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Updated: Sep 21, 2025
Multiparametric Tumor Organoid Drug Screening Using Widefield Live-Cell Imaging for Bulk and Single-Organoid Analysis
Published on: December 23, 2022
Protocol for drug screening of patient-derived tumor organoids using high-content fluorescent imaging
Brian M Larsen1, Andrea Cancino1, Jenna M Shaxted1
1Tempus Labs, Inc, Chicago, IL 60654, USA.
Abstract:
High-content imaging of tumor organoids (TOs) treated with therapeutic agents provides detailed cell viability readouts at the organoid level. In contrast, most used protocols provide one number per well. While requiring the use of inverted microscopy with an automated stage, this protocol can provide critical information about heterogeneous responses of TOs to various treatments. This protocol describes a technique for culturing and drug testing TOs using fluorescent indicators of cell viability with high reproducibility. For complete details on the use and execution of this protocol, please refer to Larsen et al. (2021).
Insights
This study presents a high-content imaging method for analyzing tumor organoids (TOs) treated with drugs. This approach offers detailed cell viability data, revealing heterogeneous responses to therapeutic agents with high reproducibility.
Area of Science:
- Oncology
- Cell Biology
- Drug Discovery
Background:
- Traditional methods for assessing tumor organoids (TOs) often yield single data points per sample.
- Tumor organoids (TOs) are valuable models for studying cancer biology and drug responses.
- Understanding heterogeneous responses of TOs to therapeutic agents is crucial for personalized medicine.
Purpose of the Study:
- To introduce a high-content imaging protocol for detailed analysis of tumor organoids (TOs).
- To enable the assessment of heterogeneous cell viability in TOs upon drug treatment.
- To provide a reproducible method for drug testing using fluorescent viability indicators.
Main Methods:
- Utilizing high-content imaging with inverted microscopy and an automated stage.
- Employing fluorescent indicators to assess cell viability within TOs.
- Developing a protocol for culturing and drug testing of TOs.
Main Results:
- The protocol provides detailed, organoid-level cell viability readouts.
- It captures heterogeneous responses of TOs to various therapeutic agents.
- The method demonstrates high reproducibility in culturing and drug testing TOs.
Conclusions:
- High-content imaging offers a significant advancement over traditional methods for TO analysis.
- This technique provides critical insights into individual TO responses to drug treatments.
- The protocol facilitates reproducible and detailed drug screening in a 3D tumor model.
