甘氨酰替甘氨酸浸泡法在冷冻割断扫描电镜术中的应用  

Observation on the Ultrastructure of Cell Freeze fractured and Rinsed in Glycylglycine with Scanning Electron Microscopy

作　　者：江一平[1,2] 赵玉珍[1,2] 李向印 雷建章 

机构地区：[1]福建医科大学组织胚胎学教研室 [2]河北医科大学电镜中心

出　　处：《福建医科大学学报》1999年第1期5-9,共5页Journal of Fujian Medical University

摘　　要：目的运用甘氨酰替甘氨酸浸泡法对常规的ＤＭＳＯ法冷冻－割断扫描电镜制样技术加以改良。方法经过多聚甲醛－戊二醛预固定的组织冷冻割断，１％ＯｓＯ４后固定，用甘氨酰替甘氨酸溶液代替常规的低浓度锇酸液浸泡作细胞基质软化。后续步骤按常规方法，扫描电镜观察。结果细胞内部膜系和丝状结构的超微形态保持完好，暴露明显，图像清晰，立体感强。各种膜系结构之间可见三维的丝状网络联系。结论醛类预固定－甘氨酰替甘氨酸浸泡法对细胞骨架的保存和显示较好。Objective\ To modified the conventional DMSO procedure of freeze fracture for scanning electron microscopy(SEM) using glycylglycine(G G) method which was developed by Pinto da Silva(1981).\ Methods\ Glutaraldehyde polyformaldehyde fixed tissue block was frozen in liquid nitrogen and fractured.\ After thawed in PBS,the fractured blocks were postfixed with 1% OsO 4 and rinsed in glycylglycine(G G) for up to 24 h.\ Dehydrate and the succeeded steps were the same with ordinary method.\ Results\ The observation with SEM showed that both the membrane system (including cell surface) and the filament structure of the fractured cells were well preserved and excellent exposed.\ The picture attained with SEM was clear and steric.\ The three dimensional filament network in cell plasma and its co relation with nuclear, mitochondria and RER was viewed.\ Conclusion\ It might be more advantageous to the study on the relationship between membrane system and cytoskeleton using this modified method.

关 键 词：SEM 冷冻割断 甘氨酰替甘氨酸 细胞骨架 

分 类 号：Q954.6[生物学—动物学] R329－33[医药卫生—人体解剖和组织胚胎学]