Multiphoton microscopy is an interesting optical technique, which allows for non-invasive imaging of highly light scattering media such as human skin. Recent reports have showed the potential of applying this technique for 3D visualisation of cell structures of biological tissue without previous sectioning of the tissue samples. In this study, we have applied two-photon microscopy on excised lesions of human non-melanoma skin cancer ex vivo in order to find diagnostic criteria using this technique. The skin samples have been investigated by a multiphoton microscopy system based on a fs-pulsed Ti:sapphire laser connected to a confocal microscope. The autofluorescence of the skin was detected using excitation at 780 nm. The cell nuclei distribution turned out to be one important parameter, which can be used for discriminating between tumour and normal tissue. We are now developing a technique for automatic detection and characterisation of tissue, based on an image analysis algorithm. The detection of cell nuclei has been found crucial for this purpose. The goal is to develop a fast characterisation algorithm that can be used on line in connection to in vivo investigations. This would allow for a true non-invasive biopsy technique in the future.

Date Published: 13 July 2007
PDF: 8 pages
Proc. SPIE 6630, Confocal, Multiphoton, and Nonlinear Microscopic Imaging III, 66300U (13 July 2007); doi: 10.1117/12.725407

Published in SPIE Proceedings Vol. 6630:
Confocal, Multiphoton, and Nonlinear Microscopic Imaging III
Tony Wilson; Ammasi Periasamy, Editor(s)