Funds are requested for the purchase of a BD FACSAria(TM) flow cytometer and cell sorter, which will serve to establish a flow cytometry core facility at the University of California, Santa Barbara. The flow cytometry core facility will provide shared instrument access to a high-end cell sorter, to support the NIH funded research of the major and minor user groups identified herein. This new instrument will provide the investigators with advanced capabilities in high speed data acquisition, high speed cell sorting, and resolution and sensitivity, while providing a platform which is relatively simple to operate and maintain, when compared to traditional droplet sorters. This particular instrument is unique among commercial flow cytometers in that it integrates true high-speed cell sorting and single cell deposition, and non-standard light sources with the sensitivity that is generally associated with 'cuvette' sorters. This combination of specifications is precisely what is necessary at UCSB to perform a variety of microbial analysis and sorting applications, for studies in bacterial pathogenesis and the screening of protein libraries produced in bacteria. The five major users sponsoring this proposal represent five different academic departments and two colleges on the UCSB campus including Chemistry, the Neuroscience Research Institute, Molecular Cellular and Developmental Biology, Chemical Engineering, the Marine Science Institute, and Biomolecular Science and Engineering. These investigators are Principal Investigators on NIH supported research projects that are in need of a cell sorting facility. These NIH projects are focused on biomedical problems that include the genetic regulation of bacterial adhesion in disease, genetic studies of bacterial virulence factors, molecular mechanisms of neuronal degenerative diseases including Alzheimer's and Parkinson's, and engineering of intracellular sensors of enzyme activity and ligand receptor engineering. The requested cell sorter is urgently needed at UCSB to support these biological and medically oriented research projects. This new sorter will provide accurate, high-speed cell sorting (which will enable applications involving rare event sorting), single cell cloning, violet diode excitation for use with engineered fluorescent proteins and quantum dots, and gel coupled optics enabling discrimination of even dimly fluorescent bacteria and lipid vesicles, and consequently will enhance and accelerate ongoing biomedical research at UCSB.
