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  • 1Department of Biomedical Engineering, Tufts University, Medford, MA 02155, USA.

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This study introduces a noninvasive method using endogenous two-photon excited fluorescence (TPEF) to monitor cellular metabolism. The approach quantifies metabolic changes in real-time, aiding disease research.

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Area of Science:

  • Biophotonics
  • Cellular Metabolism
  • Biomarker Discovery

Background:

  • Monitoring cellular metabolism is crucial for understanding diseases like cancer and diabetes.
  • Current methods are often destructive or require external agents.
  • Endogenous two-photon excited fluorescence (TPEF) shows promise for label-free metabolic monitoring, but lacks mechanistic understanding.

Purpose of the Study:

  • To develop a quantitative, noninvasive approach for detecting functional and structural metabolic biomarkers.
  • To establish a link between endogenous TPEF signals and specific metabolic pathways.
  • To enable sensitive, label-free identification of metabolic changes in living cells and tissues.

Main Methods:

  • Utilized endogenous TPEF from NADH (nicotinamide adenine dinucleotide) and FAD (flavin adenine dinucleotide).
  • Performed multiparametric analysis of cellular redox state, NADH fluorescence lifetime, and mitochondrial clustering.
  • Studied metabolic perturbations including glycolysis, glutaminolysis, mitochondrial uncoupling, and fatty acid metabolism in cells and tissues.

Main Results:

  • Demonstrated complementary insights from combined optical biomarkers.
  • Showcased the ability to monitor specific metabolic pathway changes noninvasively.
  • Validated the method's sensitivity in identifying metabolic shifts with single-cell resolution.

Conclusions:

  • The developed quantitative TPEF approach provides a valuable tool for label-free metabolic analysis.
  • This method allows for sensitive detection and characterization of metabolic pathway alterations.
  • It offers a promising strategy for studying diseases linked to metabolic dysfunction.