Selection of a fixative for identifying T cell subsets, B cells, and macrophages in paraffin-embedded mouse spleen

J Immunol Methods. 1983 Dec 16;65(1-2):137-45. doi: 10.1016/0022-1759(83)90310-1.


Fixation techniques were investigated for their ability to preserve morphology, esterase activity and cell surface antigens in paraffin-embedded mouse lymphoid tissue. The avidin-biotin-peroxidase system was used to stain antigens Thy-1.2, Lyt-1, Lyt-2, RA32C2 and F4/80. Conventional fixatives were compared with fixatives containing periodate and lysine plus paraformaldehyde and/or glutaraldehyde. Conventional fixatives preserved esterase activity but not many cell surface antigens. Periodate-lysine fixatives allowed better preservation of membrane antigens, but esterase activity was often lost at antigen-preserving concentrations of paraformaldehyde or glutaraldehyde. However, a periodate-lysine fixative containing both paraformaldehyde and glutaraldehyde preserved esterase and showed good to excellent staining of Lyt-1, Thy-1.2, RA32C2, and F4/80. Lyt-2 could not be stained with any fixative, but was well preserved in frozen material post-fixed with periodate-lysine based fixatives. We conclude that with proper fixation immune cell surface markers can be identified in paraffin-embedded tissue.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • B-Lymphocytes / classification
  • B-Lymphocytes / enzymology
  • B-Lymphocytes / immunology*
  • Fixatives*
  • Histocytochemistry
  • Immunoenzyme Techniques
  • Lysine
  • Macrophages / classification
  • Macrophages / enzymology
  • Macrophages / immunology*
  • Mice
  • Mice, Inbred BALB C
  • Paraffin
  • Periodic Acid
  • Spleen / cytology
  • T-Lymphocytes / classification
  • T-Lymphocytes / enzymology
  • T-Lymphocytes / immunology*


  • Fixatives
  • Periodic Acid
  • Paraffin
  • metaperiodate
  • Lysine