Ethidium bromide- and propidium iodide-PTA staining of nucleic acids at the electron microscopic level.
Ultra-thin sections of various tissues were stained with ethidium bromide or propidium iodide, two fluorescent markers widely used for quantitation of nucleic acids. The fluorochromes, tested at different concentrations, were then revealed by incubation of the sections with neutralized phosphotungstic acid. We showed that at the electron microscopic level only nucleic acid-containing structures are revealed. Chromatin, nucleolus, and ribosomes appear to be stained by the end-product of the reaction. Furthermore, controls with proteases and nucleases showed that the staining is related to the binding of the fluorochromes to DNA and RNA and to the subsequent detection of the dyes by neutralized PTA.
Biggiogera
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Centre de Microscopie Electronique de l'Université de Lausanne, Switzerland.
Biggiogera
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D000818:Animals; D002466:Cell Nucleolus / Q000648:ultrastructure; D002467:Cell Nucleus / Q000648:ultrastructure; D002843:Chromatin / Q000648:ultrastructure; D003851:Deoxyribonucleases / Q000494:pharmacology; D004996:Ethidium; D008099:Liver / Q000032:analysis / Q000648:ultrastructure; D008297:Male; D051379:Mice; D008854:Microscopy, Electron; D009696:Nucleic Acids / Q000032:analysis*; D010447:Peptide Hydrolases / Q000494:pharmacology; D010617:Phenanthridines; D010772:Phosphotungstic Acid; D011419:Propidium; D051381:Rats; D012260:Ribonucleases / Q000494:pharmacology; D012270:Ribosomes / Q000648:ultrastructure; D013194:Staining and Labeling; D013737:Testis / Q000032:analysis / Q000648:ultrastructure
D002843:Chromatin; D009696:Nucleic Acids; D010617:Phenanthridines; D010772:Phosphotungstic Acid; D011419:Propidium; D003851:Deoxyribonucleases; D012260:Ribonucleases; D010447:Peptide Hydrolases; D004996:Ethidium