The purpose of this study was to evaluate the pathologic changes of human breast cancer specimens ablated with high-intensity focused ultrasound (HIFU) in vitro. Twenty specimens of pathologically confirmed breast cancer tissue were ablated with ultrasound-guided HIFU. The evaluation methods include histopathologic observation using hematoxylin-eosin staining, electron microscopic imaging, enzyme histochemical and immunohistochemical examination on tumor antigens. Vacuole-like structures in cytoplasm were observed by histopathologic observation but there were no significant changes in cell morphology and nucleus karyotype. Typical phenomena related to coagulation necrosis were observed in electron microscopic studies; the contour of cell structure was still preserved but the structures of cell (all kinds of organelles and nucleus) were damaged or disappeared. Acid phosphatase and succinate dehydrogenase staining showed that tumor cells were inactivated. In immunohistochemical evaluations, estrogen receptor, progesterone receptor, cerbB-2 and P53 expression changed from 85%, 82%, 75% and 80% in nonablation tissue to no expression in ablated tumor tissue, respectively. We, therefore, conclude that breast cancer cells appear normal contour immediately after ablation with HIFU under light microscopic but they were evaluated to be dead by electron microscopic imaging, enzyme histochemical and immunohistochemical examinations.