Maximize Organoid Insight Using Live-Cell Imaging and Analysis
Advances in human disease research, specifically regenerative medication and precision therapies have resulted from ground-breaking work using in vitro 3D cell models. These advanced cell models have been shown to display structural, morphogenetic and functional properties that resemble in vivo pathophysiology, creating a complex cell model for predictive and translational research. However, to establish and accelerate organoid relevance to clinical outcomes, approaches are required that can reduce variability while also increasing insight. Innovations in culturing ex vivo human models has opened a world of scientific possibilities but are still limited by the technologies used to characterize these complex cell models.
- May introduce variability due to lack of environmental control and inadequate 3D Cell Culture protocols that compromise organoid cell health
- Low-throughput and do not scale for screening
- Subjective random assessment of organoid cell growth, death or morphology
- Involve time-consuming, expensive and manual processes for acquisition of organoid images
- Require third party software for end point analysis, or additional fluorescent marker - with limited quantitative information
The Incucyte® Organoid Assay streamlines the complexities of 3D organoid workflows by combining an optimized assay protocol with advanced software tools for real time, label-free or fluorescent analysis. Organoid measurements can be performed using label-free imaging on Incucyte S series instruments, or with enhanced fluorescence based quantification using Incucyte® CX3 spinning disk confocal acquisition, providing deeper biological insight and greater quantitative flexibility for each sample. Empower organoid research and therapeutic development with ease using the Incucyte® Organoid Analysis Software Module.
Organoid Assay Solutions
Incucyte® Organoid Assay Overview
Assess relevant data in 3D cell culture models through temporal, label-free or fluorescent image acquisition and analysis. The Incucyte® Organoid Assay uses an established protocol and the Incucyte® Organoid Analysis Software Module for continuous acquisition and objective analysis of organoid size, count and morphology in a microplate format.
Track and standardize the analysis of organoids over time, to generate reproducible, physiological relevant data long-term, all from inside your incubator.
Incucyte® Organoid Workflow
Ordering Information
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| Additional Products | Qty. | Cat. No. |
|---|---|---|
| Incucyte® Organoid Analysis Software Module (Module available for Incucyte® CX3, SX5, S3, SX1 Live-Cell Analysis System.) | 1 module | 9600-0034 |
Key Advantages
Locate and Acquire Organoids Automatically
Track organoids embedded in Matrigel® continuously to visualize and assess organoid growth and death
Quantify Organoid Growth/Death using Label-free Analysis
Perform real-time, multiparametric analysis of organoids to better predict therapeutic outcomes using the Incucyte® Organoid Analysis Software Module
Improve Reproducibility and Efficiency
Increase data consistency using an established protocol to reproducibly form organoids in a 96-well plate assay format.
Example Data for Organoid Assay
Locate and Acquire Organoids Automatically
Track organoids embedded in Matrigel® continuously and kinetically assess growth and morphology characteristics.
Figure 1. Kinetically monitor distinct organoid phenotypes using the Incucyte® Live-Cell Analysis System. Mouse intestinal, hepatic and healthy human brain organoids were seeded (50% Matrigel®) into 96-well plates and imaged in an Incucyte®. Brightfield videos taken 3d post seeding and images show cell type specific morphology & growth characteristics.
Quantify Organoid Growth/ Death using Label-free Analysis
Real-time temporal acquisition & quantification of embedded organoids in a relevant microenvironment to better predict physiological therapeutic outcomes.
Figure 2. Perform automated label-free quantification of organoids. Mouse intestinal (1:6 split) and hepatic organoids (1K cells/well) were embedded in Matrigel® (50%) in 96-well plates and imaged over 4 - 10 days. Time-lapse videos and corresponding time-courses of brightfield area demonstrate that vehicle treated organoids increase in size over time while Staurosporine exhibits inhibitory effects on organoid growth.
Vehicle (0.1% DMSO)
Intestinal Vehicle
Staurosporine (1 µM)
Intestinal Staurosporine
Hepatic Vehicle
Hepatic Staurosporine
Analyse Response to Drug Treatment Across Multiple Types of Organoids
Figure 3. Mouse Hepatic Organoids expressing Incucyte® Nuclight Orange Lentivirus were embedded in matrix and cultured for 1 day before treating with a range of Camptothecin (CMP) or Cisplatin (CIS) concentrations. Fluorescent images were captured after treatment with CMP or CIS and Incucyte Annexin Green Dye (Green fluorescence, bottom row) and compared 3 days post-treatment. An increase in the Mean Green Fluorescence Intensity in both CMP and CIS treated organoids show increased cell death. CMP and CIS EC50 from 1 day post treatment shows concentration-dependent loss of viability and increase in cell death.
Figure 4. Measure changes in organoid morphological features in response to compound treatment. Brightfield images show hepatic organoids (1K cells/well) ± protein kinase inhibitor Staurosporine (1 µM, STP). Vehicle treated organoids increase in size & number while a marked reduction is observed in the presence of STP. Images (zoomed in) and time-course data also demonstrate that in the presence of STP organoids lose distinctive rounded phenotype (increased eccentricity) and increase in darkness over time.
Improve Efficiency and Reproducibility:
Increase data consistency using an established protocol to reproducibly form organoids in 96-well plate assay format.
Figure 5. Incucyte® lab tested Organoid Assay protocol. An established simple workflow to reproducibly generate organoids for high-throughput image-based compound profiling.
Figure 6. Easy to use guided interface. Automated image and analysis tools provide a ‘set up and walk away’ experience. Track organoid growth and death in 96-well plates in real time anywhere.
