Operetta The Next Generation in High Content Screening

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Operetta – hardware and software in complete harmony Providing shared, remote and dedicated access, the Harmony software is designed to streamline and simplify the HCS process. With its intuitive user interface that leads you through your HCS experiment, Harmony suggests next steps where appropriate, simplifying the process for both new and experienced HCS users. Harmony includes all the tools you need to develop an assay, automate the hardware, analyze your data, and store and retrieve the results. Remote access to the Operetta is available via your office computer and local network for analyzing and viewing your results. Harmony for Flexible Acquisition Acquisition controls are displayed within a unified window with predefined solutions or options to create protocols to your own requirements. Multiple acquisition options are available, including channels, fields per well and z-sections. All imaging parameter metadata, such as dyes / filters, objectives, plate type, fields, z-stack or exposure, are embedded into the experiment files and stored in the database for future reference. Harmony for Rapid, Accurate Analysis Ready-Made Solutions A full range of ready-made solutions for frequently used image analysis tasks and assays is pre-installed for your convenience, making it ideal for new users. Harmony still provides the flexibility to change and modify your experiments using image analysis building blocks. Harmony Ready-Made Solution Applications (examples) Cell Counting or Nuclei Counting Live / Dead Cell Counting Quantification of Marker in Nucleus Quantification of Marker in Cytoplasm Quantification of Marker in Membrane Cytosol to Nucleus Translocation Cytosol to Membrane Translocation Fluorescence Redistribution – Cytoskeleton proliferation, cytotoxicity cell viability: live / dead cell count Protein expression in nucleus (e.g. caspase-3) • Cell cycle: S-Phase quantification (BrdU, EdU) Protein expression in cytosol • Mitochondrial mass • Loss of mitochondrial membrane potential Phospholipidosis • Hepatosteatosis • Cell cycle: Transition of G2 to M-phase (cyclin B1) Protein expression in plasma membrane • Signalling pathways: Membrane ruffling Signalling pathways: Transcription factor activation Signalling pathways: Cytoplasm to membrane translocation Cytoskeletal rearrangement Spot Analysis Peroxisome quantification • Vesicle formation • GPCR activation: arrestin recruitment (e.g. Transfluor ® ) Nuclear Spots Nuclear Analysis – Nuclear Shrinking Nuclear Fragmentation Nuclear Classification – DNA Content Cell Shape – Cell Rounding fluorescence In Situ Hybridization (FISH) Nuclear swelling and shrinking apoptosis / Nuclear fragmentation Cell cycle: DNA content Cell spreading, cell detachment, cell rounding Mitotic Index Cell cycle: Mitotic index (phosphohistone H3) Cell Cycle Classification Simultaneous phosphohistone H3, EdU, DNA content and nuclear morphology Harmony includes preconfigured image analysis sequences for common applications. Known as ready-made solutions, they are ideal for new users of HCS. 4
Custom-Designed Solutions Harmony provides a complete range of building blocks to enable simplified custom image analysis. Each clearly defined block has primary tuning parameters with optimized values that can be automatically or manually adjusted with real-time visual feedback to facilitate the set-up process. Harmony Building Block Description Find Nuclei Find Cytoplasm Find Cells Find Spots Find Image Region Select Cell Region Select Region Calculate Intensity Properties Calculate Morphology Properties Calculate Properties Select Population identifies and counts cellular nuclei based on a nuclear stain Identifies cell bodies / cytoplasmic area when nuclei have been identified before Finds whole cells based on fluorescence intensity e.g. for whole cell fluorescence assays Identifies fluorescent objects in cells of similar size, such as endosomes, vesicles, peroxisomes Segments the entire image based on fluorescence intensity Defines a region in the cytoplasm or nucleus, based on nucleus and cytoplasm identification Defines a sub-region calculates fluorescence intensity of regions / objects, provides mean, median, standard deviation, CV, max, min, contrast values Calculates morphology properties of regions / objects, such as area, length, width, axis, ratio, roundness Allows any combination of intensity and morphology properties Defines sub-populations of cells Harmony for Data Storage and Retrieval A scalable database is included for management of all images, experiment information and data analysis. Metadata information, such as exposure parameters, analysis method and users, is embedded into all data files, while a keyword browser facilitates searching using those keywords for your projects. A sortable database information tree allows quick and easy access to any desired information. Automatic parameter adjustment helps you find the best settings quickly. Find images, metadata and results easily in the Harmony database. www.perkinelmer.com/operetta 5
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Operetta The Next Generation in High Content Screening

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