Flow Cytometry

流式细胞仪

• 批准号: 8474691
• 负责人: Joanne A Lannigan
• 金额: $ 9.37万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8474691
• 关键词: Adoptive Transfer; Animals; Antibodies; Antiviral Agents; Automobile Driving; Biological Assay; Cell Communication; Cell Separation; Cells; Coculture Techniques; Data; Data Analyses; Evaluation; Flow Cytometry; Fluorochrome; Genes; Hematopoietic; Histology; Image; Immune; Immune response; In Vitro; Infection; Inflammatory; Inflammatory Response; Injury; Ligands; Liver; Lung; Plasma; Population; Preparation; Process; Production; Proteins; RNA; Reagent; Regulation; Reporter; Role; Sampling; Services; Site; Sorting - Cell Movement; Speed; Spleen; System; T-Lymphocyte; Techniques; Technology; Tissues; Viral; Virus; base; cell type; chemokine; cytokine; data acquisition; instrument; instrumentation; interest; lymph nodes; protein expression; receptor; research study; response; screening; success; tool

The Flow Cytometry Core (FCC) will provide three primary services to support the scientific aims of the four proposed projects; population purification using cell sorting, cytokine identification and quantification using bead based cytokine arrays (Luminex system), and multispectral imaging flow cytometry (MIFC) using the ImageStream system. The first of these services, cell sorting, will be accomplished using a BD DiVa FACSVantage high speed sorter or the recently acquired i-Cyt Reflection high speed sorter. Both of these instruments are full equipped to sort the populations of interest described in the aims of Projects 1, 2, and 3. Evaluation of cytokine profiles of serum/plasma, tissue homogenates and culture supernatants will be accomplished! using the Luminex bead based multiplexing technology. Screening of up to 32 analytes will be available as well as customized panels of analytes of specific interest to accommodate the aims of all four projects. The FCC will provide sample preparation, data acquisition and data analysis for these assays. Thirdly, with the use of the ImageStream imaging flow cytometer, multispectral images as well as statistically robust data will be available to evaluate and confirm protein colocalizations, cytokine production, and cellular activation, as well as provide additional morphological information on cellular populations from the lung, spleen, liver and lymph node. In addition, the FCC has available instrumentation should the need arise to support polychromatic flow cytometry in excess of the capability of the Project leaders' own instrumentation. The FCC will also provide consultative services to the Project leaders in terms of fluorochrome selection, sample preparations, data analysis, and data interpretation. Likewise, the FCC Core Director will coordinate with the Director of the Histology Core (Core B) to insure comprehensive and complimentary services for all four Projects for the evaluation of immune interactions in the process of viral clearance and tissue injury. Flow Cytometry Core will provide the tools to a)isolate specific subsets of cells to further investigate their role in this process, b) identify and quantify the milieu of secreted cytokines/chemokines driving the response, c) capture morphological and fluorescent images of specific cell types identified as key components.

流式细胞术核心 (FCC) 将提供三项主要服务来支持四个方面的科学目标 拟议项目；使用细胞分选、细胞因子鉴定和定量进行群体纯化 基于珠子的细胞因子阵列（Luminex 系统）和多光谱成像流式细胞术（MIFC） 图像流系统。第一项服务是细胞分选，将使用 BD DiVa 完成 FACSVantage 高速分选机或最近收购的 i-Cyt Reflection 高速分选机。这两个 仪器配备齐全，可以对项目 1、2 和 3 的目标中描述的感兴趣群体进行分类。 对血清/血浆、组织匀浆和培养上清液的细胞因子谱进行评估 完成了！使用基于 Luminex 珠的多重技术。将筛选多达 32 种分析物 可用以及定制的特定感兴趣的分析物组，以适应所有四个目标 项目。 FCC 将为这些检测提供样品制备、数据采集和数据分析。 第三，通过使用ImageStream成像流式细胞仪，多光谱图像以及统计数据 可靠的数据将可用于评估和确认蛋白质共定位、细胞因子产生和细胞 激活，以及提供有关肺部细胞群的额外形态信息， 脾脏、肝脏和淋巴结。此外，如果需要，FCC 还可以使用仪器 支持超出项目负责人自己仪器能力的多色流式细胞术。 FCC 还将为项目负责人提供荧光染料选择方面的咨询服务， 样品制备、数据分析和数据解释。同样，FCC 核心主任将协调 与组织学核心（核心 B）主任合作，确保为所有人提供全面且免费的服务 四个评估病毒清除和组织损伤过程中免疫相互作用的项目。 流式细胞术核心将提供工具 a) 分离特定的细胞子集以进一步研究其作用 在此过程中，b) 识别并量化驱动反应的分泌细胞因子/趋化因子的环境，c) 捕获被确定为关键成分的特定细胞类型的形态和荧光图像。