Department of Chemistry and Applied Biosciences, ETH Zurich, Zurich, Switzerland.
Lab Chip. 2012 Feb 21;12(4):765-72. doi: 10.1039/c2lc20876h. Epub 2011 Dec 20.
We present a microfluidic device that enables the determination of intracellular biomolecules in multiple single cells. The cells are individually trapped and isolated in a microchamber array. Since the microchambers can be opened and closed reversibly, the cells can be exposed to different solutions sequentially, e.g. for incubation, washing steps, labelling and finally, for lysis. The tightly sealed microchambers enable the retention and analysis of cell lysate derived from single cells. The performance of the device is demonstrated by monitoring the levels of the cofactors NADPH and NADH both in healthy mammalian cells and in cells exposed to oxidative stress. The platform was also used to determine the toxic impact of the alkaloid camptothecin on the intracellular enzyme glucose-6-phosphate dehydrogenase levels. In general, the device is applicable for the analysis of cell auto-stimulation and the detection of intracellular metabolite concentration or expression levels of proteins.
我们介绍了一种微流控装置,该装置可用于测定多个单细胞内的细胞内生物分子。细胞被单独捕获并隔离在微室阵列中。由于微室可以可逆地打开和关闭,因此可以依次将细胞暴露于不同的溶液中,例如孵育,洗涤步骤,标记,最后裂解。密封紧密的微室可保留和分析来自单细胞的细胞裂解物。该装置的性能通过监测健康哺乳动物细胞和暴露于氧化应激细胞中的辅因子 NADPH 和 NADH 的水平来证明。该平台还用于测定生物碱喜树碱对细胞内酶葡萄糖-6-磷酸脱氢酶水平的毒性影响。通常,该设备适用于分析细胞的自刺激以及检测细胞内代谢物浓度或蛋白质的表达水平。