Spatiotemporal Interrogation of Single Spheroids Using Multiplexed Nanoplasmonic-Fluorescence Imaging

Area of Science:

• Biomedical Engineering
• Cell Biology
• Advanced Imaging Technologies

Background:

• Organoid models are crucial for understanding biological processes and disease mechanisms.
• A comprehensive understanding of organoids requires simultaneous monitoring of intraorganoid and extraorganoid activities.
• Existing imaging technologies often lack the spatiotemporal resolution and multiparametric capabilities needed for detailed organoid analysis.

Purpose of the Study:

• To introduce a novel multiplexed imaging platform for simultaneous, high-resolution monitoring of organoid dynamics.
• To integrate label-free biosensing with fluorescence microscopy for comprehensive organoid characterization.
• To enable multiparametric interrogation of organoid secretions, structure, and viability over time.

Main Methods:

• Development of a multiplexed imaging platform combining nanoplasmonic biosensing and fluorescence microscopy.
• Utilizing nanoplasmonic sensors with extraordinary optical transmission for label-free detection of biomolecular secretions.
• Employing deep-learning-augmented image analysis for multiparametric data interrogation over extended periods.
• Application to tumor spheroids to monitor vascular endothelial growth factor A (VEGF-A) secretion, morphometrics, and viability.

Main Results:

• The platform successfully monitored biomolecular secretions and cellular dynamics within and around organoids with spatiotemporal resolution.
• Simultaneous tracking of VEGF-A secretion, morphometric changes, and viability in tumor spheroids was achieved.
• Variations in secretion and growth dynamics between untreated and drug-treated tumor spheroid groups were effectively captured.
• Demonstrated the platform's capability to provide comprehensive insights into organoid behavior.

Conclusions:

• The integrated imaging platform offers a powerful tool for advancing organoid research.
• This technology can accelerate discoveries in disease modeling and drug development by providing detailed insights into organoid functions.
• The platform complements existing technologies, enabling more comprehensive and efficient studies of ex vivo mini-organs.