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Oogenesis pp 19-33 | Cite as

Antibody Staining in Drosophila Germaria

  • Anette Lie-Jensen
  • Kaisa HaglundEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1457)

Abstract

Drosophila oogenesis is a powerful model for studying a wide spectrum of cellular and developmental processes in vivo. Oogenesis starts in a specialized structure called the germarium, which harbors the stem cells for both germ and somatic cells. The germarium produces egg chambers, each of which will develop into an egg. Active areas of research in Drosophila germaria include stem cell self-renewal, division, and maintenance, cell cycle control and differentiation, oocyte specification, intercellular communication, and signaling, among others. The solid knowledge base, the genetic tractability of the Drosophila model, as well as the availability and fast development of tools and imaging techniques for oogenesis research ensure that studies in this model will keep being instrumental for novel discoveries within cell and developmental biology also in the future. This chapter focuses on antibody staining in Drosophila germaria and provides a protocol for immunostaining as well as an overview of commonly used antibodies for visualization of different cell types and cellular structures. The protocol is well-suited for subsequent confocal microscopy analyses, and in addition we present key adaptations of the protocol that are useful when performing structured illumination microscopy (SIM) super-resolution imaging.

Key words

Drosophila oogenesis Germarium Germline stem cell Germline cyst Fusome Ring canal Follicle cell Antibody staining Structured illumination microscopy 

Notes

Acknowledgments

We thank Vigdis Sørensen for assistance with 3D SIM super-resolution imaging, David Glover for providing the w1118; p[w + Ub-GFP-Pav-KLP]53 fly line, and the Developmental Studies Hybridoma Bank (DSHB) for antibodies. K.H acknowledges a career researcher grant from the South-Eastern Norway Regional Health Authority, project number 2012054. A.L-J is supported by this grant. This work was partly supported by the Research Council of Norway through its Centres of Excellence funding scheme, project number 179571. The confocal microscopy core facility at Oslo University Hospital is acknowledged for access to confocal microscopes. The super-resolution microscopy core facility at Oslo University Hospital is acknowledged for access to a DeltaVision OMX microscope.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of Molecular Cell Biology, Institute for Cancer ResearchOslo University HospitalOsloNorway
  2. 2.Centre for Cancer Biomedicine, Faculty of MedicineUniversity of OsloOsloNorway

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