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Stochasticity in gene expression, protein or metabolite levels contributes to cell-cell variations, the analysis of which could lead to a better understanding of cellular processes and drug responses. Current technologies are limited in their throughput, resolution (in space, time, and tracking individual cells instead of population average) and the ability to control cellular environment. A few microfluidic tools have been developed to trap and image cells; however, in most designs available to date, there is a compromise among loading efficiency, speed, the ability to trap single cells, and density or number of trapped cells. To meet the needs of single-cell imaging studies, we developed a microfluidic platform for high-throughput capture and imaging of thousands of single cells. The optimized trapping mechanism enables 95% of the traps to be occupied with single cells, with a trap density of 860 traps/mm(2). The dense array allows up to 800 cells to be imaged simultaneously with a 4x objective and a typical camera setup. Capture occurs with low shear and 94% viability after 24 h. This platform is compatible with other upstream microfluidic components for complex cell stimulation patterns, and we show here the ability to measure heterogeneity in calcium oscillatory behavior in genetically identical cells and monitor kinetic cellular response to chemical stimuli.

Imaging Single-Cell Signaling Dynamics with a Deterministic High-Density Single-Cell Trap Array