Acquisition of BD FACSDiscovery S8 Spectral Cell Sorter

George Washington University, Washington DC

Investigators

Abstract

We request funds to purchase a Becton Dickinson (BD) FACSDiscover S8 CellView spectral flow cytometer and live cell sorter for use in The George Washington University School of Medicine and Health Sciences (GW SMHS) Flow Cytometry Core Facility. The instrument will be equipped with 5-lasers (349 nm UV, 405 nm violet, 488 nm blue, 561 nm yellow-green, and 638 nm red) and 78 fluorescent APD detectors allowing for broad detection of complex fluorescent panels. The instrument has an additional six imaging detectors generating live images of cells with up to 3 fluorescent colors used collected in the images. This is novel technology and the first live cell sorter capable flow cytometer to incorporate images of each cell into both spectral analysis and sorting. The images incorporate light scatter characteristics such as: forward and side scatter, center of mass, light loss, fluorescent intensity, correlation, diffusivity, eccentricity, and radial moment to determine physical characteristics of the cell and fluorescent labeling to render a live image of the cell. The image properties can then be used to determine which cells will be collected by sorting or analyzed as part of typical FCS files generated by the instrument. The S8 is capable of up to 6 population sorting, plate and index sorting, and standard tube sizes. We selected this instrument to replace a BD Influx 4-laser 15-color sorter that is entering end of life and will be removed from service contracts at the end of 2025. The Flow Core currently operates a Cytek Aurora 3-laser spectral analyzer capable of fluorescent panels of 25 colors. Users have come to rely on the Aurora for the enhanced analysis capabilities of larger color panels. The ability to translate the spectral panels to live cell sorting using the S8 will provide the ability to isolate and study rare/difficult to isolate cell populations while continuing to serve all core users with sorting needs. The S8 will also serve as an additional spectral analysis instrument. It utilizes user friendly software, BD Chorus, with automatic setup providing the option to train independent users to perform analysis and sorting experiments without the current limitation that only core staff may operate the Influx due to its complex setup and operation. Chorus is also fully integrated into the BD Cloud platform for easy panel design and analysis of imaging/spectral data from the instrument. BD Cloud will be fully supported by the Flow Cytometry Core Facility to allow users access to those tools. The core currently supports a FlowJo (part of BD) license and training for core users to aid in data analysis. The instrument will be immediately available to current Flow Core users that utilize the Influx and Aurora. Removing the staff requirement imposed by the current sorter will allow for higher capacity and more users on the S8. The instrument will be housed and managed in the SMHS Flow Cytometry Core Facility with all administrative tasks and user supported provide by the core. Core staff will attend BD provided training for the instrument to ensure the highest quality experience for users. Acquisition of the S8 is fully supported by the GW SMHS to provide services to a broad range of researchers from multiple disciplines.