Varndell I M, Tapia F J, Probert L, Buchan A M, Gu J, De Mey J, Bloom S R, Polak J M
Peptides. 1982 May-Jun;3(3):259-72. doi: 10.1016/0196-9781(82)90086-9.
The use of protein A- and IgG-conjugated colloidal gold staining methods for the immuno-localisation of peptide hormones and neurotransmitters at light- and electron microscope level are described and discussed. Bright-field and dark-ground illumination modes have been used to visualise the gold-labelled antigenic sites at the light microscope level. Immunogold staining procedures at the ultrastructural level using region-specific antisera have been adopted to localise specific molecular forms of peptides including gastrin (G17 and G34), glucagon and pro-glucagon, insulin and pro-insulin, in normal tissue and in tumours of the gastroenteropancreatic system. Similar methods have been used to demonstrate the heterogeneity of p-type nerves in the enteric nervous system. Vasoactive intestinal polypeptide (VIP) has been localised to granular sites (mean +/- S.D. granule diameter = 98 +/- 19 nm) in nerve terminals of the enteric plexuses and in tumour cells of diarrhoeogenic VIP-producing neoplasias (mean +/- S.D. granule diameter = 126 +/- 37 nm) using immunogold procedures applied to ultraviolet-cured ultrathin sections. Co-localisation of amines and peptides in carotid body type I cells and in chromaffin cells of normal adrenal medulla and phaeochromocytomas has also been demonstrated. Advantages of the immunogold procedures over alternative immunocytochemical techniques are discussed.
本文描述并讨论了蛋白质A和IgG偶联胶体金染色方法在光镜和电镜水平上对肽类激素和神经递质进行免疫定位的应用。在光镜水平上，采用明场和暗场照明模式来观察金标记的抗原位点。在超微结构水平上，使用区域特异性抗血清的免疫金染色程序已被用于在正常组织和胃肠胰系统肿瘤中定位包括胃泌素（G17和G34）、胰高血糖素和前胰高血糖素、胰岛素和前胰岛素在内的特定肽分子形式。类似的方法已被用于证明肠神经系统中p型神经的异质性。采用免疫金程序对紫外线固化超薄切片进行检测，已将血管活性肠肽（VIP）定位到肠丛神经末梢的颗粒位点（平均±标准差颗粒直径 = 98 ± 19 nm）以及产生腹泻性VIP的肿瘤细胞（平均±标准差颗粒直径 = 126 ± 37 nm）。还证明了胺类和肽类在颈动脉体I型细胞以及正常肾上腺髓质和嗜铬细胞瘤的嗜铬细胞中的共定位。讨论了免疫金程序相对于其他免疫细胞化学技术的优势。
