NHGRI/DIR Flow Cytometry Core

NHGRI/DIR 流式细胞术核心

• 批准号: 10672091
• 负责人: DAVID M. BODINE
• 金额: $ 142.35万
• 依托单位: NATIONAL HUMAN GENOME RESEARCH INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10672091
• 关键词: Animals; Blood Cells; Cell Nucleus; Cell Separation; Cell Size; Cells; Color; Development; Enhancers; Equipment; Flow Cytometry; Gene Expression Profile; Genes; Genetic Transcription; Hematopoiesis; Human; Human Resources; Lasers; Measures; Mus; National Human Genome Research Institute; Neurons; Population; Property; Research Personnel; Running; Sampling; Services; Sorting; Surface; Suspensions; Technology; Tissues; Training; Transplantation; Zebrafish; base; cell type; design; experimental study; high throughput screening; indexing; instrument; magnetic cell separation; particle; single-cell RNA sequencing; statistics

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.

流式细胞仪核心为NHGRI研究人员提供了高质量的流式细胞仪服务。流式细胞仪可用于分析混合群体中细胞的细胞亚群，并分类可能占总样品的0.1％的细胞分类。该技术可用于分析任何细胞类型以及许多可以作为单细胞悬浮液制备的亚细胞颗粒。可以同时测量每秒成千上万个细胞的多个参数，包括细胞大小，细胞复杂性以及表面和细胞内标记。 当前的项目包括分离用于基因概况表达研究的人神经元核，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以进行动物移植实验；分析细胞亚群，以分析功能性细胞特性和基因转录谱，以及增强子区域的高吞吐量筛选。我们的分类器上可用的索引分类功能使研究人员能够将单元格成单个板块，并在图，统计信息视图和详细的结果文件中轻松识别它们。这些特征允许将单个细胞的快速分离和表征用于单细胞RNA-seq。 该核心配备了一个BD FACS ARIA Fusion，该融合包含在生物安全柜中 - 五个激光，21彩色仪器，一个BDFACSARIA；一个索尼SH800芯片基于六种颜色分析的生物安全柜中的基于芯片的台式分系台13颜色贝克曼·库尔特·塞特弗莱克斯（Beckman Coulter Cyotflex S Analyzer），一个21颜色的贝克曼·库尔特·库尔特（Beckman Coulter）Cytoflex LX分析仪；用于磁细胞分离的Miltenyi自动MAC和用于鼠CBC/差分分析的HESKA HT5。 核心人员可用于培训，开发和项目执行。 流式细胞仪核心目前由Stacie Anderson（100％努力，GMBB）和Martha Kirby（100％努力，GMBB）组成。这些调查人员会运行分类器，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。