Abstract
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.
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Acknowledgments
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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Shibata, S. et al. (2015). Immuno-Electron Microscopy and Electron Microscopic In Situ Hybridization for Visualizing piRNA Biogenesis Bodies in Drosophila Ovaries. 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_12
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DOI: https://doi.org/10.1007/978-1-4939-2851-4_12
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