Time- And Spectral-Resolved Multiphoton Imaging Of Fresh Bladder Biopsies

Year: 2009

Authors: Cicchi R., Crisci A., Nesi G., Cosci A., Giancane S., Carini M., Pavone FS.

Autors Affiliation: European Laboratory for Non-linear Spectroscopy (L.E.N.S.), Department of Physics, University of Florence, 1 Via Nello Carrara, 50019 Sesto Fiorentino, Italy; University of Florence Medical School, Department of Surgical and Medical Critical Area, University of Florence, Florence, Italy; Division of Urology, Department of Surgical and Medical Critical Area, University of Florence, Florence, Italy

Abstract: Human tissues intrinsically contains many fluorophores, as such NADH, elastin, collagen, and flavins, that can be excited and imaged using multiphoton microscopy, up to 150 microns depth. In this work 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 ex-vivo fresh biopsies of bladder. Morphological and spectroscopic analyses allowed to characterize both healthy and pathological tissue samples in a good agreement with common routine histology. In particular, we examined tissue samples from bladder normal mucosa, and bladder carcinoma in-situ (CIS), finding both morphological and spectroscopic differences. From the morphological point of view, cancer cells appeared more elongated with respect to corresponding normal cells; they also exhibited a different nucleus to cytoplasm ratio. From the spectroscopic point of view, we found differences between the two tissue types in both spectral emission and fluorescence lifetime distribution. Even if further analysis, as well as a more significant statistics on a large number of samples would be helpful to discriminate between low and high grade cancer, our method is a promising tool to be used as diagnostic confirmation of histological results, as well as a diagnostic tool in a multiphoton endoscope or cystoscope to be used in in-vivo imaging applications.

Conference title: European Conference on Biomedical Optics – ECBO 2009

KeyWords: Bladder carcinoma; Cancer cells; Diagnostic tools; Ex-vivo; Fluorescence lifetime imaging microscopy; Fluorescence lifetimes; Human tissues; In-situ; In-Vivo imaging; Intrinsic fluorescence; Multi-photon microscopy; Multi-spectral; Multiphoton imaging; Multiphotons; Non-linear microscopy; Number of samples; Pathological tissue; Second harmonic generation microscopy; Spectral emission; Tissue samples; Tissue types; Two photon; Two-photon emission, Biopsy; Cells; Cytology; Harmonic generation; Histology; Nonlinear optics; Photons; Spectroscopic analysis; Urology, Fluorescence
DOI: 10.1117/12.831486