AHRI report No.20

Detection of Intracellular Immunoglobulin in Formalin-Paraffin Sections of the Chicken Spleen Using the Immunoperoxidase Technique

1.     Taiwan Provincia1. Research Institnte for Animal Health, Tansui, Taiwan, R. 0. C. 
2.     Department of Veterinary Medicine, National Taiwan University, Taipei, Taiwan, R. 0. C. 
3.     Department of Medical Research, Veterans General Hospital, Taipei, Taiwan, R. 0. C. 

Andrew Chang-Young Fei1 Pen-Heng Chang' And King-KiangYoung

A direct immunoperoxidase technique for the identification of immunoglobulin-containing cells in the section of chicken spleen is described. This is applicable to formalin fixed, paraffin-embedded material. It is based on an immunohistoch-emical method using peroxidase-labelled antibodies. 
Rabbits were immunized by purified chicken lgG. The lgG fraction of rabbit anti-chicken lgG antiserum was conjugated with horseradish peroxidase. The presence of immunoglobulin-containing cells in chicken spleen sections was revealed by staining the tissue-bound peroxidase-labelled antibody with diam-inobenzidine, than briefly counterstained with hematoxylin. 
The endogeneous peroxidase was satisfactorily inhibited by treating the dewaxed section with a fresh 0.5% solution of hydrogen peroxide in absolute methanol for 20 minutes. 
The structure of immunoglobulin-containing cells in chicken spleen sections is clear, preparations are permanent, and retrospective studies of stored paraffin-embedded tissue are possible.