Sipos J, McFarlane B M, Williams R, McFarlane I G
Liver Unit, King's College Hospital, London.
J Clin Pathol. 1988 Nov;41(11):1217-22. doi: 10.1136/jcp.41.11.1217.
An enzyme immunohistochemical technique for the localisation of liver membrane antigens in tissue sections by antisera raised in guinea pigs against the liver preparation known as "liver specific membrane lipoprotein (LSP)" was developed, based on the alkaline phosphatase avidin biotin complex (ABC AP) system. Of a wide range of fixatives and fixation conditions investigated, a short (five minute) exposure of cryostat sections to Bouin's fluid provided the most satisfactory results and--together with procedures to block endogenous biotin and alkaline phosphatase--yielded clear sections with no background staining or other artefacts to interfere with specific staining patterns. The sensitivity of the technique approaches that of a radioimmunoassay, as shown by the staining of the sinusoidal domains of hepatocellular plasma membranes by the guinea pig anti-LSP antisera at dilutions up to 1/50,000. Apart from its reliability and sensitivity the procedure offers additional advantages over techniques such as indirect immunofluorescence in that it provides a permanent preparation with well defined morphological details which can be seen by ordinary light microscopy.
基于碱性磷酸酶抗生物素蛋白生物素复合物（ABC AP）系统，开发了一种酶免疫组织化学技术，用于通过豚鼠针对称为“肝特异性膜脂蛋白（LSP）”的肝脏制剂产生的抗血清在组织切片中定位肝膜抗原。在研究的多种固定剂和固定条件中，将冷冻切片短时间（5分钟）暴露于Bouin液可提供最满意的结果，并且与阻断内源性生物素和碱性磷酸酶的程序一起，可产生清晰的切片，无背景染色或其他伪像干扰特异性染色模式。如豚鼠抗LSP抗血清在高达1/50,000的稀释度下对肝细胞质膜的窦状区域进行染色所示，该技术的灵敏度接近放射免疫测定法。除了可靠性和灵敏度外，该程序还具有优于间接免疫荧光等技术的其他优点，因为它提供了具有明确定义的形态细节的永久制剂，可通过普通光学显微镜观察到。
