改良透射电镜方法观察小鼠ICSI胚胎核仁超微结构  

Observation of the Nucleolar Ultrastructure of Mouse ICSI Embryos by Modified Transmission Electron Microscope Technique

作　　者：郑重[1] 胡丽丽[1] 文刚[1] 张宝东[1] 雷蕾[1] 

机构地区：[1]哈尔滨医科大学基础医学院组胚教研室,哈尔滨150081

出　　处：《中国细胞生物学学报》2011年第2期154-158,共5页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(No.30671025);黑龙江省留学归国人员科学技术专项资金(No.LC07C17)资助项目~~

摘　　要：小鼠植入前胚胎的发育过程中,核仁经历从简单到复杂、从致密结构到网状结构的变化。对核仁超微结构的观察有助于揭示早期胚胎发育过程中核仁结构的动态变化及其特定阶段的功能。但由于核仁结构微小,数目较少,并且在胚胎中只处于卵裂球细胞核的内部,难以定位,因而给核仁的超微结构观察带来很大的困难。本实验探索了透射电镜观察小鼠植入前胚胎核仁的方法:先用琼脂对小鼠胚胎进行预包埋,在经过常规的透射电镜样品制备流程后,将整个胚胎先切成半薄切片;经过甲苯胺蓝染色后,选取含核仁结构的切片进行重包埋;最后再对回收来的半薄切片进行超薄切片,醋酸铀染色后上电镜观察;最终成功获得小鼠胚胎植入前发育不同时期核仁清晰的透射电镜图像。Nucleolar structure of mouse embryo varies during pre-implantation development. Ultrastructural observation of nucleolus is necessary to reveal the dynamic changes of nucleolar structure and their important roles in pre-implantation development. However, it is relative hard to conduct the experiment for nucleoli in common TEM method as they are few in number, small in diameter and difficult to locate. In this study, we explored a way to observe the nucleolar ultra-structure of mouse pre-implantation ICSI embryos. Firstly, we embedded the embryos in agar to make it easier to operate. Then, we made semi-thin sections after routine TEM sample preparation. After stained with toluidine blue, the semi-thin sections with nucleoli structure were selected, and then they were re-embedded and recycled. Finally, the ultra-thin sections were produced and observed under TEM. By using this modified embryonic TEM sample preparation, we obtained clear images of the nucleolar ultra-structure of mouse embryos in different pre-implantation stages.

关 键 词：小鼠胚胎 核仁 透射电镜 超微结构 重包埋 

分 类 号：Q954.4[生物学—动物学]