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.