title: A microchamber array for single cell isolation and analysis of intracellular biomolecules.
abstract: 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.
authors.affiliation: Department of Chemistry and Applied Biosciences, ETH Zurich, Zurich, Switzerland.
authors.forename: Klaus
authors.initials: K
authors.lastname: Eyer
authors.forename: Phillip
authors.initials: P
authors.lastname: Kuhn
authors.forename: Conni
authors.initials: C
authors.lastname: Hanke
authors.forename: Petra S
authors.initials: PS
authors.lastname: Dittrich
mesh_terms: D002469:Cell Separation / Q000295:instrumentation / Q000379:methods; D005954:Glucosephosphate Dehydrogenase / Q000378:metabolism*; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods*; D009249:NADP / Q000378:metabolism*; D018384:Oxidative Stress / Q000502:physiology*; D020298:U937 Cells
chemical_list: D009249:NADP; D005954:Glucosephosphate Dehydrogenase