High-content Screening Research Areas

Here, we present a method for analyzing cell invasion into a 3D extracellular matrix using the Operetta® high-content analysis system and Harmony® imaging and analysis software and the Oris™ cell invasion assay from Platypus Technologies, LLC.

More than ever, researchers are turning to 3D cell cultures, microtissues and organoids to bridge the gap between 2D cell cultures and in vivo animal models. Our solutions-based approach is designed to support you at every step of the cell culture workflow so you can seamlessly culture, treat, and analyze 3D cell cultures. Begin generating more physiologically relevant data to better understand biological processes and make informed decisions with our solutions. For research use only. Not for use in diagnostic procedures

High-content assays using 3D objects such as cysts or organoids can be challenging from the perspectives of both image acquisition and image analysis. In this technical note we describe how to image and analyze epithelial Madin-Darby canine kidney (MDCK) cysts in 3D on the Operetta CLS ™ high-content analysis system. We address: How to reduce the loss of image resolution as you image deeper into the sample How to enable 3D segmentation and analysis for your 3D high-content assays How to increase throughput for 3D imaging through shorter exposure times How to eliminate long and tedious data transfer steps

Multicellular 3D “oids” (tumoroids, spheroids, organoids) have the potential to better predict the effects of drug candidates during preclinical screening. However, compared to 2D cell monolayers, assays using 3D model systems are more challenging. In this technical note we describe how to image and analyze solid spheroids in 3D using the Opera Phenix ™ and Operetta ™ CLS high-content screening systems and Harmony ® imaging and analysis software. We address: How to reduce image acquisition time and data volume How to analyse fully and partially imaged spheroids in 3D How to define zones and quantify spatial differences within spheroids

Targeted protein degradation (TPD) is an emerging drug discovery modality that offers the potential to probe biological pathways and target proteins that have previously been considered “undruggable”. Compared to traditional drug discovery approaches where small molecules bind and inhibit the function of a protein of interest (POI), TPD exploits the cell’s own degradation machinery to selectively target a protein for degradation. This offers the potential for improved therapeutic efficacy and the possibility to discover and validate novel targets. In our comprehensive review, we delve into the fundamental principles and significant advantages of TPD. Furthermore, we explore the challenges faced by the field in bringing this transformative modality into the clinic.

The detection of compound cytotoxicity is an essential part of drug discovery. In this work we describe a rapid and flexible image-based live cell approach to study cytotoxicity. Using a fluorophore dye mixture, multiple cellular phenotypic changes were analyzed following a toxic insult. In order to investigate different drug induced cellular responses, human hepatocytes (HepG2 cells) were treated with various compounds.

Explore our Technical Note unveiling a robust workflow for advanced cytotoxicity analysis in 3D cell cultures. This method addresses challenges in reproducibility, scalability, and imaging accuracy, enhancing drug testing accuracy and physiological relevance. Key Features: Reproducible 3D Models: Learn the precise creation of 3D matrix cultures using the RASTRUM platform, ensuring consistent dimensions independent of cell type or conditions. Tailored Microenvironment: Tailor matrix stiffness and biofunctionalization for physiologically relevant cell models, significantly improving drug-induced cytotoxicity predictions. Imaging Enhancement: Overcome signal attenuation and data volume challenges in 3D cell culture imaging. Utilize water immersion objectives, optical clearing, and precise positioning techniques for optimal high-content image acquisition. For a comprehensive exploration of this innovative workflow and its applications in 3D cytotoxicity analysis, download the complete Technical Note.

In this case study, we present a novel method developed by researchers at the CeMM Research Center for studying protein levels and localization within cells. Utilizing CRISPR-Cas9-based intron tagging, this scalable approach generates cell pools expressing hundreds of GFP-fusion proteins, enabling comprehensive analysis of cellular responses to perturbations. Through a combination of intron tagging with in situ sequencing, previously unrecognized proteins impacted by treatments are identified, surpassing the limitations of existing high-throughput methods. This approach can be applied to other sets of genes beyond metabolic enzymes and potentially in a genome-wide manner to study protein dynamics at scale.

The UK-based Human Induced Pluripotent Stem Cell Initiative (HipSci) aims to offer the scientific community access to a vast panel of cell lines with thorough characterization and data analysis tools. This case study details a phenotypic screen to characterize human iPSCs on diverse extracellular matrix substrates, and a method for the capture of specific phenotypes emerging upon cell-to-cell contact. This case study describes: How high-content analysis enables image-based phenotyping and the throughput for analysis of hundreds of cell lines. How data analysis tools help understand multiparametric datasets through interactive data visualization and phenotype classification.

Charter a course to biotherapeutic antibody development success The rise of bispecific antibodies for use as groundbreaking therapies continues to grow with more than 100 bispecific antibodies currently in development. This is thanks to their remarkable versatility from having dual binding sites targeting two different epitopes, providing advantages such as reducing resistance issues, increased specificity, and identifying unique combinations of drug targets. However, navigating the intricate challenges of bispecific antibodies is no small feat. That’s why we’ve created this infographic that will aid you on the journey, equipping you with the knowledge to overcome the obstacles along the way and achieve excellence in bispecific antibody development. Learn more about: Stages that make up the bispecific antibody path Challenges and considerations faced in development Various solutions and technologies available For research use only. Not for use in diagnostic procedures.

Organoids are 3D, self-assembling structures derived from stem cells which mimic the structural and functional properties of human tissues. They offer unprecedented insights into development, disease mechanisms, and drug responses. However, before utilizing them in disease modelling or drug discovery applications, a thorough characterization is needed to understand their physiological relevance and their limitations to draw robust conclusions from organoid-based experiments. In this literature review, you’ll discover recent publications that highlight advancements in organoid characterization techniques, focusing on: • Analyzing cell type diversity with flow cytometry • Assessing morphology with high-content imaging • Characterizing cell function with multiplex cytokine immunoassays

RNA-based therapeutics have garnered significant attention in recent years, fueled by the success of the mRNA COVID-19 vaccines. The development of therapeutics for a broad range of other conditions, including monogenic diseases, metabolic disorders, and cancer, are now being made possible due to advances in RNA-mediated genome editing tools and technologies. Considering the growing interest in RNA therapies, our latest article highlights several promising classes of RNA therapeutic, including RNAi, CRISPR, base editing, and mRNA vaccines, as well as the potential to combine RNA technologies with CAR-T.

Mitochondrial dynamics are essential for energy conversion and neuron survival. Understanding changes in mitochondrial dynamics is therefore crucial in developing mitochondria-based therapy options for complex pathological conditions such as cancer, neurological disorders, and metabolic syndromes. Fast kinetic live-cell imaging combined with high-content screening represents a promising strategy to quantitatively track these changes in real time and at large scale. This application note demonstrates how to investigate mitochondrial dynamics in human iPSC-derived neurons using the Operetta ® CLS ™ high-content analysis system. Fast frame rate imaging to accurately capture rapid cellular responses Reliable quantification of mitochondrial dynamics in neurons Flexible kinetic acquisition settings for each channel to avoid unnecessary data acquisition

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.

Recent progress toward regenerating pancreatic ß cells lays the foundation for continued advancement in diabetes research. Find out more about how enhanced GABAA signaling induces loss of a cell identity and encourages a cells to convert into insulin-producing ß-like cells.

There has been a lot of buzz around artificial intelligence, machine learning and deep learning. Is the reality living up to the hype? In the world of cellular imaging and its application to drug discovery, there is evidence of real progress against some of the critical challenges facing scientists using these technologies. In this white paper, you will learn about: Challenges in cellular imaging and drug discovery that Artificial Intelligence (AI), Machine Learning (ML) and Deep Learning (DL) are helping to overcome How these technologies are used by leading cellular imaging scientists An outlook to how AI, ML and DL in cellular imaging have the potential to further advance drug discovery and improve productivity in the future

Neuronal ceroid lipofuscinosis (NCL) is a complex, rare, and fatal disease belonging to a larger group of lysosomal storage disorders that presents with nervous system symptoms. Batten disease – or CLN3 disease – is another name given commonly to Juvenile NCL (JNCL). Dr. Gomez-Giro and her team focused on understanding the neurodevelopmental process of JNCL utilizing healthy hiPSCs introduced with a disease-causing c.1054C→T pathologic variant into the CLN3 gene. Read the whitepaper to find out more about Dr. Gomez-Giro's research.

Here, we present a high-content screening application for analyzing the cell migration of NSCLC cells in a live cell assay.

This case study illustrates how Revvity’s high-content imaging solutions support the therapeutic antibody development. Two steps in the workflow - the clonal B cell selection and the functional antibody characterization - rely on high-content screening using the Operetta ™ system. The coupling of the Operetta system to a plate::handler workstation allows the running of plates both day and night, further increasing the throughput. In addition, the EnVision ™ multimode plate reader and the JANUS™ Automated Workstation are essential parts of the workflow that contribute to the throughput of the process and can reduce the variability of liquid handling steps.

Precision biologics are playing an increasingly powerful role as part of therapeutic strategies such as monoclonal antibodies, bispecific and multispecific antibodies, recombinant proteins, vaccines, and targeted next-generation cell and gene therapies. Harnessing large molecules to create safe and effective therapies is a complex and expensive undertaking. So we help scientists like you develop and streamline your entire biologics workflow, helping you overcome the challenges to bringing consistent, high quality, biological medicines to market – faster than ever before. Explore our solutions and technologies that cover areas of: Biomolecular discovery Biologics characterization Investigating immune function Manufacturing and QA/QC Safety and efficacy

The incidence and prevalence of non-alcoholic fatty liver disease (NAFLD), which is characterized by excessive accumulation of lipids within the liver, is progressively rising due to increasing obesity and metabolic syndrome prevalence. For some individuals, NAFLD can progress to non-alcoholic steatohepatitis (NASH), which can lead to liver cirrhosis and hepatocellular carcinoma (HCC). NASH is poorly understood and therapies to treat the disease are lacking. Michele Vacca, from the University of Cambridge in the UK and colleagues sought to investigate the impact of one member of the TGFß-BMP superfamily, BMP8B, on the progression of NASH. Learn more about their study.

Phenotypic Drug Discovery Re-Imagined Cell Painting is a phenotypic screening method and a powerful application of high-content screening technology which combines cell and computational biology to elucidate the behavior of cells under the influence of perturbagens, such as chemical compounds, drugs, genes, or other entities. Download our white paper to learn about the origins of the Cell Painting technique, its potential impact on the drug discovery paradigm, together with practical hints and tips for success.

Cell painting is a powerful high-content screening method which combines cell and computational biology to unravel cells’ responses and gain a deeper understanding of the effects of chemical and genetic perturbagens. However, implementation of cell painting is not without its challenges - from choosing a cell model and labeling reagents, to optimizing instrumentation and making sense of the thousands of features that are extracted during data analysis. Download our application note to learn how to: Set up optimal imaging parameters and see the effect of different acquisition modes on compound clustering Easily perform the assay using the dyes provided in the PhenoVue ® cell painting kit Extract and analyze more than 5700 cellular features Visualize multidimensional data using the Signals ™ VitroVivo platform

Cell painting is an imaging-based phenotypic profiling method that measures phenotypic features to characterize the biological responses of cells to chemical and genetic perturbagens. It finds uses in functional genomics, drug discovery, efficacy, toxicity assessment, and in screening for insights into mechanism-of-action. In this infographic, you will learn: What cell painting is, its history, and how it works What you need to get started How to easily perform the assay using our cell painting solutions