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In Vitro Culturing and Live Imaging of Drosophila Egg Chambers: A History and Adaptable Method

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Oogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1457))

Abstract

The development of the Drosophila egg chamber encompasses a myriad of diverse germline and somatic events, and as such, the egg chamber has become a widely used and influential developmental model. Advantages of this system include physical accessibility, genetic tractability, and amenability to microscopy and live culturing, the last of which is the focus of this chapter. To provide adequate context, we summarize the structure of the Drosophila ovary and egg chamber, the morphogenetic events of oogenesis, the history of egg-chamber live culturing, and many of the important discoveries that this culturing has afforded. Subsequently, we discuss various culturing methods that have facilitated analyses of different stages of egg-chamber development and different types of cells within the egg chamber, and we present an optimized protocol for live culturing Drosophila egg chambers.

We designed this protocol for culturing late-stage Drosophila egg chambers and live imaging epithelial tube morphogenesis, but with appropriate modifications, it can be used to culture egg chambers of any stage. The protocol employs a liquid-permeable, weighted “blanket” to gently hold egg chambers against the coverslip in a glass-bottomed culture dish so the egg chambers can be imaged on an inverted microscope. This setup provides a more buffered, stable, culturing environment than previously published methods by using a larger volume of culture media, but the setup is also compatible with small volumes. This chapter should aid researchers in their efforts to culture and live-image Drosophila egg chambers, further augmenting the impressive power of this model system.

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Acknowledgements

We are grateful for support from the National Institutes of Health, 2 R01 GM079433 to CAB and 1S10 OD016240 to the W. M. Keck Imaging Center at the University of Washington. We thank the Bloomington Drosophila Stock Center and Trudi Schüpbach, Dan Kiehart, and Miriam Osterfield for fly strains.

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Authors and Affiliations

  1. Department of Genome Sciences, University of Washington, 3720 15th Ave NE, Seattle, WA, 98195-5065, USA

    Nathaniel C. Peters & Celeste A. Berg

  2. Department of Physiology and Biophysics, University of Washington, 1705 NE Pacific St, Seattle, WA, 98195-7290, USA

    Nathaniel C. Peters

Authors
  1. Nathaniel C. Peters
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  2. Celeste A. Berg
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Corresponding authors

Correspondence to Nathaniel C. Peters or Celeste A. Berg .

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Editors and Affiliations

  1. School of Life Sciences, University of Warwick, Coventry, United Kingdom

    Ioannis P. Nezis

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Peters, N.C., Berg, C.A. (2016). In Vitro Culturing and Live Imaging of Drosophila Egg Chambers: A History and Adaptable Method. In: Nezis, I. (eds) Oogenesis. Methods in Molecular Biology, vol 1457. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3795-0_4

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  • DOI: https://doi.org/10.1007/978-1-4939-3795-0_4

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3793-6

  • Online ISBN: 978-1-4939-3795-0

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