | |
| State | Published - 2015 |
Keywords
- Border cell migration
- Collective cell migration
- Drosophila stage 9 egg chambers
- Organ culture
- Time-lapse live imaging
ASJC Scopus subject areas
- Molecular Biology
- Genetics
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