MoFlo XDP Cell Sorter

描述(由申请人提供)：本提案请求支持配备3个激光器的MoFlo XDP细胞分选机。这一新仪器将利用空气喷射技术，根据稀有细胞群体独特的细胞表面标记的表达情况，对稀有细胞群体进行纯化和分类。我们对MoFlo XDP细胞分选器的要求是基于以下几点：(I)我们目前的流式细胞仪核心的运行能力超过了最佳水平，这对我们的研究节奏产生了负面影响；(Ii)Flow核心的几个主要用户需要空气中喷射的分选器才能在他们的研究中取得重大进展，(Iii)我们的初步数据表明，空气中喷射的MoFlo细胞分选器将满足这两项研究需求。该工具将支持和加强现有的NIH资助的项目，并将促进可能导致未来资助项目的研究途径。我们将概述8名由NIH资助的研究人员及其特定的研究项目，这些项目将通过收购MoFlo XDP细胞分类器而得到加强。所有这些研究人员目前都使用辛辛那提儿童医院医学中心(CCHMC)研究流式细胞术核心(RFCC)。他们的研究兴趣广泛，包括：(1)哮喘的发病机制；(2)胆道闭锁的发病机制；(3)调节性T细胞在艾滋病毒和衰老中的作用；(4)癌症干细胞在慢性髓细胞白血病中的作用；(5)新型树突状细胞群促进抗肿瘤免疫的机制；(6)T细胞效应器功能调节噬血细胞淋巴组织细胞增多症(HLH)的机制；(7)GTP酶在造血干细胞维持和血液病(白血病、淋巴瘤)发展中的作用；(8)剖析控制I型糖尿病发展的细胞间相互作用。此外，我们列出了20多名额外的研究人员，并描述了他们由NIH资助的研究项目将如何从这次仪器收购中受益。这位名为Wills-Karp的PI是免疫学领域的领导者，使用流式细胞仪已有20多年，他将负责MoFlo XDP的管理监督。她将与研究流式细胞术核心主任Sherry Thornton博士和监督委员会成员合作，确定时间表优先顺序和成本回收率。MoFlo XDP分拣机将安装在新装修的CCHMC-RFCC中。核心主任将在一名高素质的核心经理和3名技术人员的协助下监督细胞分选机的日常操作和维护。CCHMC高度致力于支持其研究核心，并将支持该仪器的长期维护和维护，以确保及时提供高质量的服务。

DESCRIPTION (provided by applicant): This proposal requests support for a MoFlo XDP Cell Sorter equipped with 3 lasers. This new instrument will be used to purify and sort rare cell populations based on their expression of unique cell surface markers using jet-in-air technology. Our request for the MoFlo XDP Cell Sorter is based on the following: (i) Our current Flow Cytometer Core is running above optimal capacity which is negatively impacting the tempo of our research; (ii) Several major users of the flow core require a jet- in-air sorter to make significant progress on their research and (iii) Our preliminary data demonstrate that the jet-in-air MoFlo Cell Sorter will meet both of these research needs. The instrument will support and enhance existing NIH-funded projects and will foster avenues of research that will likely lead to future funded projects. We will outline 8 NIH-funded investigators and their specific research projects that will be enhanced by the acquisition of the MoFlo XDP Cell Sorter. All of these investigators currently use the Cincinnati Children's Hospital Medical Center (CCHMC) Research Flow Cytometry Core (RFCC). Their research interests span a broad spectrum of biologic queries requiring the use of cell sorting, including: (1) mechanisms underlying the pathogenesis of asthma; (2) pathogenesis of biliary atresia; (3) the role of regulatory T cells in HIV and aging; (4) the role of cancer stem cells in chronic myelogenous leukemia; (5) mechanisms by which novel dendritic cell populations promote anti-tumor immunity; (6) mechanisms by which T cell effector function regulate hemophagocytic lymphohistiocytosis (HLH); (7) the role of GTPases in hematopoietic stem cell maintenance and development of hematopoietic diseases (leukemia, lymphoma); (8) dissecting the intercellular interplay that governs development of type I diabetes. Moreover, we list more than 20 additional investigators and describe how their NIH-funded research projects will benefit from this instrument acquisition. The PI, Dr. Wills-Karp, who is a leader in the field of immunology and a user of flow cytometry for over 20 years, will be responsible for the managerial oversight of the MoFlo XDP. She will work with the Director of the Research Flow Cytometry Core, Dr. Sherry Thornton, Ph.D. and the members of the Oversight Committee to establish scheduling priorities and cost- recovery rates. The MoFlo XDP sorter will be housed in the newly renovated CCHMC-RFCC. The Core Director will oversee the daily operation and maintenance of the Cell Sorter with the assistance of a highly qualified Core Manager and 3 technicians. CCHMC is highly committed to supporting its research cores and will support the long-term upkeep and maintenance of this instrument to ensure quality services are provided in a timely manner.