Visualization of micro-agents and surroundings by real-time multicolor fluorescence microscopy

Area of Science:

• Biomedical Imaging
• Optical Microscopy
• Molecular Biology

Background:

• Optical microscopy offers high spatiotemporal resolution but lacks specificity for distinguishing micro-agents.
• Current methods struggle to differentiate between micro-agents and their dynamic environments.

Purpose of the Study:

• To develop and demonstrate multicolor fluorescence microscopy for precise identification and tracking of mobile micro-agents.
• To visualize micro-agent interactions with biological structures like cancer spheroids and vasculature for drug delivery applications.

Main Methods:

• Utilized spectral unmixing with multicolor fluorescence microscopy to differentiate agents and surroundings.
• Developed a microfluidic chip for spheroid immobilization and stable microscopy.
• Employed in vitro vasculature networks and chicken chorioallantoic membranes to test visualization in vascularized environments.

Main Results:

• Successfully visualized single and cluster micro-agent attachment to cancer spheroids.
• Demonstrated spectral decomposition of micro-agents, cancer spheroids, and vasculature.
• Achieved real-time imaging (15 fps) at 1280x1024 resolution, enabling clear distinction and tracking.

Conclusions:

• Multicolor fluorescence microscopy provides enhanced understanding through color-coded visualization.
• This technique is effective for tracking micro-agents, analyzing morphology, and distinguishing surrounding media in complex biological models.
• Offers a powerful tool for targeted drug delivery research and other biomedical applications.