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Issue 4, 2014
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Quantifying genetically inserted fluorescent protein in single iPS cells to monitor Nanog expression using electroactive microchamber arrays

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Abstract

Interest in the gene expression levels of pluripotent stem cells has increased in order to precisely understand cellular differentiation. Here, we propose a method utilizing a large number of arrayed microchambers to quantitatively measure an intracellular fluorescence protein that is genetically inserted to monitor a pluripotency marker protein, Nanog, in pluripotent stem cells. Individual cells are isolated and lysed by inducing an electric potential on the cell membrane within the tightly enclosed microchambers. The microchambers have a size that is comparable to the target cells, making it possible to trap single cells and restrict the dilution of the cell lysate. The amount of intracellular fluorescence proteins in a single cell is precisely quantified inside the well-defined volume of each microchamber. Our method will be a useful tool for high-throughput and parallelized read-outs of gene expression levels in individual cells in a large population of cells.

Graphical abstract: Quantifying genetically inserted fluorescent protein in single iPS cells to monitor Nanog expression using electroactive microchamber arrays

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Publication details

The article was received on 24 Sep 2013, accepted on 25 Oct 2013 and first published on 09 Dec 2013


Article type: Paper
DOI: 10.1039/C3LC51086G
Citation: Lab Chip, 2014,14, 730-736
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    Quantifying genetically inserted fluorescent protein in single iPS cells to monitor Nanog expression using electroactive microchamber arrays

    S. H. Kim, X. He, S. Kaneda, J. Kawada, D. Fourmy, H. Noji and T. Fujii, Lab Chip, 2014, 14, 730
    DOI: 10.1039/C3LC51086G

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