title: Quantifying genetically inserted fluorescent protein in single iPS cells to monitor Nanog expression using electroactive microchamber arrays.
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.
authors: Institute of Industrial Science, The University of Tokyo, Komaba 4-6-1, Meguro-ku, Tokyo 153-8505, Japan. tfujii@iis.u-tokyo.ac.jp.
authors: Soo Hyeon
authors: SH
authors: Kim
authors: Xiaoming
authors: X
authors: He
authors: Shohei
authors: S
authors: Kaneda
authors: Jiro
authors: J
authors: Kawada
authors: Dominique
authors: D
authors: Fourmy
authors: Hiroyuki
authors: H
authors: Noji
authors: Teruo
authors: T
authors: Fujii
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line; D002469:Cell Separation; D005786:Gene Expression Regulation; D005818:Genetic Engineering; D049452:Green Fluorescent Proteins / Q000235:genetics*; D018398:Homeodomain Proteins / Q000235:genetics*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D046228:Microarray Analysis / Q000379:methods*; D059010:Single-Cell Analysis
chemical_list: D018398:Homeodomain Proteins; D049452:Green Fluorescent Proteins