Method of preparation of the specimens for combined immunocytological examinations by light and electron microscopy
Mogil'nyĭ, I.I.; Arkhipov, N.I.
Arkhiv Patologii 44(11): 69-71
1982
ISSN/ISBN: 0004-1955 PMID: 6297434 Document Number: 193245
A method for making preparations for combined immunocytological examinations in light and electron microscopes is described. An infected cell culture grown on slides (a piglet kidney cell culture infected with Teschen virus) was fixed with 2.5% glutaraldehyde in phosphate buffer, pH 7.2-7.4 for 10-15 min and incubated with a conjugate (peroxidase-labeled immune serum to the causative agent of the infection under study). After postfixation, cytochemical reaction for peroxidase was performed, then the material was fixed with 2% OsO4 solution in phosphate buffer, pH 7,2-7,4, dehydrated in an ascending series of alcohols and gradually impregnated with polymerising resins (epon, araldit, etc.). The cells were embedded in foil baths, and after polymerization and separation of the slide and foil preparations with good light microscopy properties. In the light microscope, the desired parts of cells or a cell were selected, removed, cut in an ultratome and examined in electron microscope.