Flow Cytometry & Cell Sorting Shared Resource

点击翻译按钮获取中文摘要

FLOW CYTOMETRY & CELL SORTING SHARED RESOURCE: SUMMARY The mission of the Flow Cytometry & Cell Sorting (FCCS) Shared Resource is to provide the Hollings Cancer Center (HCC) research community with an integrated platform to sort and analyze cells and evaluate pre-clini- cal and clinical experimental data in a cell-specific manner. Under leadership of Amanda C. LaRue, PhD, the FCCS offers comprehensive high-speed cell sorting services, analytic flow and mass cytometry, development of new methodologies, and an educational platform to further basic and translational research. Flow cytometric techniques allow for rapid phenotypic and functional analysis of biological processes and isolation of single cells, methodologies that are critical for detailed analyses of cell types contributing to cancer initiation and pro- gression. For example, identification and characterization of cancer initiating cells, therapy-resistant tumor cells, and immune and other cell types in the tumor microenvironment have provided key insights into how can- cer evolves, opening new opportunities for therapeutic interventions. Immune monitoring for clinical trials to examine efficacy and identify new targets is a recent development. The FCCS provides HCC members with routine and complex flow cytometry analyses, reliable cell purification, and expert guided education and train- ing in all aspects of flow cytometry. This is accomplished by providing assisted and unassisted flow cytometry analysis, consultation on experimental design, sample preparation, data analysis, and data preparation, along with hands-on training in the use of flow cytometric technology and analytical equipment. To remain at the leading edge of an evolving technology, since the last review the FCCS expanded services by investing in new equipment (Fluidigm Helios CyTOF, MoFlo Astrios EQ, LSR Fortessa X-20, and ZetaView NTA) and adding novel expertise by recruiting Carsten Krieg, PhD, an expert in time-of-flight mass cytometry and high dimen- sional single cell analysis, as FCCS CyTOF Operations Manager. The FCCS is party to university-wide efforts to advance future scientific goals of cancer research, and recently partnered with the local Ralph H. Johnson VA Medical Center to purchase the Amnis ImageStream Mark II imaging cytometry platform (to be installed Summer 2018). This instrument provides quantitative analysis of cell signaling, protein translocation, internali- zation, morphology, cell-cell interaction, cell cycle, DNA damage and repair, cell death, autophagy, and FISH- based assays at the single-cell level. During the current cycle FCCS Shared Resource was utilized by 95 prin- cipal investigators, including 60 HCC members in all research programs; and provided key data and insights to support the success of 22 NCI-funded grant applications and 75 peer-reviewed publications, including work published in high-impact journals such as JCI, Cancer Discov, Sci Signal, Sci Immunol, Lancet Oncol, Nat Med, and Nature. In summary, the FCCS Shared Resource advances, supports and provides important emerg- ing cancer research tools and methodology to the HCC community.