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为观察细胞培养中细胞凋亡的形态及数量,取4℃环境下不同时间培养的小鼠胸腺细胞,用光镜或吖啶橙(AO),溴化乙锭(EB)染色后荧光镜观察。光镜下可见凋亡小体,荧光镜下早期凋亡细胞发绿色或暗绿色荧光,伴有核结构的变化,而晚期凋亡细胞发橙红色荧光并有凋亡小体的形成。结果表明:光镜下需出现典型核形态变化时凋亡才能确认,AO和EB双重荧光染色后荧光镜检在细胞凋亡检测中既可定性,又可定量。
To observe the morphology and quantity of apoptotic cells in the cell culture, thymocytes were cultured in vitro at 4 ℃ and stained with light microscope or acridine orange (AO) and ethidium bromide (EB) . Apoptotic bodies were observed under light microscope. The early apoptotic cells under the fluorescence microscope showed green or dark green fluorescence with changes of nuclear structure, whereas late apoptotic cells developed orange-red fluorescence with the formation of apoptotic bodies. The results showed that apoptosis could be confirmed by the typical nuclear morphological changes under the light microscope. Fluorescence microscopy with AO and EB double staining could be both qualitative and quantitative in detecting apoptosis.