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目的 建立羊膜组织共聚焦激光扫描显微镜光学切片的方法 ,探讨早期人胚羊膜细胞结构、分层及细胞骨架的空间结构。 方法 采用共聚焦激光扫描光学切片和荧光探针双重标记技术对妊娠 7～ 9周早期人胚羊膜进行形态观察、细胞F 肌动蛋白表达定量分析、细胞分层及厚度计算。 结果 羊膜由两层不同类型的细胞组成 ,内层细胞为多边形扁平细胞 ,外层细胞为长梭形细胞 ;内层细胞胞质中的F 肌动蛋白表达量较低 ,外层细胞胞质中的F 肌动蛋白表达量明显高于内层细胞 ,两者差异有统计学意义 ;7～ 9周人胚羊膜厚度为 30± 2 μm。其中 ,上皮层厚度为 10 μm± 2 μm ,间充质层厚度为 14μm± 2 μm。 结论 1 早期人胚羊膜由两层细胞即内层的上皮细胞层和外层的间充质细胞层组成 ,胞质中F 肌动蛋白的表达量间充质细胞高于上皮细胞 ;2 通过共聚焦激光扫描连续光学切片可以计算出羊膜各层的厚度 ;3 共聚焦激光扫描显微镜光学切片技术结合FITC 鬼笔环肽和碘化丙啶双重标记技术是观察和分析羊膜组织细胞立体结构的良好方法。
Objective To establish an amniotic membrane confocal laser scanning microscopy optical section method to explore the early human embryonic amniotic cell structure, stratification and cytoskeleton spatial structure. Methods Confocal laser scanning optical microscopy and fluorescent probe double labeling technique were used to observe the morphological changes of human embryonic amniotic membrane in early 7-9 weeks gestation, quantitative analysis of F-actin expression, cell stratification and thickness calculation. Results The amniotic membrane consisted of two layers of different types of cells. The inner cells were polygonal flat cells and the outer cells were long spindle cells. The expression of F actin in the cytoplasm of the inner layer was low, F actin expression was significantly higher than the inner layer of cells, the difference was statistically significant; 7 to 9 weeks of human embryonic amniotic membrane thickness of 30 ± 2 μm. Among them, the thickness of epithelial layer is 10 μm ± 2 μm, and the thickness of mesenchymal layer is 14 μm ± 2 μm. Conclusion 1 The early human embryonic amniotic membrane consists of two layers of cells, the inner layer of the epithelial cell layer and the outer layer of mesenchymal cell layer, F-actin expression in the cytoplasm of mesenchymal cells higher than epithelial cells; 2 through a total of Confocal laser scanning of continuous optical sections can be calculated amniotic layer thickness; 3 confocal laser scanning microscopy optical sectioning technology combined with FITC phalloidin and propidium iodide double labeling technology is to observe and analyze the amniotic membrane cells a good method of stereoscopic structure .