Related Experiment Video
Updated: Dec 19, 2025
Analyzing Oxidative Stress in Murine Intestinal Organoids using Reactive Oxygen Species-Sensitive Fluorogenic Probe
Published on: September 17, 2021
Label-free redox imaging of patient-derived organoids using selective plane illumination microscopy
Peter F Favreau1, Jiaye He1,2,3, Daniel A Gil1,4
1Morgridge Institute for Research, Madison, WI 53715, USA.
Abstract:
High-throughput drug screening of patient-derived organoids offers an attractive platform to determine cancer treatment efficacy. Here, selective plane illumination microscopy (SPIM) was used to determine treatment response in organoids with endogenous fluorescence from the metabolic coenzymes NAD(P)H and FAD. Rapid 3-D autofluorescence imaging of colorectal cancer organoids was achieved. A quantitative image analysis approach was developed to segment each organoid and quantify changes in endogenous fluorescence caused by treatment. Quantitative analysis of SPIM volumes confirmed the sensitivity of patient-derived organoids to standard therapies. This proof-of-principle study demonstrates that SPIM is a powerful tool for high-throughput screening of organoid treatment response.
Insights
Selective Plane Illumination Microscopy (SPIM) rapidly images 3-D organoids using autofluorescence. This method quantifies drug treatment response in patient-derived cancer organoids, proving effective for high-throughput drug screening.
Area of Science:
- Biomedical Imaging
- Cancer Research
- Drug Discovery
Background:
- Patient-derived organoids are valuable for predicting cancer treatment efficacy.
- High-throughput screening methods are needed to analyze organoid responses efficiently.
Purpose of the Study:
- To demonstrate Selective Plane Illumination Microscopy (SPIM) for rapid 3-D imaging of organoids.
- To develop quantitative image analysis for assessing drug-induced changes in organoid autofluorescence.
- To validate SPIM as a tool for high-throughput drug screening in cancer research.
Main Methods:
- Utilized endogenous fluorescence from NAD(P)H and FAD coenzymes in organoids.
- Employed SPIM for rapid 3-D autofluorescence imaging of colorectal cancer organoids.
- Developed a quantitative image analysis pipeline for organoid segmentation and fluorescence change measurement.
Main Results:
- Achieved rapid 3-D autofluorescence imaging of patient-derived colorectal cancer organoids.
- Quantified treatment-induced changes in endogenous fluorescence using developed image analysis.
- Confirmed organoid sensitivity to standard therapies through quantitative SPIM analysis.
Conclusions:
- SPIM enables rapid, quantitative assessment of drug response in 3-D organoid models.
- This approach is suitable for high-throughput drug screening in personalized cancer therapy.
- SPIM represents a powerful tool for advancing organoid-based drug discovery.
