UNIVERSITY OF WISCONSIN SYSTEM
We request funds to purchase a BD Biosciences FACSAria II high-speed cell sorter. The instrument will be used by investigators studying immune response to viral infection in animal models. Viral pathogens include highly pathogenic influenza including H5N1 avian influenza and 1918 or Spanish influenza and swine-origin H1N1. To enable investigators to study Select Agents (H5N1 and 1918), the instrument will be installed in a Biological Safety Level 3 Agriculture (BSL3-AG) laboratory in the Influenza Research Institute at the University of Wisconsin-Madison that is approved for research on these organisms. BSL3-level sorting is required for sorts of unfixed infectious materials. The experiments proposed require live cell sorting to provide cellular identification by maintaining cell morphology, enabling sensitive marker detection, and providing conditions for detecting reporter gene product signals. In addition, live sorts enable investigators to conduct downstream analysis of gene or protein expression. This proposed instrument has an aerosol management system to contain aerosols. The sorter is equipped with three lasers that enable researchers to interrogate cells with up to 12 parameters, including 10 fluorescent parameters and forward- and side-scatter parameters. The system affords the ability to identify events using 5-10 fluorescent parameters for studies using multiple markers to resolve subpopulations of tissue-resident cells during infection. The FACSAria II will be used for both analytical and sorting procedures.