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Observation of the Nucleolar Ultrastructure of Mouse ICSI Embryos by Modified Transmission Electron Microscope Technique
Zhong Zheng, Li-Li Hu, Gang Wen, Bao-Dong Zhang, Lei Lei*
Department of Histology and Embryology, Harbin Medical University, Harbin 150081, China
Abstract: 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.
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