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Proceedings Paper

Three-dimensional multiphoton autofluorescence spectral imaging of live 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

We combined a homebuilt multiphoton microscope and a prism-CCD based spectrograph to develop a spectral imaging system capable of imaging deep into live tissues. The spectral images originate from the two-photon autofluorescence of the tissue and second harmonic signal from the collagen fibers. A highly penetrating near-infrared light is used to excite the endogenous fluorophores via multiphoton excitation enabling us to produce high quality images deep into the tissue. We were able to produce 100-channel (330 nm to 600 nm) autofluorescence spectral images of live skin tissues in less than 2 minutes for each xy-section. The spectral images rendered in RGB (real) colors showed green hair shafts, blue cells, and purple collagen. Analysis on the optical signal degradation with increasing depth of the collagen second-harmonic signal showed 1) exponential decay behavior of the intensity and 2) linear broadening of the spectrum. This spectral imaging system is a promising tool for both in biological applications and biomedical applications such as optical biopsy.

Paper Details

Date Published: 14 April 2006
PDF: 9 pages
Proc. SPIE 6191, Biophotonics and New Therapy Frontiers, 61910J (14 April 2006); doi: 10.1117/12.663736
Show Author Affiliations
Jonathan A. Palero, Utrecht Univ. (Netherlands)
Henriëtte S. de Bruijn, Erasmus Medical Ctr. (Netherlands)
Angélique van der Ploeg van den Heuvel, Erasmus Medical Ctr. (Netherlands)
Henricus J. C. M. Sterenborg, Erasmus Medical Ctr. (Netherlands)
Hans C. Gerritsen, Utrecht Univ. (Netherlands)

Published in SPIE Proceedings Vol. 6191:
Biophotonics and New Therapy Frontiers
Romualda Grzymala; Olivier Haeberle, Editor(s)

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