Abstract:
Immunohistochemistry (IHC) is the basis of histological or pathological analysis and is widely used to enable the detection and characterization of proteins in various organ tissues, including brain tissues. IHC is commonly performed on formalin-fixed paraffin-embedded (FFPE) tissues because of their easy storage and versatility. IHC is a key method for providing more accurate analysis of localization and function of neurons, neuroendocrine cells, and neural stem cells in the brain and other nervous systems. The related cells such as glial cells and neurovascular units have also been analyzed by IHC. Visualization of antibody-antigen interactions can be performed primarily in one of the following ways: chromogenically stained IHC and fluorescently stained IHC. In chromogenically stained IHC, an antibody is chemically conjugated to an enzyme, such as peroxidase, that can be reacted with a suitable substrate to give a colored product. In fluorescently stained IHC, the antibodies are finally tagged with fluorescent chemicals such as fluorescein isothiocyanate (FITC) or rhodamine. Here, we describe the standard methods of IHC applied to brain slice sections. Furthermore, an automated immunostainer is presented as another option for standardized immunohistochemistry.
摘要:
免疫组织化学(IHC)是组织学或病理学分析的基础,广泛用于检测和表征各种器官组织中的蛋白质。包括脑组织.IHC通常在福尔马林固定的石蜡包埋(FFPE)组织上进行,因为它们易于储存和多功能性。IHC是提供更准确的神经元定位和功能分析的关键方法。神经内分泌细胞,以及大脑和其他神经系统中的神经干细胞。相关细胞如神经胶质细胞和神经血管单位也已通过IHC分析。抗体-抗原相互作用的可视化可以主要以以下方式之一进行:显色染色的IHC和荧光染色的IHC。在显色染色的IHC中,抗体与酶化学结合,如过氧化物酶,可以与合适的底物反应得到有色产物。在荧光染色的IHC中,最后用荧光化学物质如异硫氰酸荧光素(FITC)或罗丹明标记抗体。这里,我们描述了应用于脑切片的IHC标准方法。此外,自动免疫染色仪是标准化免疫组织化学的另一种选择。