卵母细胞减数分裂异常是新生儿先天畸形和妊娠失败的主要原因之一，该过程涉及多种蛋白分子协同作用。但受现有成像方法空间分辨率低、景深小等因素限制，目前还无法对这一复杂动态过程进行实时成像。针对此现状，本项目将研究并建立一套大景深三维动态纳米分辨显微成像系统，利用片状光激发和大景深分子定位技术，同时获取卵母细胞减数分裂过程中微管蛋白缓变结构的三维超分辨图像，及动力蛋白Dynein、纺锤体检验点蛋白Mad和Bub的三维动态分子定位信息。结合传统生化分析结果，研究这四种关键蛋白分子在减数分裂过程中的分布、运动和相互作用规律，探索其在同源染色体精确分离时的时间和空间调控原理，推演染色体非整倍体发生的分子机制。项目成果将为活细胞内多分子实时动态研究提供新方法，对阐明卵母细胞减数分裂过程的多分子协同作用具有重要的学术价值，同时对预防和控制非整倍体畸变、降低新生儿出生缺陷具有重大的社会意义。

The abnormality of oocyte meiosis is one of the main reasons for congenital malformations and pregnancy failure, which involves the synergistic dysfunctions of multiple molecules. However, the inherent properties of optical imaging methods, such as poor spacial resolution and small depth of field, impedes researchers to visualize such complex dynamic process. Thus, we will study and establish a large depth of field, 3-dimensional dynamic super-resolution fluorescence microscopic imaging system to study the oocyte meiosis. The excitation by sheet light and the localization by extended large depth of field technique will be applied. The 3D super-resolution images of spindles constructed by microtubule protein during oocyte meiosis will be obtained. At the same time, the dynamic tracking and localization of 3 key proteins, motor protein dynein, and spindle assembly checkpoint proteins Mad (mitotic arrest deficient) and Bub (budding uninhibited by benzimidazole), will be recorded by using the same system. The conventional biochemical assays will also be performed and the quantitive information will be acquired for analyzing the oocyte meiosis. The distribution, motion and interaction of these proteins during the oocyte meiosis will be studied. The temporal and spacial regulation mechanism of these proteins for fine separation of homologous chromosome will be explored. The mechanism of the aneuploidy occurrence will be deduced. The achievement of the project will provide a novel method for studying on the real-time dynamics of multiple molecules in living cell, which has an important academic value for illustrating the oocyte meiosis involving multiple molecules, as well as has a great social significance for preventing and controlling the aneuploidy aberration, and reducing the birth defects has great social significance.