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Updated: Aug 16, 2026
A 3D Spheroid Model for Glioblastoma
Published on: April 9, 2020
Application of dual-view oblique plane microscopy to small-molecule compound screening across 3D glioblastoma stem
Martin Lee1, Jayne Culley1, Hugh Sparks2
1Cancer Research UK Scotland Centre (Edinburgh), Institute of Genetics and Cancer, University of Edinburgh, Western General Hospital, Edinburgh, EH4 2XU, UK.
Abstract:
Recent advances and increasing adoption of 3-dimensional (3D) model systems, such as tumour spheroids and organoids, attempt to more faithfully recapitulate the pathophysiology and heterogeneity observed in patients' tumours, with the goal of reducing high attrition rates observed in late stage drug development. While established high content imaging systems provide the spatial resolution and throughput necessary to place 3D models at the earliest stages of drug discovery, they yield limited information on disease heterogeneity or drug response at the single cell level in 3D. Improvements in single-cell RNA-Seq are transforming our understanding of disease trajectories and therapy response, however this technology is too expensive and laborious for high throughput screening. Here we demonstrate the high content screening capabilities of a compact and low cost light-sheet fluorescence microscopy platform called dual-view oblique plane microscope (dOPM). We apply the dOPM to screen a small library of compounds in a 3D glioblastoma (GBM) stem cell spheroid model expressing the FUCCI cell cycle reporter. We benchmark the performance and compare the outputs of the dOPM GBM spheroid assay with standard 2D and 3D spheroids assays using established spinning disk confocal high content platforms. In a proof-of-principle small molecule compound screen we demonstrate that the dOPM performs to accepted standards of reproducibility for high throughput screening in a 96-well plate format. We further demonstrate the ability of the dOPM to capture the heterogeneity across multiple spheroids within an individual well and provide single cell level data within each individual 3D spheroid. We propose that further development of the opensource dOPM platform will support the advancement of 3D high content phenotypic screening assays from cell population measurements to highly quantitative single cell analysis across 3D space and time dimensions.
Insights
A new dual-view oblique plane microscope (dOPM) enables high-content screening of 3D tumor models. This cost-effective platform provides single-cell resolution for drug discovery, improving on current methods for analyzing glioblastoma stem cell spheroids.
Area of Science:
- Biomedical Engineering
- Microscopy
- Drug Discovery
Background:
- 3D model systems like spheroids and organoids aim to improve cancer drug development by mimicking patient tumor complexity.
- Current high-content imaging offers throughput but limited single-cell resolution in 3D, while single-cell RNA-Seq is too costly for screening.
- There is a need for cost-effective, high-throughput methods to analyze cellular heterogeneity and drug response in 3D models.
Purpose of the Study:
- To demonstrate the high-content screening capabilities of a compact, low-cost light-sheet fluorescence microscopy platform, the dual-view oblique plane microscope (dOPM).
- To apply the dOPM to screen compounds in a 3D glioblastoma stem cell spheroid model and compare its performance against established methods.
- To showcase the dOPM's ability to capture single-cell heterogeneity and drug response within 3D structures for advanced phenotypic screening.
Main Methods:
- Development and application of a dual-view oblique plane microscope (dOPM) for light-sheet fluorescence imaging.
- Screening of a small molecule compound library using 3D glioblastoma stem cell spheroids with FUCCI cell cycle reporters.
- Benchmarking dOPM performance against spinning disk confocal microscopy using 2D and 3D spheroid assays in a 96-well plate format.
Main Results:
- The dOPM demonstrated reproducibility suitable for high-throughput screening in a 96-well plate format.
- The platform successfully captured cellular heterogeneity across multiple spheroids within wells.
- dOPM provided single-cell level data within individual 3D spheroids, offering insights into drug response and disease heterogeneity.
Conclusions:
- The open-source dOPM platform offers a cost-effective solution for high-content screening of 3D cell models.
- dOPM advances 3D phenotypic screening from population-level analysis to quantitative single-cell analysis in 3D.
- This technology has the potential to accelerate drug discovery by providing deeper insights into tumor biology and therapeutic responses at the single-cell level.
