BD FACSDiscover S8: A multi-laser spectral cell sorter with BD CellView Image Technology - Project Summary/ Abstract The aim of the Robert H. Lurie Comprehensive Cancer Center Flow Cytometry Core Facility is to provide analysis and cell sorting to enhance scientific interaction and productivity within Northwestern University’s research community. This is accomplished by allowing access to quality controlled highly specialized technology, education, and technical assistance in a cost-effective manner. All sorting demands for the entire campus are accomplished on six BD instruments (four high-end and two mid- level sorters) and one Miltenyi MACSQuant Tyto. Our high-end sorters are heavily utilized and exceed over 4900 hours, even though our total practical capacity is 4572 hours. Additionally, three of our high-end sorters are also nearing their end-of-life (over 10 years and showing increased downtimes resulting from age and repairs. This, together with the increased user demand, leads to scheduling wait times of 2-4 weeks. Since much of our user base utilizes human patient and other time-sensitive specimens, which tend to come in at short notice, we are no longer able to accommodate these sorting requests in a timely manner. Should a sorter be down due to major repairs or must be decommissioned due to extensiveness of repairs; not having a replacement sorter to provide uninterrupted service, further hinders investigators’ research. Additionally, with installation of a 5-laser 50-parameter BD FACSymphony S6 cell sorter in 2021, many of the users have transitioned or are in the process of transitioning to more complex 18-33 color assays for spectral based-cell sorting. However, the current FACSAria’s lack the ability to translate these panels directly into cell sorting for further downstream assays, thus limiting research. This proposal is for purchase of a 5-laser 78- fluorescent detector system (86-parameters total) BD FACSDiscover S8 cell sorter with CellView Image Technology, integrated in a biosafety cabinet. This is the first high-dimensional sorter combining spectral flow cytometry with real-time spatial and morphological insights, thus empowering scientists to address previously impossible-to-answer questions. The CellView Image Technology will further enhance and allow for assays including sort QC, label-free sorting, fluorescent localization, cell-cycle analysis and cell-cell interactions. The instrument, in addition to providing the acutely needed additional capacity, will accommodate the high parameter cell sorting demands and allow for better detection and efficient sorting of rarer events. The sorter will be permanently integrated in a Baker Class II Type A2 biosafety hood. This will provide state-of-the-art cell sorter instrumentation that can keep pace with the technological advances and growing needs of future. This sorter will enable safe single cell high- parameter image-capable spectral cell sorting of human and other animal specimen to meet the increasingly complex needs of our user base and advancing research in a timely manner.
