Nucleic acid detection by in situ molecular immunogold labeling procedures.
We have recently combined immunogold labeling procedures with molecular biology methods to pinpoint the precise locations of nucleic acids in biological material at the ultrastructural level. These new immunocytological approaches involve the incorporation of labeled nucleotides in the nucleic acids present at the surface of ultrathin sections prior to immunogold labeling. The antibodies used recognize a nucleoside analogue (bromodeoxyuridine) or a hapten (biotin) employed to label nucleotides. Examples of high-resolution detection include DNA or RNA present in different substructures of cell nuclei, and in particular, in adenovirus-induced intranuclear regions of HeLa cells. In addition to being highly sensitive and specific, these new methods offer the possibility of studying the spatial distribution of nucleic acids in very well preserved, readily recognizable structures.
Thiry
M
M
Laboratoire de Biologie Cellulaire et Tissulaire, Université de Liège, Belgium. mthiry@ulg.ac.be
D002467:Cell Nucleus / Q000737:chemistry* / Q000648:ultrastructure; D004247:DNA / Q000032:analysis* / Q000648:ultrastructure; D004254:DNA Nucleotidyltransferases; D006367:HeLa Cells; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D008854:Microscopy, Electron; D012313:RNA / Q000032:analysis* / Q000648:ultrastructure; D012316:RNA Nucleotidyltransferases; D014407:Tumor Cells, Cultured
D012313:RNA; D004247:DNA; D004254:DNA Nucleotidyltransferases; D012316:RNA Nucleotidyltransferases