Multidimensional two-photon imaging of diseased skin

Year: 2008

Authors: Cicchi R., Sestini S., De Giorgi V., Massi D., Lotti T., Pavone FS.

Autors Affiliation: European Laboratory for Non-linear Spectroscopy,
University of Florence, Via Nello Carrara 1, 50019,
Sesto Fiorentino, Italy;

Abstract: We used combined two photon intrinsic fluorescence (TPE), second harmonic generation microscopy (SHG), fluorescence lifetime imaging microscopy (FLIM), and multispectral two photon emission detection (MTPE) to investigate different kinds of human cutaneous ex-vivo skin lesions. Morphological and spectroscopic analyses allowed to characterize both healthy and pathological skin samples, including tumors, as well as to discriminate between healthy and diseased tissue, in a good agreement with common routine histology. In particular, we examined tissue samples from normal and pathological scar tissue (keloid), and skin tumors, including basal cell carcinoma (BCC) and malignant melanoma (MM). By using combined TPE-SHG microscopy we investigated morphological features of different skin regions, as BCC, tumor-stroma interface, healthy dermis, fibroblastic proliferation, and keloids. The SHG to autofluorescence aging index of dermis (SAAID) score was used to characterize each region, finding differences between BCC, healthy skin, tumor-stroma interface, keloids, and fibroblastic proliferation. Further comparative analysis of healthy skin and neoplastic samples was performed using FLIM. In particular, BCC showed a blue-shifted fluorescence emission, a higher absorption at 800 nm excitation wavelength, and a slightly longer mean fluorescence lifetime. MM showed a lifetime distribution similar to the corresponding melanocytic nevus (MN) lifetime distribution for the slow lifetime component, and different for the fast lifetime component.

Conference title: Photonics West – BIOS 2008
Place: San Francisco, US

More Information: We thank European Laboratory for Non-linear Spectroscopy (Contract number TOK-MC-509761) and Ente Cassa di Risparmio di Firenze for financial support to this project.
KeyWords: Basal cell carcinoma; Lifetime imaging; Multi-photon microscopy; Multispectral imaging; Optical biopsies, Biomolecules; Biopsy; Dermatology; Fluorescence; Harmonic generation; Histology; Multiphoton processes; Nonlinear optics; Photons; Spectroscopic analysis, Tumors
DOI: 10.1117/12.762614

Citations: 1
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