Alisky J M, Tolbert D L
Francis and Doris Murphy Neuroanatomy Research Laboratory, Department of Anatomy, St. Louis University School of Medicine, MO 63104.
J Neurosci Methods. 1994 Jun;52(2):143-8. doi: 10.1016/0165-0270(94)90122-8.
We report a new technique for 2-tracer anterograde labeling that permits unequivocal identification of the differentially labeled projections in the same section. One pathway is labeled with biotinylated dextran amine and is visualized as a black to dark gray diaminobenzidine (DAB)-cobalt precipitate by an avidin-biotinylated peroxidase reaction. The other pathway is labeled with cholera toxin subunit B and is visualized as a reddish-brown reaction product using DAB without cobalt as the substrate for peroxidase immunohistochemistry. To maintain serial order, sections can be processed mounted on slides without any loss of sensitivity for either tracer.
我们报告了一种用于双示踪剂顺行标记的新技术，该技术能够在同一切片中明确识别差异标记的投射。一条通路用生物素化葡聚糖胺标记，并通过抗生物素蛋白-生物素化过氧化物酶反应显示为黑色至深灰色的二氨基联苯胺（DAB）-钴沉淀。另一条通路用霍乱毒素B亚基标记，并使用不含钴的DAB作为过氧化物酶免疫组织化学的底物，显示为红棕色反应产物。为保持连续顺序，切片可以在不损失任何一种示踪剂敏感性的情况下进行载玻片固定处理。
