Summary
Two methods of demonstrating tissue antigens by ultrastructural enzyme immunohistochemistry were tested. The monoclonal antibodies Ki-M1 and Ki-M4 were chosen for testing the methods because Ki-M1 identifies a relatively stable, and Ki-M4 a very unstable antigen. The two antibodies react selectively with human macrophages and interdigitating reticulum cells or dendritic reticulum cells of lymphoid follicles. The Ki-Ml reaction product is confined to the surface membrane. Ki-M4 reactivity is located on the surface membrane and, less often and to a lesser extent, in the cytoplasm. The technical prerequisites for reliable conservation of the antigens identified by these two antibodies were standardized. The results indicated that prior fixation in 4% paraformaldehyde is preferable for optimum preservation of stable antigens. Application of the primary antibody prior to fixation was found to be the best procedure for demonstrating unstable antigens, although nonspecific reactions were seen more often with this method.
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Eldred WD, Zucker C, Karten HJ, Yazulla S (1983) Comparison of fixation and penetration fnenhancement techniques for use in ultrastructural immunocytochemistry. J Histochem Cytochem 31:285–292
Gerdes J, Hansmann M-L, Stein H, Naiem M, Mason DY (1982) Ultrastructural localization of human complement C3b receptors in the human kidney as determined by immunoperoxidase staining with the monoclonal antibody C3RTo5. Virchows Arch [Cell Pathol] 40:1–7
Graham RC Jr, Karnovsky MJ (1966) The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem 14:291–302
Kaiserling E (1984) Ultrastrukturelle Diagnostik am lymphatischen System. In: Schlote W (ed) Diagnostische Elektronenmikroskopie [in press]
Kuhlmann WD, Avrameas S, Ternynck T (1974) A comparative study for ultrastructural localization of intracellular immunoglobulins using peroxidase conjugates. J Immunol Methods 5:33–48
McLean IW, Nakane PK (1974) Periodate-lysine-paraformaldehyde fixative. A new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1083
Murphy GF, Bhan AK, Sato S, Mihm MC Jr, Harrist TJ (1981) A new immunologic marker for human Langerhans cells. [Letter to the Editor] N Engl J Med 304:791–792
Parwaresch MR, Radzun HJ, Hansmann M-L, Peters K-P (1983) Monoclonal antibody Ki-M4 specifically recognizes human dendritic reticulum cells (follicular dendritic cells) and their possible precursor in blood. Blood 62:585–590
Radzun HJ, Parwaresch MR (1983) Differential immunohistochemical resolution of the human mononuclear phagocyte system. Cell Immunol 82:174–183
Spurr AR (1969) A low-viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26:31–43
Stein H, Bonk A, Tolksdorf G, Lennert K, Rodt H, Gerdes J (1980) Immunohistologic analysis of the organization of normal lymphoid tissue and non-Hodkin’s lymphomas. J Histochem Cytochem 28:746–760
Tsunoda R, Terashima K, Takahashi K, Kojima M (1978) An ultrastructural study with enzyme-labeled antibody technique on immunoglobulin-containing cells in human tonsils, especially in germinal centers. Acta Pathol Jpn 28:53–75
van Ewijk W, Coffman RC, Weissman IL (1980) Immunoelectron microscopy of cell surface antigens: a quantitative analysis of antibody binding after different fixation protocols. Histochem J 12:349–361
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Hansmann, M.L., Radzun, H.J., Kaiserling, E. et al. Immunoelectron microscopic demonstration of tissue antigens with monoclonal antibodies. Virchows Archiv B Cell Pathol 46, 1–12 (1984). https://doi.org/10.1007/BF02890290
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DOI: https://doi.org/10.1007/BF02890290