Prediction-Based Method for Micropipette Approaching to Optimal Spindle Removal Position

During somatic cell nuclear transfer (SCNT), precise removal of the oocyte genetic material is critical. However, due to the invisibility of the genetic material (spindle) under brightfield observation and the potential displacement caused by the movement of micropipettes, accurately positioning the micropipette at the spindle poses a significant challenge. This study introduces an approach for optimal spindle removal by predicting its position. Initially, the polarization imaging system visualized the oocyte spindle, while a Multi-Feature Adaptive Kernel Correlation Filter (MFAKCF) algorithm tracked the spindle with 92.84% accuracy. Subsequently, enhancements were made to the Nonlinear Mass-Spring-Damper (NMSD) model to simulate live oocyte mechanical characteristics. Adjustments to NMSD model parameters simulated spindle displacement variations under diverse experimental conditions. Finally, the optimal spindle removal position was determined using NMSD model to simulate the micropipette approach to the spindle and the resulting position of spindle displacement. Experimental validation showed that the predictive accuracy of this model was 97.26%, with an average positional error of $0.4~\mu $ m. Using this approach method can reduce cytoplasm loss to 4.5% and have a 100% enucleation success rate. Thus, the proposed prediction based optimal spindle removal position method can effectively anticipates spindle final positions, aiding in minimizing cytoplasmic loss during spindle removal. Note to Practitioners—Accurate oocyte nucleus removal is crucial for somatic cell nuclear transfer (SCNT) but challenging due to spindle invisibility under brightfield microscopy and displacement caused by micropipette movements. This study proposes a predictive method to address these issues. The polarization imaging system was used to visualize the spindle, and a MFAKCF tracking algorithm enabled real-time tracking. An enhanced NMSD model simulated spindle displacement, enabling accurate position prediction and reducing cytoplasmic loss during removal. This method offers a reliable tool for precise spindle removal in SCNT and broader applications in biomedical micromanipulation.