Abstract

The loss of organization of the corneal collagen lattice induced by photothermal effects was analyzed by using second-harmonic generation (SHG) imaging. Porcine cornea samples were treated with low-power laser irradiation in order to get localized areas of tissue disorganization. The disorder induced within the irradiated area of corneal stroma was quantified by means of Discrete Fourier Transform, auto-correlation and entropy analyses of the SHG images. Polarization modulated SHG measurements allowed to probe the changes in the structural anisotropy of sub-micron hierarchical levels of the stromal collagen. Our results emphasize the great potential of the SHG imaging to detect subtle modifications in the collagen assembly. The proposed analytical methods may be used to track several genetic, pathologic, accidental or surgical-induced disorder states of biological tissues.

©2009 Optical Society of America

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References

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  1. W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003).
    [Crossref] [PubMed]
  2. P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms.,” Nat. Biotechnol. 21, 1356–1360 (2003).
    [Crossref] [PubMed]
  3. E. Brown, T. McKee, E. dI Tomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med. 9, 796–800 (2003).
    [Crossref] [PubMed]
  4. R. Cicchi, D. Massi, S. Sestini, P. Carli, V. De Giorgi, T. Lotti, and F. S. Pavone, “Multidimensional nonlinear laser imaging of basal cell carcinoma,” Opt. Express 15, 10135–10148 (2007).
    [Crossref] [PubMed]
  5. O. Nadiarnykh, S. Plotnikov, W. A. Mohler, I. Kalajzic, D. Redford-Badwal, and P. J. Campagnola, “Second harmonic generation imaging microscopy studies of osteogenesis imperfecta,” J. Biomed. Opt. 12, 051805 (2007).
    [Crossref] [PubMed]
  6. S. Roth and I. Freund, “Second harmonic generation in collagen,” J. Chem. Phys. 70, 1637–1643 (1979).
    [Crossref]
  7. P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, “Polarization-modulated second harmonic generation in collagen,” Biophys. J. 82, 3330–3342 (2002).
    [Crossref] [PubMed]
  8. K. M. Reiser, C. Bratton, D. R. Yankelevich, A. Knoesen, I. Rocha-Mendoza, and J. Lotz, “Quantitative analysis of structural disorder in intervertebral disks using second harmonic generation imaging: comparison with morphometric analysis,” J. Biomed. Opt. 12, 064019 (2007).
    [Crossref]
  9. R. Lacomb, O. Nadiarnykh, and P. J. Campagnola, “Quantitative second harmonic generation imaging of the diseased state osteogenesis imperfecta: experiment and simulation,” Biophys. J. 94, 4504–4014 (2008).
    [Crossref] [PubMed]
  10. Y. Komai and T. Ushiki, “The three-dimensional organization of collagen fibrils in the human cornea and sclera,” Invest. Ophtalmol. Visual. Sci. 32, 2244–2258 (1991).
  11. P. Matteini, F. Rossi, L. Menabuoni, and R. Pini, “Microscopic characterization of collagen modifications induced by low-temperature diode-laser welding of corneal tissue,” Lasers Surg. Med. 39, 597–604 (2007).
    [Crossref] [PubMed]
  12. F. Rossi, R. Pini, and L. Menabuoni, “Experimental and model analysis on the temperature dynamics during diode laser welding of the cornea,” J. Biomed. Opt. 12, 014031 (2007).
    [Crossref] [PubMed]
  13. S. J. Lin, C. Y. Hsiao, Y. Sun, W. Lo, W. C. Lin, G. J. Jan, S. H. Jee, and C. Y. Dong, “Monitoring the thermally induced structural transitions of collagen by use of second-harmonic generation microscopy,” Opt. Lett. 30, 622–624 (2005)
    [Crossref] [PubMed]
  14. T. Theodossiou, G. S. Rapti, V. Hovhannisyan, E. Georgiou, K. Politopoulos, and D. Yova, “Thermally induced irreversible conformational changes in collagen probed by optical second harmonic generation and laser-induced fluorescence,” Lasers Med. Sci. 17, 34–41 (2002)
    [Crossref] [PubMed]
  15. F. Rossi, R. Pini, L. Menabuoni, R. Mencucci, U. Menchini, S. Ambrosini, and G. Vannelli, “Experimental study on the healing process following laser welding of the cornea,” J. Biomed. Opt. 10, 024004 (2005).
    [Crossref] [PubMed]
  16. R. Cicchi, S. Sestini, V. De Giorgi, D. Massi, T. Lotti, and F. S. Pavone, “Non-linear laser imaging of skin lesions,” J. Biophotonics 1, 62–73 (2008).
    [Crossref]
  17. F. Rossi, P. Matteini, I. Bruno, P. Nesi, and R. Pini, “Monitoring thermally-induced phase-transitions in porcine cornea with the use of fluorescence micro-imaging analysis,” Opt. Express 15, 11178–11184 (2007).
    [Crossref] [PubMed]
  18. R. Holota and S. Němeček, Applied Electronics, J. Pinker, ed., (University of West Bohemia, Pilsen, 2002).
  19. J. Kuczynski and P. Mikolajczak, “Information theory based medical images processing,” Opto-Electron. Rev. 11, 253–259 (2003).
  20. R. Gauderon, P. B. Lukins, and C. J. Sheppard, “Optimization of second-harmonic generation microscopy,” Micron 32, 691–700 (2001).
    [Crossref] [PubMed]
  21. M. Han, G. Giese, and J. F. Bille, “Second harmonic generation imaging of collagen fibrils in cornea and sclera,” Opt. Express 13, 5791–5797 (2005).
    [Crossref] [PubMed]
  22. S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, “Studies of χ(2)/χ(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy,” Biophys. J 86, 3914–3922 (2004).
    [Crossref] [PubMed]
  23. F. Tiaho, G. Recher, and D. Rouède, “Estimation of helical angles of myosin and collagen by second harmonic generation imaging microscopy,” Opt. Express 15, 12286–12295 (2007).
    [Crossref] [PubMed]
  24. P. Stoller, B. M. Kim, A. M. Rubenchik, K. M. Reiser, and L B. Da Silva, “Polarization-dependent optical second-harmonic imaging of a rat-tail tendon,” J. Biomed. Opt. 7, 205–214 (2002).
    [Crossref] [PubMed]
  25. C. K. Chou, W. L Chen, P. T. Fwu, S. J. Lin, H. S. Lee, and C. Y. Dong, “Polarization ellipticity compensation in polarization second-harmonic generation microscopy without specimen rotation,” J. Biomed. Opt. 13, 014005 (2008).
    [Crossref] [PubMed]

2008 (3)

R. Lacomb, O. Nadiarnykh, and P. J. Campagnola, “Quantitative second harmonic generation imaging of the diseased state osteogenesis imperfecta: experiment and simulation,” Biophys. J. 94, 4504–4014 (2008).
[Crossref] [PubMed]

R. Cicchi, S. Sestini, V. De Giorgi, D. Massi, T. Lotti, and F. S. Pavone, “Non-linear laser imaging of skin lesions,” J. Biophotonics 1, 62–73 (2008).
[Crossref]

C. K. Chou, W. L Chen, P. T. Fwu, S. J. Lin, H. S. Lee, and C. Y. Dong, “Polarization ellipticity compensation in polarization second-harmonic generation microscopy without specimen rotation,” J. Biomed. Opt. 13, 014005 (2008).
[Crossref] [PubMed]

2007 (7)

O. Nadiarnykh, S. Plotnikov, W. A. Mohler, I. Kalajzic, D. Redford-Badwal, and P. J. Campagnola, “Second harmonic generation imaging microscopy studies of osteogenesis imperfecta,” J. Biomed. Opt. 12, 051805 (2007).
[Crossref] [PubMed]

K. M. Reiser, C. Bratton, D. R. Yankelevich, A. Knoesen, I. Rocha-Mendoza, and J. Lotz, “Quantitative analysis of structural disorder in intervertebral disks using second harmonic generation imaging: comparison with morphometric analysis,” J. Biomed. Opt. 12, 064019 (2007).
[Crossref]

P. Matteini, F. Rossi, L. Menabuoni, and R. Pini, “Microscopic characterization of collagen modifications induced by low-temperature diode-laser welding of corneal tissue,” Lasers Surg. Med. 39, 597–604 (2007).
[Crossref] [PubMed]

F. Rossi, R. Pini, and L. Menabuoni, “Experimental and model analysis on the temperature dynamics during diode laser welding of the cornea,” J. Biomed. Opt. 12, 014031 (2007).
[Crossref] [PubMed]

R. Cicchi, D. Massi, S. Sestini, P. Carli, V. De Giorgi, T. Lotti, and F. S. Pavone, “Multidimensional nonlinear laser imaging of basal cell carcinoma,” Opt. Express 15, 10135–10148 (2007).
[Crossref] [PubMed]

F. Rossi, P. Matteini, I. Bruno, P. Nesi, and R. Pini, “Monitoring thermally-induced phase-transitions in porcine cornea with the use of fluorescence micro-imaging analysis,” Opt. Express 15, 11178–11184 (2007).
[Crossref] [PubMed]

F. Tiaho, G. Recher, and D. Rouède, “Estimation of helical angles of myosin and collagen by second harmonic generation imaging microscopy,” Opt. Express 15, 12286–12295 (2007).
[Crossref] [PubMed]

2005 (3)

2004 (1)

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, “Studies of χ(2)/χ(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy,” Biophys. J 86, 3914–3922 (2004).
[Crossref] [PubMed]

2003 (4)

J. Kuczynski and P. Mikolajczak, “Information theory based medical images processing,” Opto-Electron. Rev. 11, 253–259 (2003).

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003).
[Crossref] [PubMed]

P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms.,” Nat. Biotechnol. 21, 1356–1360 (2003).
[Crossref] [PubMed]

E. Brown, T. McKee, E. dI Tomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med. 9, 796–800 (2003).
[Crossref] [PubMed]

2002 (3)

T. Theodossiou, G. S. Rapti, V. Hovhannisyan, E. Georgiou, K. Politopoulos, and D. Yova, “Thermally induced irreversible conformational changes in collagen probed by optical second harmonic generation and laser-induced fluorescence,” Lasers Med. Sci. 17, 34–41 (2002)
[Crossref] [PubMed]

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

P. Stoller, B. M. Kim, A. M. Rubenchik, K. M. Reiser, and L B. Da Silva, “Polarization-dependent optical second-harmonic imaging of a rat-tail tendon,” J. Biomed. Opt. 7, 205–214 (2002).
[Crossref] [PubMed]

2001 (1)

R. Gauderon, P. B. Lukins, and C. J. Sheppard, “Optimization of second-harmonic generation microscopy,” Micron 32, 691–700 (2001).
[Crossref] [PubMed]

1991 (1)

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

1979 (1)

S. Roth and I. Freund, “Second harmonic generation in collagen,” J. Chem. Phys. 70, 1637–1643 (1979).
[Crossref]

Ambrosini, S.

F. Rossi, R. Pini, L. Menabuoni, R. Mencucci, U. Menchini, S. Ambrosini, and G. Vannelli, “Experimental study on the healing process following laser welding of the cornea,” J. Biomed. Opt. 10, 024004 (2005).
[Crossref] [PubMed]

Bille, J. F.

Boucher, Y.

E. Brown, T. McKee, E. dI Tomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med. 9, 796–800 (2003).
[Crossref] [PubMed]

Bratton, C.

K. M. Reiser, C. Bratton, D. R. Yankelevich, A. Knoesen, I. Rocha-Mendoza, and J. Lotz, “Quantitative analysis of structural disorder in intervertebral disks using second harmonic generation imaging: comparison with morphometric analysis,” J. Biomed. Opt. 12, 064019 (2007).
[Crossref]

Brown, E.

E. Brown, T. McKee, E. dI Tomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med. 9, 796–800 (2003).
[Crossref] [PubMed]

Bruno, I.

Campagnola, P. J.

R. Lacomb, O. Nadiarnykh, and P. J. Campagnola, “Quantitative second harmonic generation imaging of the diseased state osteogenesis imperfecta: experiment and simulation,” Biophys. J. 94, 4504–4014 (2008).
[Crossref] [PubMed]

O. Nadiarnykh, S. Plotnikov, W. A. Mohler, I. Kalajzic, D. Redford-Badwal, and P. J. Campagnola, “Second harmonic generation imaging microscopy studies of osteogenesis imperfecta,” J. Biomed. Opt. 12, 051805 (2007).
[Crossref] [PubMed]

P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms.,” Nat. Biotechnol. 21, 1356–1360 (2003).
[Crossref] [PubMed]

Carli, P.

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, 3330–3342 (2002).
[Crossref] [PubMed]

Chen, S. Y.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, “Studies of χ(2)/χ(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy,” Biophys. J 86, 3914–3922 (2004).
[Crossref] [PubMed]

Chen, W. L

C. K. Chou, W. L Chen, P. T. Fwu, S. J. Lin, H. S. Lee, and C. Y. Dong, “Polarization ellipticity compensation in polarization second-harmonic generation microscopy without specimen rotation,” J. Biomed. Opt. 13, 014005 (2008).
[Crossref] [PubMed]

Chen, Y. C.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, “Studies of χ(2)/χ(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy,” Biophys. J 86, 3914–3922 (2004).
[Crossref] [PubMed]

Chern, G. W.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, “Studies of χ(2)/χ(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy,” Biophys. J 86, 3914–3922 (2004).
[Crossref] [PubMed]

Chou, C. K.

C. K. Chou, W. L Chen, P. T. Fwu, S. J. Lin, H. S. Lee, and C. Y. Dong, “Polarization ellipticity compensation in polarization second-harmonic generation microscopy without specimen rotation,” J. Biomed. Opt. 13, 014005 (2008).
[Crossref] [PubMed]

Christie, R.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003).
[Crossref] [PubMed]

Chu, S. W.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, “Studies of χ(2)/χ(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy,” Biophys. J 86, 3914–3922 (2004).
[Crossref] [PubMed]

Cicchi, R.

R. Cicchi, S. Sestini, V. De Giorgi, D. Massi, T. Lotti, and F. S. Pavone, “Non-linear laser imaging of skin lesions,” J. Biophotonics 1, 62–73 (2008).
[Crossref]

R. Cicchi, D. Massi, S. Sestini, P. Carli, V. De Giorgi, T. Lotti, and F. S. Pavone, “Multidimensional nonlinear laser imaging of basal cell carcinoma,” Opt. Express 15, 10135–10148 (2007).
[Crossref] [PubMed]

Da Silva, L B.

P. Stoller, B. M. Kim, A. M. Rubenchik, K. M. Reiser, and L B. Da Silva, “Polarization-dependent optical second-harmonic imaging of a rat-tail tendon,” J. Biomed. Opt. 7, 205–214 (2002).
[Crossref] [PubMed]

De Giorgi, V.

R. Cicchi, S. Sestini, V. De Giorgi, D. Massi, T. Lotti, and F. S. Pavone, “Non-linear laser imaging of skin lesions,” J. Biophotonics 1, 62–73 (2008).
[Crossref]

R. Cicchi, D. Massi, S. Sestini, P. Carli, V. De Giorgi, T. Lotti, and F. S. Pavone, “Multidimensional nonlinear laser imaging of basal cell carcinoma,” Opt. Express 15, 10135–10148 (2007).
[Crossref] [PubMed]

dI Tomaso, E.

E. Brown, T. McKee, E. dI Tomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med. 9, 796–800 (2003).
[Crossref] [PubMed]

Dong, C. Y.

C. K. Chou, W. L Chen, P. T. Fwu, S. J. Lin, H. S. Lee, and C. Y. Dong, “Polarization ellipticity compensation in polarization second-harmonic generation microscopy without specimen rotation,” J. Biomed. Opt. 13, 014005 (2008).
[Crossref] [PubMed]

S. J. Lin, C. Y. Hsiao, Y. Sun, W. Lo, W. C. Lin, G. J. Jan, S. H. Jee, and C. Y. Dong, “Monitoring the thermally induced structural transitions of collagen by use of second-harmonic generation microscopy,” Opt. Lett. 30, 622–624 (2005)
[Crossref] [PubMed]

Freund,, I.

S. Roth and I. Freund, “Second harmonic generation in collagen,” J. Chem. Phys. 70, 1637–1643 (1979).
[Crossref]

Fwu, P. T.

C. K. Chou, W. L Chen, P. T. Fwu, S. J. Lin, H. S. Lee, and C. Y. Dong, “Polarization ellipticity compensation in polarization second-harmonic generation microscopy without specimen rotation,” J. Biomed. Opt. 13, 014005 (2008).
[Crossref] [PubMed]

Gauderon, R.

R. Gauderon, P. B. Lukins, and C. J. Sheppard, “Optimization of second-harmonic generation microscopy,” Micron 32, 691–700 (2001).
[Crossref] [PubMed]

Georgiou, E.

T. Theodossiou, G. S. Rapti, V. Hovhannisyan, E. Georgiou, K. Politopoulos, and D. Yova, “Thermally induced irreversible conformational changes in collagen probed by optical second harmonic generation and laser-induced fluorescence,” Lasers Med. Sci. 17, 34–41 (2002)
[Crossref] [PubMed]

Giese, G.

Han, M.

Holota, R.

R. Holota and S. Němeček, Applied Electronics, J. Pinker, ed., (University of West Bohemia, Pilsen, 2002).

Hovhannisyan, V.

T. Theodossiou, G. S. Rapti, V. Hovhannisyan, E. Georgiou, K. Politopoulos, and D. Yova, “Thermally induced irreversible conformational changes in collagen probed by optical second harmonic generation and laser-induced fluorescence,” Lasers Med. Sci. 17, 34–41 (2002)
[Crossref] [PubMed]

Hsiao, C. Y.

Hyman, B. T.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003).
[Crossref] [PubMed]

Jain, R. K.

E. Brown, T. McKee, E. dI Tomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med. 9, 796–800 (2003).
[Crossref] [PubMed]

Jan, G. J.

Jee, S. H.

Kalajzic, I.

O. Nadiarnykh, S. Plotnikov, W. A. Mohler, I. Kalajzic, D. Redford-Badwal, and P. J. Campagnola, “Second harmonic generation imaging microscopy studies of osteogenesis imperfecta,” J. Biomed. Opt. 12, 051805 (2007).
[Crossref] [PubMed]

Kim, B. M.

P. Stoller, B. M. Kim, A. M. Rubenchik, K. M. Reiser, and L B. Da Silva, “Polarization-dependent optical second-harmonic imaging of a rat-tail tendon,” J. Biomed. Opt. 7, 205–214 (2002).
[Crossref] [PubMed]

Knoesen, A.

K. M. Reiser, C. Bratton, D. R. Yankelevich, A. Knoesen, I. Rocha-Mendoza, and J. Lotz, “Quantitative analysis of structural disorder in intervertebral disks using second harmonic generation imaging: comparison with morphometric analysis,” J. Biomed. Opt. 12, 064019 (2007).
[Crossref]

Komai, Y.

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

Kuczynski, J.

J. Kuczynski and P. Mikolajczak, “Information theory based medical images processing,” Opto-Electron. Rev. 11, 253–259 (2003).

Lacomb, R.

R. Lacomb, O. Nadiarnykh, and P. J. Campagnola, “Quantitative second harmonic generation imaging of the diseased state osteogenesis imperfecta: experiment and simulation,” Biophys. J. 94, 4504–4014 (2008).
[Crossref] [PubMed]

Lee, H. S.

C. K. Chou, W. L Chen, P. T. Fwu, S. J. Lin, H. S. Lee, and C. Y. Dong, “Polarization ellipticity compensation in polarization second-harmonic generation microscopy without specimen rotation,” J. Biomed. Opt. 13, 014005 (2008).
[Crossref] [PubMed]

Lin, B. L.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, “Studies of χ(2)/χ(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy,” Biophys. J 86, 3914–3922 (2004).
[Crossref] [PubMed]

Lin, S. J.

C. K. Chou, W. L Chen, P. T. Fwu, S. J. Lin, H. S. Lee, and C. Y. Dong, “Polarization ellipticity compensation in polarization second-harmonic generation microscopy without specimen rotation,” J. Biomed. Opt. 13, 014005 (2008).
[Crossref] [PubMed]

S. J. Lin, C. Y. Hsiao, Y. Sun, W. Lo, W. C. Lin, G. J. Jan, S. H. Jee, and C. Y. Dong, “Monitoring the thermally induced structural transitions of collagen by use of second-harmonic generation microscopy,” Opt. Lett. 30, 622–624 (2005)
[Crossref] [PubMed]

Lin, W. C.

Lo, W.

Loew, L. M.

P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms.,” Nat. Biotechnol. 21, 1356–1360 (2003).
[Crossref] [PubMed]

Lotti, T.

R. Cicchi, S. Sestini, V. De Giorgi, D. Massi, T. Lotti, and F. S. Pavone, “Non-linear laser imaging of skin lesions,” J. Biophotonics 1, 62–73 (2008).
[Crossref]

R. Cicchi, D. Massi, S. Sestini, P. Carli, V. De Giorgi, T. Lotti, and F. S. Pavone, “Multidimensional nonlinear laser imaging of basal cell carcinoma,” Opt. Express 15, 10135–10148 (2007).
[Crossref] [PubMed]

Lotz, J.

K. M. Reiser, C. Bratton, D. R. Yankelevich, A. Knoesen, I. Rocha-Mendoza, and J. Lotz, “Quantitative analysis of structural disorder in intervertebral disks using second harmonic generation imaging: comparison with morphometric analysis,” J. Biomed. Opt. 12, 064019 (2007).
[Crossref]

Lukins, P. B.

R. Gauderon, P. B. Lukins, and C. J. Sheppard, “Optimization of second-harmonic generation microscopy,” Micron 32, 691–700 (2001).
[Crossref] [PubMed]

Massi, D.

R. Cicchi, S. Sestini, V. De Giorgi, D. Massi, T. Lotti, and F. S. Pavone, “Non-linear laser imaging of skin lesions,” J. Biophotonics 1, 62–73 (2008).
[Crossref]

R. Cicchi, D. Massi, S. Sestini, P. Carli, V. De Giorgi, T. Lotti, and F. S. Pavone, “Multidimensional nonlinear laser imaging of basal cell carcinoma,” Opt. Express 15, 10135–10148 (2007).
[Crossref] [PubMed]

Matteini, P.

P. Matteini, F. Rossi, L. Menabuoni, and R. Pini, “Microscopic characterization of collagen modifications induced by low-temperature diode-laser welding of corneal tissue,” Lasers Surg. Med. 39, 597–604 (2007).
[Crossref] [PubMed]

F. Rossi, P. Matteini, I. Bruno, P. Nesi, and R. Pini, “Monitoring thermally-induced phase-transitions in porcine cornea with the use of fluorescence micro-imaging analysis,” Opt. Express 15, 11178–11184 (2007).
[Crossref] [PubMed]

McKee, T.

E. Brown, T. McKee, E. dI Tomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med. 9, 796–800 (2003).
[Crossref] [PubMed]

Menabuoni, L.

P. Matteini, F. Rossi, L. Menabuoni, and R. Pini, “Microscopic characterization of collagen modifications induced by low-temperature diode-laser welding of corneal tissue,” Lasers Surg. Med. 39, 597–604 (2007).
[Crossref] [PubMed]

F. Rossi, R. Pini, and L. Menabuoni, “Experimental and model analysis on the temperature dynamics during diode laser welding of the cornea,” J. Biomed. Opt. 12, 014031 (2007).
[Crossref] [PubMed]

F. Rossi, R. Pini, L. Menabuoni, R. Mencucci, U. Menchini, S. Ambrosini, and G. Vannelli, “Experimental study on the healing process following laser welding of the cornea,” J. Biomed. Opt. 10, 024004 (2005).
[Crossref] [PubMed]

Menchini, U.

F. Rossi, R. Pini, L. Menabuoni, R. Mencucci, U. Menchini, S. Ambrosini, and G. Vannelli, “Experimental study on the healing process following laser welding of the cornea,” J. Biomed. Opt. 10, 024004 (2005).
[Crossref] [PubMed]

Mencucci, R.

F. Rossi, R. Pini, L. Menabuoni, R. Mencucci, U. Menchini, S. Ambrosini, and G. Vannelli, “Experimental study on the healing process following laser welding of the cornea,” J. Biomed. Opt. 10, 024004 (2005).
[Crossref] [PubMed]

Mikolajczak, P.

J. Kuczynski and P. Mikolajczak, “Information theory based medical images processing,” Opto-Electron. Rev. 11, 253–259 (2003).

Mohler, W. A.

O. Nadiarnykh, S. Plotnikov, W. A. Mohler, I. Kalajzic, D. Redford-Badwal, and P. J. Campagnola, “Second harmonic generation imaging microscopy studies of osteogenesis imperfecta,” J. Biomed. Opt. 12, 051805 (2007).
[Crossref] [PubMed]

Nadiarnykh, O.

R. Lacomb, O. Nadiarnykh, and P. J. Campagnola, “Quantitative second harmonic generation imaging of the diseased state osteogenesis imperfecta: experiment and simulation,” Biophys. J. 94, 4504–4014 (2008).
[Crossref] [PubMed]

O. Nadiarnykh, S. Plotnikov, W. A. Mohler, I. Kalajzic, D. Redford-Badwal, and P. J. Campagnola, “Second harmonic generation imaging microscopy studies of osteogenesis imperfecta,” J. Biomed. Opt. 12, 051805 (2007).
[Crossref] [PubMed]

Nemecek, S.

R. Holota and S. Němeček, Applied Electronics, J. Pinker, ed., (University of West Bohemia, Pilsen, 2002).

Nesi, P.

Nikitin, A. Y.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003).
[Crossref] [PubMed]

Pavone, F. S.

R. Cicchi, S. Sestini, V. De Giorgi, D. Massi, T. Lotti, and F. S. Pavone, “Non-linear laser imaging of skin lesions,” J. Biophotonics 1, 62–73 (2008).
[Crossref]

R. Cicchi, D. Massi, S. Sestini, P. Carli, V. De Giorgi, T. Lotti, and F. S. Pavone, “Multidimensional nonlinear laser imaging of basal cell carcinoma,” Opt. Express 15, 10135–10148 (2007).
[Crossref] [PubMed]

Pini, R.

F. Rossi, R. Pini, and L. Menabuoni, “Experimental and model analysis on the temperature dynamics during diode laser welding of the cornea,” J. Biomed. Opt. 12, 014031 (2007).
[Crossref] [PubMed]

P. Matteini, F. Rossi, L. Menabuoni, and R. Pini, “Microscopic characterization of collagen modifications induced by low-temperature diode-laser welding of corneal tissue,” Lasers Surg. Med. 39, 597–604 (2007).
[Crossref] [PubMed]

F. Rossi, P. Matteini, I. Bruno, P. Nesi, and R. Pini, “Monitoring thermally-induced phase-transitions in porcine cornea with the use of fluorescence micro-imaging analysis,” Opt. Express 15, 11178–11184 (2007).
[Crossref] [PubMed]

F. Rossi, R. Pini, L. Menabuoni, R. Mencucci, U. Menchini, S. Ambrosini, and G. Vannelli, “Experimental study on the healing process following laser welding of the cornea,” J. Biomed. Opt. 10, 024004 (2005).
[Crossref] [PubMed]

Plotnikov, S.

O. Nadiarnykh, S. Plotnikov, W. A. Mohler, I. Kalajzic, D. Redford-Badwal, and P. J. Campagnola, “Second harmonic generation imaging microscopy studies of osteogenesis imperfecta,” J. Biomed. Opt. 12, 051805 (2007).
[Crossref] [PubMed]

Pluen, A.

E. Brown, T. McKee, E. dI Tomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med. 9, 796–800 (2003).
[Crossref] [PubMed]

Politopoulos, K.

T. Theodossiou, G. S. Rapti, V. Hovhannisyan, E. Georgiou, K. Politopoulos, and D. Yova, “Thermally induced irreversible conformational changes in collagen probed by optical second harmonic generation and laser-induced fluorescence,” Lasers Med. Sci. 17, 34–41 (2002)
[Crossref] [PubMed]

Rapti, G. S.

T. Theodossiou, G. S. Rapti, V. Hovhannisyan, E. Georgiou, K. Politopoulos, and D. Yova, “Thermally induced irreversible conformational changes in collagen probed by optical second harmonic generation and laser-induced fluorescence,” Lasers Med. Sci. 17, 34–41 (2002)
[Crossref] [PubMed]

Recher, G.

Redford-Badwal, D.

O. Nadiarnykh, S. Plotnikov, W. A. Mohler, I. Kalajzic, D. Redford-Badwal, and P. J. Campagnola, “Second harmonic generation imaging microscopy studies of osteogenesis imperfecta,” J. Biomed. Opt. 12, 051805 (2007).
[Crossref] [PubMed]

Reiser, K. M.

K. M. Reiser, C. Bratton, D. R. Yankelevich, A. Knoesen, I. Rocha-Mendoza, and J. Lotz, “Quantitative analysis of structural disorder in intervertebral disks using second harmonic generation imaging: comparison with morphometric analysis,” J. Biomed. Opt. 12, 064019 (2007).
[Crossref]

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

P. Stoller, B. M. Kim, A. M. Rubenchik, K. M. Reiser, and L B. Da Silva, “Polarization-dependent optical second-harmonic imaging of a rat-tail tendon,” J. Biomed. Opt. 7, 205–214 (2002).
[Crossref] [PubMed]

Rocha-Mendoza, I.

K. M. Reiser, C. Bratton, D. R. Yankelevich, A. Knoesen, I. Rocha-Mendoza, and J. Lotz, “Quantitative analysis of structural disorder in intervertebral disks using second harmonic generation imaging: comparison with morphometric analysis,” J. Biomed. Opt. 12, 064019 (2007).
[Crossref]

Rossi, F.

F. Rossi, R. Pini, and L. Menabuoni, “Experimental and model analysis on the temperature dynamics during diode laser welding of the cornea,” J. Biomed. Opt. 12, 014031 (2007).
[Crossref] [PubMed]

P. Matteini, F. Rossi, L. Menabuoni, and R. Pini, “Microscopic characterization of collagen modifications induced by low-temperature diode-laser welding of corneal tissue,” Lasers Surg. Med. 39, 597–604 (2007).
[Crossref] [PubMed]

F. Rossi, P. Matteini, I. Bruno, P. Nesi, and R. Pini, “Monitoring thermally-induced phase-transitions in porcine cornea with the use of fluorescence micro-imaging analysis,” Opt. Express 15, 11178–11184 (2007).
[Crossref] [PubMed]

F. Rossi, R. Pini, L. Menabuoni, R. Mencucci, U. Menchini, S. Ambrosini, and G. Vannelli, “Experimental study on the healing process following laser welding of the cornea,” J. Biomed. Opt. 10, 024004 (2005).
[Crossref] [PubMed]

Roth, S.

S. Roth and I. Freund, “Second harmonic generation in collagen,” J. Chem. Phys. 70, 1637–1643 (1979).
[Crossref]

Rouède, D.

Rubenchik, A. M.

P. Stoller, B. M. Kim, A. M. Rubenchik, K. M. Reiser, and L B. Da Silva, “Polarization-dependent optical second-harmonic imaging of a rat-tail tendon,” J. Biomed. Opt. 7, 205–214 (2002).
[Crossref] [PubMed]

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

Seed, B.

E. Brown, T. McKee, E. dI Tomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med. 9, 796–800 (2003).
[Crossref] [PubMed]

Sestini, S.

R. Cicchi, S. Sestini, V. De Giorgi, D. Massi, T. Lotti, and F. S. Pavone, “Non-linear laser imaging of skin lesions,” J. Biophotonics 1, 62–73 (2008).
[Crossref]

R. Cicchi, D. Massi, S. Sestini, P. Carli, V. De Giorgi, T. Lotti, and F. S. Pavone, “Multidimensional nonlinear laser imaging of basal cell carcinoma,” Opt. Express 15, 10135–10148 (2007).
[Crossref] [PubMed]

Sheppard, C. J.

R. Gauderon, P. B. Lukins, and C. J. Sheppard, “Optimization of second-harmonic generation microscopy,” Micron 32, 691–700 (2001).
[Crossref] [PubMed]

Stoller, P.

P. Stoller, B. M. Kim, A. M. Rubenchik, K. M. Reiser, and L B. Da Silva, “Polarization-dependent optical second-harmonic imaging of a rat-tail tendon,” J. Biomed. Opt. 7, 205–214 (2002).
[Crossref] [PubMed]

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

Sun, C. K.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, “Studies of χ(2)/χ(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy,” Biophys. J 86, 3914–3922 (2004).
[Crossref] [PubMed]

Sun, Y.

Theodossiou, T.

T. Theodossiou, G. S. Rapti, V. Hovhannisyan, E. Georgiou, K. Politopoulos, and D. Yova, “Thermally induced irreversible conformational changes in collagen probed by optical second harmonic generation and laser-induced fluorescence,” Lasers Med. Sci. 17, 34–41 (2002)
[Crossref] [PubMed]

Tiaho, F.

Tsai, T. H.

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, “Studies of χ(2)/χ(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy,” Biophys. J 86, 3914–3922 (2004).
[Crossref] [PubMed]

Ushiki, T.

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

Vannelli, G.

F. Rossi, R. Pini, L. Menabuoni, R. Mencucci, U. Menchini, S. Ambrosini, and G. Vannelli, “Experimental study on the healing process following laser welding of the cornea,” J. Biomed. Opt. 10, 024004 (2005).
[Crossref] [PubMed]

Webb, W. W.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003).
[Crossref] [PubMed]

Williams, R. M.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003).
[Crossref] [PubMed]

Yankelevich, D. R.

K. M. Reiser, C. Bratton, D. R. Yankelevich, A. Knoesen, I. Rocha-Mendoza, and J. Lotz, “Quantitative analysis of structural disorder in intervertebral disks using second harmonic generation imaging: comparison with morphometric analysis,” J. Biomed. Opt. 12, 064019 (2007).
[Crossref]

Yova, D.

T. Theodossiou, G. S. Rapti, V. Hovhannisyan, E. Georgiou, K. Politopoulos, and D. Yova, “Thermally induced irreversible conformational changes in collagen probed by optical second harmonic generation and laser-induced fluorescence,” Lasers Med. Sci. 17, 34–41 (2002)
[Crossref] [PubMed]

Zipfel, W. R.

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003).
[Crossref] [PubMed]

Biophys. J (1)

S. W. Chu, S. Y. Chen, G. W. Chern, T. H. Tsai, Y. C. Chen, B. L. Lin, and C. K. Sun, “Studies of χ(2)/χ(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy,” Biophys. J 86, 3914–3922 (2004).
[Crossref] [PubMed]

Biophys. J. (2)

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

R. Lacomb, O. Nadiarnykh, and P. J. Campagnola, “Quantitative second harmonic generation imaging of the diseased state osteogenesis imperfecta: experiment and simulation,” Biophys. J. 94, 4504–4014 (2008).
[Crossref] [PubMed]

Invest. Ophtalmol. Visual. Sci. (1)

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

J. Biomed. Opt. (6)

F. Rossi, R. Pini, and L. Menabuoni, “Experimental and model analysis on the temperature dynamics during diode laser welding of the cornea,” J. Biomed. Opt. 12, 014031 (2007).
[Crossref] [PubMed]

K. M. Reiser, C. Bratton, D. R. Yankelevich, A. Knoesen, I. Rocha-Mendoza, and J. Lotz, “Quantitative analysis of structural disorder in intervertebral disks using second harmonic generation imaging: comparison with morphometric analysis,” J. Biomed. Opt. 12, 064019 (2007).
[Crossref]

O. Nadiarnykh, S. Plotnikov, W. A. Mohler, I. Kalajzic, D. Redford-Badwal, and P. J. Campagnola, “Second harmonic generation imaging microscopy studies of osteogenesis imperfecta,” J. Biomed. Opt. 12, 051805 (2007).
[Crossref] [PubMed]

P. Stoller, B. M. Kim, A. M. Rubenchik, K. M. Reiser, and L B. Da Silva, “Polarization-dependent optical second-harmonic imaging of a rat-tail tendon,” J. Biomed. Opt. 7, 205–214 (2002).
[Crossref] [PubMed]

C. K. Chou, W. L Chen, P. T. Fwu, S. J. Lin, H. S. Lee, and C. Y. Dong, “Polarization ellipticity compensation in polarization second-harmonic generation microscopy without specimen rotation,” J. Biomed. Opt. 13, 014005 (2008).
[Crossref] [PubMed]

F. Rossi, R. Pini, L. Menabuoni, R. Mencucci, U. Menchini, S. Ambrosini, and G. Vannelli, “Experimental study on the healing process following laser welding of the cornea,” J. Biomed. Opt. 10, 024004 (2005).
[Crossref] [PubMed]

J. Biophotonics (1)

R. Cicchi, S. Sestini, V. De Giorgi, D. Massi, T. Lotti, and F. S. Pavone, “Non-linear laser imaging of skin lesions,” J. Biophotonics 1, 62–73 (2008).
[Crossref]

J. Chem. Phys. (1)

S. Roth and I. Freund, “Second harmonic generation in collagen,” J. Chem. Phys. 70, 1637–1643 (1979).
[Crossref]

Lasers Med. Sci. (1)

T. Theodossiou, G. S. Rapti, V. Hovhannisyan, E. Georgiou, K. Politopoulos, and D. Yova, “Thermally induced irreversible conformational changes in collagen probed by optical second harmonic generation and laser-induced fluorescence,” Lasers Med. Sci. 17, 34–41 (2002)
[Crossref] [PubMed]

Lasers Surg. Med. (1)

P. Matteini, F. Rossi, L. Menabuoni, and R. Pini, “Microscopic characterization of collagen modifications induced by low-temperature diode-laser welding of corneal tissue,” Lasers Surg. Med. 39, 597–604 (2007).
[Crossref] [PubMed]

Micron (1)

R. Gauderon, P. B. Lukins, and C. J. Sheppard, “Optimization of second-harmonic generation microscopy,” Micron 32, 691–700 (2001).
[Crossref] [PubMed]

Nat. Biotechnol. (1)

P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms.,” Nat. Biotechnol. 21, 1356–1360 (2003).
[Crossref] [PubMed]

Nat. Med. (1)

E. Brown, T. McKee, E. dI Tomaso, A. Pluen, B. Seed, Y. Boucher, and R. K. Jain, “Dynamic imaging of collagen and its modulation in tumors in vivo using second-harmonic generation,” Nat. Med. 9, 796–800 (2003).
[Crossref] [PubMed]

Opt. Express (4)

Opt. Lett. (1)

Opto-Electron. Rev. (1)

J. Kuczynski and P. Mikolajczak, “Information theory based medical images processing,” Opto-Electron. Rev. 11, 253–259 (2003).

Proc. Natl. Acad. Sci. USA (1)

W. R. Zipfel, R. M. Williams, R. Christie, A. Y. Nikitin, B. T. Hyman, and W. W. Webb, “Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation,” Proc. Natl. Acad. Sci. USA 100, 7075–7080 (2003).
[Crossref] [PubMed]

Other (1)

R. Holota and S. Němeček, Applied Electronics, J. Pinker, ed., (University of West Bohemia, Pilsen, 2002).

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Figures (7)

Fig. 1.
Fig. 1. Temperature development within a ~400 μm-radius spot area of a corneal slice subjected to diode laser irradiation. The data are the result of a 3D model (obtained with the software Comsol Multiphysics 3.4 (Comsol AB, Sweden)), based on the solution of the bio-heat equations, which describes the photothermal effects induced by the diode laser irradiation (40 mW power output, 2 s irradiation time) inside the tissue [12].

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Fig. 2.
Fig. 2. FFT of a polarization-modulated SHG profile. Upper: frequency (f) decomposition of the experimental data (◆) into zero (엯), low (◇) and high (◻) frequency contributions. Lower: frequency distribution (log scale).

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Fig. 3.
Fig. 3. SHG image of an intact porcine corneal stroma.

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Fig. 4.
Fig. 4. Stack of SHG images (60×60 μm2 ROIs) taken from the periphery to the center of a laser-irradiated spot area. The radial distance of the ROIs from the center of the irradiation spot area are 600 μm (a); 400 μm (b); 200 μm (c); 0 μm (d). The DFT diagram of each ROI is also shown in the lower panels.

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Fig. 5.
Fig. 5. Plot of the mean value (± SD) of: (a) the Aspect Ratio (AR) of the ellipse approximating the DFT magnitude and (b) the Disorganization Parameter (DP) of ROIs at different distances from the irradiated spot area, as represented in Fig. 4. Each data point is the average over values from different regions of the same sample.

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Fig. 6.
Fig. 6. Analysis of the sub-lamellar arrangement. SHG images (8×8 μm2) (a,d,g), maps of the orientations of the collagen fibrils (b,e,h), and auto-correlation matrices (c,f,i) taken at a distance of 600 μm (a,b,c); 300 μm (d,e,f); 0 μm (g,h,i) from the center of the laser-irradiated spot area. L is taken to represent the typical distance over which the regular sub-lamellar arrangement is preserved.

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Fig. 7.
Fig. 7. Polarized-SHG and TEM analyses of the interfibrillar alignment. Left: SHG images (12×12 μm2) of a control (a) and a laser-irradiated (d) region taken at a radial distance of 600 and 0 μm from the center of a laser-irradiated spot area, respectively. Middle: (b,e) average polarization profiles of single points (0.5 μm size, n = 1000) taken randomly from control (a) and laser-irradiated (d) regions, respectively. Right: TEM micrographs (bar = 300 nm) showing the nanometric assembly of collagen fibrils in a control (c) and laser-irradiated (f) region.

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Equations (4)

Equations on this page are rendered with MathJax. Learn more.

E = − ∑ i = 1 n p ( g i ) log ⁡ ( p ( g i ) )
C ( d x , d y ) ≡ ∑ i , j α ( i + d x , j + d y ) α ( i , j ) ∑ i , j α ( i , j ) 2 ,
C ( d x , d y ) ≡ 10 d x 2 + d y 2 L ,
MP = 〈 I 〉 LF − 〈 I 〉 HF I 0 .

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