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Issue 5, 2015
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Real-time tracking, retrieval and gene expression analysis of migrating human T cells

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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.

Graphical abstract: Real-time tracking, retrieval and gene expression analysis of migrating human T cells

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Supplementary files

Article information


Submitted
03 Sep 2014
Accepted
04 Dec 2014
First published
05 Dec 2014

Lab Chip, 2015,15, 1276-1283
Article type
Paper
Author version available

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

M. Mehling, T. Frank, C. Albayrak and S. Tay, Lab Chip, 2015, 15, 1276
DOI: 10.1039/C4LC01038H

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