Two-photon imaging and spectroscopy of fresh human colon biopsies

Year: 2012

Authors: Cicchi R., Sturiale A., Nesi G., Tonelli F., Pavone FS.

Autors Affiliation: European Laboratory for Non-linear Spectroscopy (LENS), University of Florence, 1 Via Nello Carrara, Sesto Fiorentino, Italy;
Department of Clinical Physiopathology, Surgical Unit, University of Florence, Florence, Italy;
Division of Human Pathology and Oncology, Department of Surgical and Medical Clinical Care, University of Florence, Florence, Italy

Abstract: Two-photon fluorescence (TPEF) microscopy is a powerful tool to image human tissues up to 200 microns depth without any exogenously added probe. TPEF can take advantage of the autofluorescence of molecules intrinsically contained in a biological tissue, as such NADH, elastin, collagen, and flavins. Two-photon microscopy has been already successfully used to image several types of tissues, including skin, muscles, tendons, bladder. Nevertheless, its usefulness in imaging colon tissue has not been deeply investigated yet. In this work we have used combined two-photon excited fluorescence (TPEF), 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 colon. Morphological and spectroscopic analyses allowed to characterize both healthy mucosa, polyp, and colon samples in a good agreement with common routine histology. Even if further analysis, as well as a more significant statistics on a large 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 as a diagnostic tool in a multiphoton endoscope or colonoscope to be used in in-vivo imaging applications.

Conference title: Photonics West – BIOS 2012
Place: San Francisco, US

More Information: The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement 228334. This research project has been also supported by Human Frontier Science Program (HFSP) under grant agreement RGP0027/2009.
KeyWords: Autofluorescence; Biological tissues; Colon; Colon tissues; Diagnostic tools; Ex-vivo; Fluorescence lifetime imaging microscopy; Human colon; Human tissues; In-Vivo imaging; Multi-spectral; Multiphotons; Number of samples; Second harmonic generation microscopies (SHG); Two photon fluorescence; Two photon imaging; Two photon microscopy; Two-photon; Two-photon emission; Two-photon excited fluorescence, Biopsy; Fluorescence; Histology; Spectroscopic analysis; Spectroscopy; Tissue, Photons
DOI: 10.1117/12.907822

Citations: 3
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