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目的：用缺口末端标记技术（TUNEL）对肝硬变、肝细胞肝癌及培养肝癌细胞系的细胞调亡进行形态学观察．方法：采用德国宝灵曼公司生产的原位末端标记试剂盒对福尔马林固定、石蜡包埋的肝组织和培养肝癌细胞进行TUNEL染色，激光共聚焦显微镜观察肝细胞凋亡的形态特征．结果：TUNEL阳性信号为黄绿色或黄色强荧光，位于胞核，呈环行、小圆形或颗粒状，提示凋亡细胞有典型的新月体状染色质边集、核浓缩以及有大量微核体的形成．在阿霉素诱导的HCC-9204细胞系还可见调亡细胞的出泡现象．TUNEL阳性细胞大多HE染色表现为调亡的细胞．结论：TUNEL技术加激光共聚焦显微镜观察凋亡细胞是目前原位观察调亡的理想方法，特别适用于调亡细胞的早期检测．
OBJECTIVE: To observe the morphological changes of cirrhosis, hepatocellular carcinoma and cultured hepatocarcinoma cell lines by nick end labeling (TUNEL). METHODS: Formalin-fixed, paraffin-embedded liver tissue and cultured hepatocellular carcinoma cells were stained by TUNEL using the in situ end-labeling kit manufactured by Bolinger Mannheim GmbH. Confocal laser scanning microscopy was used to observe the morphology of hepatocyte apoptosis. RESULTS: The positive signal of TUNEL was yellow-green or yellow fluorescence, located in the nucleus. The TUNEL positive signal was circular, small round or granular, suggesting that the apoptotic cells had typical crescent-like chromatin edge sets, nuclear condensation, and a large number of micronuclei. Body formation. Apoptosis was also observed in the adriamycin-induced HCC-9204 cell line. Most of the TUNEL positive cells stained with HE stained as apoptotic cells. Conclusion: TUNEL and laser confocal microscopy are the ideal methods to observe apoptotic cells in situ, especially for the early detection of apoptotic cells.