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免疫胶体在光学显微镜中的应用。III. 半薄树脂切片中抗原位点和凝集素结合位点的显示

Applications of immunocolloids in light microscopy. III. Demonstration of antigenic and lectin-binding sites in semithin resin sections.

作者信息

Lucocq J M, Roth J

出版信息

J Histochem Cytochem. 1984 Oct;32(10):1075-83. doi: 10.1177/32.10.6384362.

Abstract

Previous studies have demonstrated that antigens or lectin-binding sites can be localized in sections from paraffin-embedded tissues with protein A or lectins bound to colloidal gold or colloidal silver (Roth J: J Histochem Cytochem 30:691, 1982 and 31:547, 1983). In the present study the protein A-gold technique and lectin-gold complexes have been applied to semithin sections (0.5-1.5 micron) of Epon- or low temperature Lowicryl K4M-embedded rat pancreas, kidney and submandibular gland. The results show that an increase in resolution and, therefore, in amount of information can be obtained. The optimal mode of imaging was determined on sections without counterstaining. Bright-field illumination gives the maximum information about the staining signal, while phase-contrast and Nomarski differential interference contrast give predominantly structural and, to a lesser extent, staining information. Polarization epi- and transillumination microscopy is inferior in all aspects. The application of a battery of lectin-gold complexes to rat submandibular gland revealed a specific staining pattern for each lectin in acinar and excretory duct cells.

摘要

以往的研究表明,抗原或凝集素结合位点可以在石蜡包埋组织的切片中定位,其中蛋白A或凝集素与胶体金或胶体银结合(罗斯·J:《组织化学与细胞化学杂志》30:691,1982年和31:547,1983年)。在本研究中,蛋白A-金技术和凝集素-金复合物已应用于环氧树脂或低温Lowicryl K4M包埋的大鼠胰腺、肾脏和下颌下腺的半薄切片(0.5-1.5微米)。结果表明,可以提高分辨率,从而增加信息量。在不进行复染的切片上确定了最佳成像模式。明场照明提供了关于染色信号的最大信息,而相差和诺马斯基微分干涉对比主要提供结构信息,在较小程度上提供染色信息。偏振落射和透射照明显微镜在各方面都较差。将一系列凝集素-金复合物应用于大鼠下颌下腺,揭示了每个凝集素在腺泡和排泄管细胞中的特异性染色模式。

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