Mass Imaging Analysis of Organoids on iMScope QT

Importance of the Topic

Organoids derived from stem cells replicate human tissue architecture and function, enabling advanced disease modeling, drug screening, and personalized medicine workflows.
MALDI imaging mass spectrometry provides label-free, spatially resolved molecular data of proteins, lipids, and metabolites, offering critical insights into tissue heterogeneity and biochemical processes.

Objectives and Study Overview

This study established a standardized workflow using Shimadzu’s integrated imaging solution to perform high-resolution mass spectrometry imaging of human lung organoids.
Key goals included developing a reproducible sample preparation protocol, achieving uniform matrix deposition, and demonstrating detailed lipid mapping within organoid structures.

Methodology and Instrumentation

Sample Preparation:

• Fixed lung organoids in 2% paraformaldehyde and 1% glutaraldehyde.
• Embedded samples in 10% gelatin, cryosectioned, and thaw-mounted on ITO glass slides.
• Applied DHB matrix via a two-step process: sublimation with iMLayer and automated spraying with iMLayer AERO for consistent crystal formation.

Instrumentation:

• iMLayer: matrix vapor deposition system for sublimation.
• iMLayer AERO: automated sprayer for fine matrix coating.
• iMScope QT: imaging mass microscope operating in positive ion mode (m/z 500–1000), 20 µm spatial resolution.
• IMAGEREVEAL MS: data analysis platform using PCA, PLS, and p-value analysis for ROI classification and lipid mapping.

Key Results and Discussion

High-resolution imaging revealed distinct spatial distributions of lipid species:

• Fatty acylcarnitines localized to alveolar-like regions, indicating active β-oxidation.
• Diacylglycerols enriched in extracellular matrix–associated zones, reflecting membrane turnover and signaling.
• Phosphatidylcholine concentrated in club cell areas, linked to airway surfactant function.
• Phosphatidylethanolamine mapped across epithelial membranes, consistent with membrane composition.

Optimized high-resolution conditions (10 µm pitch, increased laser irradiation) allowed cell-level visualization of fatty acylcarnitine (26:4) and diacylglycerol (34:2) distributions, demonstrating metabolic and signaling heterogeneity within organoids.

Benefits and Practical Applications

The described workflow offers:

• Label-free, high-content molecular imaging without staining artifacts.
• Reproducible sample preparation and matrix application for consistent results.
• Rapid data acquisition and automated analysis for efficient profiling of organoid models.
• Functional assessment of tissue differentiation and metabolic status, valuable for drug screening and toxicity studies.

Future Trends and Potential Applications

Advancements in MALDI imaging and organoid models will enable:

• Integration of patient-derived organoids in personalized medicine to map individual molecular profiles.
• Higher spatial resolution and multimodal imaging for comprehensive tissue characterization.
• Combined omics analyses (proteomics, metabolomics) for deeper functional insights.
• Clinical translation in diagnostics, biomarker discovery, and therapeutic monitoring.

Conclusion

Shimadzu’s end-to-end MALDI imaging solution, comprising iMLayer, iMLayer AERO, iMScope QT, and IMAGEREVEAL MS, delivers robust, label-free spatial mapping of lipid distributions within human lung organoids.
This approach enhances the functional evaluation of complex 3D tissues, supporting advanced research in disease modeling and personalized therapies.

References

1. Mohit Jain, Soeun Ngoy, Sunil A. Sheth, Raymond A. Swanson, Eugene P. Rhee, Ronglih Liao, Clary B. Clish, Vamsi K. Mootha, Roland Nilsson. A systematic survey of lipids across mouse tissues. American Journal of Physiology-Endocrinology and Metabolism, 2014, 306(8):E854–E868.