Delecolle B, Womdim R N, Marchoux G
INRA, Station de Pathologie Végétale, Domaine St-Maurice, Montfavet, France.
Cell Mol Biol (Noisy-le-grand). 1992 Aug-Sep;38(5-6):495-511.
An immunogold labeling technique was carried out on plants infected with CMV, fixed with glutaraldehyde and osmium tetroxide and embedded in araldite CY 212. The effect of the type of support-film used, the resin and manipulations of the grids during immunogold steps, were studied and are discussed. The antigenic activity of virus was restored by treating the sections with sodium metaperiodate. The very high non-specific reactions observed with the support-film or with the resin were eliminated by adding powdered skimmed milk or non-purified albumin into the buffers. Purified bovine serum albumin (grade V) or chicken albumin (grade III to V) were inefficient in reducing this non-specific background.
对感染黄瓜花叶病毒（CMV）的植物进行免疫金标记技术实验，这些植物先用戊二醛和四氧化锇固定，然后包埋在CY 212环氧树脂中。研究并讨论了免疫金步骤中所用支撑膜类型、树脂以及网格操作的影响。用偏高碘酸钠处理切片可恢复病毒的抗原活性。通过在缓冲液中加入脱脂奶粉或未纯化的白蛋白，消除了在支撑膜或树脂上观察到的非常高的非特异性反应。纯化的V级牛血清白蛋白或III至V级鸡白蛋白在降低这种非特异性背景方面效果不佳。
