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This paper explores the numerous challenges materials scientists and engineers face, from the time it takes to bring new materials to market to the difficulties delivering suitable formulations and testing against specified criteria, and how those can be resolved.
Drug induced liver injury (DILI) is a major reason for late stage termination of drug discovery research projects, highlighting the importance of early integration of liver safety assessment in the drug development process. A technical approach for in vivo toxicology determination was developed using Acetaminophen (APAP), a commonly used over-the-counter analgesic and antipyretic drug, to induce acute hepatocellular liver injury.
Extracellular signal-regulated kinase (ERK) is a key component in the regulation of embryogenesis, cell differentiation, cell proliferation, and cell death. The ERK signaling pathway is altered in various cancer types and is frequently investigated as a target for therapeutic intervention. This application note describes how a live cell FRET assay to study ERK signaling was performed on the Operetta CLS™ high-content analysis system. The optimized design of the FRET-based biosensor, the high-quality imaging of the Operetta CLS system and the easy-to-use image analysis tools of the Harmony® software contribute to the robustness of the high-content assay.
Fluorescent dyes have been used for many years to label cells for microscopy studies, and a variety of dyes in the visible fluorescence spectrum are available to label different cellular compartments and organelles. Efficient delivery of the fluorophore to the cell without excessively modifying surface proteins or perturbing cell function is the major biotechnological challenge. In addition, researchers have taken on the challenge of in vivo imaging, focusing on near infrared (NIR) dyes that fluoresce in a spectral region better suited for in vivo imaging due to reduced background and higher tissue penetration.
Cells constantly sense their environment and their response is a spatio-temporal summation of all signals. To maintain physiological stability, cells need to adjust to environmental changes, a process called homeostasis. One of the most important processes involved in maintaining homeostasis is autophagy, and its significance was recognized by the award of the Nobel Prize for Physiology in 2016 to Yoshinori Ohsumi for the discovery of its underlying mechanisms. Although this is not fully understood, it is believed that autophagy can prevent tumor development by degrading, for example, damaged organelles and protein aggregates.
Epifluorescence (2D) imaging of superficially implanted mouse tumor xenograft models offers a fast and simple method for assessing tumor progression or response to therapy. This approach for tumor assessment requires the use of near infrared (NIR) imaging agents specific for different aspects of tumor biology, and this Application Note highlights the ease and utility of multiplex NIR fluorescence imaging to characterize the complex biology within tumors growing in a living mouse.
Optical-based in vivo imaging of vascular changes and vascular leak is an emerging modality for studying altered physiology in a variety of different cancers and inflammatory states. A number of fluorescent imaging probes that circulate with the blood, but have no target selectivity, have been used to detect tumor leakiness as an indication of abnormal tumor vasculature. Inflammation is also characterized by distinct vascular changes, including vasodilation and increased vascular permeability, which are induced by the actions of various inflammatory mediators. This process is essential for facilitating access for appropriate cells, cytokines, and other factors to tissue sites in need of healing or protection from infection. This application note investigates the use of three fluorescent imaging probes, to detect and monitor vascular leak and inflammation in preclinical mouse breast cancer models.
Analyzing transport of biliary metabolites is essential to predict pharmacokinetics and hepatotoxicity during drug development. A functional impairment of hepatobilary transporters, such as bile salt export pump (BSEP) and multidrug resistance-associated protein 2 (MRP-2), is strongly associated with an increased risk of cholestatic liver injury. Here, we describe a 3D high-content screening assay to study hepatobiliary transporter function in InSphero human liver microtissues. Confocal imaging and automated image analysis were used to quantify BSEP and MRP-2-mediated efflux of fluorescent substrates into bile canaliculi.
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. As a result, valuable information from precious cells and disease models is not utilized. As nearly all screening approaches require a nuclear counterstain such as Hoechst to facilitate segmentation, phenotypic profiling of the nuclei can offer new and additional perspectives on assays at no extra cost.
Quantitative pre-clinical fluorescence imaging transcends the boundaries of traditional optical imaging of biological structures and physiology by providing information at the molecular level about disease states and therapeutic response. Fluorescent Pre-clinical Imaging Agents and FMT® (Fluorescence Molecular Tomography) Quantitative Pre-clinical Imaging Systems represent powerful tools for research and drug development in the imaging of biological processes and pharmaceutical activity in living animals.
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.
Learn 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. Learn how: The Opera Phenix system has been applied in the field of neurodegenerative diseases. In this assay, the Opera Phenix system is 4 times faster than the previous Opera® system. Primary neuron morphology is analyzed in a straightforward approach using Harmony software. Careful assay optimization can increase throughput, and minimize the data burden, without compromising assay performance.