Abstract

The first in vivo optical virtual biopsy based on epi-third-harmonic-generation (THG) microscopy is successfully demonstrated using Syrian hamster oral mucosa as a model system. Without complex physical biopsy procedures, epi-THG microscopy can provide high spatial resolution dynamic images of oral mucosa and sub-mucosa in all three dimensions. The demonstrated intra-vital epi-THG microscopy provide high resolution observation of blood flow in the capillary and could be a promising tool to image angiogenesis, which is an important feature for many human diseases including malignancies. The system setup of epi-THG microscopy can be easily integrated with other nonlinear optical microscopy such as second-harmonic generation and multi-photon fluorescence microscopy by using the same laser system to provide better integrated molecular and structural information for future clinical diagnosis. By adding 6% acetic acid solution on the mucosa, THG contrast on the borders of nuclei was found to be greatly enhanced due to the alterations of their linear and nonlinear THG susceptibilities. With a virtual-transition-based technology without using fluorescence, the optical epi-THG biopsy we demonstrated shows promise for future noninvasive in vivo diseases examinations.

© 2006 Optical Society of America

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2006 (2)

D. Debarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, "Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy," Nat. Methods 3, 47-53 (2006).
[CrossRef]

T.-H Tsai, S.-P. Tai, W.-J. Lee, H.-Y. Huang, Y.-H. Liao and C.-K. Sun, "Optical signal degradation study in fixed human skin using confocal microscopy and higher-harmonic optical microscopy," Opt. Express 14, 749-758 (2006).
[CrossRef] [PubMed]

2005 (6)

S.-P. Tai, T.-H. Tsai, W.-J. Lee, D.-B. Shieh, Y.-H. Liao, H.-Y. Huang, K. Y.-J. Zhang, H.-L. Liu, and C.-K. Sun, "Optical biopsy of fixed human skin with backward-collected optical harmonics signals," Opt. Express 13, 8231-8242 (2005).
[CrossRef] [PubMed]

V. Barzda, C. Greenhalgh, J. Aus der Au, S. Elmore, J. H. van Beek, and J. Squier, "Visualization of mitochondria in cardiomyocytes by simultaneous harmonic generation andfluorescence microscopy," Opt. Express 13, 8263-8276 (2005).
[CrossRef] [PubMed]

J. F. Rey, H. Inoue, and M. Guelrud, "Magnification endoscopy with acetic acid for Barrett’s esophagus," Endoscopy 37, 583-586, (2005).
[CrossRef] [PubMed]

A. Volkmer, "Vibrational imaging and microspectroscopies based on coherent anti-Stoke Raman scattering microscopy," J. Phys. D: Appl. Phys. 38, R59-R81 (2005).
[CrossRef]

A. N. Yaroslavsky, J. Barbosa, V. Neel, C. DiMarzio, and R. R. Anderson, "Combining multispectral polarized light imaging and confocal microscopy for localization of nonmelanoma skin cancer," J. of Biomed. Opt. 10, 014011 (2005).
[CrossRef]

K. Carlson, I. Pavlova, M. Descour. M. Follen, and R. R.-Kortum, "Confocal microscopy: Imaging cervical precancerous lesions, " Gynecol. Oncol. 99, S84-S88 (2005).
[CrossRef] [PubMed]

2004 (2)

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, "Higher harmonic generation microscopy for developmental biology," J. Struct. Biol. 147, 19-30 (2004).
[CrossRef] [PubMed]

A. B. MacLean, "Acetowhite epithelium," Gynecol. Oncol. 95, 691-694 (2004).
[CrossRef] [PubMed]

2003 (2)

R. Lambert, J. F. Rey, and R. Sankaranarayanan, "Magnification and chromoscopy with the acetic cid test," Endoscopy 35, 437-445, (2003).
[CrossRef] [PubMed]

C.-K. Sun, C.-C. Chen, S.-W. Chu, T.-H. Tsai, Y.-C. Chen and B.-L. Lin, "Multi-harmonic-generation biopsy of skin," Opt. Lett. 28, 2488-2490 (2003).
[CrossRef] [PubMed]

2002 (8)

J. X. Cheng, A. Volkmer, and X. S. Xie, "Theoretical and experimental characterization of coherent anti-Stokes Raman scattering microscopy," J. Opt. Soc. Am. B 19, 1363-1375 (2002).
[CrossRef]

J. X. Cheng, and X. S. Xie, "Green's function formulation for third-harmonic generation microscopy," J. Opt. Soc. Am. B 19, 1604-1610 (2002).
[CrossRef]

M. Rajadhyaksha, S. Gonzalez, J. M. Zavislan, "Detectability of contrast agents for confocal reflectance imaging of skin and microcirculation," J. of Biomed. Opt. 9, 323-331 (2002).
[CrossRef]

I-H. Chen, S.-W. Chu, C.-K. Sun, P.-C. Cheng, and B.-L. Lin, "Wavelength dependent cell damages in biological multi-photon confocal microscopy: A micro-spectroscopic comparison between femtosecond Ti:sapphire and Cr:forsterite laser sources," Opt. Quantum. Electron. 34, 1251-1266 (2002).
[CrossRef]

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

A. A. Lacy, T. Collier, J. E. Price, S. Dharmawardhane and R. R. Richards-Kortum, "Near real-time in vivo confocal imaging of mouse mammary tumors," Front. Biosci. 7, F1-F7 (2002).
[CrossRef] [PubMed]

A. F. Zuluaga, R. Drzek, T. Collier, R. Lotan, and M. Follen, "Contrast agents for confocal microscopy: how simple chemicals affect confocal images of normal and cancer cells in suspension," J. of Biomed. Opt. 7, 398-403 (2002).
[CrossRef]

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, "Nonlinear bio-photonic crystal effects revealed with multi-modal nonlinear microscopy," J. Microsc. 208, Part 3, 190-200 (2002).
[CrossRef] [PubMed]

2001 (6)

M. Rajadhyaksha, G. Menaker, T. Flotte, P. J. Dwyer, and S. Gonzalez, "Confocal examinazation of nonmelanoma cancers in thick skin excisions to potentially guide Mohs micrographic surgery without frozen histopathology," J. Invest. Dermatol. 117,1137-1143 (2001).
[CrossRef] [PubMed]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, "An epi-detected coherent anti-stokes Raman scattering (E-CARS) microscope with high spectral resolution and high sensitivity," J. Phys. Chem. B 105, 1277-1280 (2001).
[CrossRef]

A. Volkmer, J. X. Cheng, and X. S. Xie, "Vibrational imaging with high sensitivity via epidetected coherent anti-Stokes Raman scattering microscopy," Phys. Rev. Lett. 87, 023901 (2001).
[CrossRef]

T.-M. Liu, S.-W Chiu, C.-K Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, "Multi-photon scanning microscopy using a femtosecond Cr:forsterite laser," Scanning 23, 249-254 (2001).
[CrossRef] [PubMed]

B. W. Pogue, H. B. Kaufman, A. Zelnechuk, W. Harper, G. C. Burke, E. E. Burke, and D. M. Harper, "Analysis of acetic acid-induced whitening of high-grade squamous intraepithelial lesions," J. Biomed. Opt. 6, 397-403 (2001).
[CrossRef]

L. Canioni, S. Rivet, L. Sarger, R. Barille, P. Vacher, and P. Voisin, "Imaging of Ca2+ intracellular dynamics with a third-harmonic generation microscope," Opt. Lett. 26, 515-517 (2001).).
[CrossRef]

2000 (2)

T. Collier, P. Shen, B. de Pradier, K. -B. Sung, R. Richards-Kortum, M. Follen, and A. Malpica, "Near real time confocal microscopy of amelanotic tissue: Dynamics of aceto-whitening enable nuclear segmentation," Opt. Express 6, 40-48 (2000).
[CrossRef] [PubMed]

R. A. Drezek, T. Collier, C. K. Brookner, A. Malpica, R. Lotan, R. R. Richards-Kortum, and M. Follen, "Laser scanning confocal microscopy of cervical tissue before and after application of acetic acid," Am. J. Obstet. Gynecol. 182, 1135-1139 (2000).
[CrossRef] [PubMed]

1999 (3)

G. Peleg, A. Lewis, M. Linial, and L. M. Loew "Nonlinear optical measurement of membrane potential around single molecules at selected celluar sites," Proc. Natl. Acad. Sci. 96, 6700-6704 (1999).
[CrossRef] [PubMed]

D. Yelin and Y. Silberberg, "Laser scanning third-harmonic-generation microscopy in biology," Opt. Express 5, 169-175 (1999.
[CrossRef] [PubMed]

K. H. Kim, C. Buehler, and P. T. C. So, "High speed, two-photon scanning microscope," Appl. Opt. 38, 6004-6009 (1999).
[CrossRef]

1998 (2)

J. A. Squier, M. Muller, G. J. Brakenhoff, and K. R. Wilson, "Third harmonic generation microscopy," Opt. Express 3, 315-324 (1998).
[CrossRef] [PubMed]

C.-C. ChangChien, H. Lin, S. W. Leung, C.-Y. Hsu, and C.-L. Cho, "Effect of acetic acid on telomerse acitivity in cervical intraepithelial neoplasia," Gynecol. Oncol. 71, 99-103 (1998).
[CrossRef] [PubMed]

Anderson, R. R.

A. N. Yaroslavsky, J. Barbosa, V. Neel, C. DiMarzio, and R. R. Anderson, "Combining multispectral polarized light imaging and confocal microscopy for localization of nonmelanoma skin cancer," J. of Biomed. Opt. 10, 014011 (2005).
[CrossRef]

Barbosa, J.

A. N. Yaroslavsky, J. Barbosa, V. Neel, C. DiMarzio, and R. R. Anderson, "Combining multispectral polarized light imaging and confocal microscopy for localization of nonmelanoma skin cancer," J. of Biomed. Opt. 10, 014011 (2005).
[CrossRef]

Barille, R.

Barzda, V.

Beaurepaire, E.

D. Debarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, "Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy," Nat. Methods 3, 47-53 (2006).
[CrossRef]

Book, L. D.

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, "An epi-detected coherent anti-stokes Raman scattering (E-CARS) microscope with high spectral resolution and high sensitivity," J. Phys. Chem. B 105, 1277-1280 (2001).
[CrossRef]

Brakenhoff, G. J.

Brookner, C. K.

R. A. Drezek, T. Collier, C. K. Brookner, A. Malpica, R. Lotan, R. R. Richards-Kortum, and M. Follen, "Laser scanning confocal microscopy of cervical tissue before and after application of acetic acid," Am. J. Obstet. Gynecol. 182, 1135-1139 (2000).
[CrossRef] [PubMed]

Buehler, C.

Burke, E. E.

B. W. Pogue, H. B. Kaufman, A. Zelnechuk, W. Harper, G. C. Burke, E. E. Burke, and D. M. Harper, "Analysis of acetic acid-induced whitening of high-grade squamous intraepithelial lesions," J. Biomed. Opt. 6, 397-403 (2001).
[CrossRef]

Burke, G. C.

B. W. Pogue, H. B. Kaufman, A. Zelnechuk, W. Harper, G. C. Burke, E. E. Burke, and D. M. Harper, "Analysis of acetic acid-induced whitening of high-grade squamous intraepithelial lesions," J. Biomed. Opt. 6, 397-403 (2001).
[CrossRef]

Campagnola, P. J.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

Canioni, L.

Carlson, K.

K. Carlson, I. Pavlova, M. Descour. M. Follen, and R. R.-Kortum, "Confocal microscopy: Imaging cervical precancerous lesions, " Gynecol. Oncol. 99, S84-S88 (2005).
[CrossRef] [PubMed]

Chen, C.-C.

Chen, I.-S.

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, "Nonlinear bio-photonic crystal effects revealed with multi-modal nonlinear microscopy," J. Microsc. 208, Part 3, 190-200 (2002).
[CrossRef] [PubMed]

Chen, I-H.

I-H. Chen, S.-W. Chu, C.-K. Sun, P.-C. Cheng, and B.-L. Lin, "Wavelength dependent cell damages in biological multi-photon confocal microscopy: A micro-spectroscopic comparison between femtosecond Ti:sapphire and Cr:forsterite laser sources," Opt. Quantum. Electron. 34, 1251-1266 (2002).
[CrossRef]

Chen, S.-Y.

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, "Higher harmonic generation microscopy for developmental biology," J. Struct. Biol. 147, 19-30 (2004).
[CrossRef] [PubMed]

Chen, Y.-C.

Cheng, J. X.

J. X. Cheng, and X. S. Xie, "Green's function formulation for third-harmonic generation microscopy," J. Opt. Soc. Am. B 19, 1604-1610 (2002).
[CrossRef]

J. X. Cheng, A. Volkmer, and X. S. Xie, "Theoretical and experimental characterization of coherent anti-Stokes Raman scattering microscopy," J. Opt. Soc. Am. B 19, 1363-1375 (2002).
[CrossRef]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, "An epi-detected coherent anti-stokes Raman scattering (E-CARS) microscope with high spectral resolution and high sensitivity," J. Phys. Chem. B 105, 1277-1280 (2001).
[CrossRef]

A. Volkmer, J. X. Cheng, and X. S. Xie, "Vibrational imaging with high sensitivity via epidetected coherent anti-Stokes Raman scattering microscopy," Phys. Rev. Lett. 87, 023901 (2001).
[CrossRef]

Cheng, P. C.

T.-M. Liu, S.-W Chiu, C.-K Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, "Multi-photon scanning microscopy using a femtosecond Cr:forsterite laser," Scanning 23, 249-254 (2001).
[CrossRef] [PubMed]

Cheng, P.-C.

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, "Nonlinear bio-photonic crystal effects revealed with multi-modal nonlinear microscopy," J. Microsc. 208, Part 3, 190-200 (2002).
[CrossRef] [PubMed]

I-H. Chen, S.-W. Chu, C.-K. Sun, P.-C. Cheng, and B.-L. Lin, "Wavelength dependent cell damages in biological multi-photon confocal microscopy: A micro-spectroscopic comparison between femtosecond Ti:sapphire and Cr:forsterite laser sources," Opt. Quantum. Electron. 34, 1251-1266 (2002).
[CrossRef]

Chiu, S.-W

T.-M. Liu, S.-W Chiu, C.-K Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, "Multi-photon scanning microscopy using a femtosecond Cr:forsterite laser," Scanning 23, 249-254 (2001).
[CrossRef] [PubMed]

Chu, S.-W.

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, "Higher harmonic generation microscopy for developmental biology," J. Struct. Biol. 147, 19-30 (2004).
[CrossRef] [PubMed]

C.-K. Sun, C.-C. Chen, S.-W. Chu, T.-H. Tsai, Y.-C. Chen and B.-L. Lin, "Multi-harmonic-generation biopsy of skin," Opt. Lett. 28, 2488-2490 (2003).
[CrossRef] [PubMed]

I-H. Chen, S.-W. Chu, C.-K. Sun, P.-C. Cheng, and B.-L. Lin, "Wavelength dependent cell damages in biological multi-photon confocal microscopy: A micro-spectroscopic comparison between femtosecond Ti:sapphire and Cr:forsterite laser sources," Opt. Quantum. Electron. 34, 1251-1266 (2002).
[CrossRef]

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, "Nonlinear bio-photonic crystal effects revealed with multi-modal nonlinear microscopy," J. Microsc. 208, Part 3, 190-200 (2002).
[CrossRef] [PubMed]

Collier, T.

A. A. Lacy, T. Collier, J. E. Price, S. Dharmawardhane and R. R. Richards-Kortum, "Near real-time in vivo confocal imaging of mouse mammary tumors," Front. Biosci. 7, F1-F7 (2002).
[CrossRef] [PubMed]

A. F. Zuluaga, R. Drzek, T. Collier, R. Lotan, and M. Follen, "Contrast agents for confocal microscopy: how simple chemicals affect confocal images of normal and cancer cells in suspension," J. of Biomed. Opt. 7, 398-403 (2002).
[CrossRef]

R. A. Drezek, T. Collier, C. K. Brookner, A. Malpica, R. Lotan, R. R. Richards-Kortum, and M. Follen, "Laser scanning confocal microscopy of cervical tissue before and after application of acetic acid," Am. J. Obstet. Gynecol. 182, 1135-1139 (2000).
[CrossRef] [PubMed]

T. Collier, P. Shen, B. de Pradier, K. -B. Sung, R. Richards-Kortum, M. Follen, and A. Malpica, "Near real time confocal microscopy of amelanotic tissue: Dynamics of aceto-whitening enable nuclear segmentation," Opt. Express 6, 40-48 (2000).
[CrossRef] [PubMed]

Combettes, L.

D. Debarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, "Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy," Nat. Methods 3, 47-53 (2006).
[CrossRef]

de Pradier, B.

Debarre, D.

D. Debarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, "Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy," Nat. Methods 3, 47-53 (2006).
[CrossRef]

Descour, M.

K. Carlson, I. Pavlova, M. Descour. M. Follen, and R. R.-Kortum, "Confocal microscopy: Imaging cervical precancerous lesions, " Gynecol. Oncol. 99, S84-S88 (2005).
[CrossRef] [PubMed]

Dharmawardhane, S.

A. A. Lacy, T. Collier, J. E. Price, S. Dharmawardhane and R. R. Richards-Kortum, "Near real-time in vivo confocal imaging of mouse mammary tumors," Front. Biosci. 7, F1-F7 (2002).
[CrossRef] [PubMed]

DiMarzio, C.

A. N. Yaroslavsky, J. Barbosa, V. Neel, C. DiMarzio, and R. R. Anderson, "Combining multispectral polarized light imaging and confocal microscopy for localization of nonmelanoma skin cancer," J. of Biomed. Opt. 10, 014011 (2005).
[CrossRef]

Drezek, R. A.

R. A. Drezek, T. Collier, C. K. Brookner, A. Malpica, R. Lotan, R. R. Richards-Kortum, and M. Follen, "Laser scanning confocal microscopy of cervical tissue before and after application of acetic acid," Am. J. Obstet. Gynecol. 182, 1135-1139 (2000).
[CrossRef] [PubMed]

Drzek, R.

A. F. Zuluaga, R. Drzek, T. Collier, R. Lotan, and M. Follen, "Contrast agents for confocal microscopy: how simple chemicals affect confocal images of normal and cancer cells in suspension," J. of Biomed. Opt. 7, 398-403 (2002).
[CrossRef]

Dwyer, P. J.

M. Rajadhyaksha, G. Menaker, T. Flotte, P. J. Dwyer, and S. Gonzalez, "Confocal examinazation of nonmelanoma cancers in thick skin excisions to potentially guide Mohs micrographic surgery without frozen histopathology," J. Invest. Dermatol. 117,1137-1143 (2001).
[CrossRef] [PubMed]

Fabre, A.

D. Debarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, "Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy," Nat. Methods 3, 47-53 (2006).
[CrossRef]

Flotte, T.

M. Rajadhyaksha, G. Menaker, T. Flotte, P. J. Dwyer, and S. Gonzalez, "Confocal examinazation of nonmelanoma cancers in thick skin excisions to potentially guide Mohs micrographic surgery without frozen histopathology," J. Invest. Dermatol. 117,1137-1143 (2001).
[CrossRef] [PubMed]

Follen, M.

A. F. Zuluaga, R. Drzek, T. Collier, R. Lotan, and M. Follen, "Contrast agents for confocal microscopy: how simple chemicals affect confocal images of normal and cancer cells in suspension," J. of Biomed. Opt. 7, 398-403 (2002).
[CrossRef]

R. A. Drezek, T. Collier, C. K. Brookner, A. Malpica, R. Lotan, R. R. Richards-Kortum, and M. Follen, "Laser scanning confocal microscopy of cervical tissue before and after application of acetic acid," Am. J. Obstet. Gynecol. 182, 1135-1139 (2000).
[CrossRef] [PubMed]

T. Collier, P. Shen, B. de Pradier, K. -B. Sung, R. Richards-Kortum, M. Follen, and A. Malpica, "Near real time confocal microscopy of amelanotic tissue: Dynamics of aceto-whitening enable nuclear segmentation," Opt. Express 6, 40-48 (2000).
[CrossRef] [PubMed]

Gonzalez, S.

M. Rajadhyaksha, S. Gonzalez, J. M. Zavislan, "Detectability of contrast agents for confocal reflectance imaging of skin and microcirculation," J. of Biomed. Opt. 9, 323-331 (2002).
[CrossRef]

M. Rajadhyaksha, G. Menaker, T. Flotte, P. J. Dwyer, and S. Gonzalez, "Confocal examinazation of nonmelanoma cancers in thick skin excisions to potentially guide Mohs micrographic surgery without frozen histopathology," J. Invest. Dermatol. 117,1137-1143 (2001).
[CrossRef] [PubMed]

Greenhalgh, C.

Guelrud, M.

J. F. Rey, H. Inoue, and M. Guelrud, "Magnification endoscopy with acetic acid for Barrett’s esophagus," Endoscopy 37, 583-586, (2005).
[CrossRef] [PubMed]

Harper, D. M.

B. W. Pogue, H. B. Kaufman, A. Zelnechuk, W. Harper, G. C. Burke, E. E. Burke, and D. M. Harper, "Analysis of acetic acid-induced whitening of high-grade squamous intraepithelial lesions," J. Biomed. Opt. 6, 397-403 (2001).
[CrossRef]

Harper, W.

B. W. Pogue, H. B. Kaufman, A. Zelnechuk, W. Harper, G. C. Burke, E. E. Burke, and D. M. Harper, "Analysis of acetic acid-induced whitening of high-grade squamous intraepithelial lesions," J. Biomed. Opt. 6, 397-403 (2001).
[CrossRef]

Hoppe, P. E.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

Huang, H.-Y.

Inoue, H.

J. F. Rey, H. Inoue, and M. Guelrud, "Magnification endoscopy with acetic acid for Barrett’s esophagus," Endoscopy 37, 583-586, (2005).
[CrossRef] [PubMed]

Johnson, I.

T.-M. Liu, S.-W Chiu, C.-K Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, "Multi-photon scanning microscopy using a femtosecond Cr:forsterite laser," Scanning 23, 249-254 (2001).
[CrossRef] [PubMed]

Kaufman, H. B.

B. W. Pogue, H. B. Kaufman, A. Zelnechuk, W. Harper, G. C. Burke, E. E. Burke, and D. M. Harper, "Analysis of acetic acid-induced whitening of high-grade squamous intraepithelial lesions," J. Biomed. Opt. 6, 397-403 (2001).
[CrossRef]

Kim, K. H.

Kuo, M.-X.

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, "Nonlinear bio-photonic crystal effects revealed with multi-modal nonlinear microscopy," J. Microsc. 208, Part 3, 190-200 (2002).
[CrossRef] [PubMed]

Lacy, A. A.

A. A. Lacy, T. Collier, J. E. Price, S. Dharmawardhane and R. R. Richards-Kortum, "Near real-time in vivo confocal imaging of mouse mammary tumors," Front. Biosci. 7, F1-F7 (2002).
[CrossRef] [PubMed]

Lambert, R.

R. Lambert, J. F. Rey, and R. Sankaranarayanan, "Magnification and chromoscopy with the acetic cid test," Endoscopy 35, 437-445, (2003).
[CrossRef] [PubMed]

Lee, S.-P.

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, "Nonlinear bio-photonic crystal effects revealed with multi-modal nonlinear microscopy," J. Microsc. 208, Part 3, 190-200 (2002).
[CrossRef] [PubMed]

Lee, W.-J.

Lewis, A.

G. Peleg, A. Lewis, M. Linial, and L. M. Loew "Nonlinear optical measurement of membrane potential around single molecules at selected celluar sites," Proc. Natl. Acad. Sci. 96, 6700-6704 (1999).
[CrossRef] [PubMed]

Liao, Y.-H.

Lin, B.-L.

C.-K. Sun, C.-C. Chen, S.-W. Chu, T.-H. Tsai, Y.-C. Chen and B.-L. Lin, "Multi-harmonic-generation biopsy of skin," Opt. Lett. 28, 2488-2490 (2003).
[CrossRef] [PubMed]

I-H. Chen, S.-W. Chu, C.-K. Sun, P.-C. Cheng, and B.-L. Lin, "Wavelength dependent cell damages in biological multi-photon confocal microscopy: A micro-spectroscopic comparison between femtosecond Ti:sapphire and Cr:forsterite laser sources," Opt. Quantum. Electron. 34, 1251-1266 (2002).
[CrossRef]

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, "Nonlinear bio-photonic crystal effects revealed with multi-modal nonlinear microscopy," J. Microsc. 208, Part 3, 190-200 (2002).
[CrossRef] [PubMed]

T.-M. Liu, S.-W Chiu, C.-K Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, "Multi-photon scanning microscopy using a femtosecond Cr:forsterite laser," Scanning 23, 249-254 (2001).
[CrossRef] [PubMed]

Lin, C.-Y.

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, "Higher harmonic generation microscopy for developmental biology," J. Struct. Biol. 147, 19-30 (2004).
[CrossRef] [PubMed]

Lin, D.-J.

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, "Nonlinear bio-photonic crystal effects revealed with multi-modal nonlinear microscopy," J. Microsc. 208, Part 3, 190-200 (2002).
[CrossRef] [PubMed]

Linial, M.

G. Peleg, A. Lewis, M. Linial, and L. M. Loew "Nonlinear optical measurement of membrane potential around single molecules at selected celluar sites," Proc. Natl. Acad. Sci. 96, 6700-6704 (1999).
[CrossRef] [PubMed]

Liu, H.-L.

S.-P. Tai, T.-H. Tsai, W.-J. Lee, D.-B. Shieh, Y.-H. Liao, H.-Y. Huang, K. Y.-J. Zhang, H.-L. Liu, and C.-K. Sun, "Optical biopsy of fixed human skin with backward-collected optical harmonics signals," Opt. Express 13, 8231-8242 (2005).
[CrossRef] [PubMed]

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, "Nonlinear bio-photonic crystal effects revealed with multi-modal nonlinear microscopy," J. Microsc. 208, Part 3, 190-200 (2002).
[CrossRef] [PubMed]

Liu, T.-M.

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, "Higher harmonic generation microscopy for developmental biology," J. Struct. Biol. 147, 19-30 (2004).
[CrossRef] [PubMed]

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, "Nonlinear bio-photonic crystal effects revealed with multi-modal nonlinear microscopy," J. Microsc. 208, Part 3, 190-200 (2002).
[CrossRef] [PubMed]

T.-M. Liu, S.-W Chiu, C.-K Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, "Multi-photon scanning microscopy using a femtosecond Cr:forsterite laser," Scanning 23, 249-254 (2001).
[CrossRef] [PubMed]

Loew, L. M.

G. Peleg, A. Lewis, M. Linial, and L. M. Loew "Nonlinear optical measurement of membrane potential around single molecules at selected celluar sites," Proc. Natl. Acad. Sci. 96, 6700-6704 (1999).
[CrossRef] [PubMed]

Lotan, R.

A. F. Zuluaga, R. Drzek, T. Collier, R. Lotan, and M. Follen, "Contrast agents for confocal microscopy: how simple chemicals affect confocal images of normal and cancer cells in suspension," J. of Biomed. Opt. 7, 398-403 (2002).
[CrossRef]

R. A. Drezek, T. Collier, C. K. Brookner, A. Malpica, R. Lotan, R. R. Richards-Kortum, and M. Follen, "Laser scanning confocal microscopy of cervical tissue before and after application of acetic acid," Am. J. Obstet. Gynecol. 182, 1135-1139 (2000).
[CrossRef] [PubMed]

MacLean, A. B.

A. B. MacLean, "Acetowhite epithelium," Gynecol. Oncol. 95, 691-694 (2004).
[CrossRef] [PubMed]

Malone, C. J.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

Malpica, A.

T. Collier, P. Shen, B. de Pradier, K. -B. Sung, R. Richards-Kortum, M. Follen, and A. Malpica, "Near real time confocal microscopy of amelanotic tissue: Dynamics of aceto-whitening enable nuclear segmentation," Opt. Express 6, 40-48 (2000).
[CrossRef] [PubMed]

R. A. Drezek, T. Collier, C. K. Brookner, A. Malpica, R. Lotan, R. R. Richards-Kortum, and M. Follen, "Laser scanning confocal microscopy of cervical tissue before and after application of acetic acid," Am. J. Obstet. Gynecol. 182, 1135-1139 (2000).
[CrossRef] [PubMed]

Menaker, G.

M. Rajadhyaksha, G. Menaker, T. Flotte, P. J. Dwyer, and S. Gonzalez, "Confocal examinazation of nonmelanoma cancers in thick skin excisions to potentially guide Mohs micrographic surgery without frozen histopathology," J. Invest. Dermatol. 117,1137-1143 (2001).
[CrossRef] [PubMed]

Millard, A. C.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

Mohler, W. A.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

Muller, M.

Neel, V.

A. N. Yaroslavsky, J. Barbosa, V. Neel, C. DiMarzio, and R. R. Anderson, "Combining multispectral polarized light imaging and confocal microscopy for localization of nonmelanoma skin cancer," J. of Biomed. Opt. 10, 014011 (2005).
[CrossRef]

Pavlova, I.

K. Carlson, I. Pavlova, M. Descour. M. Follen, and R. R.-Kortum, "Confocal microscopy: Imaging cervical precancerous lesions, " Gynecol. Oncol. 99, S84-S88 (2005).
[CrossRef] [PubMed]

Peleg, G.

G. Peleg, A. Lewis, M. Linial, and L. M. Loew "Nonlinear optical measurement of membrane potential around single molecules at selected celluar sites," Proc. Natl. Acad. Sci. 96, 6700-6704 (1999).
[CrossRef] [PubMed]

Pena, A.-M.

D. Debarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, "Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy," Nat. Methods 3, 47-53 (2006).
[CrossRef]

Pogue, B. W.

B. W. Pogue, H. B. Kaufman, A. Zelnechuk, W. Harper, G. C. Burke, E. E. Burke, and D. M. Harper, "Analysis of acetic acid-induced whitening of high-grade squamous intraepithelial lesions," J. Biomed. Opt. 6, 397-403 (2001).
[CrossRef]

Price, J. E.

A. A. Lacy, T. Collier, J. E. Price, S. Dharmawardhane and R. R. Richards-Kortum, "Near real-time in vivo confocal imaging of mouse mammary tumors," Front. Biosci. 7, F1-F7 (2002).
[CrossRef] [PubMed]

Rajadhyaksha, M.

M. Rajadhyaksha, S. Gonzalez, J. M. Zavislan, "Detectability of contrast agents for confocal reflectance imaging of skin and microcirculation," J. of Biomed. Opt. 9, 323-331 (2002).
[CrossRef]

M. Rajadhyaksha, G. Menaker, T. Flotte, P. J. Dwyer, and S. Gonzalez, "Confocal examinazation of nonmelanoma cancers in thick skin excisions to potentially guide Mohs micrographic surgery without frozen histopathology," J. Invest. Dermatol. 117,1137-1143 (2001).
[CrossRef] [PubMed]

Rey, J. F.

J. F. Rey, H. Inoue, and M. Guelrud, "Magnification endoscopy with acetic acid for Barrett’s esophagus," Endoscopy 37, 583-586, (2005).
[CrossRef] [PubMed]

R. Lambert, J. F. Rey, and R. Sankaranarayanan, "Magnification and chromoscopy with the acetic cid test," Endoscopy 35, 437-445, (2003).
[CrossRef] [PubMed]

Richards-Kortum, R.

Richards-Kortum, R. R.

A. A. Lacy, T. Collier, J. E. Price, S. Dharmawardhane and R. R. Richards-Kortum, "Near real-time in vivo confocal imaging of mouse mammary tumors," Front. Biosci. 7, F1-F7 (2002).
[CrossRef] [PubMed]

R. A. Drezek, T. Collier, C. K. Brookner, A. Malpica, R. Lotan, R. R. Richards-Kortum, and M. Follen, "Laser scanning confocal microscopy of cervical tissue before and after application of acetic acid," Am. J. Obstet. Gynecol. 182, 1135-1139 (2000).
[CrossRef] [PubMed]

Rivet, S.

Sankaranarayanan, R.

R. Lambert, J. F. Rey, and R. Sankaranarayanan, "Magnification and chromoscopy with the acetic cid test," Endoscopy 35, 437-445, (2003).
[CrossRef] [PubMed]

Sarger, L.

Schanne-Klein, M.-C.

D. Debarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, "Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy," Nat. Methods 3, 47-53 (2006).
[CrossRef]

Shen, P.

Shieh, D.-B.

Silberberg, Y.

So, P. T. C.

Squier, J. A.

Sun, C.-K

T.-M. Liu, S.-W Chiu, C.-K Sun, B.-L. Lin, P. C. Cheng, and I. Johnson, "Multi-photon scanning microscopy using a femtosecond Cr:forsterite laser," Scanning 23, 249-254 (2001).
[CrossRef] [PubMed]

Sun, C.-K.

T.-H Tsai, S.-P. Tai, W.-J. Lee, H.-Y. Huang, Y.-H. Liao and C.-K. Sun, "Optical signal degradation study in fixed human skin using confocal microscopy and higher-harmonic optical microscopy," Opt. Express 14, 749-758 (2006).
[CrossRef] [PubMed]

S.-P. Tai, T.-H. Tsai, W.-J. Lee, D.-B. Shieh, Y.-H. Liao, H.-Y. Huang, K. Y.-J. Zhang, H.-L. Liu, and C.-K. Sun, "Optical biopsy of fixed human skin with backward-collected optical harmonics signals," Opt. Express 13, 8231-8242 (2005).
[CrossRef] [PubMed]

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, "Higher harmonic generation microscopy for developmental biology," J. Struct. Biol. 147, 19-30 (2004).
[CrossRef] [PubMed]

C.-K. Sun, C.-C. Chen, S.-W. Chu, T.-H. Tsai, Y.-C. Chen and B.-L. Lin, "Multi-harmonic-generation biopsy of skin," Opt. Lett. 28, 2488-2490 (2003).
[CrossRef] [PubMed]

I-H. Chen, S.-W. Chu, C.-K. Sun, P.-C. Cheng, and B.-L. Lin, "Wavelength dependent cell damages in biological multi-photon confocal microscopy: A micro-spectroscopic comparison between femtosecond Ti:sapphire and Cr:forsterite laser sources," Opt. Quantum. Electron. 34, 1251-1266 (2002).
[CrossRef]

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, "Nonlinear bio-photonic crystal effects revealed with multi-modal nonlinear microscopy," J. Microsc. 208, Part 3, 190-200 (2002).
[CrossRef] [PubMed]

Sung, K. -B.

Supatto, W.

D. Debarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, "Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy," Nat. Methods 3, 47-53 (2006).
[CrossRef]

Tai, S.-P.

Terasaki, M.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
[CrossRef]

Tordjmann, T.

D. Debarre, W. Supatto, A.-M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M.-C. Schanne-Klein, and E. Beaurepaire, "Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy," Nat. Methods 3, 47-53 (2006).
[CrossRef]

Tsai, H.-J.

C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, "Higher harmonic generation microscopy for developmental biology," J. Struct. Biol. 147, 19-30 (2004).
[CrossRef] [PubMed]

Tsai, T.-H

Tsai, T.-H.

Vacher, P.

Voisin, P.

Volkmer, A.

A. Volkmer, "Vibrational imaging and microspectroscopies based on coherent anti-Stoke Raman scattering microscopy," J. Phys. D: Appl. Phys. 38, R59-R81 (2005).
[CrossRef]

J. X. Cheng, A. Volkmer, and X. S. Xie, "Theoretical and experimental characterization of coherent anti-Stokes Raman scattering microscopy," J. Opt. Soc. Am. B 19, 1363-1375 (2002).
[CrossRef]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, "An epi-detected coherent anti-stokes Raman scattering (E-CARS) microscope with high spectral resolution and high sensitivity," J. Phys. Chem. B 105, 1277-1280 (2001).
[CrossRef]

A. Volkmer, J. X. Cheng, and X. S. Xie, "Vibrational imaging with high sensitivity via epidetected coherent anti-Stokes Raman scattering microscopy," Phys. Rev. Lett. 87, 023901 (2001).
[CrossRef]

Wilson, K. R.

Xie, X. S.

J. X. Cheng, A. Volkmer, and X. S. Xie, "Theoretical and experimental characterization of coherent anti-Stokes Raman scattering microscopy," J. Opt. Soc. Am. B 19, 1363-1375 (2002).
[CrossRef]

J. X. Cheng, and X. S. Xie, "Green's function formulation for third-harmonic generation microscopy," J. Opt. Soc. Am. B 19, 1604-1610 (2002).
[CrossRef]

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, "An epi-detected coherent anti-stokes Raman scattering (E-CARS) microscope with high spectral resolution and high sensitivity," J. Phys. Chem. B 105, 1277-1280 (2001).
[CrossRef]

A. Volkmer, J. X. Cheng, and X. S. Xie, "Vibrational imaging with high sensitivity via epidetected coherent anti-Stokes Raman scattering microscopy," Phys. Rev. Lett. 87, 023901 (2001).
[CrossRef]

Yaroslavsky, A. N.

A. N. Yaroslavsky, J. Barbosa, V. Neel, C. DiMarzio, and R. R. Anderson, "Combining multispectral polarized light imaging and confocal microscopy for localization of nonmelanoma skin cancer," J. of Biomed. Opt. 10, 014011 (2005).
[CrossRef]

Yelin, D.

Zavislan, J. M.

M. Rajadhyaksha, S. Gonzalez, J. M. Zavislan, "Detectability of contrast agents for confocal reflectance imaging of skin and microcirculation," J. of Biomed. Opt. 9, 323-331 (2002).
[CrossRef]

Zelnechuk, A.

B. W. Pogue, H. B. Kaufman, A. Zelnechuk, W. Harper, G. C. Burke, E. E. Burke, and D. M. Harper, "Analysis of acetic acid-induced whitening of high-grade squamous intraepithelial lesions," J. Biomed. Opt. 6, 397-403 (2001).
[CrossRef]

Zhang, K. Y.-J.

Zuluaga, A. F.

A. F. Zuluaga, R. Drzek, T. Collier, R. Lotan, and M. Follen, "Contrast agents for confocal microscopy: how simple chemicals affect confocal images of normal and cancer cells in suspension," J. of Biomed. Opt. 7, 398-403 (2002).
[CrossRef]

Am. J. Obstet. Gynecol. (1)

R. A. Drezek, T. Collier, C. K. Brookner, A. Malpica, R. Lotan, R. R. Richards-Kortum, and M. Follen, "Laser scanning confocal microscopy of cervical tissue before and after application of acetic acid," Am. J. Obstet. Gynecol. 182, 1135-1139 (2000).
[CrossRef] [PubMed]

Appl. Opt. (1)

Biophys. J. (1)

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, "Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 81, 493-508 (2002).
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Endoscopy (2)

R. Lambert, J. F. Rey, and R. Sankaranarayanan, "Magnification and chromoscopy with the acetic cid test," Endoscopy 35, 437-445, (2003).
[CrossRef] [PubMed]

J. F. Rey, H. Inoue, and M. Guelrud, "Magnification endoscopy with acetic acid for Barrett’s esophagus," Endoscopy 37, 583-586, (2005).
[CrossRef] [PubMed]

Front. Biosci. (1)

A. A. Lacy, T. Collier, J. E. Price, S. Dharmawardhane and R. R. Richards-Kortum, "Near real-time in vivo confocal imaging of mouse mammary tumors," Front. Biosci. 7, F1-F7 (2002).
[CrossRef] [PubMed]

Gynecol. Oncol. (3)

K. Carlson, I. Pavlova, M. Descour. M. Follen, and R. R.-Kortum, "Confocal microscopy: Imaging cervical precancerous lesions, " Gynecol. Oncol. 99, S84-S88 (2005).
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C.-C. ChangChien, H. Lin, S. W. Leung, C.-Y. Hsu, and C.-L. Cho, "Effect of acetic acid on telomerse acitivity in cervical intraepithelial neoplasia," Gynecol. Oncol. 71, 99-103 (1998).
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A. B. MacLean, "Acetowhite epithelium," Gynecol. Oncol. 95, 691-694 (2004).
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J. Biomed. Opt. (1)

B. W. Pogue, H. B. Kaufman, A. Zelnechuk, W. Harper, G. C. Burke, E. E. Burke, and D. M. Harper, "Analysis of acetic acid-induced whitening of high-grade squamous intraepithelial lesions," J. Biomed. Opt. 6, 397-403 (2001).
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J. Invest. Dermatol. (1)

M. Rajadhyaksha, G. Menaker, T. Flotte, P. J. Dwyer, and S. Gonzalez, "Confocal examinazation of nonmelanoma cancers in thick skin excisions to potentially guide Mohs micrographic surgery without frozen histopathology," J. Invest. Dermatol. 117,1137-1143 (2001).
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J. Microsc. (1)

S.-W. Chu, I.-S. Chen, T.-M. Liu, C.-K. Sun, S.-P. Lee, B.-L. Lin, P.-C. Cheng, M.-X. Kuo, D.-J. Lin, and H.-L. Liu, "Nonlinear bio-photonic crystal effects revealed with multi-modal nonlinear microscopy," J. Microsc. 208, Part 3, 190-200 (2002).
[CrossRef] [PubMed]

J. of Biomed. Opt. (3)

A. F. Zuluaga, R. Drzek, T. Collier, R. Lotan, and M. Follen, "Contrast agents for confocal microscopy: how simple chemicals affect confocal images of normal and cancer cells in suspension," J. of Biomed. Opt. 7, 398-403 (2002).
[CrossRef]

A. N. Yaroslavsky, J. Barbosa, V. Neel, C. DiMarzio, and R. R. Anderson, "Combining multispectral polarized light imaging and confocal microscopy for localization of nonmelanoma skin cancer," J. of Biomed. Opt. 10, 014011 (2005).
[CrossRef]

M. Rajadhyaksha, S. Gonzalez, J. M. Zavislan, "Detectability of contrast agents for confocal reflectance imaging of skin and microcirculation," J. of Biomed. Opt. 9, 323-331 (2002).
[CrossRef]

J. Opt. Soc. Am. B (2)

J. Phys. Chem. B (1)

J. X. Cheng, A. Volkmer, L. D. Book, and X. S. Xie, "An epi-detected coherent anti-stokes Raman scattering (E-CARS) microscope with high spectral resolution and high sensitivity," J. Phys. Chem. B 105, 1277-1280 (2001).
[CrossRef]

J. Phys. D: Appl. Phys. (1)

A. Volkmer, "Vibrational imaging and microspectroscopies based on coherent anti-Stoke Raman scattering microscopy," J. Phys. D: Appl. Phys. 38, R59-R81 (2005).
[CrossRef]

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C.-K. Sun, S.-W. Chu, S.-Y. Chen, T.-H. Tsai, T.-M. Liu, C.-Y. Lin, and H.-J. Tsai, "Higher harmonic generation microscopy for developmental biology," J. Struct. Biol. 147, 19-30 (2004).
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W.-J. Lee, C.-H. Yu, S-P. Tai, H.-Y. Huang, and C.-K. Sun, "Functional third-harmonic generation microscopy of cell nuclei by using acetic acid as a contrast agent," in preparation.

S.-Y. Chen, S.-P. Tai, T.-H. Tsai and C.-K. Sun, "Direct backward-emitted third-harmonic generation and its application to clinical microscopy", in Technical Digest of Conference on Laser and Electro-optics/Quantum Electronics and Laser Science Conference (CLEO/QELS 2005), Baltimore, MD, USA, paper QMI3 (2005).

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Supplementary Material (2)

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Figures (7)

Fig. 1.
Fig. 1.

System setup of harmonics optical biopsy.

Fig. 2.
Fig. 2.

Measured lateral resolution of in vivo epi-THG microscopy vs. depth in hamster oral cavity.

Fig. 3.
Fig. 3.

In vivo horizontally sectioned epi-SHG (green) and epi-THG (red) images of hamster mucosa and sub-mucosa taken at different depths. From (a) and (b), we can identify the morphology of stratum corneum (SC), stratum granulosa (SG), stratum basale (SB), and red blood cells (RBC) in the oral cavity by using epi-THG microscopy. Besides, epi-SHG microscopy can provide the distribution and density information of collagen fibrils in the sub-mucosa. At higher magnification, (c) and (d) show more detailed morphologies of stratum granulosa and stratum basale. Clear cell-cell junctions (indicated by yellow arrows) can be picked up by epi-THG microscopy. Scale bar: 40µm.

Fig. 4.
Fig. 4.

(1.67 MB) An example movie of the in vivo horizontally sectioned epi-THG microscopy showing the blood flow in the capillary in the sub-mucosa layer of the hamster oral cavity. Image size: 80µm×80µm

Fig. 5.
Fig. 5.

(1.85 MB) This movie shows a stack of depth-resolved in vivo horizontal sections in the hamster oral cavity. Using epi-SHG and epi-THG microscopy, a series of horizontal sections from mucosa to sub-mucosa is demonstrated. This movie is composed of 30 horizontal images. The optical depth difference between adjacent images is 2.8 µm. Image size: 240µm×240µm

Fig. 6.
Fig. 6.

Vertical sections taken in the live hamster oral cavity by epi-THG microscopy (a) before and (b) after adding 6% acetic acid solution on mucosa. After adding 6% acetic acid solution, the enhanced epi-THG intensity of epithelial cells, except stratum corneum (SC), can be observed. Scale bar: 30µm

Fig. 7.
Fig. 7.

In vivo horizontal sections taken in the mucosa layer of the live hamster oral cavity at different depths by epi-THG microscopy are presented after adding 6% acetic acid solution. The nuclear morphology of squamous cells can be clearly observed by epi-THG microscopy (indicated by green arrow). The yellow arrow indicates the cell membrane. Scale bar: 10µm

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