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Proceedings Paper

Spectrally resolved multiphoton imaging of post-mortem biopsy and in-vivo mouse skin tissues
Author(s): Jonathan A. Palero; Henriëtte S. de Bruijn; Angélique van der Ploeg van den Heuvel; Henricus J. C. M. Sterenborg; Hans C. Gerritsen
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Paper Abstract

The deep-tissue penetration and submicron spatial resolution of multi-photon microscopy and the high-detection efficiency and nanometer spectral resolution capability of a spectrograph were combined to study the intrinsic emission of mouse skin post mortem biopsy and section, and in vivo tissue samples. The different layers of skin could be clearly distinguished based on both their spectral signature and morphology. Auto fluorescence could be detected from both cellular and extra cellular structures. In addition SHG from collagen and a narrowband spectral emission band related to collagen were observed. Visualization of the spectral images in RGB color allowed us to identify tissue structures such as epidermal cells, lipid-rich keratinocytes and intercellular structures, hair follicles, collagen, elastin, and dermal fibroblasts. The results also showed morphological and spectral differences between the mouse skin post mortem biopsy and in vivo samples which explained by biochemical differences, specifically of NAD(P)H. Overall, spectral imaging provided a wealth of information not easily obtainable with present conventional multi-photon imaging methods.

Paper Details

Date Published: 10 February 2007
PDF: 10 pages
Proc. SPIE 6442, Multiphoton Microscopy in the Biomedical Sciences VII, 64421B (10 February 2007); doi: 10.1117/12.702059
Show Author Affiliations
Jonathan A. Palero, Utrecht Univ. (Netherlands)
Henriëtte S. de Bruijn, Erasmus Medical Ctr., Rotterdam (Netherlands)
Angélique van der Ploeg van den Heuvel, Erasmus Medical Ctr., Rotterdam (Netherlands)
Henricus J. C. M. Sterenborg, Erasmus Medical Ctr., Rotterdam (Netherlands)
Hans C. Gerritsen, Utrecht Univ. (Netherlands)

Published in SPIE Proceedings Vol. 6442:
Multiphoton Microscopy in the Biomedical Sciences VII
Ammasi Periasamy; Peter T. C. So, Editor(s)

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