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

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.