This proposal is for a high capacity flow cytometry sorter BD FACSAria Fusion Cell Sorter to allow 6 Major Users (Merit Award funded PIs) and 7 Minor Users at the VA Ann Arbor Health Care System (VAAAHS) to conduct Multi-parameter flow cytometry cell sorting to isolate highly purified cell subsets for subsequent analysis, experimental manipulations, and studies. Our research focuses on cellular and molecular mechanisms of diseases that preferentially affect veterans: COPD, neurodegenerative disorders and CNS trauma, solid cancers and leukemias, the deployment-related lung disease and group of infectious complications for which veterans have higher risk. Involvement of specific and frequently rare cell subsets and their unique responses are increasingly found to influence these conditions and need to be studied. New experimental approaches, such as single-cell RNA sequencing, cellular vector deliveries for immunotherapeutic approaches, and adoptive transfers, all rely on high-purity cell sorting that must be performed rapidly, precisely, and aseptically to acquire specific cell populations with high viability. These needs are not being adequately met at the VAAAHS. Requested FACS sorter, presently unavailable at our VA station, will allow for collection of rare cell populations based on their expression of a unique set of multiple specific markers that would otherwise be impossible or very difficult to obtain. The high capacity sorter will allow for sorting in a timely fashion (to maintain cell viability, sterility and unaltered activation state) while working on multiple samples. Our diverse group of users will be able to 1) effectively collect quantitative data from small and precious samples; 2) use of highly specific subsets of cells in further assays like RNA transcriptome analysis, single-cell sequencing, proteomics, gene array analysis, etc; 3) generate cells for adoptive transfer experiments and preclinical therapy tests; 4) isolate and study purified cell subsets from clinical samples and implanted devices to discover new biomarkers, therapeutic targets, and to advance design of therapeutic implants. Introducing these new capabilities will enhance quality of current studies and increase efficiency and throughput. It will also enhance future grant applications of our faculty and create new learning opportunities for trainees at the VAAAHS site to ultimately benefit Veterans? health.
A shared BD FACSAria Fusion Cell Sorter is requested to provide capability to VA Ann Arbor Health Care Research Service to isolate highly purified cell subsets from a complex mixture isolated from various clinical and laboratory specimens. We will be able to separate these cells into viable, homogenous fractions with a very high purity that approaches 100%. It will enhance current studies by providing new data and improved data collection and analyses as well as increase efficiency and throughput. It will extend current studies into new directions by allowing identifications and assessments that would not otherwise be possible. The requested system will advance research on disorders effecting our veteran population towards developing therapeutic interventions to ultimately benefit our patients.