Cells are complex entities that must integrate internal and external signals in order to coordinate diverse functions. Living cells are also dynamic, and this dynamism is key to understanding the mechanisms between cause and effect for biological processes. Our lab develops new technologies to interrogate signaling cascades in cells to understand how the actions of specific proteins, molecules, and ions contribute to cellular function. We combine in vitro spectroscopic and biophysical techniques with protein design and engineering to develop novel fluorescent probes, and use long-term time-lapse fluorescence microscopy to elucidate the mechanisms of cellular signaling pathways. Longitudinal single-cell analyses also allow us to explore the origins and consequences of heterogeneity in biological systems. In this talk I will discuss approaches we have developed to elucidate how pathogens alter mammalian host cell biology and for defining and quantifying heterogeneous phenotypes. Specifically we have established strategies for tagging bacterial effector proteins with fluorescent tags, enabling tracking of translocation and dynamics within infected cells, pipelines for long-term live-cell imaging of infected mammalian cells and subsequent single-cell image analysis methods. We apply these methods to infection of epithelial cells and primary macrophage by Salmonella and Listeria.

Development of approaches for long-term time lapse imaging of host-pathogen interactions

251st National Meeting (2016)

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