title: The AMeX method. A simplified technique of tissue processing and paraffin embedding with improved preservation of antigens for immunostaining.
abstract: A new simplified tissue processing method for immunostaining was devised. Tissues were fixed in acetone at -20 C overnight, then cleared in methyl benzoate and xylene, consecutively, and embedded in ordinary paraffin at 58-60 C. Thin paraffin sections were deparaffinized with xylene, immersed in acetone and then phosphate buffered saline, and immunostained with various monoclonal and conventional antibodies, which have only been used on fresh-frozen or PLP-fixed frozen sections. As with PLP-fixed frozen sections, the following antigens were clearly demonstrated in the tissue sections processed with the present method: T (Leu-1, Leu-2a, Leu-3a, Leu-4, OKT3, OKT4, OKT8), B (B1, B2, Leu-14, IgM, IgD) lymphocyte surface markers and other antigens (Leu-7, OKT6, OKT9, OKM1, OKI1, J5, Ki-1, Ki-67, TdT, oncogene Ha-ras P21). Sections prepared by the present method demonstrated much better histologic and cytologic preservation than possible in frozen sections.
mesh_terms: D000096:Acetone; D000941:Antigens / Q000032:analysis*; D001565:Benzoates; D005404:Fixatives; D005629:Frozen Sections; D006652:Histological Techniques; D006801:Humans; D010232:Paraffin; D013194:Staining and Labeling; D014992:Xylenes
chemical_list: D000941:Antigens; D001565:Benzoates; D005404:Fixatives; D014992:Xylenes; D000096:Acetone; C044605:methyl benzoate; D010232:Paraffin