Skip to main content
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1986 Dec;125(3):431–435.

The AMeX method. A simplified technique of tissue processing and paraffin embedding with improved preservation of antigens for immunostaining.

Y Sato, K Mukai, S Watanabe, M Goto, Y Shimosato
PMCID: PMC1888473  PMID: 2432790

Abstract

A new simplified tissue processing method for immunostaining was devised. Tissues were fixed in acetone at -20 C overnight, then cleared in methyl benzoate and xylene, consecutively, and embedded in ordinary paraffin at 58-60 C. Thin paraffin sections were deparaffinized with xylene, immersed in acetone and then phosphate buffered saline, and immunostained with various monoclonal and conventional antibodies, which have only been used on fresh-frozen or PLP-fixed frozen sections. As with PLP-fixed frozen sections, the following antigens were clearly demonstrated in the tissue sections processed with the present method: T (Leu-1, Leu-2a, Leu-3a, Leu-4, OKT3, OKT4, OKT8), B (B1, B2, Leu-14, IgM, IgD) lymphocyte surface markers and other antigens (Leu-7, OKT6, OKT9, OKM1, OKI1, J5, Ki-1, Ki-67, TdT, oncogene Ha-ras P21). Sections prepared by the present method demonstrated much better histologic and cytologic preservation than possible in frozen sections.

Full text

PDF
431

Images in this article

Selected References

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

  1. McMillan E. M., Wasik R., Everett M. A. Identification of T-lymphocytes and T-subsets in human tonsil using monoclonal antibodies and the immunoperoxidase technic. Am J Clin Pathol. 1981 Dec;76(6):737–744. doi: 10.1093/ajcp/76.6.737. [DOI] [PubMed] [Google Scholar]
  2. McMillan E. M., Wasik R., Everett M. A. OKT 6-positive cells: their demonstration in human thymus, and the effect of fixation on the immunoperoxidase reaction. Arch Pathol Lab Med. 1982 Jan;106(1):9–12. [PubMed] [Google Scholar]
  3. 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]
  4. Poppema S., Bhan A. K., Reinherz E. L., McCluskey R. T., Schlossman S. F. Distribution of T cell subsets in human lymph nodes. J Exp Med. 1981 Jan 1;153(1):30–41. doi: 10.1084/jem.153.1.30. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. 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]
  6. Tanaka M., Tanaka H., Ishikawa E. Immunohistochemical demonstration of surface antigen of human lymphocytes with monoclonal antibody in acetone-fixed paraffin-embedded sections. J Histochem Cytochem. 1984 Apr;32(4):452–454. doi: 10.1177/32.4.6368682. [DOI] [PubMed] [Google Scholar]
  7. Watanabe S., Sato Y., Kodama T., Shimosato Y. Immunohistochemical study with monoclonal antibodies on immune response in human lung cancers. Cancer Res. 1983 Dec;43(12 Pt 1):5883–5889. [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES