We have firstly succeeded in enucleation of oocyte by using micro-electric knife. The discharged output power and conductive area were controlled to adapt the cellular-order ablation under water and under atmospheric pressure environment. The local area ablation was obtained by the glass shell insulation around the copper wire with silver paste, and which prevent the bubble generation and protein contamination when it was discharged under the water. The width of the ablation region was successfully reduced to more than a half by using the glass shell insulation compare to the conventional electrode. Moreover, the nucleus dyed by Hoechst was also removed successfully with limited damage region which is the difficult task for manual operation with glass capillary. This low cost micro-electric knife has a possibility to extend to fabricate any objective material under various environments and contribute to a new top-down fabrication method in the micro-nano bioengineering field.