The autofluorescence under ultraviolet excitation arising from normal squamous and columnar esophageal mucosa is investigated using multispectral microscopy. The results suggest that the autofluorescence signal arises from the superficial tissue layer due to the short penetration depth of the ultraviolet excitation. As a result, visualization of esophageal epithelial cells and their organization can be attained using wide-field autofluorescence microscopy. Our results show tryptophan to be the dominant source of emission under 266 nm excitation, while emission from NADH and collagen are dominant under 355 nm excitation. The analysis of multispectral microscopy images reveals that tryptophan offers the highest image contrast due to its non-uniform distribution in the sub-cellular matrix. This technique can simultaneously provide functional and structural imaging of the microstructure using only the intrinsic tissue fluorophores.

© 2010 OSA

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Am. J. Gastroenterol.

• B. Mayinger, P. Horner, M. Jordan, C. Gerlach, T. Horbach, W. Hohenberger, and E. G. Hahn, “Endoscopic fluorescence spectroscopy in the upper GI tract for the detection of GI cancer: initial experience,” Am. J. Gastroenterol. 96(9), 2616–2621 (2001). [CrossRef] [PubMed]

Bull. N. Y. Acad. Med.

• R. R. Alfano, B. B. Das, J. Cleary, R. Prudente, and E. J. Celmer, “Light sheds light on cancer--distinguishing malignant tumors from benign tissues and tumors,” Bull. N. Y. Acad. Med. 67(2), 143–150 (1991). [PubMed]

Gastroenterology

• M. B. Wallace and P. Fockens, “Probe-based confocal laser endomicroscopy,” Gastroenterology 136(5), 1509–1513 (2009). [CrossRef] [PubMed]

Gastrointest. Endosc.

• T. J. Muldoon, S. Anandasabapathy, D. Maru, and R. Richards-Kortum, “High-resolution imaging in Barrett’s esophagus: a novel, low-cost endoscopic microscope,” Gastrointest. Endosc. 68(4), 737–744 (2008). [CrossRef] [PubMed]
• S. V. Kantsevoy, D. G. Adler, J. D. Conway, D. L. Diehl, F. A. Farraye, V. Kaul, S. R. Kethu, R. S. Kwon, P. Mamula, S. A. Rodriguez, W. M. Tierney, and ASGE Technology Committee, “Confocal laser endomicroscopy,” Gastrointest. Endosc. 70(2), 197–200 (2009). [CrossRef] [PubMed]

IEEE J. Sel. Top. Quantum Electron.

• K. Gono, “Multifunctional endoscopic imaging system for support of early cancer diagnosis,” IEEE J. Sel. Top. Quantum Electron. 14(1), 62–69 (2008). [CrossRef]

Int. J. Cancer

• W. Zheng, W. Lau, C. Cheng, K. C. Soo, and M. Olivo, “Optimal excitation-emission wavelengths for autofluorescence diagnosis of bladder tumors,” Int. J. Cancer 104(4), 477–481 (2003). [CrossRef] [PubMed]

J. Biomed. Opt.

• W. L. Rice, D. L. Kaplan, and I. Georgakoudi, “Quantitative biomarkers of stem cell differentiation based on intrinsic two-photon excited fluorescence,” J. Biomed. Opt. 12(6), 060504 (2007). [CrossRef]
• J. D. Rogers, S. Landau, T. S. Tkaczyk, M. R. Descour, M. S. Rahman, R. Richards-Kortum, A. H. Kärkäinen, and T. Christenson, “Imaging performance of a miniature integrated microendoscope,” J. Biomed. Opt. 13(5), 054020 (2008). [CrossRef] [PubMed]

J. Mammary Gland Biol. Neoplasia

• S. G. Demos, A. J. Vogel, and A. H. Gandjbakhche, “Advances in optical spectroscopy and imaging of breast lesions,” J. Mammary Gland Biol. Neoplasia 11(2), 165–181 (2006). [CrossRef] [PubMed]

Lasers Surg. Med.

• T. J. Pfefer, D. Y. Paithankar, J. M. Poneros, K. T. Schomacker, and N. S. Nishioka, “Temporally and spectrally resolved fluorescence spectroscopy for the detection of high grade dysplasia in Barrett’s esophagus,” Lasers Surg. Med. 32(1), 10–16 (2003). [CrossRef] [PubMed]

Nature

• P. Hsiung, J. Hardy, S. Friedland, R. Soetikno, C. Du, A. Wu, P. Sahbaie, J. Crawford, A. Lowe, and C. Contag, “Detection of colonic dysplasia in vivo using a targeted heptapeptide and confocal microendoscopy,” Nature 200, 8 (2007).

Opt. Express

• B. Lin, S. Urayama, R. M. G. Saroufeem, D. L. Matthews, and S. G. Demos, “Real-time microscopic imaging of esophageal epithelial disease with autofluorescence under ultraviolet excitation,” Opt. Express 17(15), 12502–12509 (2009). [CrossRef] [PubMed]
• C. Li, R. K. Pastila, C. Pitsillides, J. M. Runnels, M. Puoris’haag, D. Côté, and C. P. Lin, “Imaging leukocyte trafficking in vivo with two-photon-excited endogenous tryptophan fluorescence,” Opt. Express 18(2), 988–999 (2010). [CrossRef] [PubMed]

Opt. Lett.

• D. Li, W. Zheng, and J. Y. Qu, “Imaging of epithelial tissue in vivo based on excitation of multiple endogenous nonlinear optical signals,” Opt. Lett. 34(18), 2853–2855 (2009). [CrossRef] [PubMed]

Otolaryngol. Head Neck Surg.

• J. Meier, D. Farwell, Y. Sun, N. Hatami, L. Marcu, and H. Xie, “Fluorescence spectroscopy as a diagnostic tool in HNSCC,” Otolaryngol. Head Neck Surg. 141(3), P51–P52 (2009). [CrossRef]

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