In situ hybridization of cytospin preparations: a rapid nonisotopic screening method for isolated cells.
A simple and rapid method for light microscopic in situ hybridization on cytospin preparations is described and demonstrated for detection of viral nucleic acid in a virus-infected cell line. Cells were fixed by acetone followed by chloroform, denatured by heat, hybridized at 37 C, and hybridized sites detected with a multiple step procedure (primary anti-biotin antibody, biotinylated second antibody, streptavidin-peroxidase). This method can be used for screening studies at the light microscope level, and offers a useful and simple way to determine optimum hybridization conditions for subsequent electron microscopic investigations.
Mandry
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P
GSF-Institute of Pathology, Oberschleissheim, Germany.
Murray
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AB
Höfler
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H
D000818:Animals; D002460:Cell Line; D002498:Centrifugation; D004279:DNA, Viral / Q000032:analysis; D006566:Herpesviridae Infections / Q000382:microbiology; D004854:Herpesvirus 4, Human / Q000235:genetics; D016591:Histocytological Preparation Techniques; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D008027:Light; D008853:Microscopy; D008854:Microscopy, Electron
D004279:DNA, Viral