Two-photon microscopy has been successfully used to image several types of tissues, including skin, muscles, tendons. Nevertheless, its usefulness in imaging bladder tissue has not been investigated yet. In this work we used combined twophoton excited fluorescence, second-harmonic generation microscopy, fluorescence lifetime imaging microscopy, and multispectral two-photon emission detection to investigate different kinds of human ex-vivo fresh biopsies of bladder. Morphological and spectroscopic analyses allowed to characterize both healthy mucosa and carcinoma in-situ samples in a good agreement with common routine histology. Cancer cells showed different morphology with respect to the corresponding healthy cells: they appeared more elongated and with a larger nucleus to cytoplasm ratio. From the spectroscopic point of view, differences between the two tissue types in both spectral emission and fluorescence lifetime distribution were found. Even if further analysis, as well as a more significant statistics on a larger number of samples would be helpful to discriminate between low, mild, and high grade cancer, our method is a promising tool to be used as diagnostic confirmation of histological results, as well to be implemented in a multi-photon endoscope or in a spectroscopic for in in-vivo imaging applications.

Date Published: 2 March 2010
PDF: 8 pages
Proc. SPIE 7548, Photonic Therapeutics and Diagnostics VI, 75480V (2 March 2010); doi: 10.1117/12.841694

Published in SPIE Proceedings Vol. 7548:
Photonic Therapeutics and Diagnostics VI
Anita Mahadevan-Jansen; Andreas Mandelis; Brian Jet-Fei Wong M.D.; Nikiforos Kollias; Henry Hirschberg M.D.; Kenton W. Gregory M.D.; Reza S. Malek; E. Duco Jansen; Guillermo J. Tearney; Steen J. Madsen; Bernard Choi; Justus F. R. Ilgner; Haishan Zeng; Laura Marcu, Editor(s)