High-throughput Full Spectrum Cell Sorter

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Project Summary / Abstract The University of California, San Francisco (UCSF) Parnassus Flow Cytometry CoLab (PFCC) is requesting funds to purchase a next generation Full Spectrum Cell Sorter. We have chosen the ThermoFisher Invitrogen™ BigFoot full spectrum cell sorter as the instrument that best fulfills the needs of the unsatiable demand for high resolution multidimension single cell data by our highly esteemed NIH-funded faculty. The BigFoot will complement our current arsenal of high-parameter analysis equipment such as the Fluidigm CyTOFs, Cytek Aurora and Becton Dickson FACSymphony X50 while most importantly adding the highly desired cell sorting capability. The BigFoot maintains the standard cell processing workflow PFCC users are accustomed to in conventional bandpass-based flow cytometers as well with the Aurora Full Spectrum Analyzer at PFCC. We will take advantage of the BigFoot's versatility and powerful spectral technology by enabling collection of fluorescent emission profiles through most of the visible spectra to near-infrared ranges making the number of parameters that can be investigated on this instrument approaching 35 to 40. The UCSF community has rapidly taken advantage of the availability of the full spectrum technology for their high-parameter research, the BigFoot will uniquely add cell sorting capabilities to aid the quest to understand the complex dynamics of changing cell population in disease progression, therapy and remission. Our Researchers have extensive experience in high-parameter data acquisition/analysis using Mass Cytometry resulting in 29 high impact publications over the last 4 years at UCSF, with at least 5 more in review. However, full spectrum flow cytometry is becoming a formidable technology to multiparameter analysis using mass cytometry; transition to the Aurora full spectrum analyzer has gained momentum due to the ease of sample preparations, simplicity of use, high-throughput (50 times faster) and especially the cost savings compared to mass cytometry. Within 1 year of installation, a peer review paper using the Aurora has been published. The high-throughput capabilities coupled with minimal sample loss during preparation and acquisition makes the technology amenable to samples with extremely low cell numbers. Hence, sorting on high-parameter panels of over 25 fluorescent markers has been in demand; however, no cell sorter is currently available at UCSF that allows panels of more than 18 fluorescent markers to be used. The BigFoot will augment the Full Spectrum Flow Cytometry portfolio at UCSF and the ability to simultaneously sort cell populations of interest which will be foundational in providing further information on cellular dynamics during multi-drug clinical research. Due to the COVID-19 lockdown, and limited accessibility at PFCC an ever- increasing number of processed clinical samples has been accumulating adding to the already pre-existing bank of samples ready for high-parameter analysis and sorting. At PFCC, the BigFoot will be a user-controlled device available 24/7, with minimal setup or daily maintenance requirements, and it will be a major high-throughput and high-performance tool in the Core. The BigFoot will help alleviate the bottleneck of machine availability continually experienced by our cell sorting and high-parameter researchers at UCSF.