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J Biomed Opt. 2017 Dec;22(12):1-10. doi: 10.1117/1.JBO.22.12.125004.

Non-Euclidean phasor analysis for quantification of oxidative stress in ex vivo human skin exposed to sun filters using fluorescence lifetime imaging microscopy.

Author information

1
Harvard Medical School, Massachusetts General Hospital, Wellman Center for Photomedicine, Charlestow, United States.
2
Massachusetts Institute of Technology, Harvard-MIT Division of Health Sciences and Technology, Cambr, United States.
3
Harvard Medical School, Massachusetts General Hospital, Cutaneous Biology Research Center, Charlesto, United States.

Abstract

Chemical sun filters are commonly used as active ingredients in sunscreens due to their efficient absorption of ultraviolet (UV) radiation. Yet, it is known that these compounds can photochemically react with UV light and generate reactive oxygen species and oxidative stress in vitro, though this has yet to be validated in vivo. One label-free approach to probe oxidative stress is to measure and compare the relative endogenous fluorescence generated by cellular coenzymes nicotinamide adenine dinucleotides and flavin adenine dinucleotides. However, chemical sun filters are fluorescent, with emissive properties that contaminate endogenous fluorescent signals. To accurately distinguish the source of fluorescence in ex vivo skin samples treated with chemical sun filters, fluorescence lifetime imaging microscopy data were processed on a pixel-by-pixel basis using a non-Euclidean separation algorithm based on Mahalanobis distance and validated on simulated data. Applying this method, ex vivo samples exhibited a small oxidative shift when exposed to sun filters alone, though this shift was much smaller than that imparted by UV irradiation. Given the need for investigative tools to further study the clinical impact of chemical sun filters in patients, the reported methodology may be applied to visualize chemical sun filters and measure oxidative stress in patients' skin.

KEYWORDS:

flavin adenine dinucleotides; fluorescence lifetime imaging microscopy; nicotinamide adenine dinucleotides; oxidative stress; sun filters; two-photon excitation fluorescence

PMID:
29222855
PMCID:
PMC5722918
[Available on 2018-12-09]
DOI:
10.1117/1.JBO.22.12.125004

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