Herter P, Laube G, Gronczewski J, Minuth W W
Max-Planck-Institut für Molekulare Physiologie, Dortmund, Germany.
J Microsc. 1993 Aug;171(Pt 2):107-15. doi: 10.1111/j.1365-2818.1993.tb03364.x.
The luminal cell surfaces of rabbit kidney cortical collecting-duct cells were labelled with peanut lectin (PNA) and investigated by scanning electron microscopy. Labelling was performed either on 20-microns-thick cryostat sections from prefixed and cryoprotected rabbit kidney tissue or on cultured collecting-duct epithelium using biotinylated PNA and a 6-nm colloidal-gold-coupled antibody against biotin. Colloidal-gold labels were detected at low magnification (2000-4000x) using silver enhancement. Coating with chromium allowed simultaneous imaging of both cell-surface morphology and labelling topography in the backscattered electron imaging mode. Our results show that PNA binding is specific for a subtype of intercalated cells equipped with microvilli on the luminal surface. The presented method promises to be useful for the identification of specific cell types in heterogeneous tissues.
用花生凝集素（PNA）标记兔肾皮质集合管细胞的管腔细胞表面，并通过扫描电子显微镜进行研究。标记操作既可以在来自预先固定和冷冻保护的兔肾组织的20微米厚的低温恒温器切片上进行，也可以使用生物素化的PNA和针对生物素的6纳米胶体金偶联抗体在培养的集合管上皮细胞上进行。使用银增强技术在低倍放大（2000 - 4000倍）下检测胶体金标记。用铬涂层可在背散射电子成像模式下同时对细胞表面形态和标记拓扑结构进行成像。我们的结果表明，PNA结合对管腔表面配备微绒毛的闰细胞亚型具有特异性。所提出的方法有望用于鉴定异质组织中的特定细胞类型。
