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Clinical and Diagnostic Laboratory Immunology logoLink to Clinical and Diagnostic Laboratory Immunology
. 1994 May;1(3):299–303. doi: 10.1128/cdli.1.3.299-303.1994

A comparative study of frozen-section immunoperoxidase and flow cytometry for immunophenotypic analysis of lymph node biopsies.

K W Biesemier 1, G A Dent 1, K B Pryzwansky 1, J D Folds 1
PMCID: PMC368252  PMID: 7496966

Abstract

Immunophenotyping by flow cytometry and frozen-section immunoperoxidase was compared on 21 consecutive lymph node biopsy specimens, of which a diagnosis of lymphoma was made for 11 specimens. Samples for flow cytometry were obtained by a fine-needle aspiration technique. Concordance between frozen-section immunoperoxidase and flow cytometry for all routine markers on all specimens ranged from 76 to 100%. In general, B-cell markers showed poorer concordance than T-cell markers, with kappa and lambda light chains having the poorest concordance, at 76% each. Flow cytometry was significantly more sensitive (90 versus 30%; P < 0.006) and had a significantly higher negative predictive value (100 versus 63%; P < 0.006) than frozen-section immunoperoxidase for demonstrating light-chain restriction. There was no significant difference in the specificities (100 versus 91%) or positive predictive values (100% each) between the two methods. Both methods demonstrated characteristic immunophenotypes for intermediate cell lymphomas, small lymphocytic lymphomas, and T-cell lymphoblastic lymphomas. Frozen-section immunoperoxidase and flow cytometry appear to be significantly concordant methods for immunophenotypic analysis of lymph node biopsies. Light-chain restriction is more readily demonstrated by flow cytometry than frozen-section immunoperoxidase. We believe that ex vivo fine-needle aspiration is a simple and reliable method of obtaining cell suspensions of lymph nodes for flow cytometry.

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Selected References

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  1. Bach B. A., Knape W. A., Edinger M. G., Tubbs R. R. Improved sensitivity and resolution in the flow cytometric DNA analysis of human solid tumor specimens. Use of in vitro fine-needle aspiration and uniform staining reagents. Am J Clin Pathol. 1991 Nov;96(5):615–627. doi: 10.1093/ajcp/96.5.615. [DOI] [PubMed] [Google Scholar]
  2. Braylan R. C., Benson N. A. Flow cytometric analysis of lymphomas. Arch Pathol Lab Med. 1989 Jun;113(6):627–633. [PubMed] [Google Scholar]
  3. Diamond L. W., Nathwani B. N., Rappaport H. Flow cytometry in the diagnosis and classification of malignant lymphoma and leukemia. Cancer. 1982 Sep 15;50(6):1122–1135. doi: 10.1002/1097-0142(19820915)50:6<1122::aid-cncr2820500616>3.0.co;2-q. [DOI] [PubMed] [Google Scholar]
  4. Eliasen C. A., Opitz L. M., Vamvakas E. C., Espiritu E. C., Marsh E. R., Roses D. F., Harris M. N., Feiner H. D. Flow cytometric analysis of DNA ploidy and S-phase fraction in breast cancer using cells obtained by ex vivo fine-needle aspiration: an optimal method for sample collection. Mod Pathol. 1991 Mar;4(2):196–200. [PubMed] [Google Scholar]
  5. Foon K. A., Todd R. F., 3rd Immunologic classification of leukemia and lymphoma. Blood. 1986 Jul;68(1):1–31. [PubMed] [Google Scholar]
  6. Foucar K., Chen I. M., Crago S. Organization and operation of a flow cytometric immunophenotyping laboratory. Semin Diagn Pathol. 1989 Feb;6(1):13–36. [PubMed] [Google Scholar]
  7. Kadin M. E., Said J. W. Pathology of malignant lymphomas. Curr Opin Oncol. 1990 Oct;2(5):822–831. doi: 10.1097/00001622-199010000-00004. [DOI] [PubMed] [Google Scholar]
  8. La Via M. F., Self S. E. Immunophenotypic analysis of non-Hodgkin's lymphomas. Recenti Prog Med. 1990 Oct;81(10):629–634. [PubMed] [Google Scholar]
  9. Levy N., Nelson J., Meyer P., Lukes R. J., Parker J. W. Reactive lymphoid hyperplasia with single class (monoclonal) surface immunoglobulin. Am J Clin Pathol. 1983 Sep;80(3):300–308. doi: 10.1093/ajcp/80.3.300. [DOI] [PubMed] [Google Scholar]
  10. Levy R., Warnke R., Dorfman R. F., Haimovich J. The monoclonality of human B-cell lymphomas. J Exp Med. 1977 Apr 1;145(4):1014–1028. doi: 10.1084/jem.145.4.1014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Little J. V., Foucar K., Horvath A., Crago S. Flow cytometric analysis of lymphoma and lymphoma-like disorders. Semin Diagn Pathol. 1989 Feb;6(1):37–54. [PubMed] [Google Scholar]
  12. Nemes Z., Thomázy T., Szeifert G. Demonstration of light chain monotypia in B cell non-Hodgkin's lymphomas using unfixed freeze-dried and formalin-fixed trypsinised paraffin sections. J Clin Pathol. 1983 Aug;36(8):883–893. doi: 10.1136/jcp.36.8.883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Norton A. J., Isaacson P. G. Lymphoma phenotyping in formalin-fixed and paraffin wax-embedded tissues. I. Range of antibodies and staining patterns. Histopathology. 1989 May;14(5):437–446. doi: 10.1111/j.1365-2559.1989.tb02180.x. [DOI] [PubMed] [Google Scholar]
  14. Norton A. J., Isaacson P. G. Lymphoma phenotyping in formalin-fixed and paraffin wax-embedded tissues: II. Profiles of reactivity in the various tumour types. Histopathology. 1989 Jun;14(6):557–579. doi: 10.1111/j.1365-2559.1989.tb02198.x. [DOI] [PubMed] [Google Scholar]
  15. Picker L. J., Weiss L. M., Medeiros L. J., Wood G. S., Warnke R. A. Immunophenotypic criteria for the diagnosis of non-Hodgkin's lymphoma. Am J Pathol. 1987 Jul;128(1):181–201. [PMC free article] [PubMed] [Google Scholar]
  16. Ratech H., Litwin S. Surface immunoglobulin light chain restriction in B-cell non-Hodgkin's malignant lymphomas. Am J Clin Pathol. 1989 May;91(5):583–586. doi: 10.1093/ajcp/91.5.583. [DOI] [PubMed] [Google Scholar]
  17. Sheibani K., Nathwani B. N., Swartz W. G., Ben-Ezra J., Brownell M. D., Burke J. S., Kennedy J. L., Koo C. H., Winberg C. D. Variability in interpretation of immunohistologic findings in lymphoproliferative disorders by hematopathologists. A comprehensive statistical analysis of interobserver performance. Cancer. 1988 Aug 15;62(4):657–664. doi: 10.1002/1097-0142(19880815)62:4<657::aid-cncr2820620402>3.0.co;2-m. [DOI] [PubMed] [Google Scholar]
  18. Sheibani K., Tubbs R. R. Enzyme immunohistochemistry: technical aspects. Semin Diagn Pathol. 1984 Nov;1(4):235–250. [PubMed] [Google Scholar]
  19. Sneige N., Dekmezian R., el-Naggar A., Manning J. Cytomorphologic, immunocytochemical, and nucleic acid flow cytometric study of 50 lymph nodes by fine-needle aspiration. Comparison with results obtained by subsequent excisional biopsy. Cancer. 1991 Feb 15;67(4):1003–1007. doi: 10.1002/1097-0142(19910215)67:4<1003::aid-cncr2820670424>3.0.co;2-w. [DOI] [PubMed] [Google Scholar]
  20. Taylor C. R. Results of multiparameter studies of B-cell lymphomas. Am J Clin Pathol. 1979 Oct;72(4 Suppl):687–698. [PubMed] [Google Scholar]
  21. Tubbs R. R., Sheibani K. Immunohistology of lymphoproliferative disorders. Semin Diagn Pathol. 1984 Nov;1(4):272–284. [PubMed] [Google Scholar]
  22. Tubbs R. R., Sheibani K., Sebek B. A., Weiss R. A. Immunohistochemistry versus immunofluorescence for non-Hodgkin's lymphomas. Am J Clin Pathol. 1980 Jan;73(1):144–145. doi: 10.1093/ajcp/73.1.144. [DOI] [PubMed] [Google Scholar]
  23. Tubbs R. R., Sheibani K., Weiss R. A., Sebek B. A., Deodhar S. D. Tissue immunomicroscopic evaluation of monoclonality of B-cell lymphomas: comparison with cell suspension studies. Am J Clin Pathol. 1981 Jul;76(1):24–28. doi: 10.1093/ajcp/76.1.24. [DOI] [PubMed] [Google Scholar]
  24. Tubbs R. R., Sheibani K., Weiss R. A., Sebek B. A. Immunohistochemistry of fresh-frozen lymphoid tissue with the direct immunoperoxidase technic. Am J Clin Pathol. 1981 Feb;75(2):172–174. doi: 10.1093/ajcp/75.2.172. [DOI] [PubMed] [Google Scholar]
  25. Têtu B., Manning J. T., Jr, Ordóez N. G. Comparison of monoclonal and polyclonal antibodies directed against immunoglobulin light and heavy chains in non-Hodgkin's lymphoma. Am J Clin Pathol. 1986 Jan;85(1):25–31. doi: 10.1093/ajcp/85.1.25. [DOI] [PubMed] [Google Scholar]
  26. Vaickus L., Ball E. D., Foon K. A. Immune markers in hematologic malignancies. Crit Rev Oncol Hematol. 1991 Dec;11(4):267–297. doi: 10.1016/1040-8428(91)90029-c. [DOI] [PubMed] [Google Scholar]
  27. Warzynski M. J., Podgurski A. E., Boldt D. M., Otto R. N. An automated method to prepare cell suspensions from human biopsy samples for immunophenotyping by flow cytometry. Am J Clin Pathol. 1990 Jan;93(1):104–108. doi: 10.1093/ajcp/93.1.104. [DOI] [PubMed] [Google Scholar]
  28. de Martini R. M., Turner R. R., Boone D. C., Lukes R. J., Parker J. W. Lymphocyte immunophenotyping of B-cell lymphomas: a flow cytometric analysis of neoplastic and nonneoplastic cells in 271 cases. Clin Immunol Immunopathol. 1988 Dec;49(3):365–379. doi: 10.1016/0090-1229(88)90126-2. [DOI] [PubMed] [Google Scholar]

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