Cell Sorting Facility

Research Inst Of Fox Chase Can Ctr, Philadelphia PA

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Abstract

PROJECT SUMMARY/ABSTRACT – SHARED RESOURCE: CELL SORTING FACILITY The Cell Sorting Facility (CSF) provides FCCC members access to and technical support for the technology of flow cytometry, a method for analyzing and isolating (sorting) cells in a complex mixture, based on a variety of parameters, including light scatter and fluorescence. The CSF is under the direction of Co-Directors Kerry Campbell, PhD (CSM) and Joan Font-Burgada, PhD (CSM) and supported by four staff members. The primary goal of the CSF is to provide FCCC members with the highest quality analytic and sorting capabilities. In addition, the flow cytometry-based services of the Human Immune Monitoring component (previously a developing Shared Resource under the co-direction of Campbell) were recently incorporated into the CSF to provide human immunology expertise and a dedicated team performing correlative analyses of immune phenotype and function on samples from patients to support immunotherapy clinical trials and research at FCCC. As flow cytometry technology has matured, it has become an essential component of research, utilized in a diverse array of laboratories. 42 FCCC members used the CSF in the 12 months ending 10/31/23 with 74% of service hours supporting peer-review funded research. 57 FCCC members from all three Research Programs used the CSF from 2019-2023 (October). The CSF services are best facilitated though a centralized facility, due to the complexity, cost and caliber of the instrumentation, as well as the need to quickly analyze perishable live cells. The combined 42 years of experience and expertise of Campbell and Font-Burgada is key in implementing state-of-the-art technology and innovative approaches. Since 2019, the Institution provided over $1.5 million to purchase new equipment and $122,580 to renovate a new CSF laboratory. The new equipment will deliver next generation service for many years to provide multi-omic single cell analysis capabilities. The CSF currently comprises an impressive collection of six flow cytometers: 1) a user-operated BD-LSR II analyzer, 2) an operator- run BD FACSAria II sorter, 3) another operator-run BD FACSAria II sorter in a biological safety cabinet to analyze unfixed human samples, 4) a user-operated Symphony A5 analyzer, 5) a user-operated BD FACSMelody sorter, and 6) an operator-run BD FACSymphony S6 spectral sorter/analyzer housed in a biological safety cabinet for biohazardous samples. In addition, a new Isoplexis IsoSpark unit performs single cell proteomics assays. The Flow-LIMS and Path-LIMS data management systems were developed by the PSF to allow all FCCC members to annotate, access, and manage flow cytometry data remotely and to facilitate sharing of experimental data, which fosters collaboration between laboratories. The CSF is advised by a dedicated Facility Advisory Committee and the Facility Parent Oversight Committee, which together serve to ensure that the CSF continues to have the necessary resources to provide the highest quality services.