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Nuclear Bodies and Noncoding RNAs pp 105–118Cite as

Ultrastructural Analysis of Nuclear Bodies Using Electron Microscopy

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1262))

Abstract

Recent immunofluorescent (IF) studies have discovered a variety of nuclear foci that have no known ultrastructurally defined counterpart. Using antibodies as ligands, immuno-electron microscopy (I-EM) is the method of choice for high-resolution recognition of these newly described nuclear compartments. However, noncoding RNAs (ncRNAs) have also been shown to be frequent components, sometimes essential, of nuclear bodies so that electron microscopic in situ hybridization (EM-ISH) can be used as an alternative means to characterize nuclear foci at the EM level. Among the array of protocols available, Lowicryl embedding of chemically fixed cells allows for high preservation of both nuclear structures and antigenicity and provides stable cell and tissue samples that can be re-probed whenever new antibodies or probes become available. Rapid and robust protocols are available for both I-EM and EM-ISH post-embedding techniques so that they can be combined on the same sections, providing ultrastructural and molecular insights into newly “emerging” nuclear bodies.

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Acknowledgment

We thank Archa Fox for the kind gift of PSPC1 antibody, Christian Lavialle for critical reading, and the CNRS and the Association pour la Recherche sur le Cancer (ARC) for funding.

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Authors and Affiliations

  1. Functional Organization of the Cell, CNRS-UMR8122, Institut Gustave Roussy, 114, rue Edouard-Vaillant, 94805, Villejuif-Cedex, France

    Sylvie Souquere & Gérard Pierron

Authors
  1. Sylvie Souquere
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  2. Gérard Pierron
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Corresponding author

Correspondence to Gérard Pierron .

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Editors and Affiliations

  1. RNA Biology Laboratory, RIKEN, Saitama, Japan

    Shinichi Nakagawa

  2. Institute for Genetic Medicine, Hokkaido University, Sapporo, Japan

    Tetsuro Hirose

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Souquere, S., Pierron, G. (2015). Ultrastructural Analysis of Nuclear Bodies Using Electron Microscopy. In: Nakagawa, S., Hirose, T. (eds) Nuclear Bodies and Noncoding RNAs. Methods in Molecular Biology, vol 1262. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2253-6_7

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  • DOI: https://doi.org/10.1007/978-1-4939-2253-6_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2252-9

  • Online ISBN: 978-1-4939-2253-6

  • eBook Packages: Springer Protocols

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