Live-Cell Imaging of Chromosome Segregation During Mouse Oocyte Meiosis.

Accurate chromosome segregation during oocyte meiosis is essential for ensuring proper embryonic development and preventing aneuploidy-related disorders. Live-cell imaging combined with fluorescence labelling techniques have become a powerful approach for studying meiotic chromosome dynamics with high spatiotemporal resolution. In this protocol, we summarize key methodologies for visualizing chromosome segregation in live mouse oocytes, focusing on the use of histone H2B-RFP for chromatin labelling and SiR-Tubulin for spindle tracking. We describe the procedures for sample preparation, mRNA microinjection, oocyte culture, live-cell imaging chamber setup, and confocal microscopy settings, all optimized to achieve high-resolution imaging while minimizing phototoxicity. Furthermore, we highlight critical experimental considerations such as phototoxicity, image processing, and quantitative analysis of meiotic events. By providing a comprehensive evaluation of current methodologies, this protocol serves as a practical guide for researchers seeking to investigate chromosome dynamics in live oocytes and improve the accuracy and reproducibility of meiotic studies.