Immunohistochemical antigen demonstration in plastic-embedded lymphoid tissue.
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.
van Goor
H
H
Department of Pathology, University of Groningen, The Netherlands.
Harms
G
G
Gerrits
P O
PO
Kroese
F G
FG
Poppema
S
S
Grond
J
J
D000179:Acrylates; D000818:Animals; D000941:Antigens / Q000032:analysis*; D001402:B-Lymphocytes / Q000276:immunology; D002072:Butylene Glycols; D005404:Fixatives; D006651:Histocytochemistry; D007124:Immunoenzyme Techniques; D008264:Macrophages / Q000276:immunology; D008297:Male; D008689:Methacrylates; D051381:Rats; D011919:Rats, Inbred Strains; D013154:Spleen / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology; D013950:Thymus Gland / Q000276:immunology*
D000179:Acrylates; D000941:Antigens; D002072:Butylene Glycols; D005404:Fixatives; D008689:Methacrylates; C005044:hydroxyethyl methacrylate