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. 1985 Jun;38(6):671–676. doi: 10.1136/jcp.38.6.671

Modified immunocytochemical slide technique for demonstrating surface antigens on viable cells.

N Frickhofen, K J Bross, W Heit, H Heimpel
PMCID: PMC499266  PMID: 3891791

Abstract

Further developments of an immunoenzymatic slide technique to demonstrate cell surface antigens with monoclonal antibodies are described. In this method cells are attached to polylysine coated glass slides in order to facilitate the handling of low cell numbers and to save antibodies and time for washing the cells. The technique has been modified for the labelling of viable cells. Endogenous peroxidase is used as an additional cell marker which does not interfere with the demonstration of antigens on the cell surface by immunoperoxidase methods. Damaged cells can be identified reliably, thereby minimising interpretation errors due to non-specific antibody uptake. A double labelling technique employing peroxidase and alkaline phosphatase coupled reagents is presented. Results of this slide technique are clear cut, so that evaluation can be performed by trained technicians.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aisenberg A. C., Wilkes B. M., Harris N. L. Monoclonal antibody studies in non-Hodgkin's lymphoma. Blood. 1983 Mar;61(3):469–475. [PubMed] [Google Scholar]
  2. Bross K. J., Pangalis G. A., Staatz C. G., Blume K. G. Demonstration of cell surface antigens and their antibodies by the peroxidase-antiperoxidase method. Transplantation. 1978 Jun;25(6):331–334. doi: 10.1097/00007890-197806000-00011. [DOI] [PubMed] [Google Scholar]
  3. Brown W. R., Burns F. A. Application of peroxidase-labeled antibody methods to identification of immunoglobulin-containing cells in gastrointestinal tissues. J Immunol Methods. 1973 Apr;2(3):303–307. doi: 10.1016/0022-1759(73)90056-2. [DOI] [PubMed] [Google Scholar]
  4. Cordell J. L., Falini B., Erber W. N., Ghosh A. K., Abdulaziz Z., MacDonald S., Pulford K. A., Stein H., Mason D. Y. Immunoenzymatic labeling of monoclonal antibodies using immune complexes of alkaline phosphatase and monoclonal anti-alkaline phosphatase (APAAP complexes). J Histochem Cytochem. 1984 Feb;32(2):219–229. doi: 10.1177/32.2.6198355. [DOI] [PubMed] [Google Scholar]
  5. Farr A. G., Nakane P. K. Immunohistochemistry with enzyme labeled antibodies: a brief review. J Immunol Methods. 1981;47(2):129–144. doi: 10.1016/0022-1759(81)90114-9. [DOI] [PubMed] [Google Scholar]
  6. Feller A. C., Parwaresch M. R. Simultaneous enzyme-immunocytochemical detection of antigens in monocellular specimens with monoclonal antibodies. J Immunol Methods. 1983 Oct 14;63(2):273–279. doi: 10.1016/0022-1759(83)90431-3. [DOI] [PubMed] [Google Scholar]
  7. Guesdon J. L., Ternynck T., Avrameas S. The use of avidin-biotin interaction in immunoenzymatic techniques. J Histochem Cytochem. 1979 Aug;27(8):1131–1139. doi: 10.1177/27.8.90074. [DOI] [PubMed] [Google Scholar]
  8. Hsu S. M., Raine L., Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem. 1981 Apr;29(4):577–580. doi: 10.1177/29.4.6166661. [DOI] [PubMed] [Google Scholar]
  9. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  10. Landay A., Gartland G. L., Abo T., Cooper M. D. Enumeration of human lymphocyte subpopulations by immunofluorescence: a comparative study using automated flow microfluorometry and fluorescence microscopy. J Immunol Methods. 1983 Mar 25;58(3):337–347. doi: 10.1016/0022-1759(83)90361-7. [DOI] [PubMed] [Google Scholar]
  11. Mason D. Y., Sammons R. Alkaline phosphatase and peroxidase for double immunoenzymatic labelling of cellular constituents. J Clin Pathol. 1978 May;31(5):454–460. doi: 10.1136/jcp.31.5.454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mason D. Y., Stein H., Naiem M., Abdulaziz Z. Immunohistological analysis of human lymphoid tissue by double immunoenzymatic labelling. J Cancer Res Clin Oncol. 1981;101(1):13–22. doi: 10.1007/BF00405059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Mehta M. M., Munker R., Kranz B., Thiel E., Thierfelder S. Immunofluorescence with monoclonal antibodies on poly-L-lysine coated slides: an alternative to conventional methods. Blut. 1983 Oct;47(4):237–242. doi: 10.1007/BF00320843. [DOI] [PubMed] [Google Scholar]
  14. Nadler L. M., Stashenko P., Hardy R., Pesando J. M., Yunis E. J., Schlossman S. F. Monoclonal antibodies defining serologically distinct HLA-D/DR related Ia-like antigens in man. Hum Immunol. 1981 Feb;2(1):77–90. doi: 10.1016/0198-8859(81)90009-4. [DOI] [PubMed] [Google Scholar]
  15. Patterson J. A., Griffiths-Chu S., Berger C. L., Edelson R. L., Chu A. C. Recognition of immature T-cell phenotypes in neonatal blood using monoclonal antibodies. Diagn Immunol. 1983;1(3):205–210. [PubMed] [Google Scholar]
  16. Ponder B. A., Wilkinson M. M. Inhibition of endogenous tissue alkaline phosphatase with the use of alkaline phosphatase conjugates in immunohistochemistry. J Histochem Cytochem. 1981 Aug;29(8):981–984. doi: 10.1177/29.8.7024402. [DOI] [PubMed] [Google Scholar]
  17. Ritz J., Schlossman S. F. Utilization of monoclonal antibodies in the treatment of leukemia and lymphoma. Blood. 1982 Jan;59(1):1–11. [PubMed] [Google Scholar]
  18. Schroff R. W., Foon K. A., Billing R. J., Fahey J. L. Immunologic classification of lymphocytic leukemias based on monoclonal antibody-defined cell surface antigens. Blood. 1982 Feb;59(2):207–215. [PubMed] [Google Scholar]
  19. Schwab R., Staiano-Coico L., Weksler M. E. Immunological studies of aging. IX. Quantitative differences in T lymphocyte subsets in young and old individuals. Diagn Immunol. 1983;1(3):195–198. [PubMed] [Google Scholar]
  20. Stashenko P., Nadler L. M., Hardy R., Schlossman S. F. Characterization of a human B lymphocyte-specific antigen. J Immunol. 1980 Oct;125(4):1678–1685. [PubMed] [Google Scholar]
  21. Straus W. Peroxidase procedures. Technical problems encountered during their application. J Histochem Cytochem. 1979 Oct;27(10):1349–1351. doi: 10.1177/27.10.92491. [DOI] [PubMed] [Google Scholar]
  22. Tidman N., Janossy G., Bodger M., Granger S., Kung P. C., Goldstein G. Delineation of human thymocyte differentiation pathways utilizing double-staining techniques with monoclonal antibodies. Clin Exp Immunol. 1981 Sep;45(3):457–467. [PMC free article] [PubMed] [Google Scholar]

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