BD FACS ARIA II CELL SORTER

描述(由申请人提供)：这份共享仪器赠款申请是为了购买Becton Dickinson公司的一台高速荧光激活细胞分类器FACSAria II。该仪器在其用户友好的设计上是无与伦比的，并提供了最大的灵活性来适应各种实验需求；因此，它是用户操作的共享设备的理想选择。我们定制了FACSAria II，配置了405 nm、488 nm、561 nm和640 nm四个激光器和12个光电倍增管，用于同时分析10个荧光参数。定制的FACSAria II还配备了完全封闭的流体系统和先进的气溶胶管理选项，符合处理受人类病原体污染的样本的安全标准。推动这一应用的原因是目前可用的细胞分选能力不足，细胞分选需求增加，以及加州大学旧金山分校(UCSF)的新研究开发缺乏适当配置的设备。新仪器预计每年由15名确定的调查人员使用超过2500个小时，表明对该仪器的真正需求。其中六名调查人员是新的和职业生涯早期的调查人员，两名是加州大学帕纳苏斯分校的新用户，三名是由于新项目开发而新使用的细胞分选机；这些新活动加起来占该仪器预计使用的70%以上。这些新的活动不能完全由加州大学旧金山分拣中心现有的、已经超额订阅的分拣机构来满足。此外，临床和人体样本处理占该机器估计使用量的40%以上。最后，除了分选机的普遍短缺之外，由于现有分选机缺乏绿色激光，基于红色荧光对细胞进行分选的能力非常有限。为了满足这些需求，我们配置了新的分选机，配备了改进的气溶胶容器，用于人体样本处理，以及额外的黄绿色激光，用于分选带有红色荧光标签的细胞。因此，新的分选机是为最大限度地满足加州大学旧金山分校的新研究需求而量身定做的。直接受益于该仪器的研究人员来自不同领域的五个部门，从动物模型的分子和细胞研究到翻译和临床项目。这一工具将直接支持由9名研究人员和6名新的职业生涯早期调查人员开展的研究项目。新的分类器将由细胞分析实验室(LCA)和移植研究实验室(TRL)联合管理。LCA是一个校园范围的核心项目，在流式细胞仪支持方面有60多年的经验。它将负责该文书的技术和行政支助。TRL将容纳该仪器，并提供现场后备技术支持。新仪器将被整合到LCA管理的现有分拣员网络中，以确保对仪器进行专业维护。对类似设备的协调管理还将确保分拣机充电率和整个校园访问的一致性，这可能会使加州大学旧金山分校的整个研究社区受益。 与公共健康相关：我们要求加州大学帕纳苏斯分校的15名以上研究人员共享一台高速分类器。该仪器将支持广泛研究领域的研究，如移植、糖尿病、哮喘、癌症和免疫学。

DESCRIPTION (provided by applicant): This Shared Instrument Grant application is for the purchase of a high-speed fluorescence-activated cell sorter, FACSAria II by Becton Dickinson. This instrument is peerless in its user friendly design and offers maximal flexibility to adjust to various experimental needs; thus it is an ideal choice for user-operated shared equipment. We have custom configured the FACSAria II to have four lasers at 405 nm, 488 nm, 561 nm, and 640 nm and twelve photomultiplier tubes for simultaneous analysis of ten fluorescent parameters. The custom FACSAria II also comes with a fully enclosed fluidic system and an advanced aerosol management option that meets the safety standard for processing samples contaminated with human pathogens. The impetus for this application is insufficient cell sorting capacity currently available, increased cell sorting demand, and the lack of appropriately configured devices for new research developments at the University of California, San Francisco (UCSF). The new instrument has a projected usage of over 2500 hours per year by fifteen defined investigators, demonstrating a real need for this instrument. Six of the investigators are new and early-career investigators, two are new to UCSF Parnassus campus, and three are new users of cell sorters due to new project development; together these new activities represent over 70% of the projected use of this instrument. These new activities cannot be fully accommodated by existing, already oversubscribed sorters at UCSF. Furthermore, clinical and human sample processing represent over 40% of the estimated usage of this machine. Lastly, in addition to the general shortage of the sorters, there is very limited capacity to sort cells based on red fluorescence due to the lack of green lasers on existing sorters. In response to these needs, we have configured the new sorter with improved aerosol containment for human sample processing and an additional yellow-green laser for sorting cells with red fluorescent tags. Thus, the new sorter is tailored to maximally meet new research demands at UCSF. Researchers directly benefiting from this instrument are from five departments in diverse fields ranging from molecular and cellular research in animal models to translational and clinical projects. This instrument will directly support research projects by 9 investigators with active NIH funding and 6 new early career investigators. The new sorter will be jointly managed by the Laboratory of Cell Analysis (LCA) and the Transplantation Research Laboratory (TRL). The LCA is a campus-wide core program that has over sixty years of experience in flow cytometry support. It will be responsible for technical and administrative support of the instrument. The TRL will house the instrument and provide backup onsite technical support. The new instrument will be integrated into the existing network of sorters managed by LCA to ensure expert maintenance of the instrument. Coordinated management of similar equipment will also ensure consistency in sorter recharge rate and access across the campus, which will likely benefit the UCSF research community at large. PUBLIC HEALTH RELEVANCE: We are requesting a high-speed Sorter to be shared by more than 15 researchers at UCSF Parnassus campus. This instrument will support research in a wide range of research fields such as Transplantation, Diabetes, Asthma, Cancer, and Immunology.