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      Real-time tracking, retrieval and gene expression analysis of migrating human T cells.

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      Lab on a chip

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          Abstract

          Dynamical analysis of single-cells allows assessment of the extent and role of cell-to-cell variability, however traditional dish-and-pipette techniques have hindered single-cell analysis in quantitative biology. We developed an automated microfluidic cell culture system that generates stable diffusion-based chemokine gradients, where cells can be placed in predetermined positions, monitored via single-cell time-lapse microscopy, and subsequently be retrieved based on their migration speed and directionality for further off-chip gene expression analysis, constituting a powerful platform for multiparameter quantitative studies of single-cell chemotaxis. Using this system we studied CXCL12-directed migration of individual human primary T cells. Spatiotemporally deterministic retrieval of T cell subsets in relation to their migration speed, and subsequent analysis with microfluidic droplet digital-PCR showed that the expression level of CXCR4 – the receptor of CXCL12 – underlies enhanced human T cell chemotaxis.

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          Author and article information

          Journal
          Lab Chip
          Lab on a chip
          1473-0189
          1473-0189
          Mar 7 2015
          : 15
          : 5
          Affiliations
          [1 ] Department of Biosystems Science and Engineering, ETH Zürich, Mattenstrasse 26, 4058 Basel, Switzerland. savas.tay@bsse.ethz.ch.
          Article
          10.1039/c4lc01038h
          25512266
          f8f88fd6-57a1-4adb-965e-68b1cb9443b9
          History

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