Immuno-Electron Microscopy and Electron Microscopic In Situ Hybridization for Visualizing piRNA Biogenesis Bodies in Drosophila Ovaries

  • Shinsuke ShibataEmail author
  • Yukiko Murota
  • Yoshinori Nishimoto
  • Mana Yoshimura
  • Toshihiro Nagai
  • Hideyuki Okano
  • Mikiko C. SiomiEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1328)


Immuno-electron microscopy and electron microscopic in situ hybridization are powerful tools to identify the precise subcellular localization of specific proteins and RNAs at the ultramicroscopic level. Here we describe detailed procedures for how to detect the precise location of a specific target labeled with both fluorescence and gold particles. Although they have been developed for the analysis of Drosophila ovarian somatic cells, these techniques are suitable for a wide range of biological applications including human, primate, and rodent analysis.


Yb Zuc flam PIWI-interacting RNAs piRNAs Electron microscopy Immuno-electron microscopy In situ hybridization ISH Electron microscopic in situ hybridization EM-ISH Fluorescence immunohistochemistry 



We are grateful to Dr. S. Nakagawa for providing insightful discussions and dedicated support to our project. We thank T. Yano at Electron microscope laboratory and G. Itai at Keio-med Open Access Facility for their special technical support and also thank all members of the Siomi and Okano laboratories for their invaluable comments. This work was supported by a Grant-in-Aid for Scientific Research from MEXT, Japan; a grant from Keio Gijuku Academic Development Funds to S.S.; and a grant from Brain Mapping by Integrated Neurotechnologies for Disease Studies (Brain/MINDS) to S.S. and H.O. The authors have no conflicts of interest to declare.


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Shinsuke Shibata
    • 1
    Email author
  • Yukiko Murota
    • 2
  • Yoshinori Nishimoto
    • 1
  • Mana Yoshimura
    • 1
  • Toshihiro Nagai
    • 3
  • Hideyuki Okano
    • 1
  • Mikiko C. Siomi
    • 2
    Email author
  1. 1.Department of Physiology, School of MedicineKeio UniversityTokyoJapan
  2. 2.Department of Biological Sciences, Graduate School of ScienceThe University of TokyoTokyoJapan
  3. 3.Electron Microscope Laboratory, School of MedicineKeio UniversityTokyoJapan

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