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  • Title: Fixation-dependent cytoplasmic false-positive staining with an immunoperoxidase method.
    Author: Bergroth V, Reitamo S, Konttinen YT, Tolvanen E.
    Journal: Histochemistry; 1982; 73(4):509-13. PubMed ID: 6175608.
    Abstract:
    Fixation-dependent nonspecific staining with the unlabeled immunoperoxidase (PAP) method was studied using paraffin-embedded human spleen sections fixed in various fixatives; the specific primary antiserum was omitted or nonimmunized normal rabbit serum was used. Strong cytoplasmic staining of polymorphonuclear leucocytes and macrophages was found after fixation in acetone, alcoholic formalin (94% alcohol) and absolute ethanol. This staining was mainly produced by the second layer of the PAP method. The most probable explanation of this phenomenon is nonspecific protein-immunoglobulin interaction as a result of alcoholic or acetone fixation of the sections. The present findings point to the importance of controls for each case under study to avoid false-positive interpretations.
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