Immunohistochemistry (IHC) and immunofluorescence (IF) staining techniques are important diagnostic tools of anatomic pathology in the clinical setting and widely used analytical tools in research laboratories. In diabetes research, they are routinely used for the assessment of beta- and alpha-cell mass, for assessment of endocrine cell distribution within the pancreas, for evaluation of islet composition and islet morphology. Here, we present the evaluation of IHC techniques for the detection of alpha-cells in human pancreatic tissue. We compared the Horse Radish Peroxidase (HRP)-based method utilizing DAB Peroxidase Substrate to the Alkaline Phosphatase (AP)-based method utilizing Vector Red substrate. We conclude that HRP–DAB staining is a robust and reliable method for detection of alpha-cells using either rabbit polyclonal or mouse monoclonal anti-glucagon antibodies. However, AP-Vector Red staining should be used with caution, because it is affected by the dehydration with ethanol and toluene preceding the mounting of slides with Permount mounting medium. When AP-Vector Red is a preferable method for alpha-cell labeling, slides should be mounted using aqueous mounting medium or, alternatively, they could be air-dried before permanent mounting

中文翻译：

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人胰腺组织中胰高血糖素免疫组化染色的评价

免疫组织化学 (IHC) 和免疫荧光 (IF) 染色技术是临床环境中解剖病理学的重要诊断工具，也是研究实验室中广泛使用的分析工具。在糖尿病研究中，它们通常用于评估 β 和 α 细胞质量、评估胰腺内的内分泌细胞分布、评估胰岛组成和胰岛形态。在这里，我们介绍了 IHC 技术在人类胰腺组织中检测 α 细胞的评估。我们比较了使用 DAB 过氧化物酶底物的基于辣根过氧化物酶 (HRP) 的方法与使用 Vector Red 底物的基于碱性磷酸酶 (AP) 的方法。我们得出结论，HRP-DAB 染色是使用兔多克隆或小鼠单克隆抗胰高血糖素抗体检测 α 细胞的稳健可靠的方法。但是，应谨慎使用 AP-Vector Red 染色，因为在使用 Permount 安装介质安装幻灯片之前，它会受到乙醇和甲苯脱水的影响。当 AP-Vector Red 是 alpha 细胞标记的首选方法时，应使用水性封固剂封固载玻片，或者，在永久封固前将其风干