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Update Date: [2013-11-05]

NO.: 20

AHRI report No.20


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


1 Taiwan Animal Health Research Institu
2.Deflent of Vetinary Mdicine., National Taiwan University 3.DertnentofM&icalResearch, Veterans JeneralHc€pil


AndxewC,Y. Fei 1.H, Chang2;K.K, Yang3


Adirect immunoperoxidase technique for the identification of immunoglobulincontaining cells in the section of chicken spleen is described. This is applicable to formalin fixed, paraff in- embedded material. It is based on an immunohistochemical method using peroxidase-labelled antibodies.

Rabbits were immunized by purified chicken IgG. The IgO fraction of rabbit anti -chicken Ig 0 antiserum was conjugated with horseradish peroxidase. The presence of immunoglobulinSinninary containing cells in chicken spleen sections was revealed by
staining the tissue-bound peroxidase-labelled antibody with diaminobenzidine, 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 paraffinembedded tissue are possible.


Immunceka’n inicrcGcopy ; e]ectxon rnicrcscopy