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Cancer Research Solutions

Bioanalytical solutions for understanding cellular pathways and cellular interactions in cancer research

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.

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.

 

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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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

 

Learn more

Figure 1. Rapidly screen 3D spheroids in microplates

fig-1

(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

fig-2

 

 

 

 

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.

Resources to aid in your cancer research

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Videos & Webinars

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Angiogenesis Research: High-Content Imaging Systems Help Unlock the Full Potential of 3D Tissue Models

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3D Imaging of Cancer Spheroids

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Oliver Kepp and Jayne Hesley - Hallmarks of Cancer - Detect and Quantify Cell Death Signatures with High Content Imaging

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ImageXpress Micro XLS

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High-Throughput 3D Imaging of Cancer Spheroids

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Hepatotoxicity Using iPSC-Derived Liver Cells and Beating Pattern in iPSC-Derived Cardiac Spheroids

Anti-Cancer Drugs in HCT116 Spheroids

Efficacy of Anti-Cancer Drugs in HCT116 Spheroids

Tools to Increase Bio-Relevance of 3D Assays

Tools to Increase Bio-Relevance of 3D Assays

Instrumentation and software solutions for your cancer research efforts

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