Immunohistochemical antigen demonstration in plastic-embedded lymphoid tissue

J Histochem Cytochem. 1988 Jan;36(1):115-20. doi: 10.1177/36.1.3275710.

Abstract

We describe a method for post-embedding immunohistochemical demonstration of a wide range of antigens in glycol methacrylate-embedded tissue. Rat spleen and thymus tissues were fixed by immersion in fixatives containing different concentrations of paraformaldehyde, washed in sucrose phosphate buffer, dehydrated in acetone, infiltrated in a glycol methacrylate mixture in which the commonly used softener 2-butoxyethanol was replaced by butaandiol monoacrylate, and embedded. Trypsin was used to re-expose the masked antigenicity. Excellent results were obtained with a panel of monoclonal antibodies (MoAbs) directed against T-cells, B-cells, Ia-positive cells, macrophages, follicular dendritic cells, and leucocyte common antigen-bearing cells. The method described combines exact localization of antigens with optimal tissue morphology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrylates
  • Animals
  • Antigens / analysis*
  • B-Lymphocytes / immunology
  • Butylene Glycols
  • Fixatives
  • Histocytochemistry
  • Immunoenzyme Techniques*
  • Macrophages / immunology
  • Male
  • Methacrylates
  • Rats
  • Rats, Inbred Strains
  • Spleen / immunology*
  • T-Lymphocytes / immunology
  • Thymus Gland / immunology*

Substances

  • Acrylates
  • Antigens
  • Butylene Glycols
  • Fixatives
  • Methacrylates
  • hydroxyethyl methacrylate