Characterization of Single-Spheroid Oxygen Consumption Using a Microfluidic Platform and Fluorescence Lifetime...

Area of Science:

• Biomedical Engineering
• Cellular Metabolism
• Microfluidics

Background:

• Three-dimensional (3D) spheroids are valuable in vitro models mimicking in vivo environments.
• Current methods for monitoring oxygen consumption in spheroids are often invasive or technically limited.
• Accurate measurement of spheroid oxygen consumption is crucial for understanding cellular activities and drug efficacy.

Purpose of the Study:

• To develop a non-invasive platform for spheroid formation, handling, and oxygen consumption analysis.
• To integrate microfluidics with frequency domain fluorescence lifetime imaging microscopy (FD-FLIM) for enhanced characterization.
• To accurately assess spheroid oxygen consumption in response to therapeutic interventions.

Main Methods:

• Development of a microfluidic platform for uniform spheroid generation and culture.
• Integration of the microfluidic platform with widefield frequency domain fluorescence lifetime imaging microscopy (FD-FLIM).
• Utilized an oxygen-sensitive dye for fluorescence lifetime measurements to quantify oxygen consumption in osteosarcoma (MG-63) spheroids.

Main Results:

• Demonstrated the successful formation and culture of uniform-sized spheroids within the microfluidic platform.
• Successfully characterized oxygen consumption of individual spheroids using FD-FLIM.
• Accurately quantified spheroid oxygen consumption changes in response to drug treatments.

Conclusions:

• The developed microfluidic-FD-FLIM approach provides a sensitive and non-invasive method for spheroid oxygen consumption studies.
• This technology enables single-spheroid resolution analysis of cellular metabolism.
• The platform holds significant potential for advancing drug discovery and personalized medicine through spheroid-based assays.