Publication: Real-time tracking, retrieval and gene expression analysis of migrating human T cells
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Date
2015-01-01T00:00:00Z
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Mehling, Matthias
Frank, Tino
Albayrak, CEM
Tay, Savas
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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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Mehling M., Frank T., Albayrak C., Tay S., -Real-time tracking, retrieval and gene expression analysis of migrating human T cells-, LAB ON A CHIP, cilt.15, sa.5, ss.1276-1283, 2015