Abstract

Collagen, as the most abundant protein in the human body, determines the unique physiological and optical properties of the connective tissues including cornea and sclera. The ultrastructure of collagen, which conventionally can only be resolved by electron microscopy, now can be probed by optical second harmonic generation (SHG) imaging. SHG imaging revealed that corneal collagen fibrils are regularly packed as a polycrystalline lattice, accounting for the transparency of cornea. In contrast, scleral fibrils possess inhomogeneous, tubelike structures with thin hard shells, maintaining the high stiffness and elasticity of the sclera.

© 2005 Optical Society of America

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  1. Y. Komai and T. Ushiki, “The three-dimensional organization of collagen fibrils in the human cornea and sclera,”, Invest. Ophthalmol. Vis. Sci. 32, 2244–2258 (1991).
    [PubMed]
  2. C. Boote, S. Dennis, R.H. Newton, H. Puri, and K.M. Meek, “Collagen fibrils appear more closely packed in the prepupillary cornea: optical and biomechanical implications,” Invest. Ophthalmol. Vis. Sci. 44, 2941–2948 (2003).
    [Crossref] [PubMed]
  3. W. Denk, J.H. Strickler, and W.W. Webb, “Two-Photon Laser Scanning Fluoresence Microscopy,” Science 248, 73–76 (1990)
    [Crossref] [PubMed]
  4. W.R. Zipfel, R.M. Williams, and W.W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nature Biotech. 21, 1369–1377 (2003)
    [Crossref]
  5. A. Zumbusch, G.R. Holtom, and & X.S. Xie, “Three-Dimensional Vibrational Imaging by Coherent Anti-Stokes Raman Scattering,” Phys. Rev. Lett. 82, 4142–4145 (1999).
    [Crossref]
  6. S. Roth and I. Freund, “Coherent Optical Harmonic Generation in Rat-tail,” Opt. Commun. 33, 292–296 (1980)
    [Crossref]
  7. I. Freund, M. Deutsch, and A. Sprecher, “Connective Tissue Polarity, Optical second-harmonic microscopy, crossed-beam summation, and small-angle scattering in rat-tail tendon,” Biophy. J. 50, 693–712 (1986)
    [Crossref]
  8. 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, 277–286 (2001)
    [Crossref] [PubMed]
  9. A.T. Yeh, N. Nassif, A. Zoumi, and B.J. Tromberg, “Selective corneal imaging using combined second-harmonic generation and two-photon excited f luorescence,” Opt. Lett. 27, 2082–2084 (2002)
    [Crossref]
  10. P. J. Campagnola, H.A. Clark, W.A. Mohler, A. Lewis, and L.M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nature Biotech. 21, 1356–1360 (2003)
    [Crossref]
  11. G. Cox, E. Kable, A. Jones, I. Fraser, F. Manconi, and M. D. Gorrell, “3-Dimensional Imaging of Collagen Using Second Harmonic Generation,” J. Struct. Bio.,  141, 53–62 (2003)
    [Crossref]
  12. M. Han, L. Zickler, G. Giese, F. Loesel, M. Walter, and J. Bille, “Second Harmonic Corneal Imaging after femtosecond laser surgery”, J. Biomed. Opt, 9, 760–766 (2004)
    [Crossref]
  13. M. Han, G. Giese, L. Zickler, H. Sun, and J. F. Bille, “Mini-invasive corneal surgery and imaging with femtosecond lasers,” Opt. Express 12, 4275–4281 (2004), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-18-4275
    [Crossref] [PubMed]
  14. L. Moreaux, O. Sandre, and J. Mertz, “Membrane imaging by second-harmonic generation microscopy,” J. Opt. Soc. Am B 17, 1685–1694 (2000)
    [Crossref]
  15. J. Mertz and L. Moreaux, “Second-harmonic generation by focused excitation of inhomogeneously distributed scatterers,” Opt. Commun. 196, 325–330 (2001)
    [Crossref]
  16. J. 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]
  17. R.M. Williams, W.R. Zipfel, and W.W. Webb, “Interpreting Second-Harmonic Generation Images of Collagen I Fibrils,” Biophy. J. 88, 1377–1386 (2005)
    [Crossref]
  18. D.M. Maurice, “The structure and transparency of the cornea,” J. Physiol. 136, 263–286 (1957).
    [PubMed]
  19. R.W. Hart and & R.A. Farrell, “Light Scattering in the Cornea,” J. Opt. Soc. Am 59, 766–774 (1969).
    [Crossref] [PubMed]
  20. G.B. Benedek, “Theory of transparency of the eye,” Appl. Opt. 10, 459–473 (1971).
    [Crossref] [PubMed]
  21. T. Gutsmann, G.E. Fantner, and M. Venturoni, et al. “Evidence that Collagen Fibrils in Tendons Are Inhomoge-neously Structured in a Tubelike Manner,” Biophys. J. 84, 2593–2598 (2003).
    [Crossref] [PubMed]

2005 (1)

R.M. Williams, W.R. Zipfel, and W.W. Webb, “Interpreting Second-Harmonic Generation Images of Collagen I Fibrils,” Biophy. J. 88, 1377–1386 (2005)
[Crossref]

2004 (2)

M. Han, L. Zickler, G. Giese, F. Loesel, M. Walter, and J. Bille, “Second Harmonic Corneal Imaging after femtosecond laser surgery”, J. Biomed. Opt, 9, 760–766 (2004)
[Crossref]

M. Han, G. Giese, L. Zickler, H. Sun, and J. F. Bille, “Mini-invasive corneal surgery and imaging with femtosecond lasers,” Opt. Express 12, 4275–4281 (2004), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-18-4275
[Crossref] [PubMed]

2003 (5)

P. J. Campagnola, H.A. Clark, W.A. Mohler, A. Lewis, and L.M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nature Biotech. 21, 1356–1360 (2003)
[Crossref]

G. Cox, E. Kable, A. Jones, I. Fraser, F. Manconi, and M. D. Gorrell, “3-Dimensional Imaging of Collagen Using Second Harmonic Generation,” J. Struct. Bio.,  141, 53–62 (2003)
[Crossref]

C. Boote, S. Dennis, R.H. Newton, H. Puri, and K.M. Meek, “Collagen fibrils appear more closely packed in the prepupillary cornea: optical and biomechanical implications,” Invest. Ophthalmol. Vis. Sci. 44, 2941–2948 (2003).
[Crossref] [PubMed]

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

T. Gutsmann, G.E. Fantner, and M. Venturoni, et al. “Evidence that Collagen Fibrils in Tendons Are Inhomoge-neously Structured in a Tubelike Manner,” Biophys. J. 84, 2593–2598 (2003).
[Crossref] [PubMed]

2002 (2)

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

J. 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]

2001 (2)

J. Mertz and L. Moreaux, “Second-harmonic generation by focused excitation of inhomogeneously distributed scatterers,” Opt. Commun. 196, 325–330 (2001)
[Crossref]

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, 277–286 (2001)
[Crossref] [PubMed]

2000 (1)

L. Moreaux, O. Sandre, and J. Mertz, “Membrane imaging by second-harmonic generation microscopy,” J. Opt. Soc. Am B 17, 1685–1694 (2000)
[Crossref]

1999 (1)

A. Zumbusch, G.R. Holtom, and & X.S. Xie, “Three-Dimensional Vibrational Imaging by Coherent Anti-Stokes Raman Scattering,” Phys. Rev. Lett. 82, 4142–4145 (1999).
[Crossref]

1991 (1)

Y. Komai and T. Ushiki, “The three-dimensional organization of collagen fibrils in the human cornea and sclera,”, Invest. Ophthalmol. Vis. Sci. 32, 2244–2258 (1991).
[PubMed]

1990 (1)

W. Denk, J.H. Strickler, and W.W. Webb, “Two-Photon Laser Scanning Fluoresence Microscopy,” Science 248, 73–76 (1990)
[Crossref] [PubMed]

1986 (1)

I. Freund, M. Deutsch, and A. Sprecher, “Connective Tissue Polarity, Optical second-harmonic microscopy, crossed-beam summation, and small-angle scattering in rat-tail tendon,” Biophy. J. 50, 693–712 (1986)
[Crossref]

1980 (1)

S. Roth and I. Freund, “Coherent Optical Harmonic Generation in Rat-tail,” Opt. Commun. 33, 292–296 (1980)
[Crossref]

1971 (1)

1969 (1)

R.W. Hart and & R.A. Farrell, “Light Scattering in the Cornea,” J. Opt. Soc. Am 59, 766–774 (1969).
[Crossref] [PubMed]

1957 (1)

D.M. Maurice, “The structure and transparency of the cornea,” J. Physiol. 136, 263–286 (1957).
[PubMed]

Benedek, G.B.

Bille, J.

M. Han, L. Zickler, G. Giese, F. Loesel, M. Walter, and J. Bille, “Second Harmonic Corneal Imaging after femtosecond laser surgery”, J. Biomed. Opt, 9, 760–766 (2004)
[Crossref]

Bille, J. F.

Boote, C.

C. Boote, S. Dennis, R.H. Newton, H. Puri, and K.M. Meek, “Collagen fibrils appear more closely packed in the prepupillary cornea: optical and biomechanical implications,” Invest. Ophthalmol. Vis. Sci. 44, 2941–2948 (2003).
[Crossref] [PubMed]

Campagnola, P. J.

P. J. Campagnola, H.A. Clark, W.A. Mohler, A. Lewis, and L.M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nature Biotech. 21, 1356–1360 (2003)
[Crossref]

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, 277–286 (2001)
[Crossref] [PubMed]

Cheng, J.

J. 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]

Clark, H.A.

P. J. Campagnola, H.A. Clark, W.A. Mohler, A. Lewis, and L.M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nature Biotech. 21, 1356–1360 (2003)
[Crossref]

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, 277–286 (2001)
[Crossref] [PubMed]

Cox, G.

G. Cox, E. Kable, A. Jones, I. Fraser, F. Manconi, and M. D. Gorrell, “3-Dimensional Imaging of Collagen Using Second Harmonic Generation,” J. Struct. Bio.,  141, 53–62 (2003)
[Crossref]

Denk, W.

W. Denk, J.H. Strickler, and W.W. Webb, “Two-Photon Laser Scanning Fluoresence Microscopy,” Science 248, 73–76 (1990)
[Crossref] [PubMed]

Dennis, S.

C. Boote, S. Dennis, R.H. Newton, H. Puri, and K.M. Meek, “Collagen fibrils appear more closely packed in the prepupillary cornea: optical and biomechanical implications,” Invest. Ophthalmol. Vis. Sci. 44, 2941–2948 (2003).
[Crossref] [PubMed]

Deutsch, M.

I. Freund, M. Deutsch, and A. Sprecher, “Connective Tissue Polarity, Optical second-harmonic microscopy, crossed-beam summation, and small-angle scattering in rat-tail tendon,” Biophy. J. 50, 693–712 (1986)
[Crossref]

Fantner, G.E.

T. Gutsmann, G.E. Fantner, and M. Venturoni, et al. “Evidence that Collagen Fibrils in Tendons Are Inhomoge-neously Structured in a Tubelike Manner,” Biophys. J. 84, 2593–2598 (2003).
[Crossref] [PubMed]

Farrell, & R.A.

R.W. Hart and & R.A. Farrell, “Light Scattering in the Cornea,” J. Opt. Soc. Am 59, 766–774 (1969).
[Crossref] [PubMed]

Fraser, I.

G. Cox, E. Kable, A. Jones, I. Fraser, F. Manconi, and M. D. Gorrell, “3-Dimensional Imaging of Collagen Using Second Harmonic Generation,” J. Struct. Bio.,  141, 53–62 (2003)
[Crossref]

Freund, I.

I. Freund, M. Deutsch, and A. Sprecher, “Connective Tissue Polarity, Optical second-harmonic microscopy, crossed-beam summation, and small-angle scattering in rat-tail tendon,” Biophy. J. 50, 693–712 (1986)
[Crossref]

S. Roth and I. Freund, “Coherent Optical Harmonic Generation in Rat-tail,” Opt. Commun. 33, 292–296 (1980)
[Crossref]

Giese, G.

M. Han, L. Zickler, G. Giese, F. Loesel, M. Walter, and J. Bille, “Second Harmonic Corneal Imaging after femtosecond laser surgery”, J. Biomed. Opt, 9, 760–766 (2004)
[Crossref]

M. Han, G. Giese, L. Zickler, H. Sun, and J. F. Bille, “Mini-invasive corneal surgery and imaging with femtosecond lasers,” Opt. Express 12, 4275–4281 (2004), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-18-4275
[Crossref] [PubMed]

Gorrell, M. D.

G. Cox, E. Kable, A. Jones, I. Fraser, F. Manconi, and M. D. Gorrell, “3-Dimensional Imaging of Collagen Using Second Harmonic Generation,” J. Struct. Bio.,  141, 53–62 (2003)
[Crossref]

Gutsmann, T.

T. Gutsmann, G.E. Fantner, and M. Venturoni, et al. “Evidence that Collagen Fibrils in Tendons Are Inhomoge-neously Structured in a Tubelike Manner,” Biophys. J. 84, 2593–2598 (2003).
[Crossref] [PubMed]

Han, M.

M. Han, L. Zickler, G. Giese, F. Loesel, M. Walter, and J. Bille, “Second Harmonic Corneal Imaging after femtosecond laser surgery”, J. Biomed. Opt, 9, 760–766 (2004)
[Crossref]

M. Han, G. Giese, L. Zickler, H. Sun, and J. F. Bille, “Mini-invasive corneal surgery and imaging with femtosecond lasers,” Opt. Express 12, 4275–4281 (2004), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-18-4275
[Crossref] [PubMed]

Hart, R.W.

R.W. Hart and & R.A. Farrell, “Light Scattering in the Cornea,” J. Opt. Soc. Am 59, 766–774 (1969).
[Crossref] [PubMed]

Holtom, G.R.

A. Zumbusch, G.R. Holtom, and & X.S. Xie, “Three-Dimensional Vibrational Imaging by Coherent Anti-Stokes Raman Scattering,” Phys. Rev. Lett. 82, 4142–4145 (1999).
[Crossref]

Jones, A.

G. Cox, E. Kable, A. Jones, I. Fraser, F. Manconi, and M. D. Gorrell, “3-Dimensional Imaging of Collagen Using Second Harmonic Generation,” J. Struct. Bio.,  141, 53–62 (2003)
[Crossref]

Kable, E.

G. Cox, E. Kable, A. Jones, I. Fraser, F. Manconi, and M. D. Gorrell, “3-Dimensional Imaging of Collagen Using Second Harmonic Generation,” J. Struct. Bio.,  141, 53–62 (2003)
[Crossref]

Komai, Y.

Y. Komai and T. Ushiki, “The three-dimensional organization of collagen fibrils in the human cornea and sclera,”, Invest. Ophthalmol. Vis. Sci. 32, 2244–2258 (1991).
[PubMed]

Lewis, A.

P. J. Campagnola, H.A. Clark, W.A. Mohler, A. Lewis, and L.M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nature Biotech. 21, 1356–1360 (2003)
[Crossref]

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, 277–286 (2001)
[Crossref] [PubMed]

Loesel, F.

M. Han, L. Zickler, G. Giese, F. Loesel, M. Walter, and J. Bille, “Second Harmonic Corneal Imaging after femtosecond laser surgery”, J. Biomed. Opt, 9, 760–766 (2004)
[Crossref]

Loew, L.M.

P. J. Campagnola, H.A. Clark, W.A. Mohler, A. Lewis, and L.M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nature Biotech. 21, 1356–1360 (2003)
[Crossref]

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, 277–286 (2001)
[Crossref] [PubMed]

Manconi, F.

G. Cox, E. Kable, A. Jones, I. Fraser, F. Manconi, and M. D. Gorrell, “3-Dimensional Imaging of Collagen Using Second Harmonic Generation,” J. Struct. Bio.,  141, 53–62 (2003)
[Crossref]

Maurice, D.M.

D.M. Maurice, “The structure and transparency of the cornea,” J. Physiol. 136, 263–286 (1957).
[PubMed]

Meek, K.M.

C. Boote, S. Dennis, R.H. Newton, H. Puri, and K.M. Meek, “Collagen fibrils appear more closely packed in the prepupillary cornea: optical and biomechanical implications,” Invest. Ophthalmol. Vis. Sci. 44, 2941–2948 (2003).
[Crossref] [PubMed]

Mertz, J.

J. Mertz and L. Moreaux, “Second-harmonic generation by focused excitation of inhomogeneously distributed scatterers,” Opt. Commun. 196, 325–330 (2001)
[Crossref]

L. Moreaux, O. Sandre, and J. Mertz, “Membrane imaging by second-harmonic generation microscopy,” J. Opt. Soc. Am B 17, 1685–1694 (2000)
[Crossref]

Mohler, W.A.

P. J. Campagnola, H.A. Clark, W.A. Mohler, A. Lewis, and L.M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nature Biotech. 21, 1356–1360 (2003)
[Crossref]

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, 277–286 (2001)
[Crossref] [PubMed]

Moreaux, L.

J. Mertz and L. Moreaux, “Second-harmonic generation by focused excitation of inhomogeneously distributed scatterers,” Opt. Commun. 196, 325–330 (2001)
[Crossref]

L. Moreaux, O. Sandre, and J. Mertz, “Membrane imaging by second-harmonic generation microscopy,” J. Opt. Soc. Am B 17, 1685–1694 (2000)
[Crossref]

Nassif, N.

Newton, R.H.

C. Boote, S. Dennis, R.H. Newton, H. Puri, and K.M. Meek, “Collagen fibrils appear more closely packed in the prepupillary cornea: optical and biomechanical implications,” Invest. Ophthalmol. Vis. Sci. 44, 2941–2948 (2003).
[Crossref] [PubMed]

Puri, H.

C. Boote, S. Dennis, R.H. Newton, H. Puri, and K.M. Meek, “Collagen fibrils appear more closely packed in the prepupillary cornea: optical and biomechanical implications,” Invest. Ophthalmol. Vis. Sci. 44, 2941–2948 (2003).
[Crossref] [PubMed]

Roth, S.

S. Roth and I. Freund, “Coherent Optical Harmonic Generation in Rat-tail,” Opt. Commun. 33, 292–296 (1980)
[Crossref]

Sandre, O.

L. Moreaux, O. Sandre, and J. Mertz, “Membrane imaging by second-harmonic generation microscopy,” J. Opt. Soc. Am B 17, 1685–1694 (2000)
[Crossref]

Sprecher, A.

I. Freund, M. Deutsch, and A. Sprecher, “Connective Tissue Polarity, Optical second-harmonic microscopy, crossed-beam summation, and small-angle scattering in rat-tail tendon,” Biophy. J. 50, 693–712 (1986)
[Crossref]

Strickler, J.H.

W. Denk, J.H. Strickler, and W.W. Webb, “Two-Photon Laser Scanning Fluoresence Microscopy,” Science 248, 73–76 (1990)
[Crossref] [PubMed]

Sun, H.

Tromberg, B.J.

Ushiki, T.

Y. Komai and T. Ushiki, “The three-dimensional organization of collagen fibrils in the human cornea and sclera,”, Invest. Ophthalmol. Vis. Sci. 32, 2244–2258 (1991).
[PubMed]

Venturoni, M.

T. Gutsmann, G.E. Fantner, and M. Venturoni, et al. “Evidence that Collagen Fibrils in Tendons Are Inhomoge-neously Structured in a Tubelike Manner,” Biophys. J. 84, 2593–2598 (2003).
[Crossref] [PubMed]

Volkmer, A.

J. 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]

Walter, M.

M. Han, L. Zickler, G. Giese, F. Loesel, M. Walter, and J. Bille, “Second Harmonic Corneal Imaging after femtosecond laser surgery”, J. Biomed. Opt, 9, 760–766 (2004)
[Crossref]

Webb, W.W.

R.M. Williams, W.R. Zipfel, and W.W. Webb, “Interpreting Second-Harmonic Generation Images of Collagen I Fibrils,” Biophy. J. 88, 1377–1386 (2005)
[Crossref]

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

W. Denk, J.H. Strickler, and W.W. Webb, “Two-Photon Laser Scanning Fluoresence Microscopy,” Science 248, 73–76 (1990)
[Crossref] [PubMed]

Williams, R.M.

R.M. Williams, W.R. Zipfel, and W.W. Webb, “Interpreting Second-Harmonic Generation Images of Collagen I Fibrils,” Biophy. J. 88, 1377–1386 (2005)
[Crossref]

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

Xie, & X.S.

A. Zumbusch, G.R. Holtom, and & X.S. Xie, “Three-Dimensional Vibrational Imaging by Coherent Anti-Stokes Raman Scattering,” Phys. Rev. Lett. 82, 4142–4145 (1999).
[Crossref]

Xie, X.S.

J. 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]

Yeh, A.T.

Zickler, L.

M. Han, L. Zickler, G. Giese, F. Loesel, M. Walter, and J. Bille, “Second Harmonic Corneal Imaging after femtosecond laser surgery”, J. Biomed. Opt, 9, 760–766 (2004)
[Crossref]

M. Han, G. Giese, L. Zickler, H. Sun, and J. F. Bille, “Mini-invasive corneal surgery and imaging with femtosecond lasers,” Opt. Express 12, 4275–4281 (2004), http://www.opticsexpress.org/abstract.cfm?URI=OPEX-12-18-4275
[Crossref] [PubMed]

Zipfel, W.R.

R.M. Williams, W.R. Zipfel, and W.W. Webb, “Interpreting Second-Harmonic Generation Images of Collagen I Fibrils,” Biophy. J. 88, 1377–1386 (2005)
[Crossref]

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

Zoumi, A.

Zumbusch, A.

A. Zumbusch, G.R. Holtom, and & X.S. Xie, “Three-Dimensional Vibrational Imaging by Coherent Anti-Stokes Raman Scattering,” Phys. Rev. Lett. 82, 4142–4145 (1999).
[Crossref]

Appl. Opt. (1)

Biophy. J. (2)

I. Freund, M. Deutsch, and A. Sprecher, “Connective Tissue Polarity, Optical second-harmonic microscopy, crossed-beam summation, and small-angle scattering in rat-tail tendon,” Biophy. J. 50, 693–712 (1986)
[Crossref]

R.M. Williams, W.R. Zipfel, and W.W. Webb, “Interpreting Second-Harmonic Generation Images of Collagen I Fibrils,” Biophy. J. 88, 1377–1386 (2005)
[Crossref]

Biophys. J. (1)

T. Gutsmann, G.E. Fantner, and M. Venturoni, et al. “Evidence that Collagen Fibrils in Tendons Are Inhomoge-neously Structured in a Tubelike Manner,” Biophys. J. 84, 2593–2598 (2003).
[Crossref] [PubMed]

Invest. Ophthalmol. Vis. Sci. (2)

Y. Komai and T. Ushiki, “The three-dimensional organization of collagen fibrils in the human cornea and sclera,”, Invest. Ophthalmol. Vis. Sci. 32, 2244–2258 (1991).
[PubMed]

C. Boote, S. Dennis, R.H. Newton, H. Puri, and K.M. Meek, “Collagen fibrils appear more closely packed in the prepupillary cornea: optical and biomechanical implications,” Invest. Ophthalmol. Vis. Sci. 44, 2941–2948 (2003).
[Crossref] [PubMed]

J. Biomed. Opt, (1)

M. Han, L. Zickler, G. Giese, F. Loesel, M. Walter, and J. Bille, “Second Harmonic Corneal Imaging after femtosecond laser surgery”, J. Biomed. Opt, 9, 760–766 (2004)
[Crossref]

J. Biomed. Opt. (1)

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Figures (5)

Fig. 1.
Fig. 1.

Schematic drawing of the SHG imaging experimental setup

Fig. 2.
Fig. 2.

SHG imaging of collagen fibrils in (a)cornea and (b)sclera. The femtosecond Ti:Sapphire laser was focused by a 40×(N.A. 0.8) water immersion objective. SHG signals were collected in the forward direction for both cornea and sclera. The image plan is parallel to the cornea/sclera surface. Bars: 20 μm

Fig. 3.
Fig. 3.

SHG imaging of corneal collagen fibrils in (a)forward and (b)backward directions. The fibrillar structures resolved in (a) correspond to collagen bundles which are composed of regularly packed collagen fibrils. Objective: 63×/1.0W, Bars: 10 μm

Fig. 4.
Fig. 4.

SHG imaging of scleral collagen fibrils in (a)forward and (b)backward directions. In contrast to cornea, the backward SHG signals from sclera are significant. Identical structures are revealed by forward and backward SHG imaging. Objective: 63×/1.0W, Bars: 10 μm

Fig. 5.
Fig. 5.

SHG imaging of single scleral collagen fibril in the scleral slice which was manually dissociated from the scleral substrate. (a) and (b) were recorded at different locations. The sharp bends of the collagen fibrils are commonly observed and are indicated by the solid triangles. Objective: 63×/1.0W, Bars: 10 μm

Equations (2)

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Δ k · l 2 Δ ϕ g π
j = 1 n E 2 ω 0 e i Δ k · r j

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