Application of Fluorescence Microscopy in Research of Oocyte Meiosis [荧光显微成像用于研究卵母细胞减数分裂过程]

作者：Deng Y.; Ye X.; Cao B.; Zhou J.; Liufu T.; Wang X.; Qu J.; Xu G.^*

来源：CHINESE JOURNAL OF LASERS-ZHONGGUO JIGUANG, 2018, 45(5): 0507002.

DOI：10.3788/CJL201845.0507002

• Biotechnology
• Chromosome
• Confocal microscopy
• Dynein
• Oocyte
• Stochastic optical reconstruction microscopy
• Tubulin

Dynein, tubulin, and chromosome play important roles in oocyte meiosis. However, the traditional fluorescence microscopy is limited by the diffraction limit and has low resolution, which cannot meet the imaging requirements of oocyte meiosis. Firstly, the normal maturation system of oocyte in vitro and abnormal maturation system under the action of sodium orthovanadate (SOV) are established. Then the confocal fluorescence microscopy and stochastic optical reconstruction microscopy (STORM) are applied to study the localization of the two important proteins of dynein and tubulin and the morphology and structure of chromosome during the oocyte meiosis. The fluorescence imaging results show that the co-localization of dynein, spindle assembled by tubulin, and the morphology and structure of chromosome can correctly reflect the different stages of oocyte meiosis in the normal system. In the SOV treated groups, the abnormality rate of spindle structure increases, and there are typical spindle structures such as barrel, slender, regiment, disorderly, piriform, and multipolar. The abnormality rate of chromosome structure also increases. The STORM results clearly present the structural information of the spindle, and the 3D STORM results reveal the disorder level of abnormal spindle. This method provides an imaging approach with higher resolution for the research of the oocyte meiosis.