Related Experiment Video
Updated: Oct 7, 2025

Multiparametric Tumor Organoid Drug Screening Using Widefield Live-Cell Imaging for Bulk and Single-Organoid Analysis
Published on: December 23, 2022
Imaging-Based Machine Learning Analysis of Patient-Derived Tumor Organoid Drug Response
Erin R Spiller1, Nolan Ung1, Seungil Kim1
1Lawrence J. Ellison Institute for Transformative Medicine of USC, Los Angeles, CA, United States.
Abstract:
Three-quarters of compounds that enter clinical trials fail to make it to market due to safety or efficacy concerns. This statistic strongly suggests a need for better screening methods that result in improved translatability of compounds during the preclinical testing period. Patient-derived organoids have been touted as a promising 3D preclinical model system to impact the drug discovery pipeline, particularly in oncology. However, assessing drug efficacy in such models poses its own set of challenges, and traditional cell viability readouts fail to leverage some of the advantages that the organoid systems provide. Consequently, phenotypically evaluating complex 3D cell culture models remains difficult due to intra- and inter-patient organoid size differences, cellular heterogeneities, and temporal response dynamics. Here, we present an image-based high-content assay that provides object level information on 3D patient-derived tumor organoids without the need for vital dyes. Leveraging computer vision, we segment and define organoids as independent regions of interest and obtain morphometric and textural information per organoid. By acquiring brightfield images at different timepoints in a robust, non-destructive manner, we can track the dynamic response of individual organoids to various drugs. Furthermore, to simplify the analysis of the resulting large, complex data files, we developed a web-based data visualization tool, the Organoizer, that is available for public use. Our work demonstrates the feasibility and utility of using imaging, computer vision and machine learning to determine the vital status of individual patient-derived organoids without relying upon vital dyes, thus taking advantage of the characteristics offered by this preclinical model system.
Insights
A new high-content imaging assay uses computer vision to assess drug efficacy in 3D patient-derived tumor organoids without vital dyes, improving preclinical drug screening and discovery.
Area of Science:
- Oncology
- Drug Discovery
- Bioimaging
Background:
- High failure rate of drug compounds in clinical trials necessitates improved preclinical screening.
- Patient-derived organoids offer a promising 3D model for drug discovery but present challenges in efficacy assessment.
- Traditional cell viability assays do not fully utilize the potential of organoid systems.
Purpose of the Study:
- To develop an image-based assay for evaluating drug efficacy in 3D patient-derived tumor organoids.
- To overcome limitations of traditional assays in phenotypically evaluating complex organoid models.
- To enable non-destructive, high-content analysis of individual organoid responses to drugs.
Main Methods:
- An image-based high-content assay utilizing computer vision for organoid segmentation and analysis.
- Acquisition of brightfield images at multiple timepoints for non-destructive, dynamic tracking.
- Development of a web-based tool (Organoizer) for simplified analysis of complex data.
Main Results:
- The assay provides object-level morphometric and textural information for individual organoids.
- Dynamic drug response of single organoids can be tracked non-destructively.
- Demonstrated feasibility of using imaging, computer vision, and machine learning to determine organoid vital status without vital dyes.
Conclusions:
- This novel assay enhances the utility of patient-derived organoids in preclinical drug discovery.
- The approach improves the translatability of compounds by providing detailed, individual organoid-level data.
- The developed methods and tools facilitate more effective drug screening and development in oncology.

