Non-linear microscopy and spectroscopy of skin tissues

Jonathan A. Palero; Gwendal Latouche; Henriëtte S. de Bruijn; Hans C. Gerritsen; Henricus J. C. M. Sterenborg

doi:10.1117/12.660069

25 October 2005 Non-linear microscopy and spectroscopy of skin tissues

Jonathan A. Palero, Gwendal Latouche, Henriëtte S. de Bruijn, Hans C. Gerritsen, Henricus J. C. M. Sterenborg

Proceedings Volume 5968, Laser Florence 2004: A Window on the Laser Medicine World; 59680H (2005) https://doi.org/10.1117/12.660069
Event: Laser Florence 2004, 2004, Florence, Italy

Abstract

We combined a non-linear microscope with a sensitive prism-based spectrograph and employed it for the imaging of the auto fluorescence of skin tissues. The system has a sub-micron spatial resolution and a spectral resolution of better than 5 nm. The spectral images contain signals arising from two-photon excited fluorescence (TPEF) of endogenous fluorophores in the skin and from second harmonic generation (SHG) produced by the collagen fibers, which have non-centrosymmetric structure. Non-linear microscopy has the potential to image deep into optically thick specimens because it uses near-infrared (NIR) laser excitation. In addition, the phototoxicity of the technique is comparatively low. Here, the technique is used for the spectral imaging of unstained skin tissue sections. We were able to image weak cellular autofluorescence as well as strong collagen SHG. The images were analyzed by spectral unmixing and the results exhibit a clear spectral signature for the different skin layers.

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Jonathan A. Palero, Gwendal Latouche, Henriëtte S. de Bruijn, Hans C. Gerritsen, and Henricus J. C. M. Sterenborg "Non-linear microscopy and spectroscopy of skin tissues", Proc. SPIE 5968, Laser Florence 2004: A Window on the Laser Medicine World, 59680H (25 October 2005); https://doi.org/10.1117/12.660069

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KEYWORDS

Skin

Tissues

Second-harmonic generation

Luminescence

Collagen

Microscopy

Charge-coupled devices

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