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 five BD instruments (three high-end and two mid- level sorters). Our high-end sorters are heavily utilized and exceed over 5712 hours, even though our total practical capacity is 4572 hours. This, together with the increased user demand, leads to scheduling wait times of 3-5 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. Due to this overutilization of our high-end sorters, the reduction in availability has negatively impacted our investigators' research. Additionally, the performance and capabilities of the current high-end sorters are not well matched to the evolving complex analytical instruments and user demands in the field of high-parameter flow cytometry. With the purchase of 6-laser 30-parameter BD FACSymphony cell analyzer in January 2018, many of the users have transitioned or are in the process of transitioning to complex 15-25 color assays for end point analysis. However, the current FACSAria's lack the ability to translate these panels directly into cell sorting for further downstream assays, thus hampering research. The FACSAria allows for collecting only from a maximum 15-16 simultaneous colors. Also, the lower speed of the electronics limits the collection of a sort population to no more than 8 sequential gates; and only a maximum of 4 populations sorted at a time. These limitations, relative to high-parameter analytical results and sorting capacity has substantially hindered user satisfaction and scientific progress of our Northwestern Research community. This proposal is for purchase of a BD FACSymphony S6 High Parameter SORP cell sorter, a bench-top high speed cell sorter equipped with five lasers (355, 405, 488, 561, and 637nm) capable of analyzing up to 28 different colors plus forward and side scatter. The instrument, in addition to providing the acutely needed added 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 with safe single cell sigh-content high-parameter cellular including human cell sorting capabilities to meet the increasingly complex needs of our user base and advancing research.
Over the past two years, we have observed a 270% increase in the number of hours of cells sorting since 2016 and now have reached maximum capacity. Moreover, with the purchase of the BD Symphony analyzer, our investigators have been able to develop more complex panels to identify heterogeneous populations of cells from patients. The proposed Special Order FACSymphony S6 High Parameter (HP) Cell Sorter (combining the BD FACSymphony high parameter cell analyzer and the BD FACSAria Fusion platform) will not only accelerate the rate of scientific discovery by providing greater capacity and capability of sorting based phenotypic information in a multi-dimensional high complexity environment (especially identification and analysis of rare cell types); but also allow for an easily accessible, yet powerful tool that is central for primary human based cell sorting. Together, this machine will meet the needs of a rapidly developing translational research initiatives at Northwestern University.