Abstract

Second harmonic generation (SHG) imaging is a well-suited multiphoton technique allowing visualization of biological tissues mainly composed of collagen with submicron resolution. Despite its inherent confocal properties, imaging of deeper layers within thick samples has still some limitations. Although the use of longer wavelengths might help to overcome this, the dependence between SHG signals and wavelength is still under discussion. We report here on the dependence with wavelength of SHG signals from collagen-based ocular tissues. The quality of SHG images for two commonly used excitation wavelengths (800 and 1045 nm) is studied. The analysis of the collagen structural information reveals that the information provided by both wavelengths is similar. It was also found that, independently of the depth location, 1045-nm SHG images presented always lower signal levels than those acquired with 800 nm. However, the contrast of the former images was higher, what may improve the visualization of certain features of interest.

© 2017 Optical Society of America

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2017 (2)

F. J. Ávila, J. M. Bueno, A. Gambín, and P. Artal, “Second harmonic generation microscopy of the human cornea and sclera in vivo,” Invest. Ophthalmol. Vis. Sci. 58, 3109 (2017).

F. J. Ávila, O. del Barco, and J. M. Bueno, “Quantifying external and internal collagen organization from Stokes-vector-based second harmonic generation imaging polarimetry,” J. Opt. 19(10), 105301 (2017).

2016 (1)

G. Lombardo, N. L. Micali, V. Villari, S. Serrao, and M. Lombardo, “All-optical method to assess stromal concentration of riboflavin in conventional and accelerated UV-A irradiation of the human cornea,” Invest. Ophthalmol. Vis. Sci. 57(2), 476–483 (2016).
[PubMed]

2015 (5)

F. J. Ávila, O. Del Barco, and J. M. Bueno, “Polarization dependence of aligned collagen tissues imaged with second harmonic generation microscopy,” J. Biomed. Opt. 20(8), 86001 (2015).
[PubMed]

F. J. Ávila and J. M. Bueno, “Analysis and quantification of collagen organization with the structure tensor in second harmonic microscopy images of ocular tissues,” Appl. Opt. 54(33), 9848–9854 (2015).
[PubMed]

J. M. Bueno, R. Palacios, A. Pennos, and P. Artal, “Second-harmonic generation microscopy of photocurable polymer intrastromal implants in ex-vivo corneas,” Biomed. Opt. Express 6(6), 2211–2219 (2015).
[PubMed]

M. Skorsetz, P. Artal, and J. M. Bueno, “Performance evaluation of a sensorless adaptive optics multiphoton microscope,” J. Microsc. 261(3), 249–258 (2015).

C. Y. Park, J. K. Lee, and R. S. Chuck, “Second Harmonic Generation Imaging Analysis of Collagen Arrangement in Human Cornea,” Invest. Ophthalmol. Vis. Sci. 56(9), 5622–5629 (2015).
[PubMed]

2014 (3)

2013 (2)

M. Shen, J. Zhao, H. Zeng, and S. Tang, “Calibrating the measurement of wavelength-dependent second harmonic generation from biological tissues with a BaB2O4 crystal,” J. Biomed. Opt. 18(3), 031109 (2013).
[PubMed]

J. M. Bueno, R. Palacios, M. K. Chessey, and H. Ginis, “Analysis of spatial lamellar distribution from adaptive-optics second harmonic generation corneal images,” Biomed. Opt. Express 4(7), 1006–1013 (2013).
[PubMed]

2012 (3)

N. Morishige, N. Yamada, X. Zhang, Y. Morita, N. Yamada, K. Kimura, A. Takahara, and K. H. Sonoda, “Abnormalities of stromal structure in the bullous keratopathy cornea identified by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 53(8), 4998–5003 (2012).
[PubMed]

W. Lo, W. L. Chen, C. M. Hsueh, A. A. Ghazaryan, S. J. Chen, D. H. K. Ma, C. Y. Dong, and H. Y. Tan, “Fast Fourier transform-based analysis of second-harmonic generation image in keratoconic cornea,” Invest. Ophthalmol. Vis. Sci. 53(7), 3501–3507 (2012).
[PubMed]

G. Latour, I. Gusachenko, L. Kowalczuk, I. Lamarre, and M.-C. Schanne-Klein, “In vivo structural imaging of the cornea by polarization-resolved second harmonic microscopy,” Biomed. Opt. Express 3(1), 1–15 (2012).
[PubMed]

2011 (3)

N. Morishige, Y. Takagi, T. Chikama, A. Takahara, and T. Nishida, “Three-dimensional analysis of collagen lamellae in the anterior stroma of the human cornea visualized by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 52(2), 911–915 (2011).
[PubMed]

J. M. Bueno, E. J. Gualda, and P. Artal, “Analysis of corneal stroma organization with wavefront optimized nonlinear microscopy,” Cornea 30(6), 692–701 (2011).
[PubMed]

J. M. Bueno, E. J. Gualda, A. Giakoumaki, P. Pérez-Merino, S. Marcos, and P. Artal, “Multiphoton microscopy of ex vivo corneas after collagen cross-linking,” Invest. Ophthalmol. Vis. Sci. 52(8), 5325–5331 (2011).
[PubMed]

2010 (3)

D. Kampik, B. Ralla, S. Keller, M. Hirschberg, P. Friedl, and G. Geerling, “Influence of corneal collagen crosslinking with riboflavin and ultraviolet-a irradiation on excimer laser surgery,” Invest. Ophthalmol. Vis. Sci. 51(8), 3929–3934 (2010).
[PubMed]

F. Aptel, N. Olivier, A. Deniset-Besseau, J.-M. Legeais, K. Plamann, M.-C. Schanne-Klein, and E. Beaurepaire, “Multimodal Nonlinear Imaging of the Human Cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[PubMed]

J. M. Bueno, E. J. Gualda, and P. Artal, “Adaptive optics multiphoton microscopy to study ex vivo ocular tissues,” J. Biomed. Opt. 15(6), 066004 (2010).
[PubMed]

2009 (1)

C. M. Hsueh, W. Lo, W. L. Chen, V. A. Hovhannisyan, G. Y. Liu, S. S. Wang, H. Y. Tan, and C. Y. Dong, “Structural characterization of edematous corneas by forward and backward second harmonic generation imaging,” Biophys. J. 97(4), 1198–1205 (2009).
[PubMed]

2008 (1)

B. G. Wang, A. Eitner, J. Lindenau, and K. J. Halbhuber, “High-resolution two-photon excitation microscopy of ocular tissues in porcine eye,” Lasers Surg. Med. 40(4), 247–256 (2008).
[PubMed]

2007 (2)

N. Morishige, A. J. Wahlert, M. C. Kenney, D. J. Brown, K. Kawamoto, T. Chikama, T. Nishida, and J. V. Jester, “Second-harmonic imaging microscopy of normal human and keratoconus cornea,” Invest. Ophthalmol. Vis. Sci. 48(3), 1087–1094 (2007).
[PubMed]

J. J. Hunter, C. J. Cookson, M. L. Kisilak, J. M. Bueno, and M. C. Campbell, “Characterizing image quality in a scanning laser ophthalmoscope with differing pinholes and induced scattered light,” J. Opt. Soc. Am. A 24(5), 1284–1295 (2007).
[PubMed]

2006 (4)

T. A. Theodossiou, C. Thrasivoulou, C. Ekwobi, and D. L. Becker, “Second harmonic generation confocal microscopy of collagen type I from rat tendon cryosections,” Biophys. J. 91(12), 4665–4677 (2006).
[PubMed]

N. Morishige, W. M. Petroll, T. Nishida, M. C. Kenney, and J. V. Jester, “Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals,” J. Cataract Refract. Surg. 32(11), 1784–1791 (2006).
[PubMed]

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

H. Y. Tan, Y. Sun, W. Lo, S. J. Lin, C. H. Hsiao, Y. F. Chen, S. C. Huang, W. C. Lin, S. H. Jee, H. S. Yu, and C. Y. Dong, “Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo,” Invest. Ophthalmol. Vis. Sci. 47(12), 5251–5259 (2006).
[PubMed]

2005 (3)

R. M. Williams, W. R. Zipfel, and W. W. Webb, “Interpreting second-harmonic generation images of collagen I fibrils,” Biophys. J. 88(2), 1377–1386 (2005).
[PubMed]

M. Han, G. Giese, and J. Bille, “Second harmonic generation imaging of collagen fibrils in cornea and sclera,” Opt. Express 13(15), 5791–5797 (2005).
[PubMed]

J. M. Bueno and B. Vohnsen, “Polarimetric high-resolution confocal scanning laser ophthalmoscope,” Vision Res. 45(28), 3526–3534 (2005).
[PubMed]

2003 (2)

I. Jalbert, F. Stapleton, E. Papas, D. F. Sweeney, and M. Coroneo, “In vivo confocal microscopy of the human cornea,” Br. J. Ophthalmol. 87(2), 225–236 (2003).
[PubMed]

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[PubMed]

2002 (3)

P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, “Polarization-modulated second harmonic generation in collagen,” Biophys. J. 82(6), 3330–3342 (2002).
[PubMed]

A. T. Yeh, N. Nassif, A. Zoumi, and B. J. Tromberg, “Selective corneal imaging using combined second-harmonic generation and two-photon excited fluorescence,” Opt. Lett. 27(23), 2082–2084 (2002).
[PubMed]

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. U.S.A. 99(17), 11014–11019 (2002).
[PubMed]

2001 (2)

P. J. Campagnola, H. A. Clark, W. A. Mohler, A. Lewis, and L. M. Loew, “Second-harmonic imaging microscopy of living cells,” J. Biomed. Opt. 6(3), 277–286 (2001).
[PubMed]

M. Oheim, E. Beaurepaire, E. Chaigneau, J. Mertz, and S. Charpak, “Two-photon microscopy in brain tissue: parameters influencing the imaging depth,” J. Neurosci. Methods 111(1), 29–37 (2001).
[PubMed]

1997 (1)

1990 (1)

1971 (1)

Alfano, R. R.

Aptel, F.

F. Aptel, N. Olivier, A. Deniset-Besseau, J.-M. Legeais, K. Plamann, M.-C. Schanne-Klein, and E. Beaurepaire, “Multimodal Nonlinear Imaging of the Human Cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[PubMed]

Artal, P.

F. J. Ávila, J. M. Bueno, A. Gambín, and P. Artal, “Second harmonic generation microscopy of the human cornea and sclera in vivo,” Invest. Ophthalmol. Vis. Sci. 58, 3109 (2017).

J. M. Bueno, R. Palacios, A. Pennos, and P. Artal, “Second-harmonic generation microscopy of photocurable polymer intrastromal implants in ex-vivo corneas,” Biomed. Opt. Express 6(6), 2211–2219 (2015).
[PubMed]

M. Skorsetz, P. Artal, and J. M. Bueno, “Performance evaluation of a sensorless adaptive optics multiphoton microscope,” J. Microsc. 261(3), 249–258 (2015).

J. M. Bueno, E. J. Gualda, A. Giakoumaki, P. Pérez-Merino, S. Marcos, and P. Artal, “Multiphoton microscopy of ex vivo corneas after collagen cross-linking,” Invest. Ophthalmol. Vis. Sci. 52(8), 5325–5331 (2011).
[PubMed]

J. M. Bueno, E. J. Gualda, and P. Artal, “Analysis of corneal stroma organization with wavefront optimized nonlinear microscopy,” Cornea 30(6), 692–701 (2011).
[PubMed]

J. M. Bueno, E. J. Gualda, and P. Artal, “Adaptive optics multiphoton microscopy to study ex vivo ocular tissues,” J. Biomed. Opt. 15(6), 066004 (2010).
[PubMed]

Ávila, F. J.

F. J. Ávila, J. M. Bueno, A. Gambín, and P. Artal, “Second harmonic generation microscopy of the human cornea and sclera in vivo,” Invest. Ophthalmol. Vis. Sci. 58, 3109 (2017).

F. J. Ávila, O. del Barco, and J. M. Bueno, “Quantifying external and internal collagen organization from Stokes-vector-based second harmonic generation imaging polarimetry,” J. Opt. 19(10), 105301 (2017).

F. J. Ávila, O. Del Barco, and J. M. Bueno, “Polarization dependence of aligned collagen tissues imaged with second harmonic generation microscopy,” J. Biomed. Opt. 20(8), 86001 (2015).
[PubMed]

F. J. Ávila and J. M. Bueno, “Analysis and quantification of collagen organization with the structure tensor in second harmonic microscopy images of ocular tissues,” Appl. Opt. 54(33), 9848–9854 (2015).
[PubMed]

Beaurepaire, E.

F. Aptel, N. Olivier, A. Deniset-Besseau, J.-M. Legeais, K. Plamann, M.-C. Schanne-Klein, and E. Beaurepaire, “Multimodal Nonlinear Imaging of the Human Cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[PubMed]

M. Oheim, E. Beaurepaire, E. Chaigneau, J. Mertz, and S. Charpak, “Two-photon microscopy in brain tissue: parameters influencing the imaging depth,” J. Neurosci. Methods 111(1), 29–37 (2001).
[PubMed]

Becker, D. L.

T. A. Theodossiou, C. Thrasivoulou, C. Ekwobi, and D. L. Becker, “Second harmonic generation confocal microscopy of collagen type I from rat tendon cryosections,” Biophys. J. 91(12), 4665–4677 (2006).
[PubMed]

Bille, J.

Brown, D. J.

N. Morishige, A. J. Wahlert, M. C. Kenney, D. J. Brown, K. Kawamoto, T. Chikama, T. Nishida, and J. V. Jester, “Second-harmonic imaging microscopy of normal human and keratoconus cornea,” Invest. Ophthalmol. Vis. Sci. 48(3), 1087–1094 (2007).
[PubMed]

Bueno, J. M.

F. J. Ávila, J. M. Bueno, A. Gambín, and P. Artal, “Second harmonic generation microscopy of the human cornea and sclera in vivo,” Invest. Ophthalmol. Vis. Sci. 58, 3109 (2017).

F. J. Ávila, O. del Barco, and J. M. Bueno, “Quantifying external and internal collagen organization from Stokes-vector-based second harmonic generation imaging polarimetry,” J. Opt. 19(10), 105301 (2017).

F. J. Ávila, O. Del Barco, and J. M. Bueno, “Polarization dependence of aligned collagen tissues imaged with second harmonic generation microscopy,” J. Biomed. Opt. 20(8), 86001 (2015).
[PubMed]

M. Skorsetz, P. Artal, and J. M. Bueno, “Performance evaluation of a sensorless adaptive optics multiphoton microscope,” J. Microsc. 261(3), 249–258 (2015).

F. J. Ávila and J. M. Bueno, “Analysis and quantification of collagen organization with the structure tensor in second harmonic microscopy images of ocular tissues,” Appl. Opt. 54(33), 9848–9854 (2015).
[PubMed]

J. M. Bueno, R. Palacios, A. Pennos, and P. Artal, “Second-harmonic generation microscopy of photocurable polymer intrastromal implants in ex-vivo corneas,” Biomed. Opt. Express 6(6), 2211–2219 (2015).
[PubMed]

J. M. Bueno, R. Palacios, M. K. Chessey, and H. Ginis, “Analysis of spatial lamellar distribution from adaptive-optics second harmonic generation corneal images,” Biomed. Opt. Express 4(7), 1006–1013 (2013).
[PubMed]

J. M. Bueno, E. J. Gualda, and P. Artal, “Analysis of corneal stroma organization with wavefront optimized nonlinear microscopy,” Cornea 30(6), 692–701 (2011).
[PubMed]

J. M. Bueno, E. J. Gualda, A. Giakoumaki, P. Pérez-Merino, S. Marcos, and P. Artal, “Multiphoton microscopy of ex vivo corneas after collagen cross-linking,” Invest. Ophthalmol. Vis. Sci. 52(8), 5325–5331 (2011).
[PubMed]

J. M. Bueno, E. J. Gualda, and P. Artal, “Adaptive optics multiphoton microscopy to study ex vivo ocular tissues,” J. Biomed. Opt. 15(6), 066004 (2010).
[PubMed]

J. J. Hunter, C. J. Cookson, M. L. Kisilak, J. M. Bueno, and M. C. Campbell, “Characterizing image quality in a scanning laser ophthalmoscope with differing pinholes and induced scattered light,” J. Opt. Soc. Am. A 24(5), 1284–1295 (2007).
[PubMed]

J. M. Bueno and B. Vohnsen, “Polarimetric high-resolution confocal scanning laser ophthalmoscope,” Vision Res. 45(28), 3526–3534 (2005).
[PubMed]

Campagnola, P. J.

Campbell, K. R.

Campbell, M. C.

Celliers, P. M.

P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, “Polarization-modulated second harmonic generation in collagen,” Biophys. J. 82(6), 3330–3342 (2002).
[PubMed]

Chaigneau, E.

M. Oheim, E. Beaurepaire, E. Chaigneau, J. Mertz, and S. Charpak, “Two-photon microscopy in brain tissue: parameters influencing the imaging depth,” J. Neurosci. Methods 111(1), 29–37 (2001).
[PubMed]

Charpak, S.

M. Oheim, E. Beaurepaire, E. Chaigneau, J. Mertz, and S. Charpak, “Two-photon microscopy in brain tissue: parameters influencing the imaging depth,” J. Neurosci. Methods 111(1), 29–37 (2001).
[PubMed]

Chen, S. J.

W. Lo, W. L. Chen, C. M. Hsueh, A. A. Ghazaryan, S. J. Chen, D. H. K. Ma, C. Y. Dong, and H. Y. Tan, “Fast Fourier transform-based analysis of second-harmonic generation image in keratoconic cornea,” Invest. Ophthalmol. Vis. Sci. 53(7), 3501–3507 (2012).
[PubMed]

Chen, W. L.

W. Lo, W. L. Chen, C. M. Hsueh, A. A. Ghazaryan, S. J. Chen, D. H. K. Ma, C. Y. Dong, and H. Y. Tan, “Fast Fourier transform-based analysis of second-harmonic generation image in keratoconic cornea,” Invest. Ophthalmol. Vis. Sci. 53(7), 3501–3507 (2012).
[PubMed]

C. M. Hsueh, W. Lo, W. L. Chen, V. A. Hovhannisyan, G. Y. Liu, S. S. Wang, H. Y. Tan, and C. Y. Dong, “Structural characterization of edematous corneas by forward and backward second harmonic generation imaging,” Biophys. J. 97(4), 1198–1205 (2009).
[PubMed]

Chen, Y. F.

H. Y. Tan, Y. Sun, W. Lo, S. J. Lin, C. H. Hsiao, Y. F. Chen, S. C. Huang, W. C. Lin, S. H. Jee, H. S. Yu, and C. Y. Dong, “Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo,” Invest. Ophthalmol. Vis. Sci. 47(12), 5251–5259 (2006).
[PubMed]

Chessey, M. K.

Chikama, T.

N. Morishige, Y. Takagi, T. Chikama, A. Takahara, and T. Nishida, “Three-dimensional analysis of collagen lamellae in the anterior stroma of the human cornea visualized by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 52(2), 911–915 (2011).
[PubMed]

N. Morishige, A. J. Wahlert, M. C. Kenney, D. J. Brown, K. Kawamoto, T. Chikama, T. Nishida, and J. V. Jester, “Second-harmonic imaging microscopy of normal human and keratoconus cornea,” Invest. Ophthalmol. Vis. Sci. 48(3), 1087–1094 (2007).
[PubMed]

Chuck, R. S.

C. Y. Park, J. K. Lee, and R. S. Chuck, “Second Harmonic Generation Imaging Analysis of Collagen Arrangement in Human Cornea,” Invest. Ophthalmol. Vis. Sci. 56(9), 5622–5629 (2015).
[PubMed]

Clark, H. A.

P. J. Campagnola, H. A. Clark, W. A. Mohler, A. Lewis, and L. M. Loew, “Second-harmonic imaging microscopy of living cells,” J. Biomed. Opt. 6(3), 277–286 (2001).
[PubMed]

Cookson, C. J.

Coroneo, M.

I. Jalbert, F. Stapleton, E. Papas, D. F. Sweeney, and M. Coroneo, “In vivo confocal microscopy of the human cornea,” Br. J. Ophthalmol. 87(2), 225–236 (2003).
[PubMed]

del Barco, O.

F. J. Ávila, O. del Barco, and J. M. Bueno, “Quantifying external and internal collagen organization from Stokes-vector-based second harmonic generation imaging polarimetry,” J. Opt. 19(10), 105301 (2017).

F. J. Ávila, O. Del Barco, and J. M. Bueno, “Polarization dependence of aligned collagen tissues imaged with second harmonic generation microscopy,” J. Biomed. Opt. 20(8), 86001 (2015).
[PubMed]

Deniset-Besseau, A.

F. Aptel, N. Olivier, A. Deniset-Besseau, J.-M. Legeais, K. Plamann, M.-C. Schanne-Klein, and E. Beaurepaire, “Multimodal Nonlinear Imaging of the Human Cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[PubMed]

Dong, C. Y.

W. Lo, W. L. Chen, C. M. Hsueh, A. A. Ghazaryan, S. J. Chen, D. H. K. Ma, C. Y. Dong, and H. Y. Tan, “Fast Fourier transform-based analysis of second-harmonic generation image in keratoconic cornea,” Invest. Ophthalmol. Vis. Sci. 53(7), 3501–3507 (2012).
[PubMed]

C. M. Hsueh, W. Lo, W. L. Chen, V. A. Hovhannisyan, G. Y. Liu, S. S. Wang, H. Y. Tan, and C. Y. Dong, “Structural characterization of edematous corneas by forward and backward second harmonic generation imaging,” Biophys. J. 97(4), 1198–1205 (2009).
[PubMed]

H. Y. Tan, Y. Sun, W. Lo, S. J. Lin, C. H. Hsiao, Y. F. Chen, S. C. Huang, W. C. Lin, S. H. Jee, H. S. Yu, and C. Y. Dong, “Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo,” Invest. Ophthalmol. Vis. Sci. 47(12), 5251–5259 (2006).
[PubMed]

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

Eitner, A.

B. G. Wang, A. Eitner, J. Lindenau, and K. J. Halbhuber, “High-resolution two-photon excitation microscopy of ocular tissues in porcine eye,” Lasers Surg. Med. 40(4), 247–256 (2008).
[PubMed]

Ekwobi, C.

T. A. Theodossiou, C. Thrasivoulou, C. Ekwobi, and D. L. Becker, “Second harmonic generation confocal microscopy of collagen type I from rat tendon cryosections,” Biophys. J. 91(12), 4665–4677 (2006).
[PubMed]

Eliceiri, K. W.

Fine, S.

Friedl, P.

D. Kampik, B. Ralla, S. Keller, M. Hirschberg, P. Friedl, and G. Geerling, “Influence of corneal collagen crosslinking with riboflavin and ultraviolet-a irradiation on excimer laser surgery,” Invest. Ophthalmol. Vis. Sci. 51(8), 3929–3934 (2010).
[PubMed]

Fukuda, S.

Gambín, A.

F. J. Ávila, J. M. Bueno, A. Gambín, and P. Artal, “Second harmonic generation microscopy of the human cornea and sclera in vivo,” Invest. Ophthalmol. Vis. Sci. 58, 3109 (2017).

Geerling, G.

D. Kampik, B. Ralla, S. Keller, M. Hirschberg, P. Friedl, and G. Geerling, “Influence of corneal collagen crosslinking with riboflavin and ultraviolet-a irradiation on excimer laser surgery,” Invest. Ophthalmol. Vis. Sci. 51(8), 3929–3934 (2010).
[PubMed]

Ghazaryan, A. A.

W. Lo, W. L. Chen, C. M. Hsueh, A. A. Ghazaryan, S. J. Chen, D. H. K. Ma, C. Y. Dong, and H. Y. Tan, “Fast Fourier transform-based analysis of second-harmonic generation image in keratoconic cornea,” Invest. Ophthalmol. Vis. Sci. 53(7), 3501–3507 (2012).
[PubMed]

Giakoumaki, A.

J. M. Bueno, E. J. Gualda, A. Giakoumaki, P. Pérez-Merino, S. Marcos, and P. Artal, “Multiphoton microscopy of ex vivo corneas after collagen cross-linking,” Invest. Ophthalmol. Vis. Sci. 52(8), 5325–5331 (2011).
[PubMed]

Giese, G.

Ginis, H.

Gualda, E. J.

J. M. Bueno, E. J. Gualda, and P. Artal, “Analysis of corneal stroma organization with wavefront optimized nonlinear microscopy,” Cornea 30(6), 692–701 (2011).
[PubMed]

J. M. Bueno, E. J. Gualda, A. Giakoumaki, P. Pérez-Merino, S. Marcos, and P. Artal, “Multiphoton microscopy of ex vivo corneas after collagen cross-linking,” Invest. Ophthalmol. Vis. Sci. 52(8), 5325–5331 (2011).
[PubMed]

J. M. Bueno, E. J. Gualda, and P. Artal, “Adaptive optics multiphoton microscopy to study ex vivo ocular tissues,” J. Biomed. Opt. 15(6), 066004 (2010).
[PubMed]

Guo, Y.

Gusachenko, I.

Halbhuber, K. J.

B. G. Wang, A. Eitner, J. Lindenau, and K. J. Halbhuber, “High-resolution two-photon excitation microscopy of ocular tissues in porcine eye,” Lasers Surg. Med. 40(4), 247–256 (2008).
[PubMed]

Hall, G.

Han, M.

Hansen, W. P.

Harris, D.

Hirschberg, M.

D. Kampik, B. Ralla, S. Keller, M. Hirschberg, P. Friedl, and G. Geerling, “Influence of corneal collagen crosslinking with riboflavin and ultraviolet-a irradiation on excimer laser surgery,” Invest. Ophthalmol. Vis. Sci. 51(8), 3929–3934 (2010).
[PubMed]

Ho, P. P.

Hovhannisyan, V. A.

C. M. Hsueh, W. Lo, W. L. Chen, V. A. Hovhannisyan, G. Y. Liu, S. S. Wang, H. Y. Tan, and C. Y. Dong, “Structural characterization of edematous corneas by forward and backward second harmonic generation imaging,” Biophys. J. 97(4), 1198–1205 (2009).
[PubMed]

Hsiao, C. H.

H. Y. Tan, Y. Sun, W. Lo, S. J. Lin, C. H. Hsiao, Y. F. Chen, S. C. Huang, W. C. Lin, S. H. Jee, H. S. Yu, and C. Y. Dong, “Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo,” Invest. Ophthalmol. Vis. Sci. 47(12), 5251–5259 (2006).
[PubMed]

Hsueh, C. M.

W. Lo, W. L. Chen, C. M. Hsueh, A. A. Ghazaryan, S. J. Chen, D. H. K. Ma, C. Y. Dong, and H. Y. Tan, “Fast Fourier transform-based analysis of second-harmonic generation image in keratoconic cornea,” Invest. Ophthalmol. Vis. Sci. 53(7), 3501–3507 (2012).
[PubMed]

C. M. Hsueh, W. Lo, W. L. Chen, V. A. Hovhannisyan, G. Y. Liu, S. S. Wang, H. Y. Tan, and C. Y. Dong, “Structural characterization of edematous corneas by forward and backward second harmonic generation imaging,” Biophys. J. 97(4), 1198–1205 (2009).
[PubMed]

Huang, S. C.

H. Y. Tan, Y. Sun, W. Lo, S. J. Lin, C. H. Hsiao, Y. F. Chen, S. C. Huang, W. C. Lin, S. H. Jee, H. S. Yu, and C. Y. Dong, “Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo,” Invest. Ophthalmol. Vis. Sci. 47(12), 5251–5259 (2006).
[PubMed]

Hunter, J. J.

Ishii, K.

Jalbert, I.

I. Jalbert, F. Stapleton, E. Papas, D. F. Sweeney, and M. Coroneo, “In vivo confocal microscopy of the human cornea,” Br. J. Ophthalmol. 87(2), 225–236 (2003).
[PubMed]

Jee, S. H.

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

H. Y. Tan, Y. Sun, W. Lo, S. J. Lin, C. H. Hsiao, Y. F. Chen, S. C. Huang, W. C. Lin, S. H. Jee, H. S. Yu, and C. Y. Dong, “Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo,” Invest. Ophthalmol. Vis. Sci. 47(12), 5251–5259 (2006).
[PubMed]

Jester, J. V.

N. Morishige, A. J. Wahlert, M. C. Kenney, D. J. Brown, K. Kawamoto, T. Chikama, T. Nishida, and J. V. Jester, “Second-harmonic imaging microscopy of normal human and keratoconus cornea,” Invest. Ophthalmol. Vis. Sci. 48(3), 1087–1094 (2007).
[PubMed]

N. Morishige, W. M. Petroll, T. Nishida, M. C. Kenney, and J. V. Jester, “Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals,” J. Cataract Refract. Surg. 32(11), 1784–1791 (2006).
[PubMed]

Kampik, D.

D. Kampik, B. Ralla, S. Keller, M. Hirschberg, P. Friedl, and G. Geerling, “Influence of corneal collagen crosslinking with riboflavin and ultraviolet-a irradiation on excimer laser surgery,” Invest. Ophthalmol. Vis. Sci. 51(8), 3929–3934 (2010).
[PubMed]

Kawamoto, K.

N. Morishige, A. J. Wahlert, M. C. Kenney, D. J. Brown, K. Kawamoto, T. Chikama, T. Nishida, and J. V. Jester, “Second-harmonic imaging microscopy of normal human and keratoconus cornea,” Invest. Ophthalmol. Vis. Sci. 48(3), 1087–1094 (2007).
[PubMed]

Keller, S.

D. Kampik, B. Ralla, S. Keller, M. Hirschberg, P. Friedl, and G. Geerling, “Influence of corneal collagen crosslinking with riboflavin and ultraviolet-a irradiation on excimer laser surgery,” Invest. Ophthalmol. Vis. Sci. 51(8), 3929–3934 (2010).
[PubMed]

Kenney, M. C.

N. Morishige, A. J. Wahlert, M. C. Kenney, D. J. Brown, K. Kawamoto, T. Chikama, T. Nishida, and J. V. Jester, “Second-harmonic imaging microscopy of normal human and keratoconus cornea,” Invest. Ophthalmol. Vis. Sci. 48(3), 1087–1094 (2007).
[PubMed]

N. Morishige, W. M. Petroll, T. Nishida, M. C. Kenney, and J. V. Jester, “Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals,” J. Cataract Refract. Surg. 32(11), 1784–1791 (2006).
[PubMed]

Kim, K. H.

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

Kimura, K.

N. Morishige, N. Yamada, X. Zhang, Y. Morita, N. Yamada, K. Kimura, A. Takahara, and K. H. Sonoda, “Abnormalities of stromal structure in the bullous keratopathy cornea identified by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 53(8), 4998–5003 (2012).
[PubMed]

Kisilak, M. L.

Kowalczuk, L.

Lamarre, I.

Latour, G.

Lee, J. K.

C. Y. Park, J. K. Lee, and R. S. Chuck, “Second Harmonic Generation Imaging Analysis of Collagen Arrangement in Human Cornea,” Invest. Ophthalmol. Vis. Sci. 56(9), 5622–5629 (2015).
[PubMed]

Legeais, J.-M.

F. Aptel, N. Olivier, A. Deniset-Besseau, J.-M. Legeais, K. Plamann, M.-C. Schanne-Klein, and E. Beaurepaire, “Multimodal Nonlinear Imaging of the Human Cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[PubMed]

Lewis, A.

P. J. Campagnola, H. A. Clark, W. A. Mohler, A. Lewis, and L. M. Loew, “Second-harmonic imaging microscopy of living cells,” J. Biomed. Opt. 6(3), 277–286 (2001).
[PubMed]

Lin, H. H.

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

Lin, S. J.

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

H. Y. Tan, Y. Sun, W. Lo, S. J. Lin, C. H. Hsiao, Y. F. Chen, S. C. Huang, W. C. Lin, S. H. Jee, H. S. Yu, and C. Y. Dong, “Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo,” Invest. Ophthalmol. Vis. Sci. 47(12), 5251–5259 (2006).
[PubMed]

Lin, W. C.

H. Y. Tan, Y. Sun, W. Lo, S. J. Lin, C. H. Hsiao, Y. F. Chen, S. C. Huang, W. C. Lin, S. H. Jee, H. S. Yu, and C. Y. Dong, “Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo,” Invest. Ophthalmol. Vis. Sci. 47(12), 5251–5259 (2006).
[PubMed]

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

Lindenau, J.

B. G. Wang, A. Eitner, J. Lindenau, and K. J. Halbhuber, “High-resolution two-photon excitation microscopy of ocular tissues in porcine eye,” Lasers Surg. Med. 40(4), 247–256 (2008).
[PubMed]

Liu, F.

Liu, G. Y.

C. M. Hsueh, W. Lo, W. L. Chen, V. A. Hovhannisyan, G. Y. Liu, S. S. Wang, H. Y. Tan, and C. Y. Dong, “Structural characterization of edematous corneas by forward and backward second harmonic generation imaging,” Biophys. J. 97(4), 1198–1205 (2009).
[PubMed]

Lo, W.

W. Lo, W. L. Chen, C. M. Hsueh, A. A. Ghazaryan, S. J. Chen, D. H. K. Ma, C. Y. Dong, and H. Y. Tan, “Fast Fourier transform-based analysis of second-harmonic generation image in keratoconic cornea,” Invest. Ophthalmol. Vis. Sci. 53(7), 3501–3507 (2012).
[PubMed]

C. M. Hsueh, W. Lo, W. L. Chen, V. A. Hovhannisyan, G. Y. Liu, S. S. Wang, H. Y. Tan, and C. Y. Dong, “Structural characterization of edematous corneas by forward and backward second harmonic generation imaging,” Biophys. J. 97(4), 1198–1205 (2009).
[PubMed]

H. Y. Tan, Y. Sun, W. Lo, S. J. Lin, C. H. Hsiao, Y. F. Chen, S. C. Huang, W. C. Lin, S. H. Jee, H. S. Yu, and C. Y. Dong, “Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo,” Invest. Ophthalmol. Vis. Sci. 47(12), 5251–5259 (2006).
[PubMed]

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

Loew, L. M.

P. J. Campagnola, H. A. Clark, W. A. Mohler, A. Lewis, and L. M. Loew, “Second-harmonic imaging microscopy of living cells,” J. Biomed. Opt. 6(3), 277–286 (2001).
[PubMed]

Lombardo, G.

G. Lombardo, N. L. Micali, V. Villari, S. Serrao, and M. Lombardo, “All-optical method to assess stromal concentration of riboflavin in conventional and accelerated UV-A irradiation of the human cornea,” Invest. Ophthalmol. Vis. Sci. 57(2), 476–483 (2016).
[PubMed]

Lombardo, M.

G. Lombardo, N. L. Micali, V. Villari, S. Serrao, and M. Lombardo, “All-optical method to assess stromal concentration of riboflavin in conventional and accelerated UV-A irradiation of the human cornea,” Invest. Ophthalmol. Vis. Sci. 57(2), 476–483 (2016).
[PubMed]

Ma, D. H. K.

W. Lo, W. L. Chen, C. M. Hsueh, A. A. Ghazaryan, S. J. Chen, D. H. K. Ma, C. Y. Dong, and H. Y. Tan, “Fast Fourier transform-based analysis of second-harmonic generation image in keratoconic cornea,” Invest. Ophthalmol. Vis. Sci. 53(7), 3501–3507 (2012).
[PubMed]

Marcos, S.

J. M. Bueno, E. J. Gualda, A. Giakoumaki, P. Pérez-Merino, S. Marcos, and P. Artal, “Multiphoton microscopy of ex vivo corneas after collagen cross-linking,” Invest. Ophthalmol. Vis. Sci. 52(8), 5325–5331 (2011).
[PubMed]

Mertz, J.

M. Oheim, E. Beaurepaire, E. Chaigneau, J. Mertz, and S. Charpak, “Two-photon microscopy in brain tissue: parameters influencing the imaging depth,” J. Neurosci. Methods 111(1), 29–37 (2001).
[PubMed]

Micali, N. L.

G. Lombardo, N. L. Micali, V. Villari, S. Serrao, and M. Lombardo, “All-optical method to assess stromal concentration of riboflavin in conventional and accelerated UV-A irradiation of the human cornea,” Invest. Ophthalmol. Vis. Sci. 57(2), 476–483 (2016).
[PubMed]

Miura, M.

Mohler, W. A.

P. J. Campagnola, H. A. Clark, W. A. Mohler, A. Lewis, and L. M. Loew, “Second-harmonic imaging microscopy of living cells,” J. Biomed. Opt. 6(3), 277–286 (2001).
[PubMed]

Morishige, N.

N. Morishige, N. Yamada, X. Zhang, Y. Morita, N. Yamada, K. Kimura, A. Takahara, and K. H. Sonoda, “Abnormalities of stromal structure in the bullous keratopathy cornea identified by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 53(8), 4998–5003 (2012).
[PubMed]

N. Morishige, Y. Takagi, T. Chikama, A. Takahara, and T. Nishida, “Three-dimensional analysis of collagen lamellae in the anterior stroma of the human cornea visualized by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 52(2), 911–915 (2011).
[PubMed]

N. Morishige, A. J. Wahlert, M. C. Kenney, D. J. Brown, K. Kawamoto, T. Chikama, T. Nishida, and J. V. Jester, “Second-harmonic imaging microscopy of normal human and keratoconus cornea,” Invest. Ophthalmol. Vis. Sci. 48(3), 1087–1094 (2007).
[PubMed]

N. Morishige, W. M. Petroll, T. Nishida, M. C. Kenney, and J. V. Jester, “Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals,” J. Cataract Refract. Surg. 32(11), 1784–1791 (2006).
[PubMed]

Morita, Y.

N. Morishige, N. Yamada, X. Zhang, Y. Morita, N. Yamada, K. Kimura, A. Takahara, and K. H. Sonoda, “Abnormalities of stromal structure in the bullous keratopathy cornea identified by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 53(8), 4998–5003 (2012).
[PubMed]

Nagase, S.

Nassif, N.

Nishida, T.

N. Morishige, Y. Takagi, T. Chikama, A. Takahara, and T. Nishida, “Three-dimensional analysis of collagen lamellae in the anterior stroma of the human cornea visualized by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 52(2), 911–915 (2011).
[PubMed]

N. Morishige, A. J. Wahlert, M. C. Kenney, D. J. Brown, K. Kawamoto, T. Chikama, T. Nishida, and J. V. Jester, “Second-harmonic imaging microscopy of normal human and keratoconus cornea,” Invest. Ophthalmol. Vis. Sci. 48(3), 1087–1094 (2007).
[PubMed]

N. Morishige, W. M. Petroll, T. Nishida, M. C. Kenney, and J. V. Jester, “Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals,” J. Cataract Refract. Surg. 32(11), 1784–1791 (2006).
[PubMed]

Oheim, M.

M. Oheim, E. Beaurepaire, E. Chaigneau, J. Mertz, and S. Charpak, “Two-photon microscopy in brain tissue: parameters influencing the imaging depth,” J. Neurosci. Methods 111(1), 29–37 (2001).
[PubMed]

Olivier, N.

F. Aptel, N. Olivier, A. Deniset-Besseau, J.-M. Legeais, K. Plamann, M.-C. Schanne-Klein, and E. Beaurepaire, “Multimodal Nonlinear Imaging of the Human Cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[PubMed]

Oshika, T.

Palacios, R.

Papas, E.

I. Jalbert, F. Stapleton, E. Papas, D. F. Sweeney, and M. Coroneo, “In vivo confocal microscopy of the human cornea,” Br. J. Ophthalmol. 87(2), 225–236 (2003).
[PubMed]

Park, C. Y.

C. Y. Park, J. K. Lee, and R. S. Chuck, “Second Harmonic Generation Imaging Analysis of Collagen Arrangement in Human Cornea,” Invest. Ophthalmol. Vis. Sci. 56(9), 5622–5629 (2015).
[PubMed]

Peli, E.

Peng, J. L.

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

Pennos, A.

Pérez-Merino, P.

J. M. Bueno, E. J. Gualda, A. Giakoumaki, P. Pérez-Merino, S. Marcos, and P. Artal, “Multiphoton microscopy of ex vivo corneas after collagen cross-linking,” Invest. Ophthalmol. Vis. Sci. 52(8), 5325–5331 (2011).
[PubMed]

Petroll, W. M.

N. Morishige, W. M. Petroll, T. Nishida, M. C. Kenney, and J. V. Jester, “Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals,” J. Cataract Refract. Surg. 32(11), 1784–1791 (2006).
[PubMed]

Plamann, K.

F. Aptel, N. Olivier, A. Deniset-Besseau, J.-M. Legeais, K. Plamann, M.-C. Schanne-Klein, and E. Beaurepaire, “Multimodal Nonlinear Imaging of the Human Cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[PubMed]

Ralla, B.

D. Kampik, B. Ralla, S. Keller, M. Hirschberg, P. Friedl, and G. Geerling, “Influence of corneal collagen crosslinking with riboflavin and ultraviolet-a irradiation on excimer laser surgery,” Invest. Ophthalmol. Vis. Sci. 51(8), 3929–3934 (2010).
[PubMed]

Reiser, K. M.

P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, “Polarization-modulated second harmonic generation in collagen,” Biophys. J. 82(6), 3330–3342 (2002).
[PubMed]

Rubenchik, A. M.

P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, “Polarization-modulated second harmonic generation in collagen,” Biophys. J. 82(6), 3330–3342 (2002).
[PubMed]

Sacks, P.

Savage, H.

Schanne-Klein, M.-C.

G. Latour, I. Gusachenko, L. Kowalczuk, I. Lamarre, and M.-C. Schanne-Klein, “In vivo structural imaging of the cornea by polarization-resolved second harmonic microscopy,” Biomed. Opt. Express 3(1), 1–15 (2012).
[PubMed]

F. Aptel, N. Olivier, A. Deniset-Besseau, J.-M. Legeais, K. Plamann, M.-C. Schanne-Klein, and E. Beaurepaire, “Multimodal Nonlinear Imaging of the Human Cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[PubMed]

Schantz, S.

Serrao, S.

G. Lombardo, N. L. Micali, V. Villari, S. Serrao, and M. Lombardo, “All-optical method to assess stromal concentration of riboflavin in conventional and accelerated UV-A irradiation of the human cornea,” Invest. Ophthalmol. Vis. Sci. 57(2), 476–483 (2016).
[PubMed]

Shen, M.

M. Shen, J. Zhao, H. Zeng, and S. Tang, “Calibrating the measurement of wavelength-dependent second harmonic generation from biological tissues with a BaB2O4 crystal,” J. Biomed. Opt. 18(3), 031109 (2013).
[PubMed]

Skorsetz, M.

M. Skorsetz, P. Artal, and J. M. Bueno, “Performance evaluation of a sensorless adaptive optics multiphoton microscope,” J. Microsc. 261(3), 249–258 (2015).

So, P. T.

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

Sonoda, K. H.

N. Morishige, N. Yamada, X. Zhang, Y. Morita, N. Yamada, K. Kimura, A. Takahara, and K. H. Sonoda, “Abnormalities of stromal structure in the bullous keratopathy cornea identified by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 53(8), 4998–5003 (2012).
[PubMed]

Stapleton, F.

I. Jalbert, F. Stapleton, E. Papas, D. F. Sweeney, and M. Coroneo, “In vivo confocal microscopy of the human cornea,” Br. J. Ophthalmol. 87(2), 225–236 (2003).
[PubMed]

Stoller, P.

P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, “Polarization-modulated second harmonic generation in collagen,” Biophys. J. 82(6), 3330–3342 (2002).
[PubMed]

Sun, Y.

H. Y. Tan, Y. Sun, W. Lo, S. J. Lin, C. H. Hsiao, Y. F. Chen, S. C. Huang, W. C. Lin, S. H. Jee, H. S. Yu, and C. Y. Dong, “Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo,” Invest. Ophthalmol. Vis. Sci. 47(12), 5251–5259 (2006).
[PubMed]

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

Sweeney, D. F.

I. Jalbert, F. Stapleton, E. Papas, D. F. Sweeney, and M. Coroneo, “In vivo confocal microscopy of the human cornea,” Br. J. Ophthalmol. 87(2), 225–236 (2003).
[PubMed]

Takagi, Y.

N. Morishige, Y. Takagi, T. Chikama, A. Takahara, and T. Nishida, “Three-dimensional analysis of collagen lamellae in the anterior stroma of the human cornea visualized by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 52(2), 911–915 (2011).
[PubMed]

Takahara, A.

N. Morishige, N. Yamada, X. Zhang, Y. Morita, N. Yamada, K. Kimura, A. Takahara, and K. H. Sonoda, “Abnormalities of stromal structure in the bullous keratopathy cornea identified by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 53(8), 4998–5003 (2012).
[PubMed]

N. Morishige, Y. Takagi, T. Chikama, A. Takahara, and T. Nishida, “Three-dimensional analysis of collagen lamellae in the anterior stroma of the human cornea visualized by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 52(2), 911–915 (2011).
[PubMed]

Tan, H. Y.

W. Lo, W. L. Chen, C. M. Hsueh, A. A. Ghazaryan, S. J. Chen, D. H. K. Ma, C. Y. Dong, and H. Y. Tan, “Fast Fourier transform-based analysis of second-harmonic generation image in keratoconic cornea,” Invest. Ophthalmol. Vis. Sci. 53(7), 3501–3507 (2012).
[PubMed]

C. M. Hsueh, W. Lo, W. L. Chen, V. A. Hovhannisyan, G. Y. Liu, S. S. Wang, H. Y. Tan, and C. Y. Dong, “Structural characterization of edematous corneas by forward and backward second harmonic generation imaging,” Biophys. J. 97(4), 1198–1205 (2009).
[PubMed]

H. Y. Tan, Y. Sun, W. Lo, S. J. Lin, C. H. Hsiao, Y. F. Chen, S. C. Huang, W. C. Lin, S. H. Jee, H. S. Yu, and C. Y. Dong, “Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo,” Invest. Ophthalmol. Vis. Sci. 47(12), 5251–5259 (2006).
[PubMed]

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

Tanaka, R.

Tang, S.

M. Shen, J. Zhao, H. Zeng, and S. Tang, “Calibrating the measurement of wavelength-dependent second harmonic generation from biological tissues with a BaB2O4 crystal,” J. Biomed. Opt. 18(3), 031109 (2013).
[PubMed]

Teng, S. W.

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

Theodossiou, T. A.

T. A. Theodossiou, C. Thrasivoulou, C. Ekwobi, and D. L. Becker, “Second harmonic generation confocal microscopy of collagen type I from rat tendon cryosections,” Biophys. J. 91(12), 4665–4677 (2006).
[PubMed]

Thrasivoulou, C.

T. A. Theodossiou, C. Thrasivoulou, C. Ekwobi, and D. L. Becker, “Second harmonic generation confocal microscopy of collagen type I from rat tendon cryosections,” Biophys. J. 91(12), 4665–4677 (2006).
[PubMed]

Tilbury, K. B.

Tromberg, B. J.

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. U.S.A. 99(17), 11014–11019 (2002).
[PubMed]

A. T. Yeh, N. Nassif, A. Zoumi, and B. J. Tromberg, “Selective corneal imaging using combined second-harmonic generation and two-photon excited fluorescence,” Opt. Lett. 27(23), 2082–2084 (2002).
[PubMed]

Villari, V.

G. Lombardo, N. L. Micali, V. Villari, S. Serrao, and M. Lombardo, “All-optical method to assess stromal concentration of riboflavin in conventional and accelerated UV-A irradiation of the human cornea,” Invest. Ophthalmol. Vis. Sci. 57(2), 476–483 (2016).
[PubMed]

Vohnsen, B.

J. M. Bueno and B. Vohnsen, “Polarimetric high-resolution confocal scanning laser ophthalmoscope,” Vision Res. 45(28), 3526–3534 (2005).
[PubMed]

Wahlert, A. J.

N. Morishige, A. J. Wahlert, M. C. Kenney, D. J. Brown, K. Kawamoto, T. Chikama, T. Nishida, and J. V. Jester, “Second-harmonic imaging microscopy of normal human and keratoconus cornea,” Invest. Ophthalmol. Vis. Sci. 48(3), 1087–1094 (2007).
[PubMed]

Wang, B. G.

B. G. Wang, A. Eitner, J. Lindenau, and K. J. Halbhuber, “High-resolution two-photon excitation microscopy of ocular tissues in porcine eye,” Lasers Surg. Med. 40(4), 247–256 (2008).
[PubMed]

Wang, S. S.

C. M. Hsueh, W. Lo, W. L. Chen, V. A. Hovhannisyan, G. Y. Liu, S. S. Wang, H. Y. Tan, and C. Y. Dong, “Structural characterization of edematous corneas by forward and backward second harmonic generation imaging,” Biophys. J. 97(4), 1198–1205 (2009).
[PubMed]

Webb, W. W.

R. M. Williams, W. R. Zipfel, and W. W. Webb, “Interpreting second-harmonic generation images of collagen I fibrils,” Biophys. J. 88(2), 1377–1386 (2005).
[PubMed]

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[PubMed]

Williams, R. M.

R. M. Williams, W. R. Zipfel, and W. W. Webb, “Interpreting second-harmonic generation images of collagen I fibrils,” Biophys. J. 88(2), 1377–1386 (2005).
[PubMed]

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[PubMed]

Yamada, N.

N. Morishige, N. Yamada, X. Zhang, Y. Morita, N. Yamada, K. Kimura, A. Takahara, and K. H. Sonoda, “Abnormalities of stromal structure in the bullous keratopathy cornea identified by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 53(8), 4998–5003 (2012).
[PubMed]

N. Morishige, N. Yamada, X. Zhang, Y. Morita, N. Yamada, K. Kimura, A. Takahara, and K. H. Sonoda, “Abnormalities of stromal structure in the bullous keratopathy cornea identified by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 53(8), 4998–5003 (2012).
[PubMed]

Yamanari, M.

Yasui, T.

Yasuno, Y.

Yeh, A.

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. U.S.A. 99(17), 11014–11019 (2002).
[PubMed]

Yeh, A. T.

Yu, H. S.

H. Y. Tan, Y. Sun, W. Lo, S. J. Lin, C. H. Hsiao, Y. F. Chen, S. C. Huang, W. C. Lin, S. H. Jee, H. S. Yu, and C. Y. Dong, “Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo,” Invest. Ophthalmol. Vis. Sci. 47(12), 5251–5259 (2006).
[PubMed]

Zeng, H.

M. Shen, J. Zhao, H. Zeng, and S. Tang, “Calibrating the measurement of wavelength-dependent second harmonic generation from biological tissues with a BaB2O4 crystal,” J. Biomed. Opt. 18(3), 031109 (2013).
[PubMed]

Zhadin, N.

Zhang, X.

N. Morishige, N. Yamada, X. Zhang, Y. Morita, N. Yamada, K. Kimura, A. Takahara, and K. H. Sonoda, “Abnormalities of stromal structure in the bullous keratopathy cornea identified by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 53(8), 4998–5003 (2012).
[PubMed]

Zhao, J.

M. Shen, J. Zhao, H. Zeng, and S. Tang, “Calibrating the measurement of wavelength-dependent second harmonic generation from biological tissues with a BaB2O4 crystal,” J. Biomed. Opt. 18(3), 031109 (2013).
[PubMed]

Zipfel, W. R.

R. M. Williams, W. R. Zipfel, and W. W. Webb, “Interpreting second-harmonic generation images of collagen I fibrils,” Biophys. J. 88(2), 1377–1386 (2005).
[PubMed]

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[PubMed]

Zoumi, A.

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. U.S.A. 99(17), 11014–11019 (2002).
[PubMed]

A. T. Yeh, N. Nassif, A. Zoumi, and B. J. Tromberg, “Selective corneal imaging using combined second-harmonic generation and two-photon excited fluorescence,” Opt. Lett. 27(23), 2082–2084 (2002).
[PubMed]

Appl. Opt. (2)

Biomed. Opt. Express (4)

Biophys. J. (4)

R. M. Williams, W. R. Zipfel, and W. W. Webb, “Interpreting second-harmonic generation images of collagen I fibrils,” Biophys. J. 88(2), 1377–1386 (2005).
[PubMed]

C. M. Hsueh, W. Lo, W. L. Chen, V. A. Hovhannisyan, G. Y. Liu, S. S. Wang, H. Y. Tan, and C. Y. Dong, “Structural characterization of edematous corneas by forward and backward second harmonic generation imaging,” Biophys. J. 97(4), 1198–1205 (2009).
[PubMed]

T. A. Theodossiou, C. Thrasivoulou, C. Ekwobi, and D. L. Becker, “Second harmonic generation confocal microscopy of collagen type I from rat tendon cryosections,” Biophys. J. 91(12), 4665–4677 (2006).
[PubMed]

P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, “Polarization-modulated second harmonic generation in collagen,” Biophys. J. 82(6), 3330–3342 (2002).
[PubMed]

Br. J. Ophthalmol. (1)

I. Jalbert, F. Stapleton, E. Papas, D. F. Sweeney, and M. Coroneo, “In vivo confocal microscopy of the human cornea,” Br. J. Ophthalmol. 87(2), 225–236 (2003).
[PubMed]

Cornea (1)

J. M. Bueno, E. J. Gualda, and P. Artal, “Analysis of corneal stroma organization with wavefront optimized nonlinear microscopy,” Cornea 30(6), 692–701 (2011).
[PubMed]

Invest. Ophthalmol. Vis. Sci. (12)

C. Y. Park, J. K. Lee, and R. S. Chuck, “Second Harmonic Generation Imaging Analysis of Collagen Arrangement in Human Cornea,” Invest. Ophthalmol. Vis. Sci. 56(9), 5622–5629 (2015).
[PubMed]

N. Morishige, Y. Takagi, T. Chikama, A. Takahara, and T. Nishida, “Three-dimensional analysis of collagen lamellae in the anterior stroma of the human cornea visualized by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 52(2), 911–915 (2011).
[PubMed]

J. M. Bueno, E. J. Gualda, A. Giakoumaki, P. Pérez-Merino, S. Marcos, and P. Artal, “Multiphoton microscopy of ex vivo corneas after collagen cross-linking,” Invest. Ophthalmol. Vis. Sci. 52(8), 5325–5331 (2011).
[PubMed]

F. Aptel, N. Olivier, A. Deniset-Besseau, J.-M. Legeais, K. Plamann, M.-C. Schanne-Klein, and E. Beaurepaire, “Multimodal Nonlinear Imaging of the Human Cornea,” Invest. Ophthalmol. Vis. Sci. 51(5), 2459–2465 (2010).
[PubMed]

D. Kampik, B. Ralla, S. Keller, M. Hirschberg, P. Friedl, and G. Geerling, “Influence of corneal collagen crosslinking with riboflavin and ultraviolet-a irradiation on excimer laser surgery,” Invest. Ophthalmol. Vis. Sci. 51(8), 3929–3934 (2010).
[PubMed]

G. Lombardo, N. L. Micali, V. Villari, S. Serrao, and M. Lombardo, “All-optical method to assess stromal concentration of riboflavin in conventional and accelerated UV-A irradiation of the human cornea,” Invest. Ophthalmol. Vis. Sci. 57(2), 476–483 (2016).
[PubMed]

N. Morishige, N. Yamada, X. Zhang, Y. Morita, N. Yamada, K. Kimura, A. Takahara, and K. H. Sonoda, “Abnormalities of stromal structure in the bullous keratopathy cornea identified by second harmonic generation imaging microscopy,” Invest. Ophthalmol. Vis. Sci. 53(8), 4998–5003 (2012).
[PubMed]

W. Lo, W. L. Chen, C. M. Hsueh, A. A. Ghazaryan, S. J. Chen, D. H. K. Ma, C. Y. Dong, and H. Y. Tan, “Fast Fourier transform-based analysis of second-harmonic generation image in keratoconic cornea,” Invest. Ophthalmol. Vis. Sci. 53(7), 3501–3507 (2012).
[PubMed]

F. J. Ávila, J. M. Bueno, A. Gambín, and P. Artal, “Second harmonic generation microscopy of the human cornea and sclera in vivo,” Invest. Ophthalmol. Vis. Sci. 58, 3109 (2017).

H. Y. Tan, Y. Sun, W. Lo, S. J. Lin, C. H. Hsiao, Y. F. Chen, S. C. Huang, W. C. Lin, S. H. Jee, H. S. Yu, and C. Y. Dong, “Multiphoton fluorescence and second harmonic generation imaging of the structural alterations in keratoconus ex vivo,” Invest. Ophthalmol. Vis. Sci. 47(12), 5251–5259 (2006).
[PubMed]

N. Morishige, A. J. Wahlert, M. C. Kenney, D. J. Brown, K. Kawamoto, T. Chikama, T. Nishida, and J. V. Jester, “Second-harmonic imaging microscopy of normal human and keratoconus cornea,” Invest. Ophthalmol. Vis. Sci. 48(3), 1087–1094 (2007).
[PubMed]

S. W. Teng, H. Y. Tan, J. L. Peng, H. H. Lin, K. H. Kim, W. Lo, Y. Sun, W. C. Lin, S. J. Lin, S. H. Jee, P. T. So, and C. Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci. 47(3), 1216–1224 (2006).
[PubMed]

J. Biomed. Opt. (4)

P. J. Campagnola, H. A. Clark, W. A. Mohler, A. Lewis, and L. M. Loew, “Second-harmonic imaging microscopy of living cells,” J. Biomed. Opt. 6(3), 277–286 (2001).
[PubMed]

F. J. Ávila, O. Del Barco, and J. M. Bueno, “Polarization dependence of aligned collagen tissues imaged with second harmonic generation microscopy,” J. Biomed. Opt. 20(8), 86001 (2015).
[PubMed]

M. Shen, J. Zhao, H. Zeng, and S. Tang, “Calibrating the measurement of wavelength-dependent second harmonic generation from biological tissues with a BaB2O4 crystal,” J. Biomed. Opt. 18(3), 031109 (2013).
[PubMed]

J. M. Bueno, E. J. Gualda, and P. Artal, “Adaptive optics multiphoton microscopy to study ex vivo ocular tissues,” J. Biomed. Opt. 15(6), 066004 (2010).
[PubMed]

J. Cataract Refract. Surg. (1)

N. Morishige, W. M. Petroll, T. Nishida, M. C. Kenney, and J. V. Jester, “Noninvasive corneal stromal collagen imaging using two-photon-generated second-harmonic signals,” J. Cataract Refract. Surg. 32(11), 1784–1791 (2006).
[PubMed]

J. Microsc. (1)

M. Skorsetz, P. Artal, and J. M. Bueno, “Performance evaluation of a sensorless adaptive optics multiphoton microscope,” J. Microsc. 261(3), 249–258 (2015).

J. Neurosci. Methods (1)

M. Oheim, E. Beaurepaire, E. Chaigneau, J. Mertz, and S. Charpak, “Two-photon microscopy in brain tissue: parameters influencing the imaging depth,” J. Neurosci. Methods 111(1), 29–37 (2001).
[PubMed]

J. Opt. (1)

F. J. Ávila, O. del Barco, and J. M. Bueno, “Quantifying external and internal collagen organization from Stokes-vector-based second harmonic generation imaging polarimetry,” J. Opt. 19(10), 105301 (2017).

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

Lasers Surg. Med. (1)

B. G. Wang, A. Eitner, J. Lindenau, and K. J. Halbhuber, “High-resolution two-photon excitation microscopy of ocular tissues in porcine eye,” Lasers Surg. Med. 40(4), 247–256 (2008).
[PubMed]

Nat. Biotechnol. (1)

W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003).
[PubMed]

Opt. Express (1)

Opt. Lett. (4)

Proc. Natl. Acad. Sci. U.S.A. (1)

A. Zoumi, A. Yeh, and B. J. Tromberg, “Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence,” Proc. Natl. Acad. Sci. U.S.A. 99(17), 11014–11019 (2002).
[PubMed]

Vision Res. (1)

J. M. Bueno and B. Vohnsen, “Polarimetric high-resolution confocal scanning laser ophthalmoscope,” Vision Res. 45(28), 3526–3534 (2005).
[PubMed]

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Figures (8)

Fig. 1
Fig. 1 Schematic diagram of the SHG imaging microscope. FM, flip mirror; LP, linear polarizer; NDF, variable neutral density filter; AP, aperture; PMT, photomultiplier.
Fig. 2
Fig. 2 Auto-scaled SHG images of a human cornea (140-μm depth, upper panels and a bovine sclera (175-μm depth, bottom panels) for 800 nm (a, d) and 1045 nm (b, e). (c, f) 1045-nm SHG images with the same color scale as (a, d). The imaged plane was randomly chosen. Scale bar: 50 μm. (g, h) SHG intensity profiles along the dashed lines indicated in Fig. 2(a) and 2(b) bovine sclera. Red line, 800 nm; blue line, 1045 nm.
Fig. 3
Fig. 3 Histograms of PO corresponding to SHG images recorded with 800 (a, c) and 1045 nm (b, d). The samples were the same as in Fig. 2: human cornea (upper plots) and bovine sclera (bottom plots).
Fig. 4
Fig. 4 SHG images of the corneal stroma of an eagle eye at different depth locations recorded with 800 (upper panels) and 1045 nm (bottom panels).
Fig. 5
Fig. 5 PO of the collagen fibers computed with the structure tensor as a function of depth for 800 (red symbols) and 1045 nm (blue symbols) in an eagle cornea. Errors bars indicate the SD.
Fig. 6
Fig. 6 Normalized SHG intensity profiles as a function of depth for 800 (red) and 1045 (blue) nm. SHG images were spaced 25 μm in depth. Data correspond to an eagle (a) and a human cornea (b). The insets correspond to the (non-normalized) raw data.
Fig. 7
Fig. 7 Map of direct comparison of SHG normalized intensity values for two wavelengths as a function of the sample’s depth.
Fig. 8
Fig. 8 Values of Michelson contrast of SHG images of a porcine cornea at different depth locations for 800 (red dots) and 1045 nm (blue dots). Images correspond to the designated locations. Bar length: 50 μm.

Tables (1)

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Table 1 Mean values of Michelson contrast of SHG images of two corneas as a function of depth (anterior, medium and posterior stroma) for the two different wavelengths.

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