Autofluorescence spectroscopy of epithelial tissues.

Autofluorescence of rabbit and human epithelial tissues were studied by using a depth-resolved fluorescence spectroscopy system with multiple excitations. Keratinization was found to be common in the squamous epithelium. Strong keratin fluorescence with excitation and emission characteristics similar to collagen were observed in the topmost layer of the keratinized squamous epithelium. The keratin signal created interference in the assessment of the endogenous fluorescence signals (NADH/FAD fluorescence in epithelium and collagen fluorescence in stroma) associated with the development of epithelial precancer. Furthermore, the keratinized epithelial layer attenuated the excitation light and reduced the fluorescence signals from underlying tissue layers. The autofluorescence of columnar epithelium was found to be dominated by NADH and FAD signals, identical to the autofluorescence measured from nonkeratinized squamous epithelium. The study also demonstrated that a fluorescence signal excited at 355 nm produced sufficient contrast to resolve the layered structure of epithelial tissue, while the signal excited at 405 nm provided the information for a good estimation of epithelial redox ratios that are directly related to tissue metabolism. Overall, the depth-resolved measurements are crucial to isolate the fluorescence signals from different sublayers of the epithelial tissue and provide more accurate information for the tissue diagnosis.

[1]  Michele Follen,et al.  Autofluorescence Patterns in Short-Term Cultures of Normal Cervical Tissue , 2000, Photochemistry and photobiology.

[2]  Peng Xi,et al.  Depth-resolved fluorescence spectroscopy reveals layered structure of tissue. , 2004, Optics express.

[3]  M. H. Ross,et al.  Histology: A Text and Atlas , 1985 .

[4]  Watt W Webb,et al.  Two-photon fluorescence spectroscopy and microscopy of NAD(P)H and flavoprotein. , 2002, Biophysical journal.

[5]  K. Shibuya,et al.  Spectroscopic analysis of the autofluorescence from human bronchus using an ultraviolet laser diode. , 2002, Journal of biomedical optics.

[6]  B F Overholt,et al.  Endoscopic fluorescence detection of high-grade dysplasia in Barrett's esophagus. , 1996, Gastroenterology.

[7]  S. Shapshay,et al.  Spectroscopic detection and evaluation of morphologic and biochemical changes in early human oral carcinoma , 2003, Cancer.

[8]  R Richards-Kortum,et al.  Understanding the contributions of NADH and collagen to cervical tissue fluorescence spectra: modeling, measurements, and implications. , 2001, Journal of biomedical optics.

[9]  G. Evan,et al.  Proliferation, cell cycle and apoptosis in cancer , 2001, Nature.

[10]  Iris Riemann,et al.  High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution. , 2003, Journal of biomedical optics.

[11]  H. Moch,et al.  Angiogenesis in cervical neoplasia: microvessel quantitation in precancerous lesions and invasive carcinomas with clinicopathological correlations. , 1997, Gynecologic oncology.

[12]  B. Chance,et al.  Intracellular Oxidation-Reduction States in Vivo , 1962, Science.

[13]  G. Zonios,et al.  Morphological model of human colon tissue fluorescence , 1996, IEEE Transactions on Biomedical Engineering.

[14]  Peng Xi,et al.  Depth-resolved fluorescence spectroscopy of normal and dysplastic cervical tissue. , 2005, Optics express.

[15]  John A. Kiernan,et al.  Histological and Histochemical Methods: Theory and Practice, 4th edition , 2008 .

[16]  T J Flotte,et al.  Ultraviolet laser‐induced fluorescence of colonic tissue: Basic biology and diagnostic potential , 1992, Lasers in surgery and medicine.

[17]  B Chance,et al.  Oxidation-reduction properties of the mitochondrial flavoprotein chain. , 1968, Biochemical and biophysical research communications.

[18]  R. Alfano,et al.  Laser induced fluorescence spectroscopy from native cancerous and normal tissue , 1984 .

[19]  A E Profio,et al.  A feasibility study of the use of fluorescence bronchoscopy for localization of small lung tumours. , 1977, Physics in medicine and biology.

[20]  Michele Follen,et al.  Sources of scattering in cervical tissue: determination of the scattering coefficient by confocal microscopy. , 2005, Applied optics.

[21]  R M Cothren,et al.  Laser-induced fluorescence microscopy of normal colon and dysplasia in colonic adenomas: implications for spectroscopic diagnosis. , 1995, The American journal of gastroenterology.

[22]  K. Badizadegan,et al.  NAD(P)H and collagen as in vivo quantitative fluorescent biomarkers of epithelial precancerous changes. , 2002, Cancer research.

[23]  R. Lotan,et al.  Autofluorescence Microscopy of Fresh Cervical-Tissue Sections Reveals Alterations in Tissue Biochemistry with Dysplasia¶ , 2001, Photochemistry and photobiology.

[24]  Rebecca Richards-Kortum,et al.  Realistic three-dimensional epithelial tissue phantoms for biomedical optics. , 2002, Journal of biomedical optics.

[25]  B. Chance,et al.  On the fluorescence of NAD(P)H in whole-cell preparations of tumours and normal tissues. , 1970, European journal of biochemistry.

[26]  H Stepp,et al.  Autofluorescence imaging and spectroscopy of normal and malignant mucosa in patients with head and neck cancer , 1999, Lasers in surgery and medicine.

[27]  Urs Utzinger,et al.  Endogenous Fluorescence Spectroscopy of Cell Suspensions for Chemopreventive Drug Monitoring¶ , 2005, Photochemistry and photobiology.

[28]  B. Schoener,et al.  Intracellular Oxidation-Reduction States in Vivo , 1962, Science.

[29]  Y. DeClerck.  Interactions between tumour cells and stromal cells and proteolytic modification of the extracellular matrix by metalloproteinases in cancer. , 2000, European journal of cancer.

[30]  B. Wilson,et al.  In Vivo Fluorescence Spectroscopy and Imaging for Oncological Applications , 1998, Photochemistry and photobiology.

[31]  N Ramanujam,et al.  In vivo diagnosis of cervical intraepithelial neoplasia using 337-nm-excited laser-induced fluorescence. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[32]  B. Chance,et al.  Oxidation-reduction ratio studies of mitochondria in freeze-trapped samples. NADH and flavoprotein fluorescence signals. , 1979, The Journal of biological chemistry.

[33]  J. V. Bacus,et al.  Properties of Intraepithelial Neoplasia Relevant to Cancer Chemoprevention and to the Development of Surrogate End Points for Clinical Trials , 1997, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[34]  E. Sevick-Muraca,et al.  Quantitative optical spectroscopy for tissue diagnosis. , 1996, Annual review of physical chemistry.

[35]  N. Ramanujam.  Fluorescence spectroscopy of neoplastic and non-neoplastic tissues. , 2000, Neoplasia.

[36]  R S Balaban,et al.  Nicotinamide adenine dinucleotide fluorescence spectroscopy and imaging of isolated cardiac myocytes. , 1989, Biophysical journal.

[37]  L. Gaboury,et al.  Steady state and time-resolved fluorescence properties of metastatic and non-metastatic malignant cells from different species. , 1995, Journal of photochemistry and photobiology. B, Biology.