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| Opera Phenix High-Content Screening System Technical Performance: Crosstalk Suppression Simultaneous multi-color imaging is a technology commonly used to increase the speed of high-content screening systems. However, one of the main problems arising from this approach is spectral crosstalk which can limit assay sensitivity. To overcome this challenge, we have developed an innovative optical concept for the Opera Phenix™ high-content screening system, known as Synchrony Optics™. In this technical note, we show how the Opera Phenix system can be equipped with up to four large field of view sCMOS cameras to maximize throughput and how Synchrony Optics enable parallel acquisition of up to four spectrally adjacent channels at the same time with minimal crosstalk. |
| Improved 3D Imaging with Opera Phenix HCS System Today’s high-content screening applications are becoming increasingly complex, as more physiologically-relevant cell models such as microtissues are being used instead of cell monolayers. Spinning disk confocal microscopy is a common tool for live cell microscopy and reduces background fluorescence from out of focus planes. However, for 3D imaging, confocal image quality can be limited by a phenomenon known as ‘pinhole crosstalk’. In this technical note, we show how the confocal scanning unit of the Opera Phenix™ high-content screening system provides an increased distance between pinholes to reduce the pinhole crosstalk, leading to improved 3D image quality. |
| How to Perform Successful Long Term Live Cell Imaging in a High-Content Analysis System Live cell imaging has gained importance within drug discovery over recent times, as researchers look for more meaningful insights into cellular behavior and function. In this technical note, we monitor evaporation rates, cell growth and morphology inside the temperature- and CO2-controlled live cell chamber of the Operetta CLS™ high-content analysis system, and compare the results with those obtained using a humidity-controlled cell culture incubator. Download our technical note to find out how you can overcome some of the challenges associated with long-term live cell imaging. |
| Phenotypic Analysis of Hypertrophy in Human iPSC-Derived Cardiomyocytes Cardiac hypertrophy, characterized by abnormal enlargement of the heart muscle, is a major health risk where new approaches to drug discovery are needed. In this application note, we outline how a phenotypic screening assay to study time-dependent effects of endothelin-1-induced hypertrophy was set up using human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes.
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| Distinguishing Cell Types by Phenotypic Profiling of the Nucleus The promise of high-content screening is the acceleration of discovery by extracting as much relevant information as possible from cells. Nevertheless, a large percentage of high-content screens analyze only a small number of image-based properties. In this application note, we show how a single nuclear stain can enable phenotypic profiling and how phenotypic profiles can be used to distinguish up to seven different cell types, without further staining or phenotypic markers.
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| Opera Phenix High-Content Screening System Technical Performance: Speed The Opera Phenix™ High-Content Screening System has been specifically developed to enable users to perform highly multi-parametric phenotypic screens effectively through combining state-of-the-art hardware to allow simultaneous acquisition of up to four fluorescent channels, with Harmony® High-Content Imaging and Analysis Software for convenient experimental set-up and analysis. Here, we outline methods of improving the speed of the Opera Phenix's technical performance through the use of the system’s Synchrony Optics and Water Immersion Lenses.
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| Fundamental processes in living cells, such as apoptosis and signal transduction are controlled by proteins, often acting in concert with other protein partners through protein-protein interactions (PPIs). Inappropriate protein-protein recognition can fundamentally contribute to many diseases, including cancer. Therefore, inhibiting protein-protein interactions represents an emerging area in drug design. |
| Analyzing transport of biliary metabolites is essential to predict pharmacokinetics and hepatotoxicity during drug development. However, the artificial models currently used for studying efflux transporter function lack the functional complexity of the natural liver environment. This application note shows how a high content assay was established in a complex 3D organotypic in vitro liver model system. |
| Phenotypic Profiling of Autophagy Using the Opera Phenix High-Content Screening System One of the most important processes involved in maintaining homeostasis is autophagy, the process of degrading cellular components such as lipids, large protein complexes or even whole organelles via the lysosomal route and altered autophagy is found in various pathological conditions. In this application note, we validate a phenotypic image and data analysis workflow provided by Harmony® High Content Imaging and Analysis and High Content Profiler™ software using an autophagy assay as an application example. |
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