Real-time tracking, retrieval and gene expression analysis of migrating human T cells. Issue 5 (16th December 2014)

• Record Type: Journal Article
• Authors: Mehling, Matthias; Frank, Tino; Albayrak, Cem; Tay, Savaş
• Is Part Of: Lab on a chip. Volume 15:Issue 5(2015)
• Page Start: 1276
• Page End: 1283
• Publication Date: 2014-12-16
• Subjects: Miniature electronic equipment -- Periodicals; Combinatorial chemistry -- Periodicals; Biotechnology -- Periodicals; 543.0813
• DOI: 10.1039/c4lc01038h
• Languages: English
• ISSNs: 1473-0197

Abstract

Abstract : A microfluidic system for the study of T-cell chemotaxis and subsequent gene expression analysis. 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.