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SELECTBIO Conferences Single Cell Analysis Europe

Single Cell Analysis Europe Poster Presentations

Poster Presentations

An integrated experimental platform to quantify oriented cell polarization in controlled morphogenetic gradients
Sung Lee, , ETH Zurich

Despite the fundamental and physiological importance, little is known at the molecular level how morphogenetic signals are sensed and amplified, in part due to the lack of suitable experimental systems to investigate these complex temporal and spatial regulatory mechanisms of gradient sensing pathways. Here, we developed a single-cell assay platform, and investigated the mating response of budding yeast S. cerevisae as a genetically-tractable model system. We engineered computer-controlled microfluidic devices that can stably regulate the desired gradient profiles at any given time, and combined this platform with multi-dimensional live cell microscopy equipped with automated image analysis programs that automatically segment, individually track and quantify non-uniform cell morphology and XFP-tagged intracellular protein localization. This integrated platform is versatile, and additional options can be implemented including altering the gradient steepness, switching of the gradient direction, or addition of specific inhibitors at any given time. Such perturbations are useful to investigate dynamic cellular feedback and control mechanisms, and in particular to probe the plasticity of polarization signaling. Finally, the experimental platform can easily be adapted to investigate chemotaxis in higher eukaryotes, and may thus have wide applications to elucidate the underlying mechanisms of cancer metastasis and inflammatory diseases.