Abstract

We explored the use of diffuse reflectance spectroscopy in the ultraviolet-visible (UV-VIS) spectrum for the diagnosis of epithelial pre-cancers and cancers in vivo. A physical model (Monte Carlo inverse model) and an empirical model (principal component analysis, (PCA)) based approach were compared for extracting diagnostic features from diffuse reflectance spectra measured in vivo from the dimethylbenz[α]anthracene-treated hamster cheek pouch model of oral carcinogenesis. These diagnostic features were input into a support vector machine algorithm to classify each tissue sample as normal (n=10) or neoplastic (dysplasia to carcinoma, n=10) and cross-validated using a leave one out method. There was a statistically significant decrease in the absorption and reduced scattering coefficient at 460 nm in neoplastic compared to normal tissues, and these two features provided 90% classification accuracy. The first two principal components extracted from PCA provided a classification accuracy of 95%. The first principal component was highly correlated with the wavelength-averaged reduced scattering coefficient. Although both methods show similar classification accuracy, the physical model provides insight into the physiological and structural features that discriminate between normal and neoplastic tissues and does not require a priori, a representative set of spectral data from which to derive the principal components.

©2007 Optical Society of America

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2007 (1)

M. C. Skala, K. M. Riching, D. K. Bird, A. Gendron-Fitzpatrick, J. Eickhoff, K. W. Eliceiri, P. J. Keely, and N. Ramanujam, “In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia,” J. Biomed. Opt. 12, 024014 (2007).
[Crossref] [PubMed]

2006 (7)

C. Zhu, G. M. Palmer, T. M. Breslin, J. Harter, and N. Ramanujam, “Diagnosis of breast cancer using diffuse reflectance spectroscopy: Comparison of a Monte Carlo versus partial least squares analysis based feature extraction technique,” Lasers Surg. Med. 38, 714–724 (2006).
[Crossref] [PubMed]

S. R. Millon, K. M. Roldan-Perez, K. M. Riching, G. M. Palmer, and N. Ramanujam, “Effect of optical clearing agents on the in vivo optical properties of squamous epithelial tissue,” Lasers Surg. Med. 38, 920–927 (2006).
[Crossref] [PubMed]

G. M. Palmer, C. Zhu, T. M. Breslin, F. Xu, K. W. Gilchrist, and N. Ramanujam, “Monte Carlo-based inverse model for calculating tissue optical properties. Part II: Application to breast cancer diagnosis,” Appl. Opt. 45, 1072–1078 (2006).
[Crossref] [PubMed]

U. Sunar, H. Quon, T. Durduran, J. Zhang, J. Du, C. Zhou, G. Yu, R. Choe, A. Kilger, R. Lustig, L. Loevner, S. Nioka, B. Chance, and A. G. Yodh, “Noninvasive diffuse optical measurement of blood flow and blood oxygenation for monitoring radiation therapy in patients with head and neck tumors: a pilot study,” J. Biomed. Opt. 11, 064021 (2006).
[Crossref]

Y. S. Fawzy, M. Petek, M. Tercelj, and H. Zeng, “In vivo assessment and evaluation of lung tissue morphologic and physiological changes from non-contact endoscopic reflectance spectroscopy for improving lung cancer detection,” J. Biomed. Opt. 11, 044003 (2006).
[Crossref] [PubMed]

N. Subhash, J. R. Mallia, S. S. Thomas, A. Mathews, P. Sebastian, and J. Madhavan, “Oral cancer detection using diffuse reflectance spectral ratio R540/R575 of oxygenated hemoglobin bands,” J. Biomed. Opt. 11, 014018 (2006).
[Crossref] [PubMed]

G. M. Palmer and N. Ramanujam, “Monte Carlo-based inverse model for calculating tissue optical properties. Part I: Theory and validation on synthetic phantoms,” Appl. Opt. 45, 1062–1071 (2006).
[Crossref] [PubMed]

2005 (5)

D. C. de Veld, M. Skurichina, M. J. Witjes, R. P. Duin, H. J. Sterenborg, and J. L. Roodenburg, “Autofluorescence and diffuse reflectance spectroscopy for oral oncology,” Lasers Surg. Med. 36, 356–364 (2005).
[Crossref] [PubMed]

D. J. Parekh, W. C. Lin, and S. D. Herrell, “Optical spectroscopy characteristics can differentiate benign and malignant renal tissues: a potentially useful modality,” J. Urol. 174, 1754–1758 (2005).
[Crossref] [PubMed]

T. Collier, M. Follen, A. Malpica, and R. Richards-Kortum, “Sources of scattering in cervical tissue: determination of the scattering coefficient by confocal microscopy,” Appl. Opt. 44, 2072–2081 (2005).
[Crossref] [PubMed]

M. C. Skala, J. M. Squirrell, K. M. Vrotsos, J. C. Eickhoff, A. Gendron-Fitzpatrick, K. W. Eliceiri, and N. Ramanujam, “Multiphoton microscopy of endogenous fluorescence differentiates normal, precancerous, and cancerous squamous epithelial tissues,” Cancer Res. 65, 1180–1186 (2005).
[Crossref] [PubMed]

C. Zhu, G. M. Palmer, T. M. Breslin, F. Xu, and N. Ramanujam, “Use of a multiseparation fiber optic probe for the optical diagnosis of breast cancer,” J. Biomed. Opt. 10, 024032 (2005).
[Crossref] [PubMed]

2004 (4)

M. C. Skala, G. M. Palmer, C. Zhu, Q. Liu, K. M. Vrotsos, C. L. Marshek-Stone, A. Gendron-Fitzpatrick, and N. Ramanujam, “Investigation of fiber-optic probe designs for optical spectroscopic diagnosis of epithelial pre-cancers,” Lasers Surg. Med. 34, 25–38 (2004).
[Crossref] [PubMed]

C. Baudelet and B. Gallez, “Effect of anesthesia on the signal intensity in tumors using BOLD-MRI: comparison with flow measurements by Laser Doppler flowmetry and oxygen measurements by luminescence-based probes,” Magn. Reson. Imaging 22, 905–912 (2004).
[Crossref] [PubMed]

A. Amelink, H. J. Sterenborg, M. P. Bard, and S. A. Burgers, “In vivo measurement of the local optical properties of tissue by use of differential path-length spectroscopy,” Opt. Lett. 29, 1087–1089 (2004).
[Crossref] [PubMed]

J. C. Finlay and T. H. Foster, “Hemoglobin oxygen saturations in phantoms and in vivo from measurements of steady-state diffuse reflectance at a single, short source-detector separation,” Med. Phys. 31, 1949–1959 (2004).
[Crossref] [PubMed]

2003 (5)

T. J. Pfefer, L. S. Matchette, C. L. Bennett, J. A. Gall, J. N. Wilke, A. J. Durkin, and M. N. Ediger, “Reflectance-based determination of optical properties in highly attenuating tissue,” J. Biomed. Opt. 8, 206–215 (2003).
[Crossref] [PubMed]

G. M. Palmer, C. Zhu, T. M. Breslin, F. Xu, K. W. Gilchrist, and N. Ramanujam, “Comparison of multiexcitation fluorescence and diffuse reflectance spectroscopy for the diagnosis of breast cancer (March 2003),” IEEE Trans. Biomed. Eng. 50, 1233–1242 (2003).
[Crossref] [PubMed]

P. Thueler, I. Charvet, F. Bevilacqua, M. St Ghislain, G. Ory, P. Marquet, P. Meda, B. Vermeulen, and C. Depeursinge, “In vivo endoscopic tissue diagnostics based on spectroscopic absorption, scattering, and phase function properties,” J. Biomed. Opt. 8, 495–503 (2003).
[Crossref] [PubMed]

M. G. Muller, T. A. Valdez, I. Georgakoudi, V. Backman, C. Fuentes, S. Kabani, N. Laver, Z. Wang, C. W. Boone, R. R. Dasari, S. M. Shapshay, and M. S. Feld, “Spectroscopic detection and evaluation of morphologic and biochemical changes in early human oral carcinoma,” Cancer 97, 1681–1692 (2003).
[Crossref] [PubMed]

I. Pavlova, K. Sokolov, R. Drezek, A. Malpica, M. Follen, and R. Richards-Kortum, “Microanatomical and biochemical origins of normal and precancerous cervical autofluorescence using laser-scanning fluorescence confocal microscopy,” Photochem. Photobiol. 77, 550–555 (2003).
[Crossref] [PubMed]

2002 (1)

I. Georgakoudi, E. E. Sheets, M. G. Muller, V. Backman, C. P. Crum, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Trimodal spectroscopy for the detection and characterization of cervical precancers in vivo,“ Am. J. Obstet. Gynecol. 186, 374–382 (2002).
[Crossref] [PubMed]

2001 (3)

R. J. Nordstrom, L. Burke, J. M. Niloff, and J. F. Myrtle, “Identification of cervical intraepithelial neoplasia (CIN) using UV-excited fluorescence and diffuse-reflectance tissue spectroscopy,” Lasers Surg. Med. 29, 118–127 (2001).
[Crossref] [PubMed]

N. Ghosh, S. K. Mohanty, S. K. Majumder, and P. K. Gupta, “Measurement of optical transport properties of normal and malignant human breast tissue,” Appl. Opt. 40, 176–184 (2001).
[Crossref]

R. Drezek, K. Sokolov, U. Utzinger, I. Boiko, A. Malpica, M. Follen, and R. Richards-Kortum, “Understanding the contributions of NADH and collagen to cervical tissue fluorescence spectra: modeling, measurements, and implications,” J. Biomed. Opt. 6, 385–396 (2001).
[Crossref] [PubMed]

1999 (2)

R. Hornung, T. H. Pham, K. A. Keefe, M. W. Berns, Y. Tadir, and B. J. Tromberg, “Quantitative near-infrared spectroscopy of cervical dysplasia in vivo,” Hum. Reprod. 14, 2908–2916 (1999).
[Crossref] [PubMed]

G. Zonios, L. Perelman, V. Backman, R. Manoharan, M. Fitzmaurice, J. Van Dam, and M. S. Feld, “Diffuse reflectance spectroscopy of human adenomatous colon polyps in vivo,” Appl. Opt. 38, 6628–6637 (1999).
[Crossref]

1998 (2)

C. T. Chen, H. K. Chiang, S. N. Chow, C. Y. Wang, Y. S. Lee, J. C. Tsai, and C. P. Chiang, “Autofluorescence in normal and malignant human oral tissues and in DMBA-induced hamster buccal pouch carcinogenesis,” J. Oral Pathol. Med. 27, 470–474 (1998).
[Crossref] [PubMed]

J. Mourant, J. Freyer, A. Hielscher, A. Eick, D. Shen, and T. Johnson, “Mechanisms of light scattering from biological cells relevant to noninvasive optical-tissue diagnostics,” Appl. Opt. 37, 3586–3593 (1998).
[Crossref]

1997 (1)

F. Steinberg, H. J. Rohrborn, T. Otto, K. M. Scheufler, and C. Streffer, “NIR reflection measurements of hemoglobin and cytochrome aa3 in healthy tissue and tumors. Correlations to oxygen consumption: preclinical and clinical data,” Adv. Exp. Med. Biol. 428, 69–77 (1997).
[Crossref] [PubMed]

1996 (2)

C. J. Gulledge and M. W. Dewhirst, “Tumor oxygenation: a matter of supply and demand,” Anticancer Res. 16, 741–749 (1996).
[PubMed]

S. Andrejevic, J. F. Savary, C. Fontolliet, P. Monnier, and H. van Den Bergh, “7,12-dimethylbenz[a]anthracene-induced ‘early’ squamous cell carcinoma in the Golden Syrian hamster: evaluation of an animal model and comparison with ‘early’ forms of human squamous cell carcinoma in the upper aero-digestive tract,” Int. J. Exp. Pathol. 77, 7–14 (1996).
[Crossref] [PubMed]

1995 (1)

1987 (1)

M. L. Ellsworth, R. N. Pittman, and C. G. Ellis, “Measurement of hemoglobin oxygen saturation in capillaries,” Am. J. Physiol. 252, H1031–1040 (1987).
[PubMed]

1981 (1)

F. H. White, K. Gohari, and C. J. Smith, “Histological and ultrastructural morphology of 7,12 dimethylbenz(alpha)-anthracene carcinogenesis in hamster cheek pouch epithelium,” Diagn. Histopathol. 4, 307–333 (1981).
[PubMed]

Amelink, A.

Andrejevic, S.

S. Andrejevic, J. F. Savary, C. Fontolliet, P. Monnier, and H. van Den Bergh, “7,12-dimethylbenz[a]anthracene-induced ‘early’ squamous cell carcinoma in the Golden Syrian hamster: evaluation of an animal model and comparison with ‘early’ forms of human squamous cell carcinoma in the upper aero-digestive tract,” Int. J. Exp. Pathol. 77, 7–14 (1996).
[Crossref] [PubMed]

Backman, V.

M. G. Muller, T. A. Valdez, I. Georgakoudi, V. Backman, C. Fuentes, S. Kabani, N. Laver, Z. Wang, C. W. Boone, R. R. Dasari, S. M. Shapshay, and M. S. Feld, “Spectroscopic detection and evaluation of morphologic and biochemical changes in early human oral carcinoma,” Cancer 97, 1681–1692 (2003).
[Crossref] [PubMed]

I. Georgakoudi, E. E. Sheets, M. G. Muller, V. Backman, C. P. Crum, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Trimodal spectroscopy for the detection and characterization of cervical precancers in vivo,“ Am. J. Obstet. Gynecol. 186, 374–382 (2002).
[Crossref] [PubMed]

G. Zonios, L. Perelman, V. Backman, R. Manoharan, M. Fitzmaurice, J. Van Dam, and M. S. Feld, “Diffuse reflectance spectroscopy of human adenomatous colon polyps in vivo,” Appl. Opt. 38, 6628–6637 (1999).
[Crossref]

Badizadegan, K.

I. Georgakoudi, E. E. Sheets, M. G. Muller, V. Backman, C. P. Crum, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Trimodal spectroscopy for the detection and characterization of cervical precancers in vivo,“ Am. J. Obstet. Gynecol. 186, 374–382 (2002).
[Crossref] [PubMed]

Bard, M. P.

Baudelet, C.

C. Baudelet and B. Gallez, “Effect of anesthesia on the signal intensity in tumors using BOLD-MRI: comparison with flow measurements by Laser Doppler flowmetry and oxygen measurements by luminescence-based probes,” Magn. Reson. Imaging 22, 905–912 (2004).
[Crossref] [PubMed]

Bennett, C. L.

T. J. Pfefer, L. S. Matchette, C. L. Bennett, J. A. Gall, J. N. Wilke, A. J. Durkin, and M. N. Ediger, “Reflectance-based determination of optical properties in highly attenuating tissue,” J. Biomed. Opt. 8, 206–215 (2003).
[Crossref] [PubMed]

Berns, M. W.

R. Hornung, T. H. Pham, K. A. Keefe, M. W. Berns, Y. Tadir, and B. J. Tromberg, “Quantitative near-infrared spectroscopy of cervical dysplasia in vivo,” Hum. Reprod. 14, 2908–2916 (1999).
[Crossref] [PubMed]

Bevilacqua, F.

P. Thueler, I. Charvet, F. Bevilacqua, M. St Ghislain, G. Ory, P. Marquet, P. Meda, B. Vermeulen, and C. Depeursinge, “In vivo endoscopic tissue diagnostics based on spectroscopic absorption, scattering, and phase function properties,” J. Biomed. Opt. 8, 495–503 (2003).
[Crossref] [PubMed]

Bird, D. K.

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N. Subhash, J. R. Mallia, S. S. Thomas, A. Mathews, P. Sebastian, and J. Madhavan, “Oral cancer detection using diffuse reflectance spectral ratio R540/R575 of oxygenated hemoglobin bands,” J. Biomed. Opt. 11, 014018 (2006).
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M. G. Muller, T. A. Valdez, I. Georgakoudi, V. Backman, C. Fuentes, S. Kabani, N. Laver, Z. Wang, C. W. Boone, R. R. Dasari, S. M. Shapshay, and M. S. Feld, “Spectroscopic detection and evaluation of morphologic and biochemical changes in early human oral carcinoma,” Cancer 97, 1681–1692 (2003).
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R. J. Nordstrom, L. Burke, J. M. Niloff, and J. F. Myrtle, “Identification of cervical intraepithelial neoplasia (CIN) using UV-excited fluorescence and diffuse-reflectance tissue spectroscopy,” Lasers Surg. Med. 29, 118–127 (2001).
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R. J. Nordstrom, L. Burke, J. M. Niloff, and J. F. Myrtle, “Identification of cervical intraepithelial neoplasia (CIN) using UV-excited fluorescence and diffuse-reflectance tissue spectroscopy,” Lasers Surg. Med. 29, 118–127 (2001).
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F. Steinberg, H. J. Rohrborn, T. Otto, K. M. Scheufler, and C. Streffer, “NIR reflection measurements of hemoglobin and cytochrome aa3 in healthy tissue and tumors. Correlations to oxygen consumption: preclinical and clinical data,” Adv. Exp. Med. Biol. 428, 69–77 (1997).
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G. M. Palmer, C. Zhu, T. M. Breslin, F. Xu, K. W. Gilchrist, and N. Ramanujam, “Monte Carlo-based inverse model for calculating tissue optical properties. Part II: Application to breast cancer diagnosis,” Appl. Opt. 45, 1072–1078 (2006).
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C. Zhu, G. M. Palmer, T. M. Breslin, J. Harter, and N. Ramanujam, “Diagnosis of breast cancer using diffuse reflectance spectroscopy: Comparison of a Monte Carlo versus partial least squares analysis based feature extraction technique,” Lasers Surg. Med. 38, 714–724 (2006).
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G. M. Palmer and N. Ramanujam, “Monte Carlo-based inverse model for calculating tissue optical properties. Part I: Theory and validation on synthetic phantoms,” Appl. Opt. 45, 1062–1071 (2006).
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C. Zhu, G. M. Palmer, T. M. Breslin, F. Xu, and N. Ramanujam, “Use of a multiseparation fiber optic probe for the optical diagnosis of breast cancer,” J. Biomed. Opt. 10, 024032 (2005).
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M. C. Skala, G. M. Palmer, C. Zhu, Q. Liu, K. M. Vrotsos, C. L. Marshek-Stone, A. Gendron-Fitzpatrick, and N. Ramanujam, “Investigation of fiber-optic probe designs for optical spectroscopic diagnosis of epithelial pre-cancers,” Lasers Surg. Med. 34, 25–38 (2004).
[Crossref] [PubMed]

G. M. Palmer, C. Zhu, T. M. Breslin, F. Xu, K. W. Gilchrist, and N. Ramanujam, “Comparison of multiexcitation fluorescence and diffuse reflectance spectroscopy for the diagnosis of breast cancer (March 2003),” IEEE Trans. Biomed. Eng. 50, 1233–1242 (2003).
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C. Zhu, G. M. Palmer, T. M. Breslin, J. Harter, and N. Ramanujam, “Diagnosis of breast cancer using diffuse reflectance spectroscopy: Comparison of a Monte Carlo versus partial least squares analysis based feature extraction technique,” Lasers Surg. Med. 38, 714–724 (2006).
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G. M. Palmer and N. Ramanujam, “Monte Carlo-based inverse model for calculating tissue optical properties. Part I: Theory and validation on synthetic phantoms,” Appl. Opt. 45, 1062–1071 (2006).
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[Crossref] [PubMed]

M. C. Skala, J. M. Squirrell, K. M. Vrotsos, J. C. Eickhoff, A. Gendron-Fitzpatrick, K. W. Eliceiri, and N. Ramanujam, “Multiphoton microscopy of endogenous fluorescence differentiates normal, precancerous, and cancerous squamous epithelial tissues,” Cancer Res. 65, 1180–1186 (2005).
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M. C. Skala, G. M. Palmer, C. Zhu, Q. Liu, K. M. Vrotsos, C. L. Marshek-Stone, A. Gendron-Fitzpatrick, and N. Ramanujam, “Investigation of fiber-optic probe designs for optical spectroscopic diagnosis of epithelial pre-cancers,” Lasers Surg. Med. 34, 25–38 (2004).
[Crossref] [PubMed]

G. M. Palmer, C. Zhu, T. M. Breslin, F. Xu, K. W. Gilchrist, and N. Ramanujam, “Comparison of multiexcitation fluorescence and diffuse reflectance spectroscopy for the diagnosis of breast cancer (March 2003),” IEEE Trans. Biomed. Eng. 50, 1233–1242 (2003).
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R. Drezek, K. Sokolov, U. Utzinger, I. Boiko, A. Malpica, M. Follen, and R. Richards-Kortum, “Understanding the contributions of NADH and collagen to cervical tissue fluorescence spectra: modeling, measurements, and implications,” J. Biomed. Opt. 6, 385–396 (2001).
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M. C. Skala, K. M. Riching, D. K. Bird, A. Gendron-Fitzpatrick, J. Eickhoff, K. W. Eliceiri, P. J. Keely, and N. Ramanujam, “In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia,” J. Biomed. Opt. 12, 024014 (2007).
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S. R. Millon, K. M. Roldan-Perez, K. M. Riching, G. M. Palmer, and N. Ramanujam, “Effect of optical clearing agents on the in vivo optical properties of squamous epithelial tissue,” Lasers Surg. Med. 38, 920–927 (2006).
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S. R. Millon, K. M. Roldan-Perez, K. M. Riching, G. M. Palmer, and N. Ramanujam, “Effect of optical clearing agents on the in vivo optical properties of squamous epithelial tissue,” Lasers Surg. Med. 38, 920–927 (2006).
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F. Steinberg, H. J. Rohrborn, T. Otto, K. M. Scheufler, and C. Streffer, “NIR reflection measurements of hemoglobin and cytochrome aa3 in healthy tissue and tumors. Correlations to oxygen consumption: preclinical and clinical data,” Adv. Exp. Med. Biol. 428, 69–77 (1997).
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M. G. Muller, T. A. Valdez, I. Georgakoudi, V. Backman, C. Fuentes, S. Kabani, N. Laver, Z. Wang, C. W. Boone, R. R. Dasari, S. M. Shapshay, and M. S. Feld, “Spectroscopic detection and evaluation of morphologic and biochemical changes in early human oral carcinoma,” Cancer 97, 1681–1692 (2003).
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I. Georgakoudi, E. E. Sheets, M. G. Muller, V. Backman, C. P. Crum, K. Badizadegan, R. R. Dasari, and M. S. Feld, “Trimodal spectroscopy for the detection and characterization of cervical precancers in vivo,“ Am. J. Obstet. Gynecol. 186, 374–382 (2002).
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Skala, M. C.

M. C. Skala, K. M. Riching, D. K. Bird, A. Gendron-Fitzpatrick, J. Eickhoff, K. W. Eliceiri, P. J. Keely, and N. Ramanujam, “In vivo multiphoton fluorescence lifetime imaging of protein-bound and free nicotinamide adenine dinucleotide in normal and precancerous epithelia,” J. Biomed. Opt. 12, 024014 (2007).
[Crossref] [PubMed]

M. C. Skala, J. M. Squirrell, K. M. Vrotsos, J. C. Eickhoff, A. Gendron-Fitzpatrick, K. W. Eliceiri, and N. Ramanujam, “Multiphoton microscopy of endogenous fluorescence differentiates normal, precancerous, and cancerous squamous epithelial tissues,” Cancer Res. 65, 1180–1186 (2005).
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M. C. Skala, G. M. Palmer, C. Zhu, Q. Liu, K. M. Vrotsos, C. L. Marshek-Stone, A. Gendron-Fitzpatrick, and N. Ramanujam, “Investigation of fiber-optic probe designs for optical spectroscopic diagnosis of epithelial pre-cancers,” Lasers Surg. Med. 34, 25–38 (2004).
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F. H. White, K. Gohari, and C. J. Smith, “Histological and ultrastructural morphology of 7,12 dimethylbenz(alpha)-anthracene carcinogenesis in hamster cheek pouch epithelium,” Diagn. Histopathol. 4, 307–333 (1981).
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I. Pavlova, K. Sokolov, R. Drezek, A. Malpica, M. Follen, and R. Richards-Kortum, “Microanatomical and biochemical origins of normal and precancerous cervical autofluorescence using laser-scanning fluorescence confocal microscopy,” Photochem. Photobiol. 77, 550–555 (2003).
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R. Drezek, K. Sokolov, U. Utzinger, I. Boiko, A. Malpica, M. Follen, and R. Richards-Kortum, “Understanding the contributions of NADH and collagen to cervical tissue fluorescence spectra: modeling, measurements, and implications,” J. Biomed. Opt. 6, 385–396 (2001).
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P. Thueler, I. Charvet, F. Bevilacqua, M. St Ghislain, G. Ory, P. Marquet, P. Meda, B. Vermeulen, and C. Depeursinge, “In vivo endoscopic tissue diagnostics based on spectroscopic absorption, scattering, and phase function properties,” J. Biomed. Opt. 8, 495–503 (2003).
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F. Steinberg, H. J. Rohrborn, T. Otto, K. M. Scheufler, and C. Streffer, “NIR reflection measurements of hemoglobin and cytochrome aa3 in healthy tissue and tumors. Correlations to oxygen consumption: preclinical and clinical data,” Adv. Exp. Med. Biol. 428, 69–77 (1997).
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D. C. de Veld, M. Skurichina, M. J. Witjes, R. P. Duin, H. J. Sterenborg, and J. L. Roodenburg, “Autofluorescence and diffuse reflectance spectroscopy for oral oncology,” Lasers Surg. Med. 36, 356–364 (2005).
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N. Subhash, J. R. Mallia, S. S. Thomas, A. Mathews, P. Sebastian, and J. Madhavan, “Oral cancer detection using diffuse reflectance spectral ratio R540/R575 of oxygenated hemoglobin bands,” J. Biomed. Opt. 11, 014018 (2006).
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U. Sunar, H. Quon, T. Durduran, J. Zhang, J. Du, C. Zhou, G. Yu, R. Choe, A. Kilger, R. Lustig, L. Loevner, S. Nioka, B. Chance, and A. G. Yodh, “Noninvasive diffuse optical measurement of blood flow and blood oxygenation for monitoring radiation therapy in patients with head and neck tumors: a pilot study,” J. Biomed. Opt. 11, 064021 (2006).
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R. Hornung, T. H. Pham, K. A. Keefe, M. W. Berns, Y. Tadir, and B. J. Tromberg, “Quantitative near-infrared spectroscopy of cervical dysplasia in vivo,” Hum. Reprod. 14, 2908–2916 (1999).
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Y. S. Fawzy, M. Petek, M. Tercelj, and H. Zeng, “In vivo assessment and evaluation of lung tissue morphologic and physiological changes from non-contact endoscopic reflectance spectroscopy for improving lung cancer detection,” J. Biomed. Opt. 11, 044003 (2006).
[Crossref] [PubMed]

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N. Subhash, J. R. Mallia, S. S. Thomas, A. Mathews, P. Sebastian, and J. Madhavan, “Oral cancer detection using diffuse reflectance spectral ratio R540/R575 of oxygenated hemoglobin bands,” J. Biomed. Opt. 11, 014018 (2006).
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P. Thueler, I. Charvet, F. Bevilacqua, M. St Ghislain, G. Ory, P. Marquet, P. Meda, B. Vermeulen, and C. Depeursinge, “In vivo endoscopic tissue diagnostics based on spectroscopic absorption, scattering, and phase function properties,” J. Biomed. Opt. 8, 495–503 (2003).
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R. Hornung, T. H. Pham, K. A. Keefe, M. W. Berns, Y. Tadir, and B. J. Tromberg, “Quantitative near-infrared spectroscopy of cervical dysplasia in vivo,” Hum. Reprod. 14, 2908–2916 (1999).
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M. G. Muller, T. A. Valdez, I. Georgakoudi, V. Backman, C. Fuentes, S. Kabani, N. Laver, Z. Wang, C. W. Boone, R. R. Dasari, S. M. Shapshay, and M. S. Feld, “Spectroscopic detection and evaluation of morphologic and biochemical changes in early human oral carcinoma,” Cancer 97, 1681–1692 (2003).
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van Den Bergh, H.

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P. Thueler, I. Charvet, F. Bevilacqua, M. St Ghislain, G. Ory, P. Marquet, P. Meda, B. Vermeulen, and C. Depeursinge, “In vivo endoscopic tissue diagnostics based on spectroscopic absorption, scattering, and phase function properties,” J. Biomed. Opt. 8, 495–503 (2003).
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M. C. Skala, J. M. Squirrell, K. M. Vrotsos, J. C. Eickhoff, A. Gendron-Fitzpatrick, K. W. Eliceiri, and N. Ramanujam, “Multiphoton microscopy of endogenous fluorescence differentiates normal, precancerous, and cancerous squamous epithelial tissues,” Cancer Res. 65, 1180–1186 (2005).
[Crossref] [PubMed]

M. C. Skala, G. M. Palmer, C. Zhu, Q. Liu, K. M. Vrotsos, C. L. Marshek-Stone, A. Gendron-Fitzpatrick, and N. Ramanujam, “Investigation of fiber-optic probe designs for optical spectroscopic diagnosis of epithelial pre-cancers,” Lasers Surg. Med. 34, 25–38 (2004).
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C. T. Chen, H. K. Chiang, S. N. Chow, C. Y. Wang, Y. S. Lee, J. C. Tsai, and C. P. Chiang, “Autofluorescence in normal and malignant human oral tissues and in DMBA-induced hamster buccal pouch carcinogenesis,” J. Oral Pathol. Med. 27, 470–474 (1998).
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M. G. Muller, T. A. Valdez, I. Georgakoudi, V. Backman, C. Fuentes, S. Kabani, N. Laver, Z. Wang, C. W. Boone, R. R. Dasari, S. M. Shapshay, and M. S. Feld, “Spectroscopic detection and evaluation of morphologic and biochemical changes in early human oral carcinoma,” Cancer 97, 1681–1692 (2003).
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F. H. White, K. Gohari, and C. J. Smith, “Histological and ultrastructural morphology of 7,12 dimethylbenz(alpha)-anthracene carcinogenesis in hamster cheek pouch epithelium,” Diagn. Histopathol. 4, 307–333 (1981).
[PubMed]

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T. J. Pfefer, L. S. Matchette, C. L. Bennett, J. A. Gall, J. N. Wilke, A. J. Durkin, and M. N. Ediger, “Reflectance-based determination of optical properties in highly attenuating tissue,” J. Biomed. Opt. 8, 206–215 (2003).
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Figures (4)

Fig. 1.
Fig. 1. Measured diffuse reflectance spectra of one normal and neoplastic (squamous cell carcinoma) tissue site within the same animal, and the corresponding fits to the physical model (c.u. = calibrated units). The measured and fitted data were multiplied by the reference phantom after the fitting procedure, so that the original diffuse reflectance spectra and its corresponding fit are shown in this figure. The residuals for the normal and neoplastic sample are also shown.

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Fig. 3.
Fig. 3. Scatter plot of the two of the most diagnostic tissue parameters obtained from the physical model based analysis (mean reduced scattering coefficient and absorption coefficient at 460 nm). The samples include normal (•), dysplasia and CIS (+), and SCC (*). The decision line (—) was obtained from the SVM classifier. (SCC=squamous cell carcinoma, CIS = carcinoma in situ).

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Fig. 4.
Fig. 4. Average phantom normalized spectra for normal (n=10, —) and neoplastic (dysplasia/CIS/SCC, n=10, ╍) tissues, and spectral re-projections of PC1 + PC2 for normal (•) and neoplastic (*) tissues. The difference between the re-projected spectra and the phantom-normalized spectra are shown as residuals for normal (Δ) and neoplastic (∇) tissues.

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Fig. 5.
Fig. 5. Scatter plot of two diagnostic parameters (PC1 and PC2) extracted from the diffuse reflectance spectra using the empirical model (PCA) based analysis for phantom-normalized spectra. The samples include normal (•), dysplasia and CIS (+), and SCC (*). The decision line (—) was obtained from the SVM classifier.

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Tables (4)

Tables Icon

Table 1. Mean and standard deviation of the reduced scattering coefficient (μs’) and absorption coefficient (μa) calculated across all normal (n=10) and neoplastic (dysplasia/CIS/SCC, n=10) tissues. Significant differences between normal vs. neoplastic tissues were found for the variables marked with an asterisk (*), based on unpaired Wilcoxon tests.

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Table 2. Results from the SVM classifier for the physical model based analysis. The two variables shown in Fig. 3 were used to classify normal (n=10) and neoplastic (dysplasia/CIS/SCC, n=10) samples. The mean and standard deviation of the overall classification rate, sensitivity and specificity are shown for the training sets, and the “leave one out” cross validation result is also shown.

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Table 3. Principal components obtained from the empirical (PCA) analysis, the percent variance accounted for by each PC, and the p-value from unpaired Wilcoxon tests of normal (n=10) vs. neoplastic (dysplasia/CIS/SCC, n=10) tissues. These PCs were derived from phantom-normalized diffuse reflectance spectra.

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Table 4: Results from the SVM classifier for the empirical model based analysis. PC1 and PC2 from phantom-normalized spectra were used to classify normal (n=10) and neoplastic (dysplasia/CIS/SCC n=10) samples. The mean and standard deviation of the overall classification rate, sensitivity and specificity are shown for the training sets, and the “leave one out” cross validation result is also shown.

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