Abstract

Fourier transform-second harmonic generation (FT-SHG) imaging is used as a technique for evaluating collagenase-induced injury in horse tendons. The differences in collagen fiber organization between normal and injured tendon are quantified. Results indicate that the organization of collagen fibers is regularly oriented in normal tendons and randomly organized in injured tendons. This is further supported through the use of additional metrics, in particular, the number of dark (no/minimal signal) and isotropic (no preferred fiber orientation) regions in the images, and the ratio of forward-to-backward second-harmonic intensity. FT-SHG microscopy is also compared with the conventional polarized light microscopy and is shown to be more sensitive to assessing injured tendons than the latter. Moreover, sample preparation artifacts that affect the quantitative evaluation of collagen fiber organization can be circumvented by using FT-SHG microscopy. The technique has potential as an assessment tool for evaluating the impact of various injuries that affect collagen fiber organization.

© 2010 OSA

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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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2010

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

2009

2007

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).
[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(5), 051805 (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(19), 12286–12295 (2007).
[CrossRef] [PubMed]

2005

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).
[CrossRef]

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

2004

T. A. H. Järvinen, T. L. N. Järvinen, P. Kannus, L. Józsa, and M. Järvinen, “Collagen fibres of the spontaneously ruptured human tendons display decreased thickness and crimp angle,” J. Orthop. Res. 22(6), 1303–1309 (2004).
[CrossRef] [PubMed]

2003

P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nat. Biotechnol. 21(11), 1356–1360 (2003).
[CrossRef] [PubMed]

G. Cox, E. Kable, A. Jones, I. K. Fraser, F. Manconi, and M. D. Gorrell, “3-dimensional imaging of collagen using second harmonic generation,” J. Struct. Biol. 141(1), 53–62 (2003).
[CrossRef] [PubMed]

2001

G. P. Riley, M. J. Goddard, and B. L. Hazleman, “Histopathological assessment and pathological significance of matrix degeneration in supraspinatus tendons,” Rheumatology (Oxford) 40(2), 229–230 (2001).
[CrossRef]

2000

K. M. Khan and J. L. Cook, “Overuse tendon injuries: Where does the pain come from?” Sports Med. Arthrosc. Rev. 8(1), 17–31 (2000).
[CrossRef]

B. A. Dowling, A. J. Dart, D. R. Hodgson, and R. K. W. Smith, “Superficial digital flexor tendonitis in the horse,” Equine Vet. J. 32(5), 369–378 (2000).
[CrossRef] [PubMed]

1999

K. M. Khan, J. L. Cook, F. Bonar, P. Harcourt, and M. Astrom, “Histopathology of common tendinopathies. Update and implications for clinical management,” Sports Med. 27(6), 393–408 (1999).
[CrossRef] [PubMed]

1998

L. C. Almekinders and J. D. Temple, “Etiology, diagnosis, and treatment of tendonitis: an analysis of the literature,” Med. Sci. Sports Exerc. 30(8), 1183–1190 (1998).
[CrossRef] [PubMed]

1984

D. B. Clement, J. E. Taunton, and G. W. Smart, “Achilles tendinitis and peritendinitis: etiology and treatment,” Am. J. Sports Med. 12(3), 179–184 (1984).
[CrossRef] [PubMed]

Almekinders, L. C.

L. C. Almekinders and J. D. Temple, “Etiology, diagnosis, and treatment of tendonitis: an analysis of the literature,” Med. Sci. Sports Exerc. 30(8), 1183–1190 (1998).
[CrossRef] [PubMed]

Astrom, M.

K. M. Khan, J. L. Cook, F. Bonar, P. Harcourt, and M. Astrom, “Histopathology of common tendinopathies. Update and implications for clinical management,” Sports Med. 27(6), 393–408 (1999).
[CrossRef] [PubMed]

Bille, J. F.

Bonar, F.

K. M. Khan, J. L. Cook, F. Bonar, P. Harcourt, and M. Astrom, “Histopathology of common tendinopathies. Update and implications for clinical management,” Sports Med. 27(6), 393–408 (1999).
[CrossRef] [PubMed]

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).
[CrossRef] [PubMed]

Campagnola, P. J.

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(5), 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(11), 1356–1360 (2003).
[CrossRef] [PubMed]

Chen, H. C.

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

Chen, W. L.

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

Chen, Y. F.

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

Chikama, T.

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).
[CrossRef] [PubMed]

Chiou, L. L.

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

Clement, D. B.

D. B. Clement, J. E. Taunton, and G. W. Smart, “Achilles tendinitis and peritendinitis: etiology and treatment,” Am. J. Sports Med. 12(3), 179–184 (1984).
[CrossRef] [PubMed]

Cook, J. L.

K. M. Khan and J. L. Cook, “Overuse tendon injuries: Where does the pain come from?” Sports Med. Arthrosc. Rev. 8(1), 17–31 (2000).
[CrossRef]

K. M. Khan, J. L. Cook, F. Bonar, P. Harcourt, and M. Astrom, “Histopathology of common tendinopathies. Update and implications for clinical management,” Sports Med. 27(6), 393–408 (1999).
[CrossRef] [PubMed]

Cox, G.

G. Cox, E. Kable, A. Jones, I. K. Fraser, F. Manconi, and M. D. Gorrell, “3-dimensional imaging of collagen using second harmonic generation,” J. Struct. Biol. 141(1), 53–62 (2003).
[CrossRef] [PubMed]

Dart, A. J.

B. A. Dowling, A. J. Dart, D. R. Hodgson, and R. K. W. Smith, “Superficial digital flexor tendonitis in the horse,” Equine Vet. J. 32(5), 369–378 (2000).
[CrossRef] [PubMed]

Dong, C. Y.

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

Dowling, B. A.

B. A. Dowling, A. J. Dart, D. R. Hodgson, and R. K. W. Smith, “Superficial digital flexor tendonitis in the horse,” Equine Vet. J. 32(5), 369–378 (2000).
[CrossRef] [PubMed]

Fraser, I. K.

G. Cox, E. Kable, A. Jones, I. K. Fraser, F. Manconi, and M. D. Gorrell, “3-dimensional imaging of collagen using second harmonic generation,” J. Struct. Biol. 141(1), 53–62 (2003).
[CrossRef] [PubMed]

Giese, G.

Goddard, M. J.

G. P. Riley, M. J. Goddard, and B. L. Hazleman, “Histopathological assessment and pathological significance of matrix degeneration in supraspinatus tendons,” Rheumatology (Oxford) 40(2), 229–230 (2001).
[CrossRef]

Gorrell, M. D.

G. Cox, E. Kable, A. Jones, I. K. Fraser, F. Manconi, and M. D. Gorrell, “3-dimensional imaging of collagen using second harmonic generation,” J. Struct. Biol. 141(1), 53–62 (2003).
[CrossRef] [PubMed]

Han, M.

Harcourt, P.

K. M. Khan, J. L. Cook, F. Bonar, P. Harcourt, and M. Astrom, “Histopathology of common tendinopathies. Update and implications for clinical management,” Sports Med. 27(6), 393–408 (1999).
[CrossRef] [PubMed]

Hazleman, B. L.

G. P. Riley, M. J. Goddard, and B. L. Hazleman, “Histopathological assessment and pathological significance of matrix degeneration in supraspinatus tendons,” Rheumatology (Oxford) 40(2), 229–230 (2001).
[CrossRef]

Hodgson, D. R.

B. A. Dowling, A. J. Dart, D. R. Hodgson, and R. K. W. Smith, “Superficial digital flexor tendonitis in the horse,” Equine Vet. J. 32(5), 369–378 (2000).
[CrossRef] [PubMed]

Hovhannisyan, V.

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

Huang, G. T.

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

Järvinen, M.

T. A. H. Järvinen, T. L. N. Järvinen, P. Kannus, L. Józsa, and M. Järvinen, “Collagen fibres of the spontaneously ruptured human tendons display decreased thickness and crimp angle,” J. Orthop. Res. 22(6), 1303–1309 (2004).
[CrossRef] [PubMed]

Järvinen, T. A. H.

T. A. H. Järvinen, T. L. N. Järvinen, P. Kannus, L. Józsa, and M. Järvinen, “Collagen fibres of the spontaneously ruptured human tendons display decreased thickness and crimp angle,” J. Orthop. Res. 22(6), 1303–1309 (2004).
[CrossRef] [PubMed]

Järvinen, T. L. N.

T. A. H. Järvinen, T. L. N. Järvinen, P. Kannus, L. Józsa, and M. Järvinen, “Collagen fibres of the spontaneously ruptured human tendons display decreased thickness and crimp angle,” J. Orthop. Res. 22(6), 1303–1309 (2004).
[CrossRef] [PubMed]

Jeng, Y. M.

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [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).
[CrossRef] [PubMed]

Jones, A.

G. Cox, E. Kable, A. Jones, I. K. Fraser, F. Manconi, and M. D. Gorrell, “3-dimensional imaging of collagen using second harmonic generation,” J. Struct. Biol. 141(1), 53–62 (2003).
[CrossRef] [PubMed]

Józsa, L.

T. A. H. Järvinen, T. L. N. Järvinen, P. Kannus, L. Józsa, and M. Järvinen, “Collagen fibres of the spontaneously ruptured human tendons display decreased thickness and crimp angle,” J. Orthop. Res. 22(6), 1303–1309 (2004).
[CrossRef] [PubMed]

Kable, E.

G. Cox, E. Kable, A. Jones, I. K. Fraser, F. Manconi, and M. D. Gorrell, “3-dimensional imaging of collagen using second harmonic generation,” J. Struct. Biol. 141(1), 53–62 (2003).
[CrossRef] [PubMed]

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(5), 051805 (2007).
[CrossRef] [PubMed]

Kannus, P.

T. A. H. Järvinen, T. L. N. Järvinen, P. Kannus, L. Józsa, and M. Järvinen, “Collagen fibres of the spontaneously ruptured human tendons display decreased thickness and crimp angle,” J. Orthop. Res. 22(6), 1303–1309 (2004).
[CrossRef] [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).
[CrossRef] [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).
[CrossRef] [PubMed]

Khan, K. M.

K. M. Khan and J. L. Cook, “Overuse tendon injuries: Where does the pain come from?” Sports Med. Arthrosc. Rev. 8(1), 17–31 (2000).
[CrossRef]

K. M. Khan, J. L. Cook, F. Bonar, P. Harcourt, and M. Astrom, “Histopathology of common tendinopathies. Update and implications for clinical management,” Sports Med. 27(6), 393–408 (1999).
[CrossRef] [PubMed]

Kim, K. H.

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

Lee, H. S.

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

Leithem, S.

Lin, W. C.

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

Liu, Y. A.

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

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(11), 1356–1360 (2003).
[CrossRef] [PubMed]

Manconi, F.

G. Cox, E. Kable, A. Jones, I. K. Fraser, F. Manconi, and M. D. Gorrell, “3-dimensional imaging of collagen using second harmonic generation,” J. Struct. Biol. 141(1), 53–62 (2003).
[CrossRef] [PubMed]

Mehta, M. R.

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(5), 051805 (2007).
[CrossRef] [PubMed]

Morishige, N.

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).
[CrossRef] [PubMed]

Nadiarnykh, O.

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(5), 051805 (2007).
[CrossRef] [PubMed]

Nishida, T.

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).
[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(5), 051805 (2007).
[CrossRef] [PubMed]

Rao, R. A.

Recher, G.

Redford-Badwal, D.

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G. P. Riley, M. J. Goddard, and B. L. Hazleman, “Histopathological assessment and pathological significance of matrix degeneration in supraspinatus tendons,” Rheumatology (Oxford) 40(2), 229–230 (2001).
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D. B. Clement, J. E. Taunton, and G. W. Smart, “Achilles tendinitis and peritendinitis: etiology and treatment,” Am. J. Sports Med. 12(3), 179–184 (1984).
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B. A. Dowling, A. J. Dart, D. R. Hodgson, and R. K. W. Smith, “Superficial digital flexor tendonitis in the horse,” Equine Vet. J. 32(5), 369–378 (2000).
[CrossRef] [PubMed]

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T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

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T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

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T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

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D. B. Clement, J. E. Taunton, and G. W. Smart, “Achilles tendinitis and peritendinitis: etiology and treatment,” Am. J. Sports Med. 12(3), 179–184 (1984).
[CrossRef] [PubMed]

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L. C. Almekinders and J. D. Temple, “Etiology, diagnosis, and treatment of tendonitis: an analysis of the literature,” Med. Sci. Sports Exerc. 30(8), 1183–1190 (1998).
[CrossRef] [PubMed]

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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).
[CrossRef] [PubMed]

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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).
[CrossRef]

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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).
[CrossRef]

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T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
[CrossRef] [PubMed]

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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).
[CrossRef]

Am. J. Sports Med.

D. B. Clement, J. E. Taunton, and G. W. Smart, “Achilles tendinitis and peritendinitis: etiology and treatment,” Am. J. Sports Med. 12(3), 179–184 (1984).
[CrossRef] [PubMed]

Biophys. J.

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).
[CrossRef]

Equine Vet. J.

B. A. Dowling, A. J. Dart, D. R. Hodgson, and R. K. W. Smith, “Superficial digital flexor tendonitis in the horse,” Equine Vet. J. 32(5), 369–378 (2000).
[CrossRef] [PubMed]

Invest. Ophthalmol. Vis. Sci.

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).
[CrossRef] [PubMed]

J. Biomed. Opt.

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(5), 051805 (2007).
[CrossRef] [PubMed]

T. L. Sun, Y. A. Liu, M. C. Sung, H. C. Chen, C. H. Yang, V. Hovhannisyan, W. C. Lin, Y. M. Jeng, W. L. Chen, L. L. Chiou, G. T. Huang, K. H. Kim, P. T. C. So, Y. F. Chen, H. S. Lee, and C. Y. Dong, “Ex vivo imaging and quantification of liver fibrosis using second-harmonic generation microscopy,” J. Biomed. Opt. 15(3), 036002 (2010).
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Med. Sci. Sports Exerc.

L. C. Almekinders and J. D. Temple, “Etiology, diagnosis, and treatment of tendonitis: an analysis of the literature,” Med. Sci. Sports Exerc. 30(8), 1183–1190 (1998).
[CrossRef] [PubMed]

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P. J. Campagnola and L. M. Loew, “Second-harmonic imaging microscopy for visualizing biomolecular arrays in cells, tissues and organisms,” Nat. Biotechnol. 21(11), 1356–1360 (2003).
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Opt. Express

Opt. Lett.

Rheumatology (Oxford)

G. P. Riley, M. J. Goddard, and B. L. Hazleman, “Histopathological assessment and pathological significance of matrix degeneration in supraspinatus tendons,” Rheumatology (Oxford) 40(2), 229–230 (2001).
[CrossRef]

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[CrossRef]

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Figures (6)

Fig. 1
Fig. 1

Experimental setup of the SHG microscope with both forward and backward collection geometries. Details are provided in text.

Fig. 2
Fig. 2

(a) Polarization image of a normal tendon overlaid with an orientation plot showing the preferred orientation of structures. (b) 2D Fourier transform of the polarization image shown in (a). (c) Bar plot of the number of dark, anisotropic, and isotropic regions averaged between three separate images of normal tendon. (d) Histogram of orientation values obtained from three different regions of normal tendon.

Fig. 3
Fig. 3

(a) Polarization image of an injured tendon overlaid with an orientation plot showing the preferred orientation of structures. (b) 2D Fourier transform of the polarization image shown in (a). (c) Bar plot of the number of dark, anisotropic, and isotropic regions averaged between three separate images of injured tendon. (d) Histogram of orientation values obtained from three different regions of injured tendon.

Fig. 4
Fig. 4

(a) Preferred orientation of collagen fibers from the SHG image of normal tendon. (b) 2D Fourier transform of (a). The orientation of the Fourier transform (FT) is ~0° which corresponds to the orientation in real space (RS) of 90°. (c) Bar plot of number of dark, anisotropic, and isotropic regions obtained from three different regions of normal tendon. (d) Histogram of orientation from the three images of normal tendon. (e) Image of F/B ratio per pixel for normal tendon. (f) Histogram of F/B ratios from three different regions of normal tendon.

Fig. 5
Fig. 5

(a) Preferred orientation of collagen fibers from the SHG image of injured tendon. (b) 2D Fourier transform of (a). The orientation of the Fourier transform (FT) is ~40° which corresponds to the orientation in real space (RS) of 130°. (c) Bar plot of number of dark, anisotropic, and isotropic regions obtained from three different regions of injured tendon. (d) Histogram of orientation from the three images of injured tendon. (e) Image of F/B ratio per pixel for injured tendon. (f) Histogram of F/B ratios from three different regions of injured tendon.

Fig. 6
Fig. 6

(a) Polarized light image of normal tendon sample. (b) Backward SHG from the surface (c) and 6 µm into the sample. Arrows show sectioning artifacts.

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