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Opt Express. 2010 Feb 15;18(4):3840-9. doi: 10.1364/OE.18.003840.

Time- and Spectral-resolved two-photon imaging of healthy bladder mucosa and carcinoma in situ.

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LENS, European Laboratory for Non-Linear Spectroscopy, Via Nello Carrara 1, Sesto Fiorentino, I-50019, Italy.


Combined non-linear imaging techniques were used to deeply image human ex-vivo fresh biopsies of bladder as well as to discriminate between healthy bladder mucosa and carcinoma in situ. Morphological examination by two-photon excited fluorescence and second-harmonic generation has shown a good agreement with corresponding common routine histology performed on the same samples. Tumor cells appeared slightly different in shape and with a smaller cellular-to-nuclear dimension ratio with respect to corresponding normal cells. Further differences between the two tissue types were found in both spectral emission and fluorescence lifetime distribution by performing temporal- and spectral- resolved analysis of fluorescence. This method may represent a promising tool to be used in a multi-photon endoscope, in a confocal endoscope or in a spectroscopic probe for in-vivo optical diagnosis of bladder cancer.

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