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Updated: Jul 2, 2025

Author Spotlight: Standardizing Spheroid Formation Methods for Metabolic and Oxygenation Analysis Using Fluorescence Lifetime Imaging Microscopy
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Characterization of Single-Spheroid Oxygen Consumption Using a Microfluidic Platform and Fluorescence Lifetime

Santhosh Kannan1,2,3, Chien-Chung Peng1, Hsiao-Mei Wu4

  • 1Research Center for Applied Sciences, Academia Sinica, Taipei 115201, Taiwan.

Biosensors
|February 23, 2024
PubMed
Summary

This study introduces a novel microfluidic platform and frequency domain fluorescence lifetime imaging microscopy (FD-FLIM) for non-invasively measuring oxygen consumption in 3D cell spheroids, enabling sensitive drug response analysis.

Keywords:
frequency domain fluorescence lifetime imaging microscopy (FD-FLIM)microfluidicsoxygen consumptionspheroid

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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.