title: Ultrastructural analysis of nuclear bodies using electron microscopy.
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.
authors.affiliation: Functional Organization of the Cell, CNRS-UMR8122, Institut Gustave Roussy, 114, rue Edouard-Vaillant, 94805, Villejuif-Cedex, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Souquere
authors.forename: Gérard
authors.initials: G
authors.lastname: Pierron
mesh_terms: D004247:DNA / Q000648:ultrastructure; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D045586:Intranuclear Inclusion Bodies / Q000648:ultrastructure*; D008854:Microscopy, Electron / Q000379:methods*; D012313:RNA / Q000648:ultrastructure; D022661:RNA, Untranslated
chemical_list: D022661:RNA, Untranslated; D012313:RNA; D004247:DNA