1. Real-time tracking, retrieval and gene expression analysis of migrating human T cells
- Author
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Savaş Tay, Cem Albayrak, Tino Frank, Matthias Mehling, and ALBAYRAK, CEM
- Subjects
CD4-Positive T-Lymphocytes ,Receptors, CXCR4 ,Chemokine ,T cell ,Biomedical Engineering ,Bioengineering ,Biology ,Time-Lapse Imaging ,Biochemistry ,CXCR4 ,Cell Movement ,Gene expression ,medicine ,Humans ,Directionality ,RNA, Messenger ,Cells, Cultured ,Reverse Transcriptase Polymerase Chain Reaction ,Chemotaxis ,Gene Expression Profiling ,T cell chemotaxis ,General Chemistry ,Microfluidic Analytical Techniques ,Chemokine CXCL12 ,Cell biology ,medicine.anatomical_structure ,Gene Expression Regulation ,Cell culture ,biology.protein ,Single-Cell Analysis ,Fluorescein-5-isothiocyanate - 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.
- Published
- 2015
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