WVU Flow Cytometry and Single Cell Core Facility (FCSCCF)

项目概要/摘要 西弗吉尼亚大学流式细胞术和单细胞核心设施（FCSCCF）的使命是提供最先进的 生物医学研究的技术、仪器和专业知识。作为肿瘤微环境的一部分 (TME)生物医学研究卓越中心（CoBRE），WVU FCSCCF扩大了其单细胞 将Fluidigm C1单细胞自动制备系统置入堆芯，沿着 用的是Miltenyi的温和MACS Octo分离器这项新技术补充了目前的仪器， 该设施包括两台流式细胞仪（3激光BD FACSAria I分选仪和4激光LSRFortessa分析仪）， 和磁性细胞分选仪（AutoMACs）。与机构采购了一台新的4激光FACSAria III分拣机， 将于2017年初安装，以取代现有的3激光分选机。FACS或MACS分选的细胞 将在肿瘤解离后制备的细胞加载到C1集成流控芯片（IFC）上，用于捕获 从复杂的样品中分离单个细胞，特别是肿瘤。RNA（包括microRNA）或DNA将从 将单细胞分离并加工用于进一步的下游应用，包括RNA/DNA测序，或更高的 通量qRT-PCR。本研究核心应用的具体目标是：1）继续实施新的 将这些能力转化为现有的、非常成功的核心设施，以支持TME CoBRE的研究项目 以及2）为TME提供流式细胞术和单细胞分析领域的专业知识和培训 西弗吉尼亚大学由CoBR资助的研究人员和其他机构发展奖（IDEA）资助的项目。的 收购最先进的Fluidigm C1系统，使WVU FCSCCF用户能够获得最先进的 这些技术将大大促进和加强他们的研究。该提案描述了服务和 设施提供的培训、其管理计划、费用结构、质量保证计划以及 在项目的整个生命周期及以后维持和扩大该设施。

PROJECT SUMMARY/ABSTRACT The mission of the WVU Flow Cytometry and Single Cell Core Facility (FCSCCF) is to provide state-of-the-art technologies, instrumentation, and expertise for biomedical research. As part of the Tumor Microenvironment (TME) Center of Biomedical Research Excellence (CoBRE), the WVU FCSCCF has expanded its single cell analysis capabilities with the emplacement of a Fluidigm C1 Single Cell Auto Prep System into the core, along with a Miltenyi gentleMACS Octo Dissociator. This new technology complements the current instrumentation in the facility, which includes two flow cytometers (3-laser BD FACSAria I sorter and 4-laser LSRFortessa analyzer), and a magnetic cell sorter (AutoMACs). A new 4-laser FACSAria III sorter has been purchased with institutional funds and will be installed in early 2017 to replace the existing 3-laser sorter. FACS or MACS-sorted cells prepared following tumor dissociation will be loaded onto a C1 integrated fluidics chip (IFC) for the capture of single cells from complex samples, particularly tumors. RNA (including microRNAs) or DNA will be isolated from the single cells and processed for further downstream applications including RNA/DNA sequencing, or high throughput qRT-PCR. The specific aims of this research core application are to: 1) continue to implement new capabilities into an existing, highly successful core facility in support of research projects for the TME CoBRE and 2) Provide exceptional expertise and training in the area of flow cytometry and single cell analysis to TME CoBRE-funded investigators and other Institutional Development Award (IDeA)-funded projects at WVU. The acquisition of the state-of-the-art Fluidigm C1 system gives WVU FCSCCF users access to cutting edge technology that will significantly advance and enhance their research. The proposal describes the services and training provided by the facility, its management plan, fee structure, quality assurance plan, and strategies to sustain and expand the facility over the life of the project and beyond.