Aiming at the segmentation of human cytoplasm，injection pipette and egg polar body in the process of intra-cytoplasmic sperm injection (ICSI)，firstly，the operator of mathematical morphology is adopted to detect the region of interest according to the difference in shapes of segmentation targets． Secondly，according to local gray level and local variance，an improved fuzzy clustering method is used to implement the rough classification of pre-processing images with fuzzy boundaries and noises． Then，the level set algorithm is used to carry out the segmen-tation of cytoplasm，polar body and injection pipette，with its initialization location and controlling parameters be-ing set according to fuzzy clustering results． Finally，line fitting is employed to mark the whole injection pipette from the local images of injection pipette inserted in cytoplasm． Experimental results show that the proposed algo-rithm can implement the segmentation of human oocyte cytoplasm，polar body and injection pipette correctly．