Flow and Tissue Cytometry Core

摘要 提出了一种定量流动和组织细胞分析核心(FTCC，CORE 3)，用于妇幼保健合作中心 血液学杰出奖(CCEH)，以支持中心成员的研究努力。流式细胞术是 对于绝大多数CCEH调查人员来说，这是一个不可或缺的研究工具。在25名CCEH现任成员中，有16名 在他们的研究中常规使用经典的流式细胞仪服务，他们的出版物中超过65% 在过去的10年里，使用了流式细胞仪服务。然而，许多人需要解释流式细胞术。 在造血生态位空间关系的背景下的信息；例如，确定如何 在正常和应激状态下，细胞之间相互作用，并与细胞和生理成分相互作用 条件。为了满足这一需求，FTCC将用独特的成像服务来补充流式细胞术， 包括3D组织细胞术和活体显微镜。通过这种方式，核心将为CCEH成员提供 关键服务以及将他们对造血细胞的询问范围扩大到包括 不同条件下HSC与其他细胞空间分布的定量分析 非恶性造血。FTCC利用现有的共享设施和专业知识 印第安纳大学医学院(IUSM)为CCEH成员提供最新技术 以及预计将由复杂的流式细胞仪和组织细胞仪设施提供的服务。FTCC将提供成本- 为CCEH调查人员提供的有效资源，他们可以利用此核心应用强大的集成方法 非恶性血液学的研究，这将是他们研究的无价资产，以及 全国各地的血液学调查人员。FTCC的具体目标是：1-提供杰出的， 一致、及时且经济的多色分析、细胞分类、图像流动分析和单个细胞 对所有CCEH成员进行蛋白质组学分析，以促进其在造血和干细胞方面的研究 需要上述任何一项服务的细胞生物学。2-通过以下方式提高CCEH成员的生产力 集中化和标准化的流式细胞术和其他相关服务，允许交换 印第安纳大学内外研究人员之间的协议和结果，并推动分配 科学知识。3-为CCEH成员提供大规模3D和高度多元化的2D组织 细胞学服务，从研究设计、荧光标记、图像采集、定量图像 分析以及监督和非监督数据分析。4-为CCEH成员提供活体显微镜 服务，包括编制机构合规文件、研究设计、动物手术、成像 收集和定量图像分析。这一安排将使CCEH调查人员和所有调查人员 在全国范围内，为他们的非恶性血液学调查提供强大的新工具。

ABSTRACT A quantitative Flow and Tissue Cytometry core (FTCC, core 3) is proposed for the Cooperative Center of Excellence in Hematology (CCEH) to support the research efforts of members of the center. Flow cytometry is an integral research tool for the vast majority of CCEH investigators. Of the 25 current CCEH members, 16 routinely utilize classical flow cytometric services in their research, and more than 65% of their publications over the last 10 years utilized flow cytometric services. However, many need to interpret flow cytometry information in the context of the spatial relationships in the hematopoietic niche; for example, to identify how cells interact with one another and with cellular and physical components of the niche in normal and stressed conditions. To address this need, the FTCC will complement flow cytometry with unique imaging services, including 3D tissue cytometry and intravital microscopy. In this way, the core will offer CCEH members a critical service along with the opportunity to extend their interrogation of hematopoietic cells to include quantitative analyses of the spatial distribution of HSC with other cells during different conditions of nonmalignant hematopoiesis. The FTCC capitalizes on existing shared facilities and expertise present within the Indiana University School of Medicine (IUSM) to provide CCEH members with the latest in technologies and services expected form a sophisticated flow and tissue cytometry facility. The FTCC will provide a cost- effective resource to CCEH investigators who can utilize this core to apply a powerful and integrated approach to studies of non-malignant hematology that will be an invaluable asset to their research, as well as to hematology investigators across the country. The specific aims of the FTCC are: 1- Provide outstanding, consistent, timely, and economical multicolor analysis, cell sorting, image flow analysis, and single cell proteomic analysis to all CCEH members to facilitate the conduct of their research in hematopoiesis and stem cell biology that requires any of these services. 2- Enhance the productivity of CCEH members by providing centralized and standardized flow cytometric and other related services that allow for the exchange of protocols and results between investigators within and outside Indiana University and advance the distribution of scientific knowledge. 3-Provide CCEH members with large-scale 3D and highly-multiplexed 2D tissue cytometry services, extending from study design, fluorescence labeling, image collection, quantitative image analysis and supervised and unsupervised data analysis. 4- Provide CCEH members with intravital microscopy services, including preparation of institutional compliance documents, study design, animal surgery, image collection and quantitative image analysis. This arrangement will give CCEH investigators and all investigators Nationwide, powerful new tools for their nonmalignant hematology investigations.