This request is for funds to purchase a biocontained ThermoFisher Bigfoot spectral sorter (7 laser, 60 detector). There is no full-spectrum cell sorter at the University of Pittsburgh or within 100 miles. Unique as an instrument, the Bigfoot sorter combines the strengths of high-parameter, high-content imaging (multi-laser, high-detector) with modern full-spectrum technologies (algorithmic spectral unmixing). Our ongoing NIH-funded research is hampered by the inability to directly transition complex, validated panels from Pitt's three spectral analyzers to an equivalent spectral cell sorter. In spectral approaches, the full spectrum of light emitted from each cell is collected by multiple detectors followed by unmixing to separate the signal into individual markers. Spectral cytometry expands parameter space by permitting the use of new fluors and also new combinations of existing fluors that could not previously be paired together in conventionally configured cytometers (i.e., one detector, one color systems). All of the sorters at Pitt are the conventional configuration type. The consequence is that most or all of our highly complex immunophenotypic spectral panels have to be scaled down, adapted to fewer detectors, and re-validated for compatibility with existing conventional sorters, all of which fails to capture the original complexity and novel information tantalizingly revealed by the spectral analyzers. Further, our current conventional sorters are heavily used. A very attractive feature of the Bigfoot is that it that has two operational modes, spectral and conventional. Thus, the Bigfoot meets the demands of users with high-complexity spectral sorting needs, and also benefits all users of our very busy core who have conventionally compensated needs. The requested Bigfoot spectral sorter will be housed within the centralized Unified Flow Cytometry Core in Pitt's School of Medicine. The core serves over 180 Pis and 500 individual users. For the proposed instrument, Major Users include 10 Pis (9 NIH funded) with another 7 Pis as Minor Users. Together, these Pis have over $30 Min NIH-funded direct costs from at least 5 different institutes. A major research focus is in the broad area of immunology and allied immune diseases, with sister disciplines such as transplantation, dermatology, vascular biology, infectious disease, and oncology also well represented. Our preliminary and demo work proved to us that this instrument will truly allow our investigators to break new ground in their research. The Bigfoot will be well cared for by the staff of the Unified Flow Cytometry Core, which has an extensive infrastructure and culture in place to support panel design, oversee instrument operations, and ensure that it is financially self-supporting. In addition to providing unique and much needed spectral sorting capabilities, the instrument will accommodate demands for configurable sorts including 6-way sorts into tubes and direct sorting into 1 Ox ch...