中文摘要：

目的探索一种新的能长期、高效保存并研究肿瘤于细胞球囊的技术。方法从恶性胶质瘤原代肿瘤细胞中培养出肿瘤干细胞。将第5代肿瘤干细胞球囊经蛋清石蜡两次包埋、切片后，行HE染色、免疫组织化学染色和免疫组织荧光染色，并与传统的肿瘤干细胞球囊包埋技术及球囊免疫组织荧光染色实验进行比较。结果应用蛋清石蜡两次包埋处理的球囊切片经HE染色后，球囊分布均匀、结构完整，胞核、胞浆颜色鲜艳、对比度高。球囊切片的免疫组织化学染色和免疫组织荧光染色能准确定位球囊中阳性细胞和阳性蛋白表达位点，可进行阳性细胞及蛋白的半定量分析。结论肿瘤干细胞球囊蛋清石蜡两次包埋切片技术能提高实验效率，减少实验误差，节省成本，优于传统球囊包埋技术及传统球囊免疫荧光技术。

英文摘要：

Objective To explore a new long-term efficient embedding technique of tumorspheres. Methods Tumor spheres, mainly composed of cancer stem cells, were cultured from glioblastoma tissues. The fifth generation of tumor spheres was chosen for egg-white-paraffin embedding and section. Then, those tumor sphere slices were observed by HE staining, immunohistochemistry and immunofluorescence staining. And the immunofluorescence results of these tumor sphere slices were compared with those tumor spheres kept with traditional methods. Results Immunofluorescence results showed that tumor spheres kept with traditional methods looked blurry, and the positive cells and the positive protein expression sites in the cells could not be displayed. HE staining demonstrated that the tumor sphere slices had well-distributed intact spheres and coloring cells with high karyoplasm contrast. Immunohistochemistry and immunofluorescence staining of tumor sphere slices showed clear background, from which positive cells and positive locus could be easily displayed; therefore, semi-quantitative analysis of the positive cells could be performed. Conclusion Egg-white-paraffin embedding technique of the tumor sphere slices can reduce experimental errors and cut down the costs, which enjoys its advantage as compared with traditional embedding technique of the tumor sphere slices.