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Issue 1, 2011
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High-throughput single-cell quantification using simple microwell-based cell docking and programmable time-course live-cell imaging

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Abstract

Extracting single-cell information during cellular responses to external signals in a high-throughput manner is an essential step for quantitative single-cell analyses. Here, we have developed a simple yet robust microfluidic platform for measuring time-course single-cell response on a large scale. Our method combines a simple microwell-based cell docking process inside a patterned microfluidic channel, with programmable time-course live-cell imaging and software-aided fluorescent image processing. The budding yeast, Saccharomyces cerevisiae(S. cerevisiae), cells were individually captured in microwells by multiple sweeping processes, in which a cell-containing solution plug was actively migrating back and forth several times by a finger-pressure induced receding meniscus. To optimize cell docking efficiency while minimizing unnecessary flooding in subsequent steps, circular microwells of various channel dimensions (4–24 µm diameter, 8 µm depth) along with different densities of cell solution (1.5–6.0 × 109cells per mL) were tested. It was found that the microwells of 8 µm diameter and 8 µm depth allowed for an optimal docking efficiency (>90%) without notable flooding issues. For quantitative single-cell analysis, time-course (time interval 15 minute, for 2 hours) fluorescent images of the cells stimulated by mating pheromone were captured using computerized fluorescence microscope and the captured images were processed using a commercially available image processing software. Here, real-time cellular responses of the mating MAPK pathway were monitored at various concentrations (1 nM–100 µM) of mating pheromone at single-cell resolution, revealing that individual cells in the population showed non-uniform signaling response kinetics.

Graphical abstract: High-throughput single-cell quantification using simple microwell-based cell docking and programmable time-course live-cell imaging

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Publication details

The article was received on 15 Jun 2010, accepted on 06 Sep 2010 and first published on 19 Oct 2010


Article type: Paper
DOI: 10.1039/C0LC00114G
Citation: Lab Chip, 2011,11, 79-86

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    High-throughput single-cell quantification using simple microwell-based cell docking and programmable time-course live-cell imaging

    M. C. Park, J. Y. Hur, H. S. Cho, S. Park and K. Y. Suh, Lab Chip, 2011, 11, 79
    DOI: 10.1039/C0LC00114G

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