NERCE Core E - Imaging Resource: Tomas Kirchhausen, Ph.D., Immune Diseases InstituteRecent developments in fluorescence microscopy geared for single-molecule detection, optical sectioningand three-dimensional data acquisition, used in combination with object-identification algorithms, now allowsufficient temporal and spatial resolution to visualize the sequential recruitment and intracellular traffic ofproteins, lipids, and pathogens in living cells including viruses and bacteria. The Kirchhausen laboratory hasa longstanding interest in understanding the mechanisms underlying entry of toxins, viruses and bacteria intocells, and how these processes relate to the organization and intracellular traffic of vesiculo-tubularmembrane carriers. As part of this effort, we have devoted substantial efforts to developing an imaging suitecontaining state-of-the-art microscopes and supporting software suited for data collection and analysis withhigh spatial and temporal resolution. It is also a priority to maintain an imaging suite that is 100% accessibleto all users, so that after initial training, any investigator can perform their work with total flexibility andindependence.In this proposal we outline plans for a NERCE Imaging Core facility that will provide access to contemporarytools and expertise for live-cell and single molecule imaging aimed towards, but not limited to, quantitativedescriptions of mechanisms of invasion for bacterial and viral pathogens into mammalian cells, of viralreplication, and of molecular aspects related to toxin entry into cells.
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