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.
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Acknowledgements
This work was supported by the National Institute of General Medical Sciences grant GM73164 to D.J.M.
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Prasad, M., Wang, X., He, L., Cai, D., Montell, D.J. (2015). Border Cell Migration: A Model System for Live Imaging and Genetic Analysis of Collective Cell Movement. In: Bratu, D., McNeil, G. (eds) Drosophila Oogenesis. Methods in Molecular Biology, vol 1328. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2851-4_6
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DOI: https://doi.org/10.1007/978-1-4939-2851-4_6
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2850-7
Online ISBN: 978-1-4939-2851-4
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