# Robotic Precisely Oocyte Blind Enucleation Method
## 3 Results
### Enucleation Results  
A total of 30 oocytes were enucleated by the proposed robotic operation method. The typical operating process is shown in Figure 11. Figure 11a shows the process of translating the selected oocyte to the specified position near the HM. Figure 11b shows the process of reducing pressure to immobilize the oocyte. Figure 11c shows the process of rotating the oocyte. Figure 11d shows the process of penetrating the EM into the oocyte following the planned trajectory. Figure 11e shows the process of aspirating the specific volume of cytoplasm and genetic material by the proposed SMAC method. Figure 11f shows the process of withdrawing the EM from the oocyte and releasing the enucleated cytoplasm and genetic material. Figure 11: Oocyte enucleation operating process. (**a**) Translate selected oocyte to the specified position near the HM. (**b**) Reduce pressure to immobilize the oocyte. (**c**) Rotate the oocyte. (**d**) Penetrate the EM into the oocyte following the planned trajectory. (**e**) Aspirate the specific volume of cytoplasm by the proposed SMAC. (**f**) Withdraw the EM from the oocyte and remove the cytoplasm. Figure 10: Cytoplasm extracting control result. (**a**) Quadratic trajectory tracking control result. (**b**) Ramp trajectory tracking control result. In order to validate the effectiveness of the proposed enucleation method, the average enucleation time, enucleation accuracy, enucleation success rate, and the developmental potential after enucleation were counted and compared with those obtained in the manual blind aspiration method. The results are summarized in Table 1.  
From translating oocytes to the extraction of genetic material, the average costing time for the proposed method was 34.4 s (\(n=30\)), which was much smaller than the manual method (62 s). The maximum time consumed in operating on a cell was 52.8 s, and the minimum was 14.2 s. The reason for the large operation time difference is that the posture of each immobilized oocyte is random, so the rotated angle and the time consumed are different between each other. In the process of enucleation, the volume of cytoplasm inside the EM is fed back to the control system by image processing. The desired quadratic trajectory was tracked by the designed adaptive slide mode controller. Based on the fact that the removed cytoplasm and genetic material will form into a sphere in the culture medium, we counted the volume of enucleation, as shown in Figure 12a. The volume of enucleation was 8% of the cytoplasm. We calculated the standard deviation of the enucleation volume as the criterion of enucleation accuracy. The result shows that the accuracy of the proposed enucleation method was 5.2, and the accuracy of the manual method was 30.7. Then, we counted the success rate of enucleation by the oocyte fluorescent dye method. The results demonstrate that the success rate was 93.3% (28/30) by the proposed enucleation method. The accuracy of enucleation and success rate of enucleation are both significantly improved in comparison to the manual blind enucleation method [34], as shown in Table 1.  
At last, we injected the somatic cell into the enucleated oocyte and recorded the cleavage rate of the embryos. The cleavage rate was used to qualify the developmental potential after enucleation. The embryos after enucleation and somatic cell injection were cultured at 37 \({}^{\circ}\)C with 5% CO\({}_{2}\) for 24 h, as shown in Figure 12b. The cleavage rate after enucleation with the proposed method was 63.3% (19/30), which was also significantly improved in comparison to the manual blind enucleation method (41.67%) [32]. The low time-cost, high accuracy, high success rate, and high cleavage rate could demonstrate the effectiveness of the proposed robotic enucleation method.