Bohn W
J Histochem Cytochem. 1978 Apr;26(4):293-7. doi: 10.1177/26.4.77869.
A fixation method for electron microscopical immunoperoxidase staining has been developed, which (a) allows penetration of antibodies through cell membranes to intracellular antigen sites, (b) provides a reasonable cell preservation and (c) does not alter the antigenic structure in too great an extent. Penetration of the antibodies has been achieved by using saponin as a cell membrane attacking agent. The best results could be obtained after pretreatment of cell monolayers with a mixture of 0.05% saponin, 0.0125%-0.05% glutaraldehyde and 1% paraformaldehyde for 5 min at 4 degrees C, and postfixing them with the corresponding fixative without saponin for 45 min at 4 degrees C.
已开发出一种用于电子显微镜免疫过氧化物酶染色的固定方法，该方法(a) 允许抗体穿透细胞膜到达细胞内抗原位点，(b) 能实现合理的细胞保存，且(c) 不会在很大程度上改变抗原结构。通过使用皂苷作为细胞膜攻击剂实现了抗体的穿透。在4℃下用含有0.05%皂苷、0.0125%-0.05%戊二醛和1%多聚甲醛的混合物对细胞单层进行预处理5分钟，然后在4℃下用不含皂苷的相应固定剂后固定45分钟，可获得最佳结果。
