![]() The metabolic demands required by cellular processes like proliferation or macromolecule synthesis will shift the relative balance of these cofactors in oxidation/reduction reactions during glycolysis, the tricarboxylic acid (TCA) cycle, and oxidative phosphorylation 16– 21. This allows MPM to not only capture morphological features found in wounds, but also provides a means to assess cell function. Utilizing the simultaneous absorption of two near-infrared photons, MPM can detect fluorescence from reduced nicotinamide dinucleotide (NADH) and flavin adenine dinucleotide (FAD) which are key electron carriers ubiquitous in cell metabolism 13– 15. Optical imaging techniques such as multiphoton microscopy (MPM) have proven capable of measuring the natural autofluorescence in live tissues providing a means to visualize tissue microstructure without traditional histological stains 11, 12. Histology has provided many useful insights into wound pathophysiology 9, 10, but is time-intensive, reliant on subjective user input, and relies on the destructive process of biopsying, sectioning, and staining the tissue. A substantial number of wound healing products and treatment strategies have been developed in recent years 6– 8, but their efficacy is largely determined by time to closure and qualitative histological analysis. These wounds disproportionately afflict the elderly 4, who frequently suffer from comorbidities like diabetes mellitus that can further inhibit healthy wound closure 4, 5. Chronic, non-healing wounds are a serious detriment to public health, afflicting up to 2% of the global population 1 and 6 million Americans 2 while costing the US healthcare system up to $50 billion dollars annually 3. ![]()
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