A High Content Screening Platform for High Throughput, High Content Imaging and Analysis

项目摘要/摘要 佛罗里达州莫菲特癌症中心和研究所是美国国家癌症研究所唯一指定的综合癌症中心 申请资金购买Perkin Elmer的‘Opera Phenix’Plus High Content Screen(HCS)成像 系统。自动显微镜和图像分析的应用最初在肿瘤学中占有一席之地 这项研究源于早期测量细胞凋亡的应用。现在，HCS成像系统在许多领域得到了应用 与癌症研究有关的应用，包括细胞形态、细胞增殖、细胞死亡、细胞跟踪、 亚细胞定位、共定位、细胞代谢和药物发现。此外，多个单端点 以及表型结果可以同时在细胞单层、微组织、球体和 甚至是整个有机体。莫菲特癌症中心(MCC)的研究实验室使用其中许多应用程序来 研究癌症生物学的复杂、多方面的本质；然而，MCC目前的成像技术缺乏 HCS系统提供的灵敏度和实验吞吐量相结合，导致数据集缺乏 范围和/或图像质量。为了证明高通量(HT)-HCS仪器的必要性，我们提出了 对10个主要用户和3个次要用户的研究，其主要由NIH资助的研究将大量 获得这一文书的机会产生了积极的影响。Perkin Elmer，Inc.的Opera Phenix Plus HCS系统。 已经被确定为最合适的工具，因为它结合了必要的技术和软件 以满足MCC教职员工多样化的HCS需求。此HCS平台因多项指标而被认为是同类最佳 先进的硬件和软件功能组合在一起时，可提供高分辨率的图像 以无与伦比的采集速度实现频谱串扰和真正的点共焦。当与机载设备配对时 除了环境控制和机器人液体/板材处理，该系统是一个完整的HT-HCS工具。内置式 ‘Harmony’软件允许访问更强大的图像分析功能，包括3D渲染、定制 分析构建块和PhenoLogic机器学习。我们的调查人员将利用歌剧院的力量 Phenix Plus解开癌症遗传学和新陈代谢的奥秘，询问肿瘤微环境，以及 筛选潜在的治疗方法。他们将使用高含量、活细胞、细胞的多重荧光成像， 显微组织和球体生成多参数数据输出，获取更多的定量数据，更多 效率更高，同时还能降低每次实验的成本。如果获奖，凤凰歌剧院将获得 安装在MCC共享资源分析显微镜核心(AMC)中，有助于广泛采用 技术AMC工作人员拥有有效操作所需的显微镜知识和经验 仪器和管理大数据集。这项提案的主要调查者帕特里夏·麦克唐纳博士是 在HTS和HCS技术方面接受过正式培训并具有丰富经验，并将监督 乐器。PI将与AMC主管和员工一起培训用户组如何准备HCS 实验、操作仪器、分析图像和执行HTS活动。

Project Summary/Abstract The Moffitt Cancer Center & Research Institute, Florida’s only NCI-designated Comprehensive Cancer Center is requesting funds to purchase Perkin Elmer’s ‘Opera Phenix’ Plus High Content Screening (HCS) imaging System. HCS, the application of automated microscopy and image analysis initially found its place in oncology research due to early applications that measured apoptosis. Now, HCS imaging systems are used in numerous applications relevant to cancer research, including, cell morphology, cell proliferation, cell death, cell tracking, subcellular localization, colocalization, cell metabolism, and drug discovery. Furthermore, multiple single endpoints as well as phenotypic outcomes can be measured simultaneously in cell monolayers, microtissues, spheroids and even whole organisms. Research labs at Moffitt Cancer Center (MCC) use many of these applications to investigate the complex, multi-faceted nature of cancer biology; however, current imaging technology at MCC lacks the combination of sensitivity and experimental throughput afforded by HCS systems, leading to datasets that lack scope and/or image quality. To justify the need for a high throughput (HT)-HCS instrument, we present the research of 10 major users and 3 minor users whose primarily NIH-funded research would be substantially positively impacted by access to such an instrument. The Opera Phenix Plus HCS system from Perkin Elmer, Inc. has been identified as the most suitable instrument, because it incorporates the necessary technology and software to meet the diverse HCS needs of MCC faculty. This HCS platform is considered ‘best in class’ due to a number of advanced hardware and software features, that when combined, provide high resolution images with minimized spectrum cross-talk and true point confocality at unmatched acquisition speeds. When paired with onboard environmental controls and robotic liquid/plate handling, the system is a complete HT-HCS tool. The built in ‘Harmony’ software grants access to more powerful image analysis functions including, 3D rendering, customized analysis building blocks, and PhenoLogic machine learning. Our investigators will leverage the power of the Opera Phenix Plus to unlock mysteries in cancer genetics and metabolism, interrogate the tumor micro-environment, and screen for potential therapeutics. They will use high content, live-cell, multiplex fluorescence imaging of cells, microtissues, and spheroids to generate multi-parametric data outputs, acquiring more quantitative data, more efficiently, with the added benefit of reduced costs per experiment. If awarded, the Opera Phenix Plus will be installed in the MCC shared resource Analytic Microscopy Core (AMC) facilitating widespread adoption of this technology. The AMC staff possess the necessary Microscopy knowledge and experience to effectively operate the instrument and manage large datasets. The principal investigator for this proposal, Dr. Patricia McDonald, is formally trained and well experienced in HTS and HCS technology and will oversee the scientific direction of the instrument. The PI, together with the AMC director and staff will train the user group on how to prepare HCS experiments, operate the instrument, analyze images and execute HTS campaigns.