Analysis of the metabolic deterioration of ex vivo skin from ischemic necrosis through the imaging of intracellular NAD(P)H by multiphoton tomography and fluorescence lifetime imaging microscopy

Washington Y. Sanchez; Tarl W. Prow; Washington H. Sanchez; Jeffrey Grice; Michael S. Roberts

doi:10.1117/1.3466580

1 July 2010 Analysis of the metabolic deterioration of ex vivo skin from ischemic necrosis through the imaging of intracellular NAD(P)H by multiphoton tomography and fluorescence lifetime imaging microscopy

Washington Y. Sanchez, Tarl W. Prow, Washington H. Sanchez, Jeffrey Grice, Michael S. Roberts

Author Affiliations +

Journal of Biomedical Optics, Vol. 15, Issue 4, 046008 (July 2010). https://doi.org/10.1117/1.3466580

Abstract

Ex vivo human skin has been used extensively for cosmeceutical and drug delivery studies, transplantable skin allografts, or skin flaps. However, it has a half-life of a few days due to ischemic necrosis. Traditional methods of assessing viability can be time-consuming and provide limited metabolic information. Using multiphoton tomography and fluorescence lifetime imaging (MPT-FLIM) we assess ischemic necrosis of ex vivo skin by NAD(P)H autofluorescence intensity and fluorescence lifetime. Ex vivo skin is stored in the presence and absence of nutrient media (Dulbecco Modified Eagle Medium) at −20, 4, and 37 °C and room temperature over a 7-day time course to establish different rates of metabolic deterioration. At higher temperatures we observe a decrease in NAD(P)H autofluorescence, higher image noise, and a significant increase in the average fluorescence lifetime (Πm) from ~1000 to 2000 ps. Additionally, significant distortions in NAD(P)H fluorescence lifetime histograms correspond to the reduction in autofluorescence. Skin kept at 4 °C, with or without media, showed the least change. Our findings suggest that MPT-FLIM enables useful noninvasive optical biopsies to monitor the metabolic state and deterioration of human skin for research and clinical purposes.

©(2010) Society of Photo-Optical Instrumentation Engineers (SPIE)

Citation Download Citation

Washington Y. Sanchez, Tarl W. Prow, Washington H. Sanchez, Jeffrey Grice, and Michael S. Roberts "Analysis of the metabolic deterioration of ex vivo skin from ischemic necrosis through the imaging of intracellular NAD(P)H by multiphoton tomography and fluorescence lifetime imaging microscopy," Journal of Biomedical Optics 15(4), 046008 (1 July 2010). https://doi.org/10.1117/1.3466580

Published: 1 July 2010

ACCESS THE FULL ARTICLE

JOURNAL ARTICLE
11 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY