Purchase of an Operetta High Content Screening Reader

描述(由申请者提供)：请提供资金购买Perkin Elmer Operetta台式自动化高通量、高内容成像系统。目前，维斯塔尔研究所或邻近的宾夕法尼亚大学或德雷克塞尔大学的研究界还没有这样的仪器。Wistar研究所最近建立了一个分子筛选设施，以支持在微滴定板中筛选shRNA、siRNA、miRNA和小分子化学库。该设施在开发利用全井读数的分析方面取得了很大进展。然而，维斯塔尔研究所的分子筛选和显微镜设备都没有能力支持高含量的筛选。此外，对邻近的宾夕法尼亚大学和德雷克塞尔医学院的一项调查表明，在我们的研究社区中，将这项技术作为支持筛查实验的共享资源是不够的。Wistar的许多研究人员要求该设施将其能力扩展到基于高含量细胞的筛查分析。所要求的轻歌剧将设在维斯塔尔研究所分子筛查设施，该设施成立于2008年，致力于维护自动化、高通量技术，以支持系统生物学方法，以识别和验证用于治疗人类疾病的细胞靶点和/或先导分子，包括癌症、糖尿病、炎症和动脉粥样硬化。轻歌剧的独特功能将极大地扩展Wistar的成像能力，包括自动高通量采集和图像数据的定量分析。最重要的是，轻歌剧将立即影响NIH资助的8个主要用户和一群次要用户开发以细胞为基础的高通量筛查分析的能力，这些分析侧重于细胞及其分子成分的分析。一个由维斯塔尔研究所的用户和高级工作人员组成的内部咨询委员会将监督该仪器的操作、维护和公平使用。在本应用程序中，我们介绍了需要获得高通量、高内容成像的研究项目和初步数据。这些项目包括开发利用3D器官型培养进行高通量筛选的微型分析方法，评估调节伤口愈合和组织再生的细胞信号网络，识别在潜伏的疱疹病毒感染期间调节染色质结构和基因表达的细胞因子，以及参与上皮向间充质转化、迁移和肿瘤转移的细胞辅因子的核质穿梭。购买轻歌剧将大大扩展Wistar分子筛查设施和显微镜设施的能力和多功能性，以支持旨在减轻人类疾病负担的变革性研究。

DESCRIPTION (provided by applicant): Funds are requested to purchase a Perkin Elmer Operetta benchtop automated high-throughput, high- content imaging system. At present, no such instrument is available to the Wistar Institute or the neighboring research community at the University of Pennsylvania or Drexel University. The Wistar Institute has recently established a Molecular Screening Facility to support screening of shRNA, siRNA, miRNA, and small molecule chemical libraries in microtiter plates. The facility has made great progress in developing assays that utilize whole-well readouts. However neither the Molecular Screening nor Microscopy Facility at the Wistar Institute have the capability to support high-content screening. Furthermore, a survey of the neighboring University of Pennsylvania and Drexel School of Medicine indicate that access to this technology as a shared resource to support screening experiments is inadequate in our research community. Numerous investigators at Wistar have requested the facility expand its capabilities into high- content cell-based screening assays. The requested Operetta will be located in the Wistar Institute Molecular Screening Facility, which was established in 2008 and is dedicated to maintaining automated, high-throughput technologies required to support systems biology approaches to identify and validate cellular targets and/or lead molecules to treat human diseases, including cancer, diabetes, inflammation, and atherosclerosis. The unique features of the Operetta will greatly expand imaging capabilities at Wistar to include automated high-throughput acquisition and quantitative analysis of image data. Most importantly the Operetta will immediately impact the ability of 8 Major NIH-funded users and a group of Minor users to develop cell-based high-throughput screening assays that focus on the analysis of cells and their molecular components. An internal advisory committee comprised of users and senior staff from the Wistar Institute will oversee the operation, maintenance, and equitable access to this instrument. In this application, we present projects and preliminary data from studies that require access to high-throughput, high-content imaging. These projects include development of miniaturized assays that utilize 3D organotypic cultures for high-throughput screening, assessment of cellular signaling networks that regulate wound healing and tissue regeneration, identification of cellular factors that regulate the architecture of chromatin domains and gene expression during latent Herpes Virus infection, and nucleocytoplasmic shuttling of cellular cofactors involved in epithelial-to mesenchymal transition, migration and tumor metastasis. The purchase of the Operetta will greatly expand the capabilities and versatility of the Wistar Molecular Screening Facility and Microscopy facilities to support transformative research aimed to reduce the burden of human disease.