A microchamber array for single cell isolation and analysis of intracellular biomolecules

被引:80
作者
Eyer, Klaus [1 ]
Kuhn, Phillip [1 ]
Hanke, Conni [1 ]
Dittrich, Petra S. [1 ]
机构
[1] ETH, Dept Chem & Appl Biosci, CH-8093 Zurich, Switzerland
基金
欧洲研究理事会;
关键词
MICROWELL ARRAYS; GLUCOSE-6-PHOSPHATE-DEHYDROGENASE; QUANTITATION; EXPRESSION; DYNAMICS; SYSTEMS; NUMBER;
D O I
10.1039/c2lc20876h
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:765 / 772
页数:8
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