Analysis of leukocyte surface antigens on ethanol-fixed paraffin-embedded tissue material.
Ethanol-fixed paraffin-embedded specimens of human tissues were studied whether the surface antigens of leukocytes in these tissues can be stained and analyzed. Three-layer indirect immunoperoxidase staining was performed on the ethanol-fixed paraffin-embedded sections by the use of several monoclonal antibodies for whole human leukocytes (Dako LC), B cells (Dako CD-22, 4KB5, and L26; Leu 14), T cells and their subsets (Dako UCHL-1, T1, T3, T4 and T8; Leu 4, 3a and 2a) and monocyte/macrophage lineage (Dako macrophage, Leu M1, M3 and M5). The results were compared with those on fresh-frozen sections. No essential differences were obtained between the paraffin-embedded and the fresh frozen sections stained by the following antibodies; Dako LC for whole human leukocytes; Dako UCHL-1, T3 and Leu 4 for T cells; Dako CD22, 4KB5, L26 and Leu 14 for B cells; Dako macrophage, Leu M1 and M5 for monocyte/macrophage lineage. On the other hand, the subsets of T cells could only be detected on the fresh-frozen sections. The results of the leukocyte analysis on the paraffin-embedded specimens of several renal diseases were very similar to those reported by other investigators on fresh-frozen sections or PLP-fixed materials. Thus, by the use of appropriate monoclonal antibodies, the ethanol-fixed paraffin-embedded material can be used for leukocyte analysis except for the definition of T cell subsets.
Second Department of Internal Medicine, Tohoku University School of Medicine, Sendai, Japan.
T
T
Ootaka
T
T
Saito
J
J
Soma
K
K
Yoshinaga
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis*; D000431:Ethanol; D006801:Humans; D007124:Immunoenzyme Techniques; D007668:Kidney / Q000276:immunology; D007962:Leukocytes / Q000276:immunology*; D008221:Lymphoid Tissue / Q000276:immunology; D010232:Paraffin
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D000431:Ethanol; D010232:Paraffin