Effect of tissue processing on the immunostaining of inflammatory cell surface epitopes identified by monoclonal antibodies.
Optimal tissue processing conditions were defined for the immunohistochemical detection of inflammatory cell surface epitopes identified by OKT3, OKT4, OKT6, OKT8, OKIa1, OKM1, Leu7 and pan B cell antibodies. Snap-freezing in isopentan was superior to liquid nitrogen in preservation of morphology. Embedding of the tissues in Tissue-Tek II O.C.T. Compound diminished the intensity of immunostaining with all antibodies studied; however, the embedded tissues tolerated longer storage without drying. Optimal fixation with satisfactory preservation of morphology and immunogenicity was achieved with fixation of the frozen sections in acetone at 4 degrees C for 5 min. Blocking of the endogenous peroxidase with methanol-H2O2 treatment destroyed all epitopes studied except those identified with OKIa1, OKT6 and Leu7.
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D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis*; D000939:Epitopes / Q000032:analysis*; D006801:Humans; D007124:Immunoenzyme Techniques; D008198:Lymph Nodes / Q000473:pathology; D008867:Microtomy / Q000379:methods; D014066:Palatine Tonsil / Q000473:pathology; D012867:Skin / Q000473:pathology; D013194:Staining and Labeling / Q000379:methods
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D000939:Epitopes