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Ultrastructural studies conducted in situ using conventional transmission electron microscopy have had relatively little impact on defining the structural organization of chromatin. This is due to the fact that in routine transmission electron microscopy, together with the deoxyribonucleoprotein, many different intermingled substances are contrasted, masking the ultrastructure of chromatin. By selective staining of DNA in thin sections, using the Feulgen-like osmium-ammine reaction, these drawbacks have been overcome and worthwhile data have been obtained both on the gross morphology and the ultrastructural-functional organization of chromatin in situ.
View Article and Find Full Text PDFStructural and functional organization of ribosomal genes within the mammalian cell nucleolus.
J Histochem Cytochem
February 2006
Dipartimento di Patologia Sperimentale, Via S. Giacomo 14, 40126 Bologna, Italy.
Data on the in situ structural-functional organization of ribosomal genes in the mammalian cell nucleolus are reviewed here. Major findings on chromatin structure in situ come from investigations carried out using the Feulgen-like osmium ammine reaction as a highly specific electron-opaque DNA tracer. Intranucleolar chromatin shows three different levels of organization: compact clumps, fibers ranging from 11 to 30 nm, and loose agglomerates of extended DNA filaments.
View Article and Find Full Text PDFRetinoic acid induction of nuclear envelope-limited chromatin sheets in HL-60.
Exp Cell Res
November 1998
Foundation for Blood Research, 69 US Route One, Scarborough, Maine, 04070-0190, USA.
Exposure of the human leukemic cell line (HL-60) to 1 microM retinoic acid (RA) induces in vitro granulopoiesis, including the development of lobulated nuclei. Ultrastructural studies, presented here, demonstrate the formation of extensive quantities of nuclear envelope-limited chromatin sheets (ELCS), in addition to nuclear lobulation, following treatment with RA. ELCS contain DNA, as shown by the Feulgen-like electron microscope stain osmium ammine-B.
View Article and Find Full Text PDFFine structural cytochemical and immunocytochemical analysis of nucleic acids and ribonucleoprotein distribution in nuclei of pig oocytes and early preimplantation embryos.
Chromosoma
June 1996
Centre of Electron Microscopy, University of Lausanne, Bugnon 27, CH-1005 Lausanne, Switzerland.
The fine structure of pig oocytes at the germinal vesicle (GV) stage and early preimplantation embryos (one to four blastomeres) isolated at slaughter was investigated by cytochemical and immunocytochemical methods. The distribution of nucleic acids and ribonucleoproteins (RNPs) in "compact nucleoli" [denominated nucleolus-like bodies (NLB) in oocytes and nucleolus precursor bodies (NPB) in early embryos] and in intranuclear bodies or granules was investigated by staining methods preferential for nuclear RNPs or using the osmium ammine or ethidium bromide-phosphotungstic acid (EB-PTA) reactions for nucleic acids. The distributions of the Sm antigen of nucleoplasmic small nuclear RNPs (snRNPs), the methyl-3 guanosine (m3G) cap of snRNAs and the splicing factor SC-35 were detected by immunoelectron microscopy using specific antibodies.
View Article and Find Full Text PDFThe osmium ammine-SO2 staining method for studying the in situ configuration of viral genomes in ultrathin sections of DNA virus infected cells.
Biol Cell
April 1997
Laboratoire Organisation fonctionnelle du Noyau, Institut de Recherches sur le Cancer/IFC1, UPR 9044 CNRS, Villejuif, France.
The Feulgen-like osmium ammine-SO2 method developed by Cogliati and Gautier (CR Acad Sci Ser D 1973, 276, 3041-3044) stains the DNA at the ultrastructural level. Compared to several other techniques for detecting DNA, this method remains the only one revealing the configuration of the DNA molecules within the cell whatever their compactness. In the present article we summarize the results we obtained with the osmium ammine method in the study of the fate of viral genomes along the infectious cycles in several DNA virus infected cells including adenovirus, herpes simplex virus, simian virus 40 and poxvirus.
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