High-content imaging (HCI) combines high-resolution microscopy with advanced algorithmic image analysis to reveal complex cellular behaviours and phenotypic changes. At o2h discovery, we leverage the CellInsight CX7 LED Pro HCS platform to deliver customised, data-driven assays that fuel innovation in drug discovery. With decades of expertise in high-content imaging, our team provides tailored solutions to support even the most challenging discovery campaigns.

The Role of High-Content Imaging in Modern Drug Discovery

Traditional drug discovery often relies on a target-based approach, focusing on specific biological pathways. While effective in certain contexts, this method can limit the discovery of novel therapeutics. In contrast, high-content imaging enables target-agnostic drug discovery, assessing phenotypic changes without assuming a predefined molecular target. Through quantitative image analysis, this approach identifies promising drug candidates and provides a comprehensive understanding of their impact on cellular systems and clinical phenotypic endpoints.

High-content imaging goes beyond surface-level observations by systematically evaluating complex cellular processes. By offering deeper insights into phenotypic outcomes, it enhances decision-making and the development of innovative therapeutics.

Capabilities in High-Content Imaging

At o2h discovery, our high-content imaging assays are designed to address a broad spectrum of biological questions, including protein trafficking, translocation, receptor internalisation, and target recruitment to specific organelles. These assays offer critical insights into molecular interactions and sub-cellular processes, enabling a deeper understanding of cellular dynamics.

Our expertise also spans cell cycle analysis, where we investigate mitotic progression, cytokinesis, and the effects of DNA Damage Response (DDR) mechanisms. By integrating assessments of proliferation, apoptosis, and autophagy, we provide a comprehensive view of cellular stress responses and overall cellular health. Additionally, we specialise in identifying phenotypic and morphological changes that signal invasiveness, drug resistance, and other disease-relevant behaviours, offering valuable insights for therapeutic development.

Case Study 1:

o2h discovery specializes in developing cellular assays that integrate biological reporter systems, allowing for quantitative assessment of target engagement. These assays are invaluable for structure-activity relationship (SAR) studies, demonstrating dose-dependency, and validating cell-free predictions of binding potencies to differentiate candidate molecules effectively.

Our HCI phenotypic assays, utilising stable cell lines with tagged reporters, are particularly advantageous in scenarios where scale, cost-per-well, and rapid turnover are critical factors in the drug discovery screening cycle. This approach ensures efficient and informative screening, enabling more informed decision-making in your drug development process.

In the above provided example, the raw data from the HCI assay tracks the sub-cellular translocation of a GFP-tagged reporter in response to either vehicle treatment or an experimental small molecule.

Above, you can see the HCI nuclear reporter translocation data, which serves as a dose-dependent phenotypic measure of cellular target engagement. This analysis allows us to quantify how the reporter’s localisation changes with varying concentrations of the small molecule, providing insights into its mechanism of action and efficacy. Such data are crucial for understanding the dynamics of target engagement and optimising lead compounds in the drug discovery process.

Case Study 2:

In support of drug discovery campaigns, the HCI experts at o2h discovery can create assays that track cell phenotypes indicative of target engagement. One notable example is our HCI speckle assay, which detects cellular activity by monitoring the punctate versus diffuse states of endogenous spliceosome factors. In this assay, small molecules are evaluated for their ability to induce dose-dependent phenotypic shifts, reflecting their engagement with a target kinase known to regulate mRNA splicing. This approach not only provides insights into the mechanisms of action of potential drug candidates but also helps identify compounds that effectively modulate cellular processes related to splicing.

In the above example HCI data in which MCF7 cells were treated with either vehicle or a tool compound and subsequently stained with fluorescent markers, enabling algorithmic detection and characterisation of spliceosome speckles.

In the above HCI speckle assay performed in MCF7 cells, we generate dose-response curves guiding structure-activity relationship (SAR) development

Case Study 3:

Assay development on HCI systems facilitates innovative approaches to information management and study design. At o2h discovery, we integrated HCI assay development with the SemaCyte® cell microcarrier platform developed by Semarion.

Combining HCI with this microcarrier system enables novel strategies for managing cell monolayers in drug discovery workflows. Key enhancements include the miniaturisation of cell seeding, freezing of assay-ready adherent cells, and multiplexing of cell lines using subpopulation barcoding.

Powered by HCI image analysis, this methodology provides significant benefits for projects that require economical cell seeding, efficient utilisation of premium assay reagents, and advanced internal control strategies.

At o2h discovery, we bring together cutting-edge technology and decades of expertise to deliver exceptional results in high-content imaging. Our team specializes in custom assay development that is tailored to your project’s specific biological requirements. We combine phenotypic and algorithmic analysis to provide quantitative data that uncovers meaningful insights about your molecules and targets. Our platform offers scalable solutions for high-throughput screening, ensuring that your project progresses efficiently while maintaining the highest quality standards.

We are committed to innovation, constantly developing new assay designs and integrating advanced platforms like the CellInsight CX7 LED Pro and SemaCyte® microcarriers to address unique biological challenges.