Flow Cytometry and Cell Sorting Shared Resources Core

项目摘要/摘要-流式细胞术和细胞分选共享资源 流式细胞术和细胞分选共享资源(FCCSSR)的目标是通过 提供各种最先进的流式细胞仪设备、服务、专业知识和培训。 这包括提供关于实验设计和结果解释的咨询、交付 对高端流动分析仪的操作进行广泛培训，并提供4台高端分析仪，供 训练有素的用户。理查德·阿什蒙博士担任FCCSSR主任，负责磋商 配合用户、人员监督、培训、仪器操作和监督运行的方方面面 这个设施。阿什蒙拥有生物医学工程博士学位，并曾担任非常成功的 FCCSSR自1987年以来。FCCSSR的工作人员包括3名高级流式细胞仪专家和5名其他工作人员 会员。工作人员操作9台流动分析仪(4台供用户操作)和5台电池分选机。在当前 报告期间，FCCSSR购买了3台仪器：一台BD交响乐分析仪，一台BD Aria Fusion分类器，以及 一个额外的BD LSR II分析仪。FCCSSR在2017财年被47个SJCCC成员使用，并支持工作 来自5个程序中的4个(HMP、DBSTP、NBTP和CBP)。在使用该设施的SJCCC成员中，77% (n=36)拥有与癌症相关的、同行评审的资金。这项工作产生了112篇同行评议的出版物，占34.8% 来自计划内合作的占26.8%，来自计划间合作的占26.8%，其中许多来自影响较大的期刊 如《自然》、《细胞》、《柳叶刀肿瘤学》和《血液》。FCCSSR的贡献通过几个例子得到了证明 取得重大成就。例如，FCCSSR与HMP成员(Charles博士)进行了广泛的合作 穆莱兰、坦贾·格鲁伯、玛丽·雷林和威廉·埃文斯)在急性淋巴细胞白血病和 急性髓系白血病分离表型特征的肿瘤细胞用于进一步研究，跟踪肿瘤 异种移植模型的进展，并使用细胞分选分离转基因种群。此外， FCCSSR是DBSTP工作的重要部分，有助于研究细胞周期对药物的反应 骨肉瘤和其他儿科实体瘤的治疗和表型特征及分离。 未来的计划包括测试复杂的流动分析和评估增强电流的仪器 服务，包括可能扩展到其他领域，如基于图像的或质量细胞术。FCCSSR 将继续与SJCCC成员合作开发和验证新协议，优化新协议的使用 试剂，并扩大用户可用试剂的储存库。此外，由于健壮性和 新仪器产生的数据复杂性增加，FCCSSR主任和工作人员将致力于 有更多的时间与用户详细咨询Flow技术，这将扩大内部培训 计划，并将通过演示、测试和 与终端用户和FCCSSR咨询委员会的讨论。

PROJECT SUMMARY/ABSTRACT—Flow Cytometry and Cell Sorting Shared Resource The goal of the Flow Cytometry and Cell Sorting Shared Resource (FCCSSR) is to support SJCCC members by providing access to a wide variety of state-of-the-art flow cytometry equipment, services, expertise, and training. This includes provision of consultation regarding experimental design and interpretation of results, delivery of extensive training in the operation of high-end flow analyzers, and provision of 4 high-end analyzers for use by trained users. Dr. Richard Ashmun serves as the Director of the FCCSSR and is responsible for consultations with users, personnel supervision, training, instrument operation, and oversight of all aspects of the operation of this facility. Ashmun holds a PhD in biomedical engineering and has served as the highly successful Director of the FCCSSR since 1987. The FCCSSR staff includes three senior flow cytometry specialists and 5 other staff members. The staff operates 9 flow analyzers (4 provided for user operation) and 5 cell sorters. In the current reporting period the FCCSSR acquired 3 instruments: a BD Symphony analyzer, a BD Aria Fusion sorter, and an additional BD LSR II analyzer. The FCCSSR was used by 47 SJCCC members in FY17 and supported work from 4 of the 5 Programs (HMP, DBSTP, NBTP, and CBP). Among the SJCCC members using the facility, 77% (n=36) have cancer-related, peer-reviewed funding. This work resulted in 112 peer-reviewed publications, 34.8% from intraprogrammatic and 26.8% from interprogrammatic collaborations, with many in high-impact journals such as Nature, Cell, Lancet Oncology, and Blood. The contribution of the FCCSSR is exemplified by several key accomplishments. For example, the FCCSSR worked extensively with members of the HMP (Drs. Charles Mullighan, Tanja Gruber, Mary Relling, and William Evans) in studies of acute lymphoblastic leukemia and acute myeloid leukemia to isolate phenotypically characterized tumor cells for further study, track tumor progression in xenograft models, and isolate genetically modified populations using cell sorting. Additionally, the FCCSSR was an important part of work by the DBSTP, contributing to studies of cell cycle responses to drug treatment and the phenotypic characterization and isolation of osteosarcomas and other pediatric solid tumors. Future plans include testing of complex flow assays and evaluation of instrumentation for enhancement of current services, including possible expansion into additional areas such as image-based or mass cytometry. FCCSSR will continue working with SJCCC members to develop and validate new protocols, optimizing the use of new reagents, and expanding the repository of reagents available to users. Additionally, as the robustness and complexity of data emerging from the new instruments increases, the FCCSSR Director and Staff will dedicate more time for detailed consultations with users regarding flow techniques, will expand the in-house training program, and will carefully evaluate replacements for obsolete equipment through demonstrations, testing and discussions with end users and the FCCSSR Advisory Committee.