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Application of Flow Cytometry to Cell Biology

流式细胞术在细胞生物学中的应用

基本信息

批准号：
7732936
负责人：
SUSAN SHARROW
金额：
$ 147.69万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
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/7732936
关键词：
Aliquot Animals Area Arts Basic Science Cell Cycle Cell Death Cell Lineage Cell Separation Cell physiology Cell surface Cells Cellular biology Color Computer software Consultations Core Facility Cost Savings Cultured Cells Data Data Analyses Defect Development Electronics Event Experimental Designs Flow Cytometry Generations Genetic Goals Histocompatibility Antigens Human Resources Immune Immunobiology Immunology In Vitro Individual Investigation Lasers Manufacturer Name Measures Metabolic Modification NCI Center for Cancer Research National Cancer Institute Pathogenesis Problem Solving Reagent Research Personnel Research Project Grants Research Support Running Signal Transduction Source Stem cells Sterility T-Lymphocyte Techniques Technology Tissues anticancer research base cost cytokine in vivo instrument instrumentation member release of sequestered calcium ion into cytoplasm single cell analysis

项目摘要

The Experimental Immunology Branch (EIB) Flow Cytometry Core Facility currently supports multiple research projects for more than 50 investigators from within the EIB and elsewhere in the Center for Cancer Research (CCR). These investigations involve multiparametric quantitative single cell analysis of, and electronic cell separation based upon, parameters associated with cells freshly prepared from different species and/or tissues, as well as a spectrum of in vitro cultured cells. Basic research support is provided to members of the EIB and to other investigators within the Center for Cancer Research, NCI. Currently supported projects include, but are not limited to, the following areas of study: a) in vivo and in vitro analyses of intra-cellular signaling via cell surface molecules; b) analyses of cellular processes and/or defects in animals and/or cells with genetic modifications; c) studies of the mechanisms and consequences of immune pathogenesis; d) analyses of the coordinate cell surface expression of a variety of molecules; e) investigations of T cell repertoire generation; g) analyses of expression of transplantation antigens; h) investigations of mechanisms involved in T cell lineage development; i) mechanisms of cell death; and j) stem cell analyses. The facility operates and maintains 2 operator run 3-laser flow cytometers with cell sorting capabilities and 3 user-operator flow cytometers with analysis only capabilities. The facility has recently acquired a state-of-the-art 5-laser, analysis only, flow cytometer that will be used to extend and expand available applications. Facility staff provide consultation to investigators in the areas of: experimental design, problem-solving, reagent selection and data analysis and interpretation. The facility supports a wide variety of flow cytometric applications including: rare event analysis (including stem cell analysis) and cell sorting; multi-color phenotypic analyses, cell cycle analysis, proliferation analysis, metabolic analyses including calcium flux analysis, sterile cell sorting, and intra-cellular cytokine analyses. The facility also, as a cost-savings measure, maintains a reagent bank of over 150 commonly used flow cytometry reagents that are pre-titred and aliquoted by facility personnel for use by multiple EIB investigators. The reagent bank minimizes costs by buying in bulk and minimizing labor and effort involved in characterizing individual batches of reagents. The facility is developing WINDOWS-based pc software for flow cytometry analysis that will provide capabilities not currently available in software available from instrument manufacturers or 3rd party software sources.

实验免疫学分支 (EIB) 流式细胞术核心设施目前支持来自 EIB 内部和癌症研究中心 (CCR) 其他地方的 50 多名研究人员的多个研究项目。这些研究涉及对从不同物种和/或组织新鲜制备的细胞以及一系列体外培养细胞相关的参数进行多参数定量单细胞分析和电子细胞分离。向 EIB 成员和 NCI 癌症研究中心的其他研究人员提供基础研究支持。目前支持的项目包括但不限于以下研究领域：a) 通过细胞表面分子对细胞内信号传导进行体内和体外分析； b) 对动物和/或基因修饰细胞的细胞过程和/或缺陷进行分析； c) 免疫发病机制和后果的研究； d) 多种分子的坐标细胞表面表达分析； e) T细胞库生成的研究； g) 移植抗原表达分析； h) T细胞谱系发育相关机制的研究； i) 细胞死亡机制； j) 干细胞分析。该设施运营和维护 2 台操作员运行的具有细胞分选功能的 3 激光流式细胞仪和 3 台仅具有分析功能的用户操作员流式细胞仪。该设施最近购买了一台最先进的 5 激光、仅分析型流式细胞仪，将用于扩展和扩展可用的应用。设施工作人员在以下领域为研究人员提供咨询：实验设计、问题解决、试剂选择以及数据分析和解释。该设施支持多种流式细胞术应用，包括：罕见事件分析（包括干细胞分析）和细胞分选；多色表型分析、细胞周期分析、增殖分析、代谢分析（包括钙流分析）、无菌细胞分选和细胞内细胞因子分析。作为一项节省成本的措施，该机构还维护着一个包含 150 多种常用流式细胞术试剂的试剂库，这些试剂由机构人员进行预先滴定和等分，供多个 EIB 研究人员使用。试剂库通过批量购买并最大限度地减少表征各个批次试剂所涉及的劳动力和精力来最大限度地降低成本。该设施正在开发用于流式细胞术分析的基于 WINDOWS 的 PC 软件，该软件将提供仪器制造商或第三方软件源目前无法提供的功能。