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In Vivo, Label-Free Multiphoton Microscopy Is Sensitive to Altered Metabolism and Structure in Aged Skin
Marcos R Rodriguez1, Alan E Woessner2, Kyle P Quinn1,2,3
1Department of Biomedical Engineering, University of Arkansas, Fayetteville, Arkansas, USA.
Advances in Wound Care
|February 5, 2026
Summary
Multiphoton microscopy identified endogenous biomarkers in mouse skin that reveal age-related changes in metabolism and collagen. These findings offer new ways to track skin aging in vivo.
Area of Science:
- Biomedical Optics
- Dermatology
- Aging Research
Background:
- Rising healthcare costs necessitate understanding biological aging.
- Characterizing cellular and tissue aging is crucial for developing interventions.
- Identifying reliable biomarkers for skin aging is an unmet need.
Purpose of the Study:
- To utilize in vivo multiphoton microscopy (MPM) for identifying endogenous biomarkers of skin aging.
- To investigate age-related changes in skin metabolism and collagen structure.
- To establish optical markers for assessing skin aging progression.
Main Methods:
- MPM was performed on mouse skin at 2 and 21 months of age.
- Autofluorescence of nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD) was measured.
- Second-harmonic generation signal from collagen was analyzed, alongside NADH fluorescence lifetime imaging and mitochondrial fractal dimension.
Main Results:
- Aged keratinocytes exhibited fragmented mitochondria with reduced catabolism, indicated by optical redox ratio and NADH fluorescence lifetime.
- Mitochondrial organization in aged skin showed decreased fractal dimension.
- Aged collagen displayed increased nonenzymatic cross-links and fragmented organization.
Conclusions:
- Label-free MPM identified biomarkers sensitive to intrinsic skin aging.
- The study provides the first in vivo demonstration of age-dependent metabolic changes in skin using optical parameters.
- These surrogate markers of metabolic function, mitochondrial, and collagen organization can longitudinally assess aging progression.
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