NHGRI/DIR Flow Cytometry Core

流式细胞术核心为NHGRI研究人员提供高质量的流式细胞术服务。流式细胞术可用于分析、鉴定和分离混合细胞群中的细胞亚群，并对可能占总样本不到0.1%的细胞进行分类。该技术可用于分析任何细胞类型以及许多可制备为单细胞悬浮液的亚细胞颗粒。可以同时测量每秒数千个细胞的多个参数，包括细胞大小、细胞复杂性以及表面和细胞内标记物。 目前的项目包括分离人类神经元细胞核用于基因表达谱研究，许多斑马鱼造血项目以及积极参与各种应用，包括分离组织和血细胞群体用于动物移植实验;分离细胞亚群用于功能细胞特性和基因转录谱分析，以及高通量筛选增强子区域。我们的分选仪上提供的索引分选功能使研究人员能够将细胞分选到平板的单个威尔斯孔中，并在图、统计视图和详细的结果文件中轻松识别它们。这些特征允许快速分离和表征用于单细胞RNA-seq的单细胞。 Core配备了一台BD FACS ARIA Fusion，封闭在生物安全柜中-五台激光器，21色仪器，一台BDFACSAria;一台SONY SH 800芯片台式分选机，位于生物安全柜中，能够进行六色分析，一台13色Beckman Coulter CyotFLEX S分析仪，一台21色Beckman Coulter CytoFLEX LX分析仪;用于磁性细胞分离的Miltenyi Auto MACS和用于鼠CBC/差异分析的HESKA HT 5。 核心人员可用于培训，开发和项目执行。 流式细胞术核心目前由Stacie安德森（核心总监，100%努力，GMBB）和Martha Kirby（副总监，100%努力，GMBB）组成。这些研究人员负责运行分拣机，维护分析仪和其他设备，并就实验的设计和执行提供专家建议。

The Flow Cytometry Core provides NHGRI researchers access to high-quality flow cytometry services. Flow cytometry can be used to analyze, identify, and isolate subpopulations of cells from mixed populations and to classify cells that may represent less than 0.1% of the total sample. This technology can be used to analyze any cell type as well as many subcellular particles that can be prepared as a single-cell suspension. Multiple parameters on thousands of cells per second can be measured simultaneously, including cell size, cell complexity, and surface and intracellular markers. Current projects include isolation of human neuronal nuclei for gene profile expression studies, many zebrafish hematopoiesis projects as well as active involvement in a variety of applications including the isolation of tissue and blood cell populations for animal transplantation experiments; isolation of cell sub-populations for analyses of functional cellular properties and gene transcription profiles, as well as high throughput screening of enhancer regions. The index-sorting feature available on our sorters enables investigators to sort cells into single wells of plates, and easily identify them on plots, in statistics views, and in a detailed results file. These features allow the rapid isolation and characterization of single cells for use in single cell RNA-seq. The Core is equipped with one BD FACS ARIA Fusion enclosed in a biosafety cabinet - five laser, 21 color instrument, one BDFACSAria; one SONY SH800 chip based benchtop sorter in a biosafety cabinet capable of six color analysis one thirteen color Beckman Coulter CyotFLEX S analyzer, one 21 color Beckman Coulter CytoFLEX LX analyzer; a Miltenyi Auto MACS for magnetic cell separation and a HESKA HT5 for murine CBC/Differential analysis. Core personnel are available for training, development, and project execution. The Flow Cytometry Core is currently staffed by Stacie Anderson (Core Director,100% effort, GMBB), and Martha Kirby (Associate Director, 100% effort, GMBB). These investigators run the sorters, maintain the analyzers and other equipment and provide expert advice on design and execution of experiments.