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

  • Mohit Prasad
  • Xiaobo Wang
  • Li He
  • Danfeng Cai
  • Denise J. MontellEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1328)

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.

Keywords

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

Notes

Acknowledgements

This work was supported by the National Institute of General Medical Sciences grant GM73164 to D.J.M.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Mohit Prasad
    • 1
    • 2
  • Xiaobo Wang
    • 1
    • 3
  • Li He
    • 1
    • 4
  • Danfeng Cai
    • 1
    • 5
  • Denise J. Montell
    • 1
    • 5
    Email author
  1. 1.Department of Biological ChemistryJohns Hopkins School of MedicineBaltimoreUSA
  2. 2.Department of Biological SciencesIISER KolkataMohanpurIndia
  3. 3.Université P. Sabatier Toulouse IIIToulouse cedex 9France
  4. 4.Department of GeneticsHarvard Medical SchoolBostonUSA
  5. 5.Molecular, Cellular and Developmental Biology DepartmentUniversity of CaliforniaSanta BarbaraUSA

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