High Content Screening Scientific Posters
Quantification of Nf-kB Signaling in living cells: A high-content workflow for the generation of time-resolved single-cell data
Here, we present a high content analysis (HCA) based assay to study NF-kB signaling in living cells using the Operetta® High Content Imaging System and the Promega HaloTag® technology. By analyzing the acquired Operetta images with our Acapella® High Content Imaging and Analysis Software, we were able to track individual cells over time. View PDF
Enhanced assay sensitivity with HCA ImagAmp™: An enabling signal amplification high content technology for technically challenging targets
We utilized the HCA ImagAmpTM reagent kit in: (1) two typical high content analysis (HCA) assays for cytotoxicity, and in (2) a specific epigenetic assay monitoring the modulation of dimethylated lysine 9 residue on histone H3 (H3K9me2). Signal amplification with HCA ImagAmp is achieved through enzyme-mediated deposition of multiple fluorophores in close proximity to a given antigen. View PDF
Automation of both sample preparation and analysis of a cell migration assay
Here we demonstrate a successful automation of the sample preparation of Platypus Technologies’ Oris™ Pro Cell Migration Assay on a PerkinElmer JANUS® Automated Workstation. Analysis of cell migration using the Operetta® High Content Imaging System and the EnSpire® Multimode Plate Reader shows the assay to be a robust and reproducible quantification of cell motility. View PDF
Reagent-based Signal Amplification for High Content Analysis of PKCαLocalization in HeLaCells
Here we demonstrate use of TSA for quantification of PKCαin unstimulatedand stimulated HeLacells using the Operetta® High Content Imaging System. View PDF
Multiparametric Live-Cell Cytotoxicity Studies on Primary Human Hepatocytes Using the Operetta/Harmony Imaging Platform
To investigate drug-induced cytotoxicity we used cryopreserved single-donor hepatocytes seeded into microplates in a conventional monolayer and treated them with three model hepatotoxins. After applying a convenient no-wash staining protocol live cells were imaged directly on the Operetta® High Content Screening system. View PDF
Chromobody® based High Content cell cycle analysis in live cells
In this study we describe how single live cells can be followed through the entire cell cycle using a specific cell cycle protein binding cameloid antibody (Chromobody®) tagged with GFP (green fluorescent protein). View & Download Poster
Measuring of migration in a fully automated imaging based approach
Cell migration plays a major role in a variety of physiological events, such as immune response and wound healing. It also contributes to pathological processes such as metastasis. Understanding the molecular components of migration is crucial for discovering new targets to develop drugs that affect migration.View & Download Poster
Automated confocal laser microscopy for high content structure/function analysis of individual neurons in CNS slice preparations
Previous research in basic and clinical neurosciences has improved the understanding of genetic disturbances associated with human neurodegenerative disease, which has permitted the generation of an increasing number of transgenic mouse models of Alzheimers disease, Parkinson disease and amyotrophic lateral sclerosis (ALS). Although the stru ctural and functional analysis of existing models has generated important insights, the detailed investigation of pathophysiological signal pathways in each mouse model remains a time – consuming challenge.View & Download Poster
Image-based quantification of HCS cell-based assays using the Operetta and Harmony imaging platform
The characterization of agents that inhibit cell proliferation and division is particularly important for drug discovery research. Both events can be analyzed using HCS approaches by multiplexing cell cycle-specific cellular targets. One technique is EdU staining, which detects the S-phase of the cell cycle through incorporation of the nucleoside analog Uridine into newly synthesized DNA strands. Furthermore, there are well validated protein markers available that are associated with certain cell cycle phases. View & Download Poster
Image-Based Quantification of Cytotoxicity by Vital Dyes using the Opera™ High Content Screening Platform
Cytotoxicity is a very complex process affecting multiple pathways and is not manifested by determining one morphological parameter. However, the ability to measure early indicators of toxicity is an essential part of drug discovery. Cell-based High Content Applications are a powerful tool for determining several events in cytotoxicity simultaneously. We describe a rapid and flexible dye-based high content cytotoxicity assay performed with the widely-used HepG2 (human hepatocellular carcinoma) cells. View & Download Poster
Image-Based Quantification of Cyclin B1 and DNA content during Cell Cycle using the Opera™ HCS Platform
One of the most important tasks in anticancer treatment is the inhibition of cell proliferation by interruption of the cell cycle. As the cell cycle is subdivided into the four phases G1-, S-, G2- and M-phase, there are diverse targets for arresting cells either during DNA replication in S-phase or during division in mitosis. Several cytoplasmic proteins like the cyclines have been identified to be proprietary in cell cycle control whereas Cyclin B1 in particular is essential for initiation of mitosis. View & Download Poster
Image-Based Quantification of Apoptosis by Caspase-3-Activation and Nuclear Fragmentation using the Opera™ High Content Screening Platform
Apoptosis – the genetically coded program leading to the self-destruction of a cell – can be induced via two main pathways, the death receptor-mediated pathway, and the mitochondrial pathway. Induction of either finally results in the activation of caspases, a class of intracellular cytokine proteases which are considered to be the central components of the apoptotic response. By breaking down key cellular components that are required for maintaining normal cellular functions caspases are responsible for executing morphological and biochemical consequences directly or indirectly attributed to apoptosis. View & Download Poster
Direct Fluorescent Labelling Approaches for G-Protein Coupled Receptors and Imaging using the Opera™ High Content Screening Platform
G-protein coupled receptors (GPCRs) are an attractive drug target for several clinically relevant disorders. This is primarily due to their membrane location, ubiquitous expression and critical involvement in many mammalian physiological systems. While several assays targeted at downstream GPCR signalling events have allowed characterization of this receptor superfamily, there are few non-invasive techniques that allow analysis of the receptor-ligand interaction at the cellular level. View & Download Poster