3D imaging and analysis of cell models like spheroids, organoids, and organ-on-a-chip biology for screening cancer therapeutics
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.
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.

Time-lapse imaging of prostate cancer spheroids cultured on Cell-able plates and treated with 100nM anti-cancer drug paclitaxel. Cells were stained with caspase-3/7 (green, apoptosis marker) and ethidium homodimer-1 (red, necrosis marker), and imaged every 30 minutes for 72 hours.
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).
(1f) By normalizing the amount of apoptosis as compared to untreated spheroids and plotting on a graph, it can be seen that Paclitaxel (green line) induces apoptosis at a much lower concentration than either Mitomycin C or Etoposide.
Apoptotic Cell Normalized to Control
Video: Angiogenesis research for cancer therapeutics
Angiogenesis is an important field of research and a focus for cancer therapeutics. In this interview, Dr. Bas Trietsch, CTO, MIMETAS, introduces a new solution for the study of angiogenesis: the OrganoPlate® Graft, an in vitro cell culture microplate platform that allows vascularization of 3D tissues.
Hear how Molecular Devices ImageXpress® Pico Automated Cell Imaging System and ImageXpress® Micro Confocal High-Content Imaging System play a vital role in the development and analysis of 3D tissue models built on the OrganoPlate Graft.

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
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
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.
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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Live cell 이미징
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
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:
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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
Resources to aid in your cancer research
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Measure cancer cell viability using a homogeneous, stable luminescence assay
Measure cancer cell viability using a homogeneous, stable luminescence assay
Luminescent cell viability assays offer sensitivity and an easy workflow for monitoring the effects of various experimental conditions.
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Measure p53-MDM2 protein interaction with NanoBRET technology
Measure p53-MDM2 protein interaction with NanoBRET technology
We describe validation of the SpectraMax® iD5 reader with the NanoBRET™ PPI Control Pair, consisting of the interacting protein partners p53 and MDM2.
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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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High-throughput screening of 3D cell cultures with multiple high density scaffold-free spheroids for cancer toxicity studies
High-throughput screening of 3D cell cultures with multiple high density scaffold-free spheroids for cancer toxicity studies
3D spheroid models for cancer research are gaining popularity because they better mimic the in vivo tissue architecture, gene expression and metabolic profile of tumors compared to…
과학 포스터
AI-based analysis of complex biological phenotypes
AI-based analysis of complex biological phenotypes
Cell-based phenotypic assays have become an increasingly attractive alternative to traditional in vitro and in vivo testing in pharmaceutical drug development and toxicological safety ass…
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3D image analysis and characterization of angiogenesis in organ-on-a-chip model
3D image analysis and characterization of angiogenesis in organ-on-a-chip model
Angiogenesis is the physiological process of formation and remodeling of new blood vessels and capillaries from pre-existing blood vessels. This can be achieved through endothelial…
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A high-content tube formation assay using an 𝘪𝘯 𝘷𝘪𝘵𝘳𝘰 angiogenesis model
A high-content tube formation assay using an 𝘪𝘯 𝘷𝘪𝘵𝘳𝘰 angiogenesis model
Angiogenesis, the formation of new blood vessels from existing ones, is a critical step involved in various biological processes, such as endothelial sprouting, proliferation, migration,…
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Measure cell migration using a simple scratch assay with timelapse live-cell imaging
Measure cell migration using a simple scratch assay with timelapse live-cell imaging
The movement or migration of cells has long been studied to elucidate the physiological mechanisms of angiogenesis, embryogenesis, cancer metastasis, immune responses, and wound healing.
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Monitor multiple stages of apoptosis with live cell kinetic imaging
Monitor multiple stages of apoptosis with live cell kinetic imaging
The study of apoptosis is a critical aspect of drug discovery and development. Additionally, studying the relationship between apoptosis and other factors, such as oxidative stress, is…
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Monitor cell proliferation and cell cycle in real time
Monitor cell proliferation and cell cycle in real time
There is an increased need to expand the variety and complexity of cell-based assays for biological research and drug discovery. Live-cell assays allow monitoring of cell responses in real…
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Measure cell migration using discontinuous time-lapse imaging of live cells
Measure cell migration using discontinuous time-lapse imaging of live cells
Cell migration is an essential process required for many biological events including embryonic development, wound healing, cancer metastasis, and immunological responses.
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Detection of autophagy using automated imaging
Detection of autophagy using automated imaging
Detect and quantify compound effects on the process of autophagy on the ImageXpress Pico system. The PC12 human neuroblastoma cell line was used as a model for assay development.
eBook
Cellular Imaging Insights
Cellular Imaging Insights
Gain insights and expedite studies for 2D and 3D cellular structures using automated cellular imaging.
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Phenotypic characterization of anti-cancer drug effects using automated imaging
Phenotypic characterization of anti-cancer drug effects using automated imaging
Discovery and evaluation of anti-cancer therapies is an active area of research that includes development of cell-based models, screening for novel drugs, comparison of drug efficacy, and…
eBook
Acquire and analyze 3D images like a pro
Acquire and analyze 3D images like a pro
There has been significant progress in the development of 3D models and techniques during the last few years. Methods include biodegradable scaffolds, organ-on-a chip structures, or self-…
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Streamline assessment of DNA damage using the ImageXpress Nano system
Streamline assessment of DNA damage using the ImageXpress Nano system
Assessing damage to DNA or chromosomes is frequently addressed with research applications because of its implications in diverse diseases including genetic mutations, cancer, and aging. DNA…
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Evaluating cell cycle inhibitors using a live cell assay
Evaluating cell cycle inhibitors using a live cell assay
Monitoring treatment effects on the cell cycle is relevant to progressing oncology research and drug discovery. High-content screening assays using live cells have been developed to enable…
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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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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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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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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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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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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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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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