Protocol

Cell Migration

Volume 769 of the series Methods in Molecular Biology pp 277-286

Date:

Border Cell Migration: A Model System for Live Imaging and Genetic Analysis of Collective Cell Movement

  • Mohit Prasad
  • , Xiaobo Wang
  • , Li He
  • , Denise J. MontellAffiliated withDepartment of Biological Chemistry, Johns Hopkins University School of Medicine Email author 

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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 widefield or confocal microscopes in either an upright or inverted configuration.

Key words

Border cell migration Drosophila stage 9 egg chambers Organ culture Collective cell migration Time-lapse live imaging