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. 1988 May;131(2):183–189.

A simplified plastic embedding and immunohistologic technique for immunophenotypic analysis of human hematopoietic and lymphoid tissues.

T T Casey 1, J B Cousar 1, R D Collins 1
PMCID: PMC1880591  PMID: 3282442

Abstract

Routine fixation and paraffin embedding destroys many hematopoietic and lymphoid differentiation antigens detected by flow cytometry or frozen section immunohistochemistry. On the other hand, morphologic evaluation is difficult in flow cytometric or frozen section studies. A simplified three-step plastic embedding system using acetone-fixed tissues embedded in glycol-methacrylate (GMA) resin has been found to provide both excellent morphologic and antigenic preservation. With our system, a wide variety of antigens are detected in plastic sections without trypsinization or prolonged embedding procedures; pan-B (CD19, CD22), pan-T (CD7, CD5, CD3, CD2), T-subset (CD4, CD8, CD1, CD25) markers as well as surface immunoglobulin and markers for myeloid and mononuclear-phagocyte cells are preserved. In summary, modifications of plastic embedding techniques used in this study simplify the procedure, apparently achieve excellent antigenic preservation, and facilitate evaluation of morphologic details in relation to immunocytochemical markers.

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Selected References

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  1. Beckstead J. H., Halverson P. S., Ries C. A., Bainton D. F. Enzyme histochemistry and immunohistochemistry on biopsy specimens of pathologic human bone marrow. Blood. 1981 Jun;57(6):1088–1098. [PubMed] [Google Scholar]
  2. Beckstead J. H. Optimal antigen localization in human tissues using aldehyde-fixed plastic-embedded sections. J Histochem Cytochem. 1985 Sep;33(9):954–958. doi: 10.1177/33.9.4020104. [DOI] [PubMed] [Google Scholar]
  3. Holgate C. S., Jackson P., Pollard K., Lunny D., Bird C. C. Effect fixation on T and B lymphocyte surface membrane antigen demonstration in paraffin processed tissue. J Pathol. 1986 Aug;149(4):293–300. doi: 10.1002/path.1711490405. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Linder J., Ye Y. L., Harrington D. S., Armitage J. O., Weisenburger D. D. Monoclonal antibodies marking T lymphocytes in paraffin-embedded tissue. Am J Pathol. 1987 Apr;127(1):1–8. [PMC free article] [PubMed] [Google Scholar]
  6. Marder R. J., Variakojis D., Silver J., Epstein A. L. Immunohistochemical analysis of human lymphomas with monoclonal antibodies to B cell and Ia antigens reactive in paraffin sections. Lab Invest. 1985 May;52(5):497–504. [PubMed] [Google Scholar]
  7. Moir D. J., Ghosh A. K., Abdulaziz Z., Knight P. M., Mason D. Y. Immunoenzymatic staining of haematological samples with monoclonal antibodies. Br J Haematol. 1983 Nov;55(3):395–410. doi: 10.1111/j.1365-2141.1983.tb02154.x. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Pinkus G. S., Kurtin P. J. Epithelial membrane antigen--a diagnostic discriminant in surgical pathology: immunohistochemical profile in epithelial, mesenchymal, and hematopoietic neoplasms using paraffin sections and monoclonal antibodies. Hum Pathol. 1985 Sep;16(9):929–940. doi: 10.1016/s0046-8177(85)80132-5. [DOI] [PubMed] [Google Scholar]
  10. Pollard K., Lunny D., Holgate C. S., Jackson P., Bird C. C. Fixation, processing, and immunochemical reagent effects on preservation of T-lymphocyte surface membrane antigens in paraffin-embedded tissue. J Histochem Cytochem. 1987 Nov;35(11):1329–1338. doi: 10.1177/35.11.3309048. [DOI] [PubMed] [Google Scholar]
  11. Sato Y., Mukai K., Watanabe S., Goto M., Shimosato Y. The AMeX method. A simplified technique of tissue processing and paraffin embedding with improved preservation of antigens for immunostaining. Am J Pathol. 1986 Dec;125(3):431–435. [PMC free article] [PubMed] [Google Scholar]
  12. Smith S. H., Brown M. H., Rowe D., Callard R. E., Beverley P. C. Functional subsets of human helper-inducer cells defined by a new monoclonal antibody, UCHL1. Immunology. 1986 May;58(1):63–70. [PMC free article] [PubMed] [Google Scholar]
  13. Stein H., Bonk A., Tolksdorf G., Lennert K., Rodt H., Gerdes J. Immunohistologic analysis of the organization of normal lymphoid tissue and non-Hodgkin's lymphomas. J Histochem Cytochem. 1980 Aug;28(8):746–760. doi: 10.1177/28.8.7003001. [DOI] [PubMed] [Google Scholar]
  14. Stein H., Gatter K. C., Heryet A., Mason D. Y. Freeze-dried paraffin-embedded human tissue for antigen labelling with monoclonal antibodies. Lancet. 1984 Jul 14;2(8394):71–73. doi: 10.1016/s0140-6736(84)90243-5. [DOI] [PubMed] [Google Scholar]
  15. Strickler J. G., Weiss L. M., Copenhaver C. M., Bindl J., McDaid R., Buck D., Warnke R. Monoclonal antibodies reactive in routinely processed tissue sections of malignant lymphoma, with emphasis on T-cell lymphomas. Hum Pathol. 1987 Aug;18(8):808–814. doi: 10.1016/s0046-8177(87)80055-2. [DOI] [PubMed] [Google Scholar]
  16. Takamiya H., Batsford S. R., Tokunaga J., Vogt A. Immunohistological staining of antigens on semithin sections of specimens embedded in plastic (GMA-Quetol 523). J Immunol Methods. 1979;30(3):277–288. doi: 10.1016/0022-1759(79)90102-9. [DOI] [PubMed] [Google Scholar]
  17. Takamiya H., Batsford S., Vogt A. An approach to postembedding staining of protein (immunoglobulin) antigen embedded in plastic: prerequisites and limitations. J Histochem Cytochem. 1980 Oct;28(10):1041–1049. doi: 10.1177/28.10.6158534. [DOI] [PubMed] [Google Scholar]
  18. Takamiya H., Bodemer W., Vogt A. Masking of protein antigen by modification of amino groups with carbobenzoxychloride (benzyl chloroformate) and demasking by treatment with nonspecific protease. J Histochem Cytochem. 1978 Nov;26(11):914–920. doi: 10.1177/26.11.82572. [DOI] [PubMed] [Google Scholar]
  19. Warnke R., Levy R. Detection of T and B cell antigens hybridoma monoclonal antibodies: a biotin-avidin-horseradish peroxidase method. J Histochem Cytochem. 1980 Aug;28(8):771–776. doi: 10.1177/28.8.7003003. [DOI] [PubMed] [Google Scholar]

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