Flow Cytometry & Cell Sorting Shared Resource

流式细胞术和细胞分选共享资源：总结 流式细胞术和细胞分选（FCCS）共享资源的使命是为Hollings癌症提供 中心（HCC）研究社区，拥有一个集成平台，可对细胞进行分类和分析，并评估临床前 校准和临床实验数据。在阿曼达C. LaRue博士， FCCS提供全面的高速细胞分选服务，分析流式细胞术和质谱仪， 新的方法，和一个教育平台，以进一步基础和转化研究。流式细胞术 技术允许生物过程的快速表型和功能分析以及单个 细胞，方法学是关键的细胞类型的详细分析有助于癌症的启动和促 格雷辛。例如，鉴定和表征癌症起始细胞、治疗抗性肿瘤细胞、肿瘤生长抑制剂和肿瘤细胞。 肿瘤微环境中的细胞、免疫细胞和其他细胞类型提供了关键的见解， cer的发展为治疗干预提供了新的机会。临床试验的免疫监测， 检查疗效和确定新的靶点是最近的发展。FCCS为HCC成员提供 常规和复杂的流式细胞仪分析，可靠的细胞纯化，以及专家指导的教育和培训， 在流式细胞术的各个方面。这是通过提供辅助和非辅助流式细胞术来实现的 分析、实验设计咨询、样品制备、数据分析和数据准备，沿着 在使用流式细胞技术和分析设备方面进行了实践培训。保持在 作为一项不断发展的技术的领先优势，自上次审查以来，FCCS通过投资新的 设备（Fluidigm Helios CyTOF、MoFlo Astrios EQ、LSR Fortessa X-20和ZetaView NTA）并添加 新的专业知识，通过招募卡斯滕克里格，博士，在飞行时间质谱细胞计数和高维度专家， 作为FCCS CyTOF操作经理，进行单细胞分析。FCCS是整个大学努力的一部分 以推进癌症研究的未来科学目标，最近与当地的拉尔夫·H·约翰逊 VA Medical Center购买Amnis ImageStream Mark II成像细胞仪平台（待安装 2018年夏季）。该仪器提供了细胞信号传导、蛋白质易位、细胞内信号传导和细胞外信号传导的定量分析。 形态学，细胞间相互作用，细胞周期，DNA损伤和修复，细胞死亡，自噬和FISH- 基于单细胞水平的测定。在本周期内，95个印刷厂利用了FCCS共享资源， 联合国研究人员，包括所有研究项目中的60名HCC成员;并提供了关键数据和见解， 支持22个国家癌症研究所资助的赠款申请和75个同行评审的出版物，包括工作的成功 发表在高影响力期刊上，如JCI，Cancer Discov，Sci Signal，Sci Immunol，Lancet Oncol，Nat 医学和自然总之，FCCS共享资源推进、支持并提供了重要的应急资源， 向HCC社区提供癌症研究工具和方法。

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.