Hisano S, Adachi T, Daikoku S
J Histochem Cytochem. 1985 May;33(5):485-90. doi: 10.1177/33.5.3989275.
Postembedding immunocytochemical labeling was performed on sections of rat neurohypophysis prepared by either freeze-drying, vapor fixation and Spurr resin embedding, or conventional aqueous fixation and Spurr resin embedding. Arginine vasopressin (AVP) and oxytocin (OXT) were immunolabeled with protein A-gold-anti-AVP and protein A-gold-anti-OXT complexes, respectively. The freeze-drying procedure (FD) resulted in excellent preservation of ultrastructure and greater antigenicity than the conventional procedure (Con). More gold particles were seen over secretory granules in FD sections than in Con sections. In addition, in FD sections, the gold label was restricted to secretory granules while in Con sections, both the granules and the extragranular axoplasm exhibited label. The two antigens in FD sections could be labeled simultaneously with protein A-small gold particle-anti-OXT complex and protein A-large gold particles-anti-AVP complex. In this way the two antigens were seen to be present in secretory granules within different axon terminals. Thus FD preparations should be useful for demonstrating the presence of multiple antigens in the same granules of nerve terminals.
对通过冷冻干燥、蒸汽固定和Spurr树脂包埋或传统水固定和Spurr树脂包埋制备的大鼠神经垂体切片进行包埋后免疫细胞化学标记。分别用蛋白A-金-抗精氨酸加压素（AVP）和蛋白A-金-抗催产素（OXT）复合物对精氨酸加压素（AVP）和催产素（OXT）进行免疫标记。冷冻干燥程序（FD）比传统程序（Con）能更好地保存超微结构和抗原性。在FD切片的分泌颗粒上比在Con切片上看到更多的金颗粒。此外，在FD切片中，金标记仅限于分泌颗粒，而在Con切片中，颗粒和颗粒外轴浆均显示有标记。FD切片中的两种抗原可以用蛋白A-小金颗粒-抗OXT复合物和蛋白A-大金颗粒-抗AVP复合物同时标记。通过这种方式，可以看到两种抗原存在于不同轴突终末的分泌颗粒中。因此，FD制剂对于证明神经终末同一颗粒中多种抗原的存在应该是有用的。
