Taatjes D J, Schaub U, Roth J
Histochem J. 1987 Apr;19(4):235-45. doi: 10.1007/BF01680634.
In the present study, we have investigated the applicability of semi-thin sections from low temperature Lowicryl K4M-embedded tissues for cytochemical labelling with protein A-gold and lectin-gold complexes. In order to ensure the best possible signal-to-noise ratio antibodies, protein A-gold and lectin-gold were applied in concentrations used for labelling at the electron microscope level. Furthermore, due to the lack of an appropriate chemical procedure for resin removal, untreated semi-thin sections were incubated. Under such conditions, semi-thin sections displayed either no visible staining or only a faint incomplete staining. However, following photochemical silver reaction, the latent or faint incomplete staining was rendered visible in most cases. It is concluded that the same block of Lowicryl K4M-embedded tissue and the same labelling reagents can be used for both light and electron microscopical cytochemical studies. At the light microscopical level, a high degree of structural and specific staining information is obtained. The reactivity of cellular components with antibodies or lectins is preserved even after years of storage of the blocks or slides containing semi-thin sections.
在本研究中,我们研究了低温Lowicryl K4M包埋组织的半薄切片用于蛋白A-金和凝集素-金复合物细胞化学标记的适用性。为确保获得最佳信噪比,抗体、蛋白A-金和凝集素-金均采用用于电子显微镜水平标记的浓度。此外,由于缺乏去除树脂的合适化学方法,对未处理的半薄切片进行孵育。在这种条件下,半薄切片要么没有可见染色,要么只有微弱的不完全染色。然而,经过光化学银反应后,大多数情况下潜在的或微弱的不完全染色变得可见。得出的结论是,同一Lowicryl K4M包埋组织块和相同的标记试剂可用于光镜和电镜细胞化学研究。在光镜水平上,可获得高度的结构和特异性染色信息。即使含有半薄切片的组织块或玻片保存多年后,细胞成分与抗体或凝集素的反应性仍得以保留。