Ellis I O, Bell J, Bancroft J D
Department of Histopathology, Queen's Medical Centre, University Hospital, Nottingham, United Kingdom.
J Histochem Cytochem. 1988 Jan;36(1):121-4. doi: 10.1177/36.1.3335767.
We investigated the optimal gold particle size for use with polarized incident light (epi polarization) microscopy with immunogold immunohistological preparation in both immunogold indirect (IGS) and silver-enhanced immunogold-silver staining (IGSS) techniques. A range of gold particle sizes from 5 nm-40 nm was used along with tissue of known immunoreactivity with a well-characterized primary monoclonal antibody. Checkerboard titrations were carried out for each technique and for each particle size. The preparations were viewed using a standard polarized incident light microscope and assessed in a semi-quantitative manner. Adequate visualization of gold particles was achieved using the indirect staining method only with a particle size of 40 nm. With silver enhancement (IGSS), particles of all sizes were clearly seen. However, 5-nm particles were considered optimal for this method because of reduced background staining, high titration of antisera possible, and crisp localization of the visual signal.
我们研究了在免疫金免疫组织学制备中，用于偏振入射光（落射偏振）显微镜检查的免疫金间接法（IGS）和银增强免疫金银染色法（IGSS）的最佳金颗粒大小。使用了一系列粒径从5纳米到40纳米的金颗粒，以及与一种特征明确的一级单克隆抗体具有已知免疫反应性的组织。对每种技术和每种颗粒大小都进行了棋盘滴定法。使用标准偏振入射光显微镜观察制备物，并进行半定量评估。仅使用粒径为40纳米的颗粒通过间接染色法可实现金颗粒的充分可视化。通过银增强（IGSS），所有大小的颗粒都能清晰看到。然而，5纳米的颗粒被认为是该方法的最佳选择，因为背景染色减少、抗血清滴定度高且视觉信号定位清晰。
