Special Section on Optical Methods of Imaging in the Skin

Raman spectroscopic analysis of human skin tissue sections ex-vivo: evaluation of the effects of tissue processing and dewaxing

[+] Author Affiliations
Syed M. Ali, Franck Bonnier, Fiona M. Lyng

Dublin Institute of Technology, Radiation and Environmental Science Centre, Focas Research Institute, Kevin Street, Dublin 8, Ireland

Ali Tfayli

Univ Paris-Sud, GCAPS “Groupe de Chimie Analytique de Paris-Sud”, Faculty of Pharmacy, 92290 Chatenay Malabry, France

Helen Lambkin, Kathleen Flynn

Institute of Technology, School of Biological Sciences, Dublin Kevin Street, Dublin 8, Ireland

Vincent McDonagh, Claragh Healy, T. Clive Lee

Royal College of Surgeons in Ireland, Department of Anatomy, 123 St. Stephen’s Green, Dublin 2, Ireland

Hugh J. Byrne

Dublin Institute of Technology, Focas Research Institute, Kevin Street, Dublin 8, Ireland

J. Biomed. Opt. 18(6), 061202 (Nov 02, 2012). doi:10.1117/1.JBO.18.6.061202
History: Received May 2, 2012; Revised July 11, 2012; Accepted July 12, 2012
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Abstract.  Raman spectroscopy coupled with K-means clustering analysis (KMCA) is employed to elucidate the biochemical structure of human skin tissue sections and the effects of tissue processing. Both hand and thigh sections of human cadavers were analyzed in their unprocessed and formalin-fixed, paraffin-processed (FFPP), and subsequently dewaxed forms. In unprocessed sections, KMCA reveals clear differentiation of the stratum corneum (SC), intermediate underlying epithelium, and dermal layers for sections from both anatomical sites. The SC is seen to be relatively rich in lipidic content; the spectrum of the subjacent layers is strongly influenced by the presence of melanin, while that of the dermis is dominated by the characteristics of collagen. For a given anatomical site, little difference in layer structure and biochemistry is observed between samples from different cadavers. However, the hand and thigh sections are consistently differentiated for all cadavers, largely based on lipidic profiles. In dewaxed FFPP samples, while the SC, intermediate, and dermal layers are clearly differentiated by KMCA of Raman maps of tissue sections, the lipidic contributions to the spectra are significantly reduced, with the result that respective skin layers from different anatomical sites become indistinguishable. While efficient at removing the fixing wax, the tissue processing also efficiently removes the structurally similar lipidic components of the skin layers. In studies of dermatological processes in which lipids play an important role, such as wound healing, dewaxed samples are therefore not appropriate. Removal of the lipids does however accentuate the spectral features of the cellular and protein components, which may be more appropriate for retrospective analysis of disease progression and biochemical analysis using tissue banks.

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© 2013 Society of Photo-Optical Instrumentation Engineers

Topics

Skin ; Tissues

Citation

Syed M. Ali ; Franck Bonnier ; Ali Tfayli ; Helen Lambkin ; Kathleen Flynn, et al.
"Raman spectroscopic analysis of human skin tissue sections ex-vivo: evaluation of the effects of tissue processing and dewaxing", J. Biomed. Opt. 18(6), 061202 (Nov 02, 2012). ; http://dx.doi.org/10.1117/1.JBO.18.6.061202


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