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


Mehling M., Frank T., Albayrak C. , Tay S.

LAB ON A CHIP, vol.15, no.5, pp.1276-1283, 2015 (Journal Indexed in SCI) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 15 Issue: 5
  • Publication Date: 2015
  • Doi Number: 10.1039/c4lc01038h
  • Title of Journal : LAB ON A CHIP
  • Page Numbers: pp.1276-1283

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.