【摘要】 目的为更客观地观察胚胎脑神经祖细胞的增殖、分化和迁移,建立胚胎脑片免疫荧光组织化学双重漂染技术。方法灌流固定,取胚胎14天(E14)大鼠脑,低熔点琼脂糖包埋,振动切片机连续冠状切片,免疫荧光组织化学双重漂染,激光扫描共聚焦显微镜下观察。结果波形蛋白(Vimentin)和乙酰胆碱转移酶(ChAT)双阳性细胞呈黄色荧光,胞体位于脑室区,长突起呈放射状。ChAT阳性细胞呈红色荧光,除胞体位于脑室区,长突起呈放射状伸展外,可见皮层板区也有阳性细胞集聚。结论此项技术可直接观察到胚胎脑神经祖细胞和成神经细胞的完整形态,并通过免疫荧光组织化学方法鉴定其表型,从而阐明相互之间的关系。更多还原

【Abstract】 Objective To establish the double immunofluorescent staining technique for the embryonic brain slices to objectively observe the proliferation,differentiation and migration of neural progenitor cells in rat embryo brains.Methods Rat embryos of 14 days(E14) were fixed by perfusion.The brains were taken and embedded in low-melting agarose,then sliced with a vibratome,stained by double immunofluorescence and observed by using confocal laser scanning microscopy.Results Vimentin and choline acetyltransferase(ChAT) double positive cells showed yellow fluorescence their bodies located in the ventricle zone(VZ),and the radial long processes in the subventricle zone(SVZ).Only ChAT positive cells with red fluorescence were found in the cortex plate of dorsal telencephalon.Conclusion The morphology and identity of neural progenitor cells and neuroblasts are identified by using the technique,and their relationship can be analyzed by double immunofluorescent staining.更多还原