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免疫细胞化学的一些最新进展。

Some recent developments in immunocytochemistry.

作者信息

Bosman F T

出版信息

Histochem J. 1983 Mar;15(3):189-200. doi: 10.1007/BF01006235.

Abstract

Immunocytochemistry has become an indispensable tool both in basic biomedical research and in diagnostic histopathology. Several recent innovations have led to improvements in the sensitivity; specificity and precision of these techniques. Numerous modifications of the original methods have been developed, many with increased sensitivity. In particular, methods using protein A or the avidin-biotin complex as second steps appear to be promising. New methods for the quantitative determination of sensitivity have become available. The introduction of monoclonal antibodies as immunocytochemical reagents appears to be a major improvement. New methods of immunochemical analysis, such as immunoblotting and immunospotting of antigens extracted from tissue specimens, allow the molecular composition of immunoreactive antigenic sites in a tissue to be analysed. The accuracy of localization in immunoelectron microscopy has been improved significantly through the use of ultracryotomy of unfixed tissue in combination with colloidal gold particles as a label. In addition, gold particles can be counted and thus allow relatively simple quantification of the immune reaction. Using flow cytometry, especially in combination with monoclonal antibodies, quantitative immunofluorescence has become feasible.

摘要

免疫细胞化学已成为基础生物医学研究和诊断组织病理学中不可或缺的工具。最近的几项创新已使这些技术在敏感性、特异性和精确性方面得到改进。人们已开发出对原始方法的多种改进,其中许多改进提高了敏感性。特别是,将蛋白A或抗生物素蛋白-生物素复合物用作第二步的方法似乎很有前景。已出现用于定量测定敏感性的新方法。引入单克隆抗体作为免疫细胞化学试剂似乎是一项重大改进。新的免疫化学分析方法,如对从组织标本中提取的抗原进行免疫印迹和免疫斑点分析,能够分析组织中免疫反应性抗原位点的分子组成。通过将未固定组织的超薄冷冻切片术与作为标记物的胶体金颗粒相结合,免疫电子显微镜中的定位准确性已得到显著提高。此外,可以对金颗粒进行计数,从而实现对免疫反应相对简单的定量。利用流式细胞术,尤其是与单克隆抗体结合使用时,定量免疫荧光已变得可行。

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