title: Real-time tracking, retrieval and gene expression analysis of migrating human T cells.
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.
authors.affiliation: Department of Biosystems Science and Engineering, ETH Zürich, Mattenstrasse 26, 4058 Basel, Switzerland. savas.tay@bsse.ethz.ch.
authors.forename: Matthias
authors.initials: M
authors.lastname: Mehling
authors.forename: Tino
authors.initials: T
authors.lastname: Frank
authors.forename: Cem
authors.initials: C
authors.lastname: Albayrak
authors.forename: Savaş
authors.initials: S
authors.lastname: Tay
mesh_terms: D015496:CD4-Positive T-Lymphocytes / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D002465:Cell Movement; D002478:Cells, Cultured; D054377:Chemokine CXCL12 / Q000737:chemistry / Q000378:metabolism; D002633:Chemotaxis / Q000502:physiology; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods; D012333:RNA, Messenger / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059010:Single-Cell Analysis; D059008:Time-Lapse Imaging
chemical_list: D054377:Chemokine CXCL12; D012333:RNA, Messenger; D019718:Receptors, CXCR4; D016650:Fluorescein-5-isothiocyanate