Laser-assisted microdissection of membrane-mounted paraffin sections for polymerase chain reaction analysis: Identification of cell populations using immunohistochemistry and in situ hybridization

Lise Mette Gjerdrum*, Ilze Lielpetere, Lars Melholt Rasmussen, Knud Bendix, Stephen Hamilton-Dutoit

*Corresponding author af dette arbejde

    Publikation: Bidrag til tidsskriftArtikelForskningpeer review

    Abstrakt

    Laser microbeam microdissection (LMM) is an increasingly important method for obtaining pure cell samples for genetic and proteomic analysis. Immuno-histochemistry (IHC) and in situ hybridization (ISH) are useful techniques for targeting specific cell populations for microdissection but are difficult to apply with the tissue support membranes often used during LMM. Using detection of cytokeratins and Epstein-Barr virus gene products in head and neck carcinoma as a model, we describe optimized protocols for membrane and section preparation and for low temperature antigen retrieval that allow IHC and ISH to be used reliably on membrane mounted paraffin tissue sections. Visualization of cellular targets was markedly improved by staining and this could be further improved using a variety of optical media before microdissection. Tissue fragments thus stained were suitable for subsequent polymerase chain reaction analysis of extracted DNA using standard techniques. These IHC and ISH procedures are generally applicable and will be useful for detecting a wide range of antigens and nucleic acids in paraffin sections in conjunction with LMM.

    OriginalsprogEngelsk
    Artikelnummer60659
    Sider (fra-til)105-110
    Antal sider6
    TidsskriftJournal of Molecular Diagnostics
    Vol/bind3
    Udgave nummer3
    DOI
    StatusUdgivet - 1 jan. 2001

    Fingeraftryk

    Udforsk hvilke forskningsemner 'Laser-assisted microdissection of membrane-mounted paraffin sections for polymerase chain reaction analysis: Identification of cell populations using immunohistochemistry and in situ hybridization' indeholder.

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