Johansson O, Virtanen M, Hilliges M, Yang Q
Department of Histology and Neurobiology, Karolinska Institutet, Stockholm, Sweden.
Histochem J. 1992 May;24(5):283-7. doi: 10.1007/BF01046843.
Mast cells have characteristic granulae containing various glucoseaminoglycans, proteases and amines (predominantly histamine). The conventional histological methods for studying mast cells are based upon acidic ortho- and metachromatic routine stains of the glucoseaminoglycans. However, the success of these procedures is dependent upon both the fixatives and the tissues used. In this study, we wanted to find out whether an immunohistochemical procedure could overcome some of these difficulties. Normal human skin was fixed in five different types of fixative and processed for indirect immunofluorescence, using an antiserum to histamine. Only one, 4% carbodiimide in 0.1 M phosphate buffer (pH 7.4), resulted in immunostaining. The quality of the staining was good, with a high signal-to-noise ratio, and was located on the mast cells. The method made it possible to visualize small structures such as a single secreted granula, the thin cytoplasmatic extension of some cells, and a previously undescribed dendritic morphology of some of the mast cells. We therefore recommend this procedure for cellular studies of mast cells when accuracy is needed.
肥大细胞具有特征性颗粒,其中含有各种葡糖胺聚糖、蛋白酶和胺类物质(主要是组胺)。研究肥大细胞的传统组织学方法基于葡糖胺聚糖的酸性正染和异染常规染色。然而,这些方法的成功与否取决于所使用的固定剂和组织。在本研究中,我们想弄清楚免疫组织化学方法是否能够克服其中的一些困难。将正常人皮肤用五种不同类型的固定剂固定,并使用抗组胺抗血清进行间接免疫荧光处理。只有一种固定剂,即0.1M磷酸盐缓冲液(pH 7.4)中的4%碳二亚胺,能产生免疫染色。染色质量良好,信噪比高,且位于肥大细胞上。该方法使得可视化小结构成为可能,如单个分泌颗粒、某些细胞的细细胞质延伸以及一些肥大细胞以前未描述过的树突状形态。因此,当需要准确性时,我们推荐此方法用于肥大细胞的细胞研究。