BD FACSymphony A3: A multi-laser analytical flow cytometer for the Princeton University Flow Cytometry Resource Facility

项目摘要/摘要 这项提案要求提供资金，并表明有必要更换一匹13岁的主力马 在普林斯顿大学使用高通量采样器(HTS)的LSRII流式细胞仪 流式细胞术资源设备(FCRF)，配备最先进的FAC Symphone A3 Flow 细胞仪，带HTS。现有的LSRII是在2007年购买的，Cytek Biosciences和 到2020年底，BD生物科学公司将为该系统提供年度服务合同，并 分别为2022年。这台利用良好的流式细胞仪由FCRF的工作人员进行维护和操作 由研究实验室客户进行培训，以便在系统上运行他们的实验 独立的。FCRF是一个久负盛名并得到财政补贴的共享资源 普林斯顿大学分子生物学系实验室运作31年 好几年了。FCRF有两名全职工作人员，为普林斯顿大学的研究提供服务 社区；它是校园内唯一的流式细胞仪核心设施。流式细胞术和 FCRF提供的荧光激活细胞分选(FACS)服务已成为 用于描述和隔离混合生物中特定种群的主要工具 样本，因此对越来越多的研究人员来说是必不可少的。更换 LSRII采用最先进的FAC Symphony A3流式细胞仪的HTS将加强研究 普林斯顿大学的能力，并将允许我们的研究人员继续不间断地 正在进行的研究项目。这款5激光、23色检测系统可提供极大的灵活性 在提高灵敏度的情况下在流式细胞术实验中使用不同的荧光 由凝胶耦合流动池、多条激发激光谱线、高效的荧光采集光学元件组成 以及该系统的新的低噪声VPX电子设备。FAC交响乐团A3流程的添加 细胞仪将对普林斯顿的多个由NIH资助的研究实验室产生影响 研究领域包括：对转移性乳腺癌的免疫系统反应，病毒感染和 免疫、细菌毒力和生物被膜形成、抗生素耐药性和细菌持久性， 发现具有抗生素特性的小分子，识别糖酵解细胞类型，以及细胞 信令网络动态等。这些项目中有许多明显与健康有关 这意味着什么。FAC交响乐A3将极大地提高NIH的研究能力- 资助研究实验室，并将对质量和 普林斯顿大学进行的研究项目的数量。

Project Summary/Abstract This proposal requests funding and demonstrates the need to replace a 13-year old workhorse LSRII w/ High Throughput Sampler (HTS) flow cytometer operating within the Princeton University Flow Cytometry Resource Facility (FCRF) with a new state-of-the art FACSymphony A3 Flow Cytometer w/ HTS. The existing LSRII was purchased in 2007 and neither Cytek Biosciences nor BD Biosciences will be providing annual service contracts on this system by the end of 2020 and 2022, respectively. This well-utilized flow cytometer is maintained by FCRF staff and is operated by research laboratory clients who are staff trained to run their experiments on the system independently. The FCRF is a well-established and financially subsidized Shared Resource Laboratory operating at Princeton University within the Department of Molecular Biology for 31 years. The FCRF operates with two full time staff and provides service to the Princeton research community; it is the only flow cytometry core facility on campus. The flow cytometry and fluorescence activated cell sorting (FACS) services provided by the FCRF have become the primary tools for the characterization and isolation of specific populations within a mixed biological sample and are therefore essential to an ever-growing number of researchers. Replacement of the LSRII w/HTS by the state-of-the-art FACSymphony A3 Flow Cytometer will enhance the research capabilities at Princeton University and will allow our researchers to continue uninterrupted their ongoing research projects. This 5-laser, 23-color detection system allows for great flexibility in employing diverse fluorescence in flow cytometry experiments with the increased sensitivity offered by the gel-coupled flow cell, multiple excitation laser lines, efficient fluorescence collection optics and the new low-noise VPX electronics of the system. The addition of the FACSymphony A3 Flow Cytometer will have an impact on multiple NIH-funded research laboratories at Princeton in diverse research areas including: immune system response to metastatic breast cancer, viral infection and immunity, bacterial virulence and biofilm formation, antibiotic resistance and bacterial persisters, discovery of small molecules with antibiotic properties, identifying glycolytic cell types, and cell signaling network dynamics, among others. Many of these projects have obvious health-related implications. The FACSymphony A3 will dramatically improve the research capabilities of NIH- funded research laboratories and will have a sustained and powerful impact on both the quality and quantity of the research projects conducted at Princeton University.