We have developed a flow cytometric assay to measure the oxidative capacity of cultured lymphoblasts as a possible screening test for patients suspected of having a defect of the mitochondrial respiratory chain. Cells were incubated overnight in serum free media, followed by incubation with dihydroethidium with and without rotenone, and then analysed using flow cytometry to measure fluorescence. Inhibition with rotenone gave an increase in fluorescence compared to uninhibited cells. The change in fluorescence was significantly lower in four of the six patient cell lines, with a correlation between the activity of complex I and change in fluorescence. This method may be applicable to cell lines with defects in other complexes of the respiratory chain.

中文翻译：

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流式细胞术在线粒体呼吸链疾病研究中的应用

我们开发了一种流式细胞仪测定法来测量培养的淋巴母细胞的氧化能力，作为对怀疑有线粒体呼吸链缺陷的患者的一种可能的筛查试验。将细胞在无血清培养基中孵育过夜，然后与含有和不含鱼藤酮的二氢乙锭一起孵育，然后使用流式细胞术进行分析以测量荧光。与未抑制的细胞相比，鱼藤酮的抑制导致荧光增加。六个患者细胞系中的四个细胞系的荧光变化显着降低，复合物 I 的活性与荧光变化之间存在相关性。该方法可能适用于呼吸链其他复合体存在缺陷的细胞系。