The BD FACSDiscover S8 Cell Sorter for Flow Cytometric Sorting

PROJECT SUMMARY The goal of this Shared Instrumentation Grant application is to provide high-parameter flow cytometric and image-based cell sorting to the Penn State College of Medicine research community through the well- established Flow Cytometry Core Facility by purchase of the BD FACSDiscover S8 Cell Sorter. Our current high-parameter sorter will soon become obsolete by the manufacturer, creating an urgent need for its replacement to avoid interruptions to current NIH-funded projects. Cell sorting by high-parameter flow cytometry enables isolation of pure target cell populations from a complex mixture of cell types derived from a variety of sources, an essential technology for our NIH-funded users. These cell populations are identified using a strategically developed panel of fluorochrome-tagged biomarkers which are excited by lasers and the resulting emission captured by sensitive photodetectors to yield intensity signals linked to each cell. Through electronic gating of the resulting cell subpopulations, each cell of interest is electrostatically sorted into a receptacle of choice for further experiments. Multiple populations and large numbers of cells can be simultaneously identified and collected in a short time due to processing of thousands of events per second. Our current heavily used BD Biosciences FACSAria instrument will become obsolete by the manufacturer in 2025. This instrument is a workhorse for our users to purify cell populations for a wide array of downstream studies investigating biological and genomic cell properties. In this proposal, we describe the new BD FACSDiscover S8 Cell Sorter and how its state-of-the-art technology is essential for the NIH funded projects of our users. The S8 sorter uses full spectrum fluorescence detection to increase the sensitivity and expand the number of fluorochromes that can be simultaneously detected on single cells. In addition, this instrument provides novel image-based cell sorting, not previously achievable, so dramatically expands the cell sorting capabilities that can be offered to support our investigators. This new instrument will support the funded research of 11 major users and three minor users addressing key questions in medical science that include cancer, infectious diseases, immunology, heart disease and neuroscience. The added benefit of imaging-based quality control combined with full spectrum fluorescence detection is expected to increase the rigor and reproducibility of cell sorting experiments. Penn State College of Medicine provides consistently strong institutional support in the form of a fully staffed Flow Cytometry Core and purchase of yearly service contracts to maintain state-of-the-art instrumentation in top working order. The studies proposed by our investigators match well with the public health goals established by the NIH and Penn State College of Medicine. This instrument will increase the achievable discoveries supported by existing NIH funding and will provide a powerful research tool to support new NIH funded studies in cancer, immunology, virology, genomics, cardiovascular disease and neuroscience.