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首页> 外文期刊>Clinical and diagnostic laboratory immunology >Detection of apoptotic cells by selective precipitation of [3H]thymidine-labelled DNA.
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Detection of apoptotic cells by selective precipitation of [3H]thymidine-labelled DNA.

机译：通过选择性沉淀[3H]胸苷标记的DNA来检测凋亡细胞。

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Apoptosis is characterized by systematic fragmentation of high-molecular-weight DNA into oligonucleosome fragments. A sensitive method for detection of apoptotic cells involving [3H]thymidine-labelled DNA is presented. Cells from mid-log-phase cultures were labelled with [3H]thymidine for 15 to 18 h and then exposed to gamma irradiation to induce apoptosis. A modified Hirt method was used to separate low-molecular-weight DNA from high-molecular-weight DNA. The percentage of fragmented DNA and high-molecular-weight DNA were measured by scintillation spectrometry. This method was compared with an established flow cytometric method for detection of apoptotic cells by using propidium iodide staining of DNA. We observed a good correlation between these two methods in detecting apoptosis. Hence, expensive flow cytometric assays for detection of apoptosis in dividing cells may be replaceable by a method involving [3H]thymidine labelling of DNA and separation of low-molecular-weight DNA from high-molecular-weight DNA by precipitation.
机译：凋亡的特征是高分子量DNA系统性断裂为寡核小体片段。提出了一种敏感的方法来检测涉及[3 H]胸苷标记的DNA的凋亡细胞。用[3 H]胸苷标记来自对数中期培养物的细胞15至18小时，然后暴露于γ射线以诱导凋亡。使用改良的Hirt方法从高分子量DNA分离低分子量DNA。通过闪烁光谱法测量片段化DNA和高分子量DNA的百分比。将该方法与已建立的流式细胞术方法进行了比较，该方法通过使用碘化丙啶对DNA进行染色来检测凋亡细胞。我们观察到这两种方法在检测细胞凋亡中有良好的相关性。因此，用于检测分裂细胞凋亡的昂贵的流式细胞术测定方法可以通过涉及[3H]胸苷标记DNA并通过沉淀从高分子量DNA分离出低分子量DNA的方法来代替。

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