Flow Cytometry

流式细胞术核心(FCC)将提供三项主要服务，以支持四项科学目标 拟议的项目；使用细胞分选、细胞因子鉴定和使用 基于微珠的细胞因子阵列(Luminex系统)，以及使用 ImageStream系统。这些服务中的第一项，细胞分类，将使用BD DIVA完成 FACSVantage高速分选机或最近收购的I-Cyt反射高速分选机。这两个都是 仪器设备齐全，可以对项目1、2和3的目标中描述的感兴趣的人群进行分类。 将对血清/血浆、组织匀浆和培养上清的细胞因子谱进行评估 完成了！使用基于Luminex珠子的多路复用技术。将对多达32种分析物进行筛选 可提供特定感兴趣的分析物的定制面板，以满足所有四个方面的目标 项目。FCC将为这些检测提供样品准备、数据采集和数据分析。 第三，利用ImageStream成像流式细胞仪，对多光谱图像进行统计 强大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，以及提供来自肺的细胞群体的额外形态信息， 脾、肝、淋巴结。此外，如果需要，FCC有可用的仪器 对多色流式细胞术的支持超出了项目负责人自己的仪器能力。 FCC还将向项目负责人提供氟铬选择方面的咨询服务， 样品准备、数据分析和数据解释。同样，FCC核心主管将协调 与组织学核心(核心B)主任合作，确保为所有人提供全面和免费的服务 评估病毒清除和组织损伤过程中的免疫相互作用的四个项目。 Flow Cytotics Core将提供工具来a)分离特定的细胞亚群，以进一步研究它们的作用 在这一过程中，b)确定和量化分泌的细胞因子/趋化因子驱动反应的环境，c) 捕捉被识别为关键组件的特定细胞类型的形态和荧光图像。

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.