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| Adding Depth: 3-D Cell Culture Physiologically relevant models were, in recent history, thought of as the Holy Grail in life science research. However, with three-dimensional (3-D) cell culture models now better able to mimic in vivo conditions, living organism replicas are within sight. The ability of 3-D cultures to mimic the microenvironment of in vivo tissues sets the technique apart. Compared to 2-D cultures, cellular models containing three-dimensions better represent intricate cell-cell interactions, more closely resemble the exquisite nature of cellular communication, and have matrices that are analogous to in vivo structures. Within this eBook, we address four sections entitled:
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| In the Spotlight: Focus in on High-Content Analysis High-Content Analysis (HCA) merges the benefits of high-throughput automation with microscopic imaging. This powerful technique allows researchers, whether in small academic or large commercial labs, to collect and quantify reproducible multiparametric cellular data. The combination of high sample throughput with multiparametric image and multivariate data analysis enables the detection of even subtle phenotypic changes that isn’t possible with traditional laboratory techniques. In this poster, we show how HCA techniques are used widely and benefit a diverse range of scientific applications and research areas including oncology, neuroscience, infectious disease, and toxicology.
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| High-Content Analysis: Advancing Your Knowledge to Help Combat Infectious Disease High-content screening (HCS) plays a significant role in infectious disease research as it enables high throughput functional and phenotypic assays that can be adapted to a wide range of pathogens including virus, bacteria and eukaryotic parasites. High-content analysis enables, for example, intracellular tracking of viral particles to profile the antiviral mechanisms of each compound and sensitive measurements of bacterial infection rates. Adaptation to high-throughput screening in bacteriology and parasitology has already led to the discovery of new types of host-specific inhibitors that differ from those inhibitors that act directly on microbes.
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| One of the greatest challenges in multiple sclerosis (MS) therapy is the halting or reversal of the failure of remyelination in the brain in order to reverse disabilities in MS patients. This case study highlights the recent work of Dr. Paul Tesar and colleagues at the Case Western Reserve University School of Medicine, which could potentially lead to such novel treatments, as it aims to control the function of stem cells in the body and thereby to help the body repair itself. The case study focusses on the development of a high-content phenotypic assay to identify bioactive small molecules enhancing oligodendrocyte maturation.
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| In this case study, we highlight how the neuroprotection assays described in the paper by Rudhard et al. (2015), using the Opera® QEHS system, can easily and directly be transferred to the new Opera Phenix™ High-Content Screening System, focusing upon the benefits — including a four-fold decrease in acquisition time — of performing an imaging assay for axon degeneration on the Opera Phenix High-Content System.
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| A Workflow to Characterize and Benchmark Human Induced Pluripotent Stem Cells Human induced pluripotent stem cells (iPSCs) offer tremendous opportunities for disease modeling and discovery of novel therapeutics. To advance iPSC technology, the Human Induced Pluripotent Stem Cell Initiative (HipSci) offers the scientific community access to a large panel of cell lines with thorough characterization and data analysis tools. This case study details a phenotypic screen used to characterize human iPSCs on diverse extracellular matrix substrates.
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| Phenotypic Drug Discovery with High Content Screening Today’s drug discovery strategies require candidate compounds to fail early and cheaply in the discovery stage, rather than late and expensively in the clinical phase. This white paper explains why phenotypic and target-based approaches are being combined, some challenges of phenotypic assays, and how to maximize the return on your phenotypic assays.
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| Inhibition of MicroRNA Sensitizes 3D Breast Cancer Microtissues to Radiation Therapy This case study describes an investigation by scientists at the Helmholtz Center Munich into the radiosensitizing potential of miR-21 down-regulation, using 3D breast cancer microtissues. To analyze the impact of radiation on microtissue growth, researchers used the Operetta® High Content Imaging System and Harmony® High Content Imaging and Analysis Software.
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| This case study illustrates how researchers at AIMM Therapeutics, including Dr. Mark Kwakkenbos and Dr. Pauline van Helden, use PerkinElmer’s High Content Imaging solutions to generate therapeutic antibodies against C. difficile infection, the most common cause of healthcare-associated diarrhea that can develop in patients after hospitalization and treatment with antibiotics.
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| In this case study, Dr. Chris Bakal and Dr. Julia Sero from the Institute of Cancer Research (ICR) in London have validated the use of advanced morphological and texture analysis features in the Columbus™ Image Data Storage and Analysis System to quantify the effects of chemotherapeutic compounds on mitochondria morphology in breast cancer cells.
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| Combining Promega HaloTag Technology with PerkinElmer High Content Imaging Systems Promega Corporation’s HaloTag® platform offers interchangeable labeling, high sensitivity and low background levels and is an ideal tool for high content assays. Combining this technology with the Opera® High Content Screening System and Operetta® High Content Imaging System allows high-quality live-cell imaging with exceptionally low phototoxicity on a large scale, enabling the generation of unbiased statistically relevant data sets. |
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