Three-dimensional multiphoton autofluorescence spectral imaging of live tissues

Jonathan A. Palero; Henriëtte S. de Bruijn; Angélique van der Ploeg van den Heuvel; Henricus J. C. M. Sterenborg; Hans C. Gerritsen

doi:10.1117/12.663736

14 April 2006 Three-dimensional multiphoton autofluorescence spectral imaging of live tissues

Jonathan A. Palero, Henriëtte S. de Bruijn, Angélique van der Ploeg van den Heuvel, Henricus J. C. M. Sterenborg, Hans C. Gerritsen

Author Affiliations +

Proceedings Volume 6191, Biophotonics and New Therapy Frontiers; 61910J (2006) https://doi.org/10.1117/12.663736
Event: SPIE Photonics Europe, 2006, Strasbourg, France

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.

Citation Download Citation

Jonathan A. Palero, Henriëtte S. de Bruijn, Angélique van der Ploeg van den Heuvel, Henricus J. C. M. Sterenborg, and Hans C. Gerritsen "Three-dimensional multiphoton autofluorescence spectral imaging of live tissues", Proc. SPIE 6191, Biophotonics and New Therapy Frontiers, 61910J (14 April 2006); https://doi.org/10.1117/12.663736

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