3D imaging and analysis of cell models like spheroids, organoids, and organ-on-a-chip biology for screening cancer therapeutics
Cancer involves changes which enable cells to grow and divide without respect to normal limits, to invade and destroy adjacent tissues, and ultimately to metastasize to distant sites in the body. Cancer researchers need tools that enable them to more easily study the complex and often poorly understood interactions between cancerous cells and their environment, and to identify points of therapeutic intervention.
Learn about our high-content imaging systems and analysis software solution that facilitate cancer research using biologically relevant 3D cellular models like spheroids, organoids, and organ-on-a-chip systems that simulate the in vivo environment of a tumor or organ.
Figure 1: A workflow for testing spheroids in a high-throughput screening environment. A single spheroid can be grown in a 96- or 384-well plate, treated with compound, and stained with a cocktail of dyes that can be imaged without washing. Spheroids may also be fixed if desired. (Right) Transmitted light images of HCT116 cells were taken over the course of 63 hours using Timelapse acquisition on ImageXpress Micro confocal System to show the formation of a spheroid (10X objective).
Advantages of 3D imaging technology for cancer spheroids
Cancer spheroids mimic tumor behavior far more effectively than standard 2D cell cultures. Such 3D spheroid models are being successfully used in screening environments for identifying potential cancer therapeutics. These culture systems can be used in multiparametric analysis to quantify different biological outputs, accelerating cancer drug development.
Key benefits include:
- The development of 3D high-content imaging signifies a major step in facilitating more relevant and accurate testing
- 3D culture systems can rapidly produce uniform human cancer cell spheroids that can be used in high-throughput format to accelerate cancer drug development
- Research with confocal 3D image analysis of cancer cells has enabled multiparametric characterization of many biological outputs
Workflow for analyzing 3D cancer spheroids in high-throughput screening environment
Spheroids can be grown in 96- or 384-well plates, treated with compounds, and stained with dyes that reveal the cellular processes and pathways at work. In some cases, spheroids can be imaged without washing; they may also be fixed if desired.
Simplify your oncology workflow with a broad range of imaging, cellular screening, and microplate reader systems.
The workflow illustrates a simplified process for analyzing spheroids and highlights systems to help you streamline research and increase your throughput.
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Culture spheroids – Cancer cells can be cultured directly in an ultra-low attachment (ULA), round bottom plate, or other labware to develop the typical morphology of a spheroid. Other labware allows one to grow multiple spheroids in a single well.
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Treat with compounds – After spheroid formation, compounds at the desired concentrations are added into the wells, and then incubated for one to several days, depending on the mechanism being studied.
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Stain for markers – After compound treatment is complete, stains are added directly to the media. Stains that require no washing can be used to avoid disturbing spheroids, but spheroids can be carefully washed, even using automation, if necessary.
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Acquire spheroid images – Images within the body of the spheroid can be captured individually or as a z-stack (multiple images taken at differing depths) using specialized imaging equipment.
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Analyze cancer cells – Use cellular imaging analysis software to run quantitative analysis of the cell images to monitor the expression of different markers and to quantify biological readouts.
High-throughput confocal imaging of spheroids for screening cancer therapeutics
In recent years, there has been significant progress in development of in vitro aggregates of tumor cells for use as models for in vivo tissue environments. When seeded into a well of a low-attachment round bottom microplate, these aggregates will form a discrete spheroid. Spheroids are believed to mimic tumor behavior more effectively than regular two dimensional (2D) cell cultures because, much like tumors, they contain both surface-exposed and deeply buried cells, proliferating and non-proliferating cells, and a hypoxic center with a well-oxygenated outer layer of cells. Such 3D spheroid models are being successfully used in screening environments for identifying potential cancer therapeutics.
While there are several challenges to developing robust spheroid assays, using automated high-throughput, high-content imaging is a significant step in facilitating more relevant testing of chemotherapeutic drug candidates
- Capture an entire spheroid in one field-of-view at 20X magnification
- Screen biologically relevant 3D spheroids in 96- or 384-well format
- Use confocal imaging to accurately detect cellular responses
- Conserve storage space by saving only 2D reconstructions of the z plane images
Figure 1. Rapidly screen 3D spheroids in microplates
(1a) Montage of image thumbnails of HCT116 spheroids in a 96-well plate treated with compounds and imaged with a 10X Plan Fluor objective. Hoechst stained nuclei (blue) are overlaid with CellEvent Caspase 3/7 apoptosis marker (green).
(1b) Untreated controls are in column 4 and (1c) a Caspase 3/7 response is evident in columns 5–7 where Paclitaxel was serially diluted 1:3 from 1 µM in Row A (replicates of 3 across).
(1d, 1e) Eleven z planes were combined into a 2D Maximum Projection image and analyzed with a simple custom module. Raw images showing low and high degree of apoptosis with their corresponding segmentation masks are shown (royal blue = nuclei, pink = apoptotic cells).
Applications and assays
Molecular Devices, an industry leader in cellular imaging, provides a wide range of tools to support life science research, drug discovery, and high-throughput screening. Our high-content imaging systems can drive the success of your bioanalytical cancer research efforts. We also provide several configurations of our multi-mode microplate readers as well as a line of easy-to-use microarray scanners.
Learn more about how our technology can help your research in cancer therapeutics.
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3D cancer cell spheroids
The rapid progress of cancer research over the past few decades has seen the rise of spheroids—3D aggregates of cells in culture—as a valuable research tool that provides more physiological relevance than traditional 2D cell culture.
Learn how spheroids formed using a variety of cancer cell types are analyzed using methods featured in our application notes.
- Acquire and analyze images of FUCCI spheroids
- 3D analysis and morphometric characterization of compound effects on cancer spheroid cultures
- High-throughput confocal imaging of spheroids for screening cancer therapeutics
- Multi-parameter imaging assay for measuring toxicity in a tumor model
- 3D Imaging of cancer cell spheroids
- High-throughput screening of 3D cell cultures with multiple high density scaffold-free spheroids for cancer toxicity studies
3D cancer cell spheroids
The rapid progress of cancer research over the past few decades has seen the rise of spheroids—3D aggregates of cells in culture—as a valuable research tool that provides more physiological relevance than traditional 2D cell culture.
Learn how spheroids formed using a variety of cancer cell types are analyzed using methods featured in our application notes.
- Acquire and analyze images of FUCCI spheroids
- 3D analysis and morphometric characterization of compound effects on cancer spheroid cultures
- High-throughput confocal imaging of spheroids for screening cancer therapeutics
- Multi-parameter imaging assay for measuring toxicity in a tumor model
- 3D Imaging of cancer cell spheroids
- High-throughput screening of 3D cell cultures with multiple high density scaffold-free spheroids for cancer toxicity studies
Angiogenesis
Angiogenesis is an important factor in blood vessel formation and cancer progression. in vitro with tube formation assays is an established method to evaluate angiogenesis. Recent newly developed methods like organ-on-chip models or in vivo using zebrafish as a model system provide different insights in studying angiogenesis. Importantly, scaling up these assays for screening poses significant challenges for image acquisition and analysis.
Learn how our ImageXpress high-content imaging system and MetaXpress software provide a streamlined workflow for the assessment of angiogenesis using models such as zebrafish, tube formation assays, and organ-on-a-chip.
Angiogenesis
Angiogenesis is an important factor in blood vessel formation and cancer progression. in vitro with tube formation assays is an established method to evaluate angiogenesis. Recent newly developed methods like organ-on-chip models or in vivo using zebrafish as a model system provide different insights in studying angiogenesis. Importantly, scaling up these assays for screening poses significant challenges for image acquisition and analysis.
Learn how our ImageXpress high-content imaging system and MetaXpress software provide a streamlined workflow for the assessment of angiogenesis using models such as zebrafish, tube formation assays, and organ-on-a-chip.
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Autophagy, DNA damage
Discovery and evaluation of anti-cancer therapies include development of cell-based models, screening for novel drugs, and understanding the relevant mechanisms of action. The cellular effects of processes like DNA damage and autophagy, a regulated process of degrading and recycling damaged proteins and organelles in response to cellular stress, can be analyzed effeciently using automated cell imaging.
Learn how automated cellular imaging provides an effecient method for analyzing the cellular effects of anti-cancer compounds.
Autophagy, DNA damage
Discovery and evaluation of anti-cancer therapies include development of cell-based models, screening for novel drugs, and understanding the relevant mechanisms of action. The cellular effects of processes like DNA damage and autophagy, a regulated process of degrading and recycling damaged proteins and organelles in response to cellular stress, can be analyzed effeciently using automated cell imaging.
Learn how automated cellular imaging provides an effecient method for analyzing the cellular effects of anti-cancer compounds.
Cell migration
Cell migration is broadly defined as the movement of cells from one location to another. It is an essential process required for many biological events including embryonic development, wound healing, and immunological responses. The invasion of tumor cells into surrounding tissues, as well as metastasis, are areas of cancer that can be studied using in vitro cell migration methods.
Learn how to measure cellular migration over time and perform real-time analysis.
Cell migration
Cell migration is broadly defined as the movement of cells from one location to another. It is an essential process required for many biological events including embryonic development, wound healing, and immunological responses. The invasion of tumor cells into surrounding tissues, as well as metastasis, are areas of cancer that can be studied using in vitro cell migration methods.
Learn how to measure cellular migration over time and perform real-time analysis.
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Disease Modeling
Understanding the biology of human diseases is crucial to find effective treatment, and cell lines derived from cancer tissues help develop better anti-cancer drugs. However, now there is a pressing need to increase complexity and relevance of cell-based models to more accurately predict the effects of new drug candidates. Genetically engineered human or animal cells that carry disease mutation display all or some of the pathological processes observed in actual human disease. Designing 3D models that mimic tissues and cell interactions allows researchers to better predict drug effects. 3D cell models, including spheroids, organoids, and organ-on-a-chip, can be built from established cell lines derived from iPSC cells or patient derived cells. Cells can be derived from patient tissue representing rare type of cancer, or genetically modified to introduce or repair disease-related genes.
- Webinar: Establishing and imaging 3D oncology models
- Webinar: Disease modeling in the 21st century: Automated organoid assays with 3D imaging
- Publication: Disease Modeling with 3D Cell-Based Assays Using a Novel Flowchip System and High-Content Imaging
- Webinar: Capturing the complexity of 3D biology: Organoids for disease modelling and toxicity research
- Poster: Novel assay methods for cancer patient derived organoids
- Customer Breakthrough: Bioneer use the ImageXpress Micro Confocal for high-throughput imaging of 3D disease models
- Webinar: High-throughput, organoid-derived organ-on-a-chip systems for drug discovery and disease modelling
Disease Modeling
Understanding the biology of human diseases is crucial to find effective treatment, and cell lines derived from cancer tissues help develop better anti-cancer drugs. However, now there is a pressing need to increase complexity and relevance of cell-based models to more accurately predict the effects of new drug candidates. Genetically engineered human or animal cells that carry disease mutation display all or some of the pathological processes observed in actual human disease. Designing 3D models that mimic tissues and cell interactions allows researchers to better predict drug effects. 3D cell models, including spheroids, organoids, and organ-on-a-chip, can be built from established cell lines derived from iPSC cells or patient derived cells. Cells can be derived from patient tissue representing rare type of cancer, or genetically modified to introduce or repair disease-related genes.
- Webinar: Establishing and imaging 3D oncology models
- Webinar: Disease modeling in the 21st century: Automated organoid assays with 3D imaging
- Publication: Disease Modeling with 3D Cell-Based Assays Using a Novel Flowchip System and High-Content Imaging
- Webinar: Capturing the complexity of 3D biology: Organoids for disease modelling and toxicity research
- Poster: Novel assay methods for cancer patient derived organoids
- Customer Breakthrough: Bioneer use the ImageXpress Micro Confocal for high-throughput imaging of 3D disease models
- Webinar: High-throughput, organoid-derived organ-on-a-chip systems for drug discovery and disease modelling
Live cell imaging
Live cell imaging is the study of cellular structure and function in living cells via microscopy. It enables the visualization and quantitation of dynamic cellular processes in real time.
Live cell imaging encompasses a broad range of biological applications, from long-term kinetic assays to fluorescently labeling live cells.
Learn how cellular processes are analyzed using methods featured in our application notes
Live cell imaging
Live cell imaging is the study of cellular structure and function in living cells via microscopy. It enables the visualization and quantitation of dynamic cellular processes in real time.
Live cell imaging encompasses a broad range of biological applications, from long-term kinetic assays to fluorescently labeling live cells.
Learn how cellular processes are analyzed using methods featured in our application notes
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Luminescence-based methods
Cancer studies often utilize luminescence-based methods for measuring parameters such as cell viability in response to drug treatment, and biomolecular interactions occurring in response to stimulation of cell signaling pathways.
Here, we describe how the SpectraMax® iD5 Multi-Mode Microplate Reader yields optimal results for luminescence assays. Find out how you can:
Luminescence-based methods
Cancer studies often utilize luminescence-based methods for measuring parameters such as cell viability in response to drug treatment, and biomolecular interactions occurring in response to stimulation of cell signaling pathways.
Here, we describe how the SpectraMax® iD5 Multi-Mode Microplate Reader yields optimal results for luminescence assays. Find out how you can:
Organoid technology / Organ-on-a-Chip
Organoid technology, like organ-on-a-chip emulates organ physiology though co-culture of cells in a supportive 3D matrix and use of microfluidic channels to perfuse nutrients or compounds over the resulting cellular structures. It is rapidly gaining popularity as a biologically relevant screening model for new drugs and toxicity.
- 3D image analysis and characterization of angiogenesis in organ-on-a-chip model
- Video: Physiologically-Relevant Tissue Models Using a High-Throughput Organ-on-a-Chip Platform
- High-content assay for morphological characterization of 3D neuronal networks in a microfluidic platform
- Webinar: Developing high-throughput organ-on-a-chip tissue models for drug discovery using high-content imaging
- Poster: Water immersion objectives for automated high-content imaging to improve precision and quality of complex biological assays
Organoid technology / Organ-on-a-Chip
Organoid technology, like organ-on-a-chip emulates organ physiology though co-culture of cells in a supportive 3D matrix and use of microfluidic channels to perfuse nutrients or compounds over the resulting cellular structures. It is rapidly gaining popularity as a biologically relevant screening model for new drugs and toxicity.
- 3D image analysis and characterization of angiogenesis in organ-on-a-chip model
- Video: Physiologically-Relevant Tissue Models Using a High-Throughput Organ-on-a-Chip Platform
- High-content assay for morphological characterization of 3D neuronal networks in a microfluidic platform
- Webinar: Developing high-throughput organ-on-a-chip tissue models for drug discovery using high-content imaging
- Poster: Water immersion objectives for automated high-content imaging to improve precision and quality of complex biological assays
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Organoids
Organoids are three-dimensional (3D) multi-cellular microtissues that are designed to closely mimic the complex structure and functionality of human organs. Organoids typically consist of a co-culture of cells which demonstrate a high order of self-assembly to allow for an even better representation of complex in vivo cell responses and interactions, as compared to traditional 2D cell cultures.
Organoids
Organoids are three-dimensional (3D) multi-cellular microtissues that are designed to closely mimic the complex structure and functionality of human organs. Organoids typically consist of a co-culture of cells which demonstrate a high order of self-assembly to allow for an even better representation of complex in vivo cell responses and interactions, as compared to traditional 2D cell cultures.
Tumoroids
A tumoroid is a special type of organoid derived from primary tumor cells harvested from cancer patients. Because tumoroids can perfectly mimic a typical tumor microenvironment (TME), they give us insights into the impact of TME on tumor progression. That’s why tumoroids boast massive potential for accurate and cost-effective anticancer drug discovery.
Learn how our ImageXpress High-Content Imaging System can help you visualize tumoroids during drug screening:
- Webinar: Establishing and imaging 3D oncology models
- Webinar: Disease modeling in the 21st century: Automated organoid assays with 3D imaging
- Publication: Disease Modeling with 3D Cell-Based Assays Using a Novel Flowchip System and High-Content Imaging
- Webinar: Capturing the complexity of 3D biology: Organoids for disease modelling and toxicity research
- Poster: Novel assay methods for cancer patient derived organoids
Tumoroids
A tumoroid is a special type of organoid derived from primary tumor cells harvested from cancer patients. Because tumoroids can perfectly mimic a typical tumor microenvironment (TME), they give us insights into the impact of TME on tumor progression. That’s why tumoroids boast massive potential for accurate and cost-effective anticancer drug discovery.
Learn how our ImageXpress High-Content Imaging System can help you visualize tumoroids during drug screening:
- Webinar: Establishing and imaging 3D oncology models
- Webinar: Disease modeling in the 21st century: Automated organoid assays with 3D imaging
- Publication: Disease Modeling with 3D Cell-Based Assays Using a Novel Flowchip System and High-Content Imaging
- Webinar: Capturing the complexity of 3D biology: Organoids for disease modelling and toxicity research
- Poster: Novel assay methods for cancer patient derived organoids
Latest Resources
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Acquire and analyze images of FUCCI spheroids on the SpectraMax MiniMax cytometer
Acquire and analyze images of FUCCI spheroids on the SpectraMax MiniMax cytometer
Spheroids are small three-dimensional (3D) cellular microenvironments grown using a variety of specialized culture methods such as low-adhesion microplates. This 3D cell culture confers a…
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Application Note
Evaluate cell migration with FluoroBlok inserts on the SpectraMax MiniMax cytometer
Evaluate cell migration with FluoroBlok inserts on the SpectraMax MiniMax cytometer
Cell migration, broadly defined as the movement of cells from one location to another, is important in diverse processes including embryonic development and wound healing. It is also a key…
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Application Note
Multi-parameter imaging assay for measuring toxicity in a tumor model
Multi-parameter imaging assay for measuring toxicity in a tumor model
There is an increasing interest in using three-dimensional (3D) spheroids for modeling cancer and tissue biology to accelerate translational research. The goal of this study was to develop…
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Application Note
3D analysis and morphometric characterization of compound effects on cancer spheroid cultures
3D analysis and morphometric characterization of compound effects on cancer spheroid cultures
Cellular transformation/tumorigenicity assays using cultures of cells in semi-solid media (soft agar or Matrigel) has been well established for cancer research1,2,5. The assay requires…
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Application Note
High-Throughput Confocal Imaging of Spheroids for Screening Cancer Therapeutics
High-Throughput Confocal Imaging of Spheroids for Screening Cancer Therapeutics
In recent years, there has been significant progress in development of in vitro aggregates of tumor cells for use as models for in vivo tissue environments. When seeded into a well of a…
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Application Note
3D Imaging of Cancer Cell Spheroids
3D Imaging of Cancer Cell Spheroids
Many cancer cell lines will form spheroids if cultured on a favorable three dimensional (3D) matrix. These spheroids are believed to represent tumor physiology more closely than cells…
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Application Note
Cell migration analysis with Oris Pro assay on the SpectraMax MiniMax cytometer
Cell migration analysis with Oris Pro assay on the SpectraMax MiniMax cytometer
Cell migration, the movement of cells from one location to another, is a critical component of both normal and abnormal biological processes. The importance of cell migration in diverse…
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Application Note
High-throughput imaging assays using zebrafish, a model organism for human disease
High-throughput imaging assays using zebrafish, a model organism for human disease
Recently, zebrafish-based screening has gained favor as an alternative to mammalian screening due to cost, throughput and reduced ethical concerns. Zebrafish are a useful model for drug…
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Publications
Phenotypic Characterization of Toxic Compound Effects on Liver Spheroids Derived from iPSC Using Confocal Imaging and Three-Dimensional Image Analysis
Phenotypic Characterization of Toxic Compound Effects on Liver Spheroids Derived from iPSC Using Confocal Imaging and Three-Dimensional Image Analysis
Cell models are becoming more complex to better mimic the in vivo environment and provide greater predictivity for compound efficacy and toxicity. There is an increasing interest in explorin…
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Publications
High-Content Assays for Characterizing the Viability and Morphology of 3D Cancer Spheroid Cultures
High-Content Assays for Characterizing the Viability and Morphology of 3D Cancer Spheroid Cultures
There is an increasing interest in using three-dimensional (3D) spheroids for modeling cancer and tissue biology to accelerate translation research. Development of higher throughput assays t…
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Publications
Abstract 5099: Automated rapid and accurate cell cycle analysis
Abstract 5099: Automated rapid and accurate cell cycle analysis
It is essential to have detailed information about compounds identified in screening campaigns during drug discovery and to know their influence on cell cycle progression or DNA repair. The…
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Publications
High-Content Assays for Hepatotoxicity Using Induced Pluripotent Stem Cell–Derived Cells
High-Content Assays for Hepatotoxicity Using Induced Pluripotent Stem Cell–Derived Cells
Abstract Development of predictive in vitro assays for early toxicity evaluation is extremely important for improving the drug development process and reducing drug attrition rates during cl…
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Instrumentation and software solutions for your cancer research efforts
IN Carta Image Analysis Software
Provides robust, quantitative results from complex biological images and datasets
