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我们最近把荧光金(FG)和WGA-HRP注入雄性Sprague-Dawley大鼠一侧卵圆核(Ov),在中央杏仁核(Ce)等许多核团看到了大量逆行标记细胞。为了了解Ce至Ov投射神经元的化学性质,本组实验把FG逆行束路追踪法和间接免疫荧光法相结合,进行双标记法研究。实验选用雄性Sprague-Dawley大鼠,把FG注入一侧Ov区域,动物存活3～4天,常规方法灌注、取材,冰冻冠状连续切片,厚40μm。荧光显微镜下(紫外线激发)选择注射部位准确,且Ce内有较多FG标记细胞的切片进行免疫荧光反应,即先用M-ENK或CCK兔血清孵育,4℃2～3天;再用F1TC结合的羊抗免IgG孵育,37℃30—40分钟,荧光显微镜下观察结果。
We recently injected fluorescent gold (FG) and WGA-HRP into the ovule of male Sprague-Dawley rats (Ov), and we observed a large number of retrogradely labeled cells in many nuclei such as central amygdaloid (Ce). In order to understand the chemical properties of Ce to Ov projection neurons, this experiment combined FG retrograde beam-tracking with indirect immunofluorescence to perform double-labeling studies. Male Sprague-Dawley rats were selected and injected into the Ov area on one side. The animals survived for 3 to 4 days. The animals were perfused and drawn by conventional method. Fluorescence microscopy (UV excitation) to select the injection site accurate, and Ce more FG labeled cells in sections for immunofluorescence, that is, first with M-ENK or CCK rabbit serum incubation, 4 ℃ 2 ~ 3 days; then F1TC The combined goat anti-IgG was incubated for 30-40 minutes at 37 ° C and the results were observed under a fluorescence microscope.