Robotic Single-Micropipette Enucleation Method for Zona-free Oocytes

Somatic cell nuclear transfer is a mammalian cloning method that involves replacing the oocyte nucleus with a somatic cell nucleus. One of the most critical steps in this process is the efficient production of large numbers of enucleated oocytes. While a single-micropipette-based manual enucleation method has been widely used in manual cloning, it is still a manual operation that requires exceptional technical skills and experience. Additionally, it is difficult to accurately control the amount of enucleation and to meet the demand for high-throughput enucleation. Therefore, we propose a robotic enucleation method using a single micropipette for zona-free oocytes, which allows for the detection and control of the amount of enucleation, as well as continuous enucleation on a large scale. A single oocyte enucleation can be completed in less than 13 seconds. This robotic enucleation method provides a solution for enhancing the operational efficiency of robotic somatic cell nuclear transfer.