Gendelman H E, Moench T R, Narayan O, Griffin D E
J Immunol Methods. 1983 Dec 16;65(1-2):137-45. doi: 10.1016/0022-1759(83)90310-1.
Fixation techniques were investigated for their ability to preserve morphology, esterase activity and cell surface antigens in paraffin-embedded mouse lymphoid tissue. The avidin-biotin-peroxidase system was used to stain antigens Thy-1.2, Lyt-1, Lyt-2, RA32C2 and F4/80. Conventional fixatives were compared with fixatives containing periodate and lysine plus paraformaldehyde and/or glutaraldehyde. Conventional fixatives preserved esterase activity but not many cell surface antigens. Periodate-lysine fixatives allowed better preservation of membrane antigens, but esterase activity was often lost at antigen-preserving concentrations of paraformaldehyde or glutaraldehyde. However, a periodate-lysine fixative containing both paraformaldehyde and glutaraldehyde preserved esterase and showed good to excellent staining of Lyt-1, Thy-1.2, RA32C2, and F4/80. Lyt-2 could not be stained with any fixative, but was well preserved in frozen material post-fixed with periodate-lysine based fixatives. We conclude that with proper fixation immune cell surface markers can be identified in paraffin-embedded tissue.
研究了固定技术在保存石蜡包埋的小鼠淋巴组织中的形态、酯酶活性和细胞表面抗原方面的能力。采用抗生物素蛋白-生物素-过氧化物酶系统对抗原Thy-1.2、Lyt-1、Lyt-2、RA32C2和F4/80进行染色。将传统固定剂与含有高碘酸盐、赖氨酸以及多聚甲醛和/或戊二醛的固定剂进行比较。传统固定剂可保存酯酶活性，但不能保存许多细胞表面抗原。高碘酸盐-赖氨酸固定剂能更好地保存膜抗原，但在多聚甲醛或戊二醛的抗原保存浓度下，酯酶活性常常丧失。然而，一种同时含有多聚甲醛和戊二醛的高碘酸盐-赖氨酸固定剂能保存酯酶，并对Lyt-1、Thy-1.2、RA32C2和F4/80显示出良好至优异的染色效果。Lyt-2不能用任何固定剂染色，但在用基于高碘酸盐-赖氨酸的固定剂进行后固定的冷冻材料中保存良好。我们得出结论，通过适当的固定，可以在石蜡包埋的组织中鉴定免疫细胞表面标志物。
