Advertisement

Live-Cell Imaging of the Adult Drosophila Ovary Using Confocal Microscopy

  • Nevine A. ShalabyEmail author
  • Michael BuszczakEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1463)

Abstract

The Drosophila ovary represents a key in vivo model used to study germline stem cell (GSC) maintenance and stem cell daughter differentiation because these cells and their somatic cell neighbors can be identified at single-cell resolution within their native environment. Here we describe a fluorescent-based technique for the acquisition of 4D datasets of the Drosophila ovariole for periods that can exceed 12 consecutive hours. Live-cell imaging facilitates the investigation of molecular and cellular dynamics that were not previously possible using still images.

Key words

Drosophila ovary Live-cell imaging Confocal microscopy Molecular dynamics Cellular dynamics 

Notes

Acknowledgement

We would like to thank P. Robin Hiesinger and Neset Ozel for their input and advice while developing this protocol.

References

  1. 1.
    Eliazer S, Buszczak M (2011) Finding a niche: studies from the Drosophila ovary. Stem Cell Res Ther 2:45CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Gilboa L (2015) Organizing stem cell units in the Drosophila ovary. Curr Opin Genet Dev 32:31–36CrossRefPubMedGoogle Scholar
  3. 3.
    Slaidina M, Lehmann R (2014) Translational control in germline stem cell development. J Cell Biol 207:13–21CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Fichelson P, Moch C, Ivanovitch K et al (2009) Live-imaging of single stem cells within their niche reveals that a U3snoRNP component segregates asymmetrically and is required for self-renewal in Drosophila. Nat Cell Biol 11:685–693CrossRefPubMedGoogle Scholar
  5. 5.
    Zhang Q, Shalaby NA, Buszczak M (2014) Changes in rRNA transcription influence proliferation and cell fate within a stem cell lineage. Science 343:298–301CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Nienhaus GU, Nienhaus K, Holzle A et al (2006) Photoconvertible fluorescent protein EosFP: biophysical properties and cell biology applications. Photochem Photobiol 82:351–358CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Molecular BiologyUniversity of Texas Southwestern Medical CenterDallasUSA
  2. 2.Institute for BiologyFreie Universität BerlinBerlinGermany

Personalised recommendations