目的 介绍一种移植预处理后组织的冰冻切片制作方法.方法 予BALB/c小鼠清髓剂量(60C0 7.5 Gy全身照射)照射后4h内输入体外培养的含有绿色荧光蛋白(GFP)内皮祖细胞(EPC),分别于移植后6、12、24、36、48、72及96 h取受鼠内脏组织制作冰冻切片,用免疫荧光观察组织内GFP阳性细胞分布.结果 荧光显微镜下可见肝、肺中GFP、大鼠抗小鼠泛内皮细胞单克隆抗体(MECA-32)阳性细胞,脾中见GFP、CD68阳性细胞；小肠中未发现明显GFP阳性、MECA-32阳性细胞.结论 对移植预处理的组织进行及时恰当的处理后,荧光显微镜下可清晰地观察到组织形态结构变化及GFP阳性细胞分布.%Objective To introduce a method of making frozen sections for implanted GFP+ endothelial progenitor cells after transplantation preconditioning. Methods BALB/c mice were subjected to 7.5 Gy 60Co irradiation(at a rate of 0.53 Gy/min). The endothelial progenitor cells (EPCs) containing green fluorescent protein positive cells (GFP+ ) were infused into mice within 4 hours. The frozen cryosections of visceral organs were made after cultured in vitro on the 6th, 12th , 24th ,36th ,48th ,72nd and 96th hour after transplantation and immunofluorescence was used to observe GFP+ in different tissues. Results Under fluorescence microscope, GFP and MECA-32 positive cells were clearly found in the liver and GFP and CD68 positive cells were observed in the spleen and lungs, but GFP and MECA-32 positive cells were not found in the intestine. Conclusion When the tissues after transplantation preconditioning are treated timely and appropriately, the structure and morphology of the tissues as well as GFP positive cells could be observed clearly under fluorescence microscope.
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