Abstract

We compare second harmonic generation (SHG) to histological and immunohistochemical techniques for the visualization and scoring of collagen in biological tissues. We show that SHG microscopy is highly specific for fibrillar collagens and that combined SHG and two-photon excited fluorescence (2PEF) imaging can provide simultaneous three-dimensional visualization of collagen synthesis and assembly sites in transgenic animal models expressing GFP constructs. Finally, we propose several scores for characterizing collagen accumulation based on SHG images and appropriate for different types of collagen distributions. We illustrate the sensitivity of these scores in a murine model of renal fibrosis using a morphological segmentation of the tissue based on endogenous 2PEF signals.

© 2007 Optical Society of America

Full Article  |  PDF Article

References

  • View by:
  • |
  • |
  • |

  1. P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone and W. A. Mohler, "Three-dimensional high-resolution Second-Harmonic Generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
    [CrossRef]
  2. A. Zoumi, A. Yeh and B. J. Tromberg, "Imaging cells and extracellular matrix in vivo by using second-harmonic generation and two-photon excited fluorescence," Proc. Natl. Acad. Sci. USA 99, 11014-11019 (2002).
    [CrossRef] [PubMed]
  3. G. Cox, E. Kable, A. Jones, I. Fraser, K. Marconi and M. D. Gorrell, "3-dimensional imaging of collagen using second harmonic generation," J. Struct. Biol. 141, 53-62 (2003).
    [CrossRef] [PubMed]
  4. E. Brown, T. Mckee, E. Ditomaso, 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]
  5. P. Stoller, P. M. Celliers, K. M. Reiser and A. M. Rubenchik, "Quantitative second-harmonic generation microscopy in collagen," Appl. Opt. 42, 5209-5219 (2003).
    [CrossRef] [PubMed]
  6. 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]
  7. K. König and I. Riemann, "High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution," J. Biomed. Opt. 8, 432-439 (2003).
    [CrossRef] [PubMed]
  8. A.-M. Pena, M. Strupler, T. Boulesteix, G. Godeau and M.-C. Schanne-Klein, "Spectroscopic analysis of keratin endogenous signal for skin multiphoton microscopy," Opt. Express 13, 6268-6274; erratum: 13 6667 (2005).
    [CrossRef] [PubMed]
  9. J. A. Palero, H. S. D. Bruijn, A. V. D. P.-V. D. Heuvel, H. J. C. M. Sterenborg and H. C. Gerritsen, "In vivo nonlinear spectral imaging in mouse skin," Opt. Express 14, 4395-4402 (2006).
    [CrossRef] [PubMed]
  10. T. Boulesteix, A. M. Pena, N. Pagès, G. Godeau, M.-P. Sauviat, E. Beaurepaire and M. C. Schanne-Klein, "Micrometer scale ex vivo multiphoton imaging of unstained arterial wall structure," Cytometry 69A, 20-26 (2006).
    [CrossRef]
  11. A.-M. Pena, T. Boulesteix, T. Dartigalongue and M.-C. Schanne-Klein, "Chiroptical effects in the second harmonic signal of collagens I and IV," J. Am. Chem. Soc. 127, 10314-10322 (2005).
    [CrossRef] [PubMed]
  12. S.-J. Lin, R.-Jr Wu, H.-Y. Tan, W. Lo, W.-C. Lin, T.-H. Young, C.-J. Hsu, J.-S. Chen, S.-H. Jee and C.-Y. Dong, "Evaluating cutaneous photoaging by use of multiphoton fluorescence and second-harmonic generation microscopy," Opt. Lett. 30, 2275-2277 (2005).
    [CrossRef] [PubMed]
  13. M. J. Koehler, K. König, P. Elsner, R. Bückle and M. Kaatz, "In vivo assessment of human skin aging by multiphoton laser scanning tomography," Opt. Lett. 31, 2879-2881 (2006).
    [CrossRef] [PubMed]
  14. A.-M. Pena, A. Fabre, D. Débarre, J. Marchal-Somme, B. Crestani, J.-L. Martin, E. Beaurepaire and M.-C. Schanne-Klein, "Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy," Microsc. Res. Tech.in press (2007).
    [CrossRef]
  15. A. Zoumi, X. Lu, G. S. Kassab and B. J. Tromberg, "Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy," Biophys. J. 87, 2778-2786 (2004).
    [CrossRef] [PubMed]
  16. K. Schenke-Layland, I. Riemann, U. A. Stock and K. König, "Imaging of cardiovascular structures using near-infrared femtosecond multiphoton laser scanning microscopy," J. Biomed. Opt. 10, 024017 (2005).
    [CrossRef] [PubMed]
  17. N. Buyukbabani and D. Droz, "Distribution of the extracellular matrix components in human glomerular lesions," J. Pathol. 172, 199-207 (1994).
    [CrossRef] [PubMed]
  18. M. S. Razzaque, M. Cheng, Y. Horita, M. Nishihara, T. Harada and T. Taguchi, "Immunohistochemical analysis of type III and IV collagens in tubulointerstitial damage in human benign nephrosclerosis," J. Int. Med. Res. 23, 480-486 (1995).
    [PubMed]
  19. L. C. U. Junqueira, G. Bignolas and R. R. Brentani, "Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections," Histochem. J. 11, 447-455 (1979).
    [CrossRef] [PubMed]
  20. P. Whittaker, R. A. Kloner, D. R. Boughner and J. G. Pickering, "Quantitative assessment of myocardial collagen with picrosirius red staining and circularly polarized light," Basic Res. Cardiol. 89, 397-410 (1994).
    [CrossRef] [PubMed]
  21. M. Oheim, E. Beaurepaire, E. Chaigneau, J. Mertz and S. Charpak, "Two-photon microscopy in brain tissue: parameters influencing the imaging depth," J. Neurosci. Methods 111, 29-37 (2001).
    [CrossRef] [PubMed]
  22. D. Débarre, W. Suppato, A. M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M. C. Schanne-Klein and E. Beaurepaire, "Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy," Nat. Methods 3,47 - 53 (2006).
    [CrossRef]
  23. K. L. Kusser and T. D. Randall, "Simultaneous detection of EGFP and cell surface markers by fluorescence Microscopy in Lymphoid Tissues," J. Histochem. Cytochem. 51, 5-14 (2003).
    [CrossRef]
  24. W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
    [PubMed]
  25. W. Lo, S-W. Teng, H-Y. Tan, K. H. Kim, H-C. Chen, H-S. Lee, Y-F. Chen, P. T. C. So and C.-Y. Dong, "Intact corneal stroma visualization of GFP mouse revealed by multiphoton imaging," Microsc. Res. Tech. 69, 973-975 (2006)
    [CrossRef] [PubMed]
  26. Y. Yata, A. Scanga, A. Gillan, L. Yang, S. Reif, M. Breindl, D. A. Brenner and R. A. Rippe, "DNase I-hypersensitive sites enhance a1(I) collagen gene expression in Hepatic Stellate Cells," Hepatology 37, 267-276 (2003).
    [CrossRef] [PubMed]
  27. J. Mertz and L. Moreaux, "Second-harmonic generation by focused excitation in inhomogeneously distributed scatterers," Opt. Commun. 196, 325-330 (2001).
    [CrossRef]
  28. A. K. Dunn, V. P. Wallace, M. Coleno, M. W. Berns and B. J. Tromberg, "Influence of optical properties on two-photon fluorescence imaging in turbid samples," Appl. Opt. 39, 1194-1201 (2000).
    [CrossRef]
  29. B. M. Kim, J. Eichler, K. M. Reiser, A. M. Rubenchik and L. B. Da Silva, "Collagen structure and nonlinear susceptibility: effects of heat, glycation, and enzymatic cleavage on second harmonic signal intensity," Lasers Surg. Med. 27, 329-335 (2000).
    [CrossRef] [PubMed]
  30. R. M. Williams, W. R. Zipfel and W. W. Webb, "Interpreting second-harmonic generation images of collagen fibrils," Biophys. J. 88, 1377-1386 (2005).
    [CrossRef]
  31. 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]
  32. D. J. S. Hulmes, "Building collagen molecules, fibrils, and suprafibrillar structures," J. Struct. Biol. 137,2-10 (2002).
    [CrossRef] [PubMed]
  33. A. Fogo, J. A. Breyer, M. C. Smith, W. H. Cleveland, L. Agodoa, K. A. Kirk and R. Glassock, "Accuracy of the diagnosis of hypertensive nephrosclerosis in African Americans: a report from the African American Study of Kidney Disease (AASK) Trial. AASK pilot study investigators," Kidney Int. 51 244-252 (1997).
    [CrossRef] [PubMed]
  34. K. A. Nath, "Tubulointerstitial changes as a major determinant in the progression of renal damage," Am. J. Kidney Dis. 20, 1-17 (1992)
    [PubMed]

2007

A.-M. Pena, A. Fabre, D. Débarre, J. Marchal-Somme, B. Crestani, J.-L. Martin, E. Beaurepaire and M.-C. Schanne-Klein, "Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy," Microsc. Res. Tech.in press (2007).
[CrossRef]

2006

J. A. Palero, H. S. D. Bruijn, A. V. D. P.-V. D. Heuvel, H. J. C. M. Sterenborg and H. C. Gerritsen, "In vivo nonlinear spectral imaging in mouse skin," Opt. Express 14, 4395-4402 (2006).
[CrossRef] [PubMed]

T. Boulesteix, A. M. Pena, N. Pagès, G. Godeau, M.-P. Sauviat, E. Beaurepaire and M. C. Schanne-Klein, "Micrometer scale ex vivo multiphoton imaging of unstained arterial wall structure," Cytometry 69A, 20-26 (2006).
[CrossRef]

M. J. Koehler, K. König, P. Elsner, R. Bückle and M. Kaatz, "In vivo assessment of human skin aging by multiphoton laser scanning tomography," Opt. Lett. 31, 2879-2881 (2006).
[CrossRef] [PubMed]

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

W. Lo, S-W. Teng, H-Y. Tan, K. H. Kim, H-C. Chen, H-S. Lee, Y-F. Chen, P. T. C. So and C.-Y. Dong, "Intact corneal stroma visualization of GFP mouse revealed by multiphoton imaging," Microsc. Res. Tech. 69, 973-975 (2006)
[CrossRef] [PubMed]

2005

R. M. Williams, W. R. Zipfel and W. W. Webb, "Interpreting second-harmonic generation images of collagen fibrils," Biophys. J. 88, 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, 5791-5797 (2005).
[CrossRef] [PubMed]

A.-M. Pena, T. Boulesteix, T. Dartigalongue and M.-C. Schanne-Klein, "Chiroptical effects in the second harmonic signal of collagens I and IV," J. Am. Chem. Soc. 127, 10314-10322 (2005).
[CrossRef] [PubMed]

S.-J. Lin, R.-Jr Wu, H.-Y. Tan, W. Lo, W.-C. Lin, T.-H. Young, C.-J. Hsu, J.-S. Chen, S.-H. Jee and C.-Y. Dong, "Evaluating cutaneous photoaging by use of multiphoton fluorescence and second-harmonic generation microscopy," Opt. Lett. 30, 2275-2277 (2005).
[CrossRef] [PubMed]

K. Schenke-Layland, I. Riemann, U. A. Stock and K. König, "Imaging of cardiovascular structures using near-infrared femtosecond multiphoton laser scanning microscopy," J. Biomed. Opt. 10, 024017 (2005).
[CrossRef] [PubMed]

2004

A. Zoumi, X. Lu, G. S. Kassab and B. J. Tromberg, "Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy," Biophys. J. 87, 2778-2786 (2004).
[CrossRef] [PubMed]

2003

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

E. Brown, T. Mckee, E. Ditomaso, 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]

P. Stoller, P. M. Celliers, K. M. Reiser and A. M. Rubenchik, "Quantitative second-harmonic generation microscopy in collagen," Appl. Opt. 42, 5209-5219 (2003).
[CrossRef] [PubMed]

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]

K. König and I. Riemann, "High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution," J. Biomed. Opt. 8, 432-439 (2003).
[CrossRef] [PubMed]

Y. Yata, A. Scanga, A. Gillan, L. Yang, S. Reif, M. Breindl, D. A. Brenner and R. A. Rippe, "DNase I-hypersensitive sites enhance a1(I) collagen gene expression in Hepatic Stellate Cells," Hepatology 37, 267-276 (2003).
[CrossRef] [PubMed]

K. L. Kusser and T. D. Randall, "Simultaneous detection of EGFP and cell surface markers by fluorescence Microscopy in Lymphoid Tissues," J. Histochem. Cytochem. 51, 5-14 (2003).
[CrossRef]

2002

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

D. J. S. Hulmes, "Building collagen molecules, fibrils, and suprafibrillar structures," J. Struct. Biol. 137,2-10 (2002).
[CrossRef] [PubMed]

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone and W. A. Mohler, "Three-dimensional high-resolution Second-Harmonic Generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

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

2001

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

J. Mertz and L. Moreaux, "Second-harmonic generation by focused excitation in inhomogeneously distributed scatterers," Opt. Commun. 196, 325-330 (2001).
[CrossRef]

2000

A. K. Dunn, V. P. Wallace, M. Coleno, M. W. Berns and B. J. Tromberg, "Influence of optical properties on two-photon fluorescence imaging in turbid samples," Appl. Opt. 39, 1194-1201 (2000).
[CrossRef]

B. M. Kim, J. Eichler, K. M. Reiser, A. M. Rubenchik and L. B. Da Silva, "Collagen structure and nonlinear susceptibility: effects of heat, glycation, and enzymatic cleavage on second harmonic signal intensity," Lasers Surg. Med. 27, 329-335 (2000).
[CrossRef] [PubMed]

1997

A. Fogo, J. A. Breyer, M. C. Smith, W. H. Cleveland, L. Agodoa, K. A. Kirk and R. Glassock, "Accuracy of the diagnosis of hypertensive nephrosclerosis in African Americans: a report from the African American Study of Kidney Disease (AASK) Trial. AASK pilot study investigators," Kidney Int. 51 244-252 (1997).
[CrossRef] [PubMed]

1995

M. S. Razzaque, M. Cheng, Y. Horita, M. Nishihara, T. Harada and T. Taguchi, "Immunohistochemical analysis of type III and IV collagens in tubulointerstitial damage in human benign nephrosclerosis," J. Int. Med. Res. 23, 480-486 (1995).
[PubMed]

1994

N. Buyukbabani and D. Droz, "Distribution of the extracellular matrix components in human glomerular lesions," J. Pathol. 172, 199-207 (1994).
[CrossRef] [PubMed]

P. Whittaker, R. A. Kloner, D. R. Boughner and J. G. Pickering, "Quantitative assessment of myocardial collagen with picrosirius red staining and circularly polarized light," Basic Res. Cardiol. 89, 397-410 (1994).
[CrossRef] [PubMed]

1992

K. A. Nath, "Tubulointerstitial changes as a major determinant in the progression of renal damage," Am. J. Kidney Dis. 20, 1-17 (1992)
[PubMed]

1979

L. C. U. Junqueira, G. Bignolas and R. R. Brentani, "Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections," Histochem. J. 11, 447-455 (1979).
[CrossRef] [PubMed]

Agodoa, L.

A. Fogo, J. A. Breyer, M. C. Smith, W. H. Cleveland, L. Agodoa, K. A. Kirk and R. Glassock, "Accuracy of the diagnosis of hypertensive nephrosclerosis in African Americans: a report from the African American Study of Kidney Disease (AASK) Trial. AASK pilot study investigators," Kidney Int. 51 244-252 (1997).
[CrossRef] [PubMed]

Beaurepaire, E.

A.-M. Pena, A. Fabre, D. Débarre, J. Marchal-Somme, B. Crestani, J.-L. Martin, E. Beaurepaire and M.-C. Schanne-Klein, "Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy," Microsc. Res. Tech.in press (2007).
[CrossRef]

T. Boulesteix, A. M. Pena, N. Pagès, G. Godeau, M.-P. Sauviat, E. Beaurepaire and M. C. Schanne-Klein, "Micrometer scale ex vivo multiphoton imaging of unstained arterial wall structure," Cytometry 69A, 20-26 (2006).
[CrossRef]

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

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

Berns, M. W.

Bignolas, G.

L. C. U. Junqueira, G. Bignolas and R. R. Brentani, "Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections," Histochem. J. 11, 447-455 (1979).
[CrossRef] [PubMed]

Bille, J. F.

Bottinger, E. P.

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

Boucher, Y.

E. Brown, T. Mckee, E. Ditomaso, 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]

Boughner, D. R.

P. Whittaker, R. A. Kloner, D. R. Boughner and J. G. Pickering, "Quantitative assessment of myocardial collagen with picrosirius red staining and circularly polarized light," Basic Res. Cardiol. 89, 397-410 (1994).
[CrossRef] [PubMed]

Boulesteix, T.

T. Boulesteix, A. M. Pena, N. Pagès, G. Godeau, M.-P. Sauviat, E. Beaurepaire and M. C. Schanne-Klein, "Micrometer scale ex vivo multiphoton imaging of unstained arterial wall structure," Cytometry 69A, 20-26 (2006).
[CrossRef]

A.-M. Pena, T. Boulesteix, T. Dartigalongue and M.-C. Schanne-Klein, "Chiroptical effects in the second harmonic signal of collagens I and IV," J. Am. Chem. Soc. 127, 10314-10322 (2005).
[CrossRef] [PubMed]

Breindl, M.

Y. Yata, A. Scanga, A. Gillan, L. Yang, S. Reif, M. Breindl, D. A. Brenner and R. A. Rippe, "DNase I-hypersensitive sites enhance a1(I) collagen gene expression in Hepatic Stellate Cells," Hepatology 37, 267-276 (2003).
[CrossRef] [PubMed]

Brenner, D. A.

Y. Yata, A. Scanga, A. Gillan, L. Yang, S. Reif, M. Breindl, D. A. Brenner and R. A. Rippe, "DNase I-hypersensitive sites enhance a1(I) collagen gene expression in Hepatic Stellate Cells," Hepatology 37, 267-276 (2003).
[CrossRef] [PubMed]

Brentani, R. R.

L. C. U. Junqueira, G. Bignolas and R. R. Brentani, "Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections," Histochem. J. 11, 447-455 (1979).
[CrossRef] [PubMed]

Breyer, J. A.

A. Fogo, J. A. Breyer, M. C. Smith, W. H. Cleveland, L. Agodoa, K. A. Kirk and R. Glassock, "Accuracy of the diagnosis of hypertensive nephrosclerosis in African Americans: a report from the African American Study of Kidney Disease (AASK) Trial. AASK pilot study investigators," Kidney Int. 51 244-252 (1997).
[CrossRef] [PubMed]

Brown, E.

E. Brown, T. Mckee, E. Ditomaso, 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]

Bruijn, H. S. D.

Bückle, R.

Buyukbabani, N.

N. Buyukbabani and D. Droz, "Distribution of the extracellular matrix components in human glomerular lesions," J. Pathol. 172, 199-207 (1994).
[CrossRef] [PubMed]

Campagnola, P. J.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone and W. A. Mohler, "Three-dimensional high-resolution Second-Harmonic Generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

Celliers, P. M.

Cermak, L.

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

Chaigneau, E.

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

Charpak, S.

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

Chen, H-C.

W. Lo, S-W. Teng, H-Y. Tan, K. H. Kim, H-C. Chen, H-S. Lee, Y-F. Chen, P. T. C. So and C.-Y. Dong, "Intact corneal stroma visualization of GFP mouse revealed by multiphoton imaging," Microsc. Res. Tech. 69, 973-975 (2006)
[CrossRef] [PubMed]

Chen, Y-F.

W. Lo, S-W. Teng, H-Y. Tan, K. H. Kim, H-C. Chen, H-S. Lee, Y-F. Chen, P. T. C. So and C.-Y. Dong, "Intact corneal stroma visualization of GFP mouse revealed by multiphoton imaging," Microsc. Res. Tech. 69, 973-975 (2006)
[CrossRef] [PubMed]

Cheng, M.

M. S. Razzaque, M. Cheng, Y. Horita, M. Nishihara, T. Harada and T. Taguchi, "Immunohistochemical analysis of type III and IV collagens in tubulointerstitial damage in human benign nephrosclerosis," J. Int. Med. Res. 23, 480-486 (1995).
[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]

Cleveland, W. H.

A. Fogo, J. A. Breyer, M. C. Smith, W. H. Cleveland, L. Agodoa, K. A. Kirk and R. Glassock, "Accuracy of the diagnosis of hypertensive nephrosclerosis in African Americans: a report from the African American Study of Kidney Disease (AASK) Trial. AASK pilot study investigators," Kidney Int. 51 244-252 (1997).
[CrossRef] [PubMed]

Coleno, M.

Combettes, L.

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

Condeelis, J. S.

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

Cox, G.

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

Crestani, B.

A.-M. Pena, A. Fabre, D. Débarre, J. Marchal-Somme, B. Crestani, J.-L. Martin, E. Beaurepaire and M.-C. Schanne-Klein, "Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy," Microsc. Res. Tech.in press (2007).
[CrossRef]

Da Silva, L. B.

B. M. Kim, J. Eichler, K. M. Reiser, A. M. Rubenchik and L. B. Da Silva, "Collagen structure and nonlinear susceptibility: effects of heat, glycation, and enzymatic cleavage on second harmonic signal intensity," Lasers Surg. Med. 27, 329-335 (2000).
[CrossRef] [PubMed]

Dartigalongue, T.

A.-M. Pena, T. Boulesteix, T. Dartigalongue and M.-C. Schanne-Klein, "Chiroptical effects in the second harmonic signal of collagens I and IV," J. Am. Chem. Soc. 127, 10314-10322 (2005).
[CrossRef] [PubMed]

Débarre, D.

A.-M. Pena, A. Fabre, D. Débarre, J. Marchal-Somme, B. Crestani, J.-L. Martin, E. Beaurepaire and M.-C. Schanne-Klein, "Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy," Microsc. Res. Tech.in press (2007).
[CrossRef]

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

Ditomaso, E.

E. Brown, T. Mckee, E. Ditomaso, 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]

Droz, D.

N. Buyukbabani and D. Droz, "Distribution of the extracellular matrix components in human glomerular lesions," J. Pathol. 172, 199-207 (1994).
[CrossRef] [PubMed]

Dunn, A. K.

Eichler, J.

B. M. Kim, J. Eichler, K. M. Reiser, A. M. Rubenchik and L. B. Da Silva, "Collagen structure and nonlinear susceptibility: effects of heat, glycation, and enzymatic cleavage on second harmonic signal intensity," Lasers Surg. Med. 27, 329-335 (2000).
[CrossRef] [PubMed]

Elsner, P.

Fabre, A.

A.-M. Pena, A. Fabre, D. Débarre, J. Marchal-Somme, B. Crestani, J.-L. Martin, E. Beaurepaire and M.-C. Schanne-Klein, "Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy," Microsc. Res. Tech.in press (2007).
[CrossRef]

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

Fogo, A.

A. Fogo, J. A. Breyer, M. C. Smith, W. H. Cleveland, L. Agodoa, K. A. Kirk and R. Glassock, "Accuracy of the diagnosis of hypertensive nephrosclerosis in African Americans: a report from the African American Study of Kidney Disease (AASK) Trial. AASK pilot study investigators," Kidney Int. 51 244-252 (1997).
[CrossRef] [PubMed]

Fraser, I.

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

Frohlich, V. C.

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

Gerritsen, H. C.

Giese, G.

Gillan, A.

Y. Yata, A. Scanga, A. Gillan, L. Yang, S. Reif, M. Breindl, D. A. Brenner and R. A. Rippe, "DNase I-hypersensitive sites enhance a1(I) collagen gene expression in Hepatic Stellate Cells," Hepatology 37, 267-276 (2003).
[CrossRef] [PubMed]

Glassock, R.

A. Fogo, J. A. Breyer, M. C. Smith, W. H. Cleveland, L. Agodoa, K. A. Kirk and R. Glassock, "Accuracy of the diagnosis of hypertensive nephrosclerosis in African Americans: a report from the African American Study of Kidney Disease (AASK) Trial. AASK pilot study investigators," Kidney Int. 51 244-252 (1997).
[CrossRef] [PubMed]

Godeau, G.

T. Boulesteix, A. M. Pena, N. Pagès, G. Godeau, M.-P. Sauviat, E. Beaurepaire and M. C. Schanne-Klein, "Micrometer scale ex vivo multiphoton imaging of unstained arterial wall structure," Cytometry 69A, 20-26 (2006).
[CrossRef]

Gorrell, M. D.

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

Han, M.

Harada, T.

M. S. Razzaque, M. Cheng, Y. Horita, M. Nishihara, T. Harada and T. Taguchi, "Immunohistochemical analysis of type III and IV collagens in tubulointerstitial damage in human benign nephrosclerosis," J. Int. Med. Res. 23, 480-486 (1995).
[PubMed]

Heuvel, A. V. D. P.-V. D.

Hoppe, P. E.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone and W. A. Mohler, "Three-dimensional high-resolution Second-Harmonic Generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

Horita, Y.

M. S. Razzaque, M. Cheng, Y. Horita, M. Nishihara, T. Harada and T. Taguchi, "Immunohistochemical analysis of type III and IV collagens in tubulointerstitial damage in human benign nephrosclerosis," J. Int. Med. Res. 23, 480-486 (1995).
[PubMed]

Hulmes, D. J. S.

D. J. S. Hulmes, "Building collagen molecules, fibrils, and suprafibrillar structures," J. Struct. Biol. 137,2-10 (2002).
[CrossRef] [PubMed]

Hüttelmaier, S.

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

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. Ditomaso, 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]

Jones, A.

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

Junqueira, L. C. U.

L. C. U. Junqueira, G. Bignolas and R. R. Brentani, "Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections," Histochem. J. 11, 447-455 (1979).
[CrossRef] [PubMed]

Kaatz, M.

Kable, E.

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

Kassab, G. S.

A. Zoumi, X. Lu, G. S. Kassab and B. J. Tromberg, "Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy," Biophys. J. 87, 2778-2786 (2004).
[CrossRef] [PubMed]

Kim, B. M.

B. M. Kim, J. Eichler, K. M. Reiser, A. M. Rubenchik and L. B. Da Silva, "Collagen structure and nonlinear susceptibility: effects of heat, glycation, and enzymatic cleavage on second harmonic signal intensity," Lasers Surg. Med. 27, 329-335 (2000).
[CrossRef] [PubMed]

Kim, K. H.

W. Lo, S-W. Teng, H-Y. Tan, K. H. Kim, H-C. Chen, H-S. Lee, Y-F. Chen, P. T. C. So and C.-Y. Dong, "Intact corneal stroma visualization of GFP mouse revealed by multiphoton imaging," Microsc. Res. Tech. 69, 973-975 (2006)
[CrossRef] [PubMed]

Kirk, K. A.

A. Fogo, J. A. Breyer, M. C. Smith, W. H. Cleveland, L. Agodoa, K. A. Kirk and R. Glassock, "Accuracy of the diagnosis of hypertensive nephrosclerosis in African Americans: a report from the African American Study of Kidney Disease (AASK) Trial. AASK pilot study investigators," Kidney Int. 51 244-252 (1997).
[CrossRef] [PubMed]

Kloner, R. A.

P. Whittaker, R. A. Kloner, D. R. Boughner and J. G. Pickering, "Quantitative assessment of myocardial collagen with picrosirius red staining and circularly polarized light," Basic Res. Cardiol. 89, 397-410 (1994).
[CrossRef] [PubMed]

Koehler, M. J.

König, K.

M. J. Koehler, K. König, P. Elsner, R. Bückle and M. Kaatz, "In vivo assessment of human skin aging by multiphoton laser scanning tomography," Opt. Lett. 31, 2879-2881 (2006).
[CrossRef] [PubMed]

K. Schenke-Layland, I. Riemann, U. A. Stock and K. König, "Imaging of cardiovascular structures using near-infrared femtosecond multiphoton laser scanning microscopy," J. Biomed. Opt. 10, 024017 (2005).
[CrossRef] [PubMed]

K. König and I. Riemann, "High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution," J. Biomed. Opt. 8, 432-439 (2003).
[CrossRef] [PubMed]

Kusser, K. L.

K. L. Kusser and T. D. Randall, "Simultaneous detection of EGFP and cell surface markers by fluorescence Microscopy in Lymphoid Tissues," J. Histochem. Cytochem. 51, 5-14 (2003).
[CrossRef]

Lee, H-S.

W. Lo, S-W. Teng, H-Y. Tan, K. H. Kim, H-C. Chen, H-S. Lee, Y-F. Chen, P. T. C. So and C.-Y. Dong, "Intact corneal stroma visualization of GFP mouse revealed by multiphoton imaging," Microsc. Res. Tech. 69, 973-975 (2006)
[CrossRef] [PubMed]

Lin, S.-J.

Lo, W.

W. Lo, S-W. Teng, H-Y. Tan, K. H. Kim, H-C. Chen, H-S. Lee, Y-F. Chen, P. T. C. So and C.-Y. Dong, "Intact corneal stroma visualization of GFP mouse revealed by multiphoton imaging," Microsc. Res. Tech. 69, 973-975 (2006)
[CrossRef] [PubMed]

Lu, X.

A. Zoumi, X. Lu, G. S. Kassab and B. J. Tromberg, "Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy," Biophys. J. 87, 2778-2786 (2004).
[CrossRef] [PubMed]

Malone, C. J.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone and W. A. Mohler, "Three-dimensional high-resolution Second-Harmonic Generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

Marchal-Somme, J.

A.-M. Pena, A. Fabre, D. Débarre, J. Marchal-Somme, B. Crestani, J.-L. Martin, E. Beaurepaire and M.-C. Schanne-Klein, "Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy," Microsc. Res. Tech.in press (2007).
[CrossRef]

Marconi, K.

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

Martin, J.-L.

A.-M. Pena, A. Fabre, D. Débarre, J. Marchal-Somme, B. Crestani, J.-L. Martin, E. Beaurepaire and M.-C. Schanne-Klein, "Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy," Microsc. Res. Tech.in press (2007).
[CrossRef]

Mckee, T.

E. Brown, T. Mckee, E. Ditomaso, 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]

Mertz, J.

J. Mertz and L. Moreaux, "Second-harmonic generation by focused excitation in inhomogeneously distributed scatterers," Opt. Commun. 196, 325-330 (2001).
[CrossRef]

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

Millard, A. C.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone and W. A. Mohler, "Three-dimensional high-resolution Second-Harmonic Generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

Mohler, W. A.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone and W. A. Mohler, "Three-dimensional high-resolution Second-Harmonic Generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

Moreaux, L.

J. Mertz and L. Moreaux, "Second-harmonic generation by focused excitation in inhomogeneously distributed scatterers," Opt. Commun. 196, 325-330 (2001).
[CrossRef]

Nath, K. A.

K. A. Nath, "Tubulointerstitial changes as a major determinant in the progression of renal damage," Am. J. Kidney Dis. 20, 1-17 (1992)
[PubMed]

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]

Nishihara, M.

M. S. Razzaque, M. Cheng, Y. Horita, M. Nishihara, T. Harada and T. Taguchi, "Immunohistochemical analysis of type III and IV collagens in tubulointerstitial damage in human benign nephrosclerosis," J. Int. Med. Res. 23, 480-486 (1995).
[PubMed]

Oheim, M.

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

Oleynikov, Y.

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

Pagès, N.

T. Boulesteix, A. M. Pena, N. Pagès, G. Godeau, M.-P. Sauviat, E. Beaurepaire and M. C. Schanne-Klein, "Micrometer scale ex vivo multiphoton imaging of unstained arterial wall structure," Cytometry 69A, 20-26 (2006).
[CrossRef]

Palero, J. A.

Pena, A. M.

T. Boulesteix, A. M. Pena, N. Pagès, G. Godeau, M.-P. Sauviat, E. Beaurepaire and M. C. Schanne-Klein, "Micrometer scale ex vivo multiphoton imaging of unstained arterial wall structure," Cytometry 69A, 20-26 (2006).
[CrossRef]

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

Pena, A.-M.

A.-M. Pena, A. Fabre, D. Débarre, J. Marchal-Somme, B. Crestani, J.-L. Martin, E. Beaurepaire and M.-C. Schanne-Klein, "Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy," Microsc. Res. Tech.in press (2007).
[CrossRef]

A.-M. Pena, T. Boulesteix, T. Dartigalongue and M.-C. Schanne-Klein, "Chiroptical effects in the second harmonic signal of collagens I and IV," J. Am. Chem. Soc. 127, 10314-10322 (2005).
[CrossRef] [PubMed]

Pickering, J. G.

P. Whittaker, R. A. Kloner, D. R. Boughner and J. G. Pickering, "Quantitative assessment of myocardial collagen with picrosirius red staining and circularly polarized light," Basic Res. Cardiol. 89, 397-410 (1994).
[CrossRef] [PubMed]

Pluen, A.

E. Brown, T. Mckee, E. Ditomaso, 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]

Randall, T. D.

K. L. Kusser and T. D. Randall, "Simultaneous detection of EGFP and cell surface markers by fluorescence Microscopy in Lymphoid Tissues," J. Histochem. Cytochem. 51, 5-14 (2003).
[CrossRef]

Razzaque, M. S.

M. S. Razzaque, M. Cheng, Y. Horita, M. Nishihara, T. Harada and T. Taguchi, "Immunohistochemical analysis of type III and IV collagens in tubulointerstitial damage in human benign nephrosclerosis," J. Int. Med. Res. 23, 480-486 (1995).
[PubMed]

Reif, S.

Y. Yata, A. Scanga, A. Gillan, L. Yang, S. Reif, M. Breindl, D. A. Brenner and R. A. Rippe, "DNase I-hypersensitive sites enhance a1(I) collagen gene expression in Hepatic Stellate Cells," Hepatology 37, 267-276 (2003).
[CrossRef] [PubMed]

Reiser, K. M.

P. Stoller, P. M. Celliers, K. M. Reiser and A. M. Rubenchik, "Quantitative second-harmonic generation microscopy in collagen," Appl. Opt. 42, 5209-5219 (2003).
[CrossRef] [PubMed]

B. M. Kim, J. Eichler, K. M. Reiser, A. M. Rubenchik and L. B. Da Silva, "Collagen structure and nonlinear susceptibility: effects of heat, glycation, and enzymatic cleavage on second harmonic signal intensity," Lasers Surg. Med. 27, 329-335 (2000).
[CrossRef] [PubMed]

Riemann, I.

K. Schenke-Layland, I. Riemann, U. A. Stock and K. König, "Imaging of cardiovascular structures using near-infrared femtosecond multiphoton laser scanning microscopy," J. Biomed. Opt. 10, 024017 (2005).
[CrossRef] [PubMed]

K. König and I. Riemann, "High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution," J. Biomed. Opt. 8, 432-439 (2003).
[CrossRef] [PubMed]

Rippe, R. A.

Y. Yata, A. Scanga, A. Gillan, L. Yang, S. Reif, M. Breindl, D. A. Brenner and R. A. Rippe, "DNase I-hypersensitive sites enhance a1(I) collagen gene expression in Hepatic Stellate Cells," Hepatology 37, 267-276 (2003).
[CrossRef] [PubMed]

Rubenchik, A. M.

P. Stoller, P. M. Celliers, K. M. Reiser and A. M. Rubenchik, "Quantitative second-harmonic generation microscopy in collagen," Appl. Opt. 42, 5209-5219 (2003).
[CrossRef] [PubMed]

B. M. Kim, J. Eichler, K. M. Reiser, A. M. Rubenchik and L. B. Da Silva, "Collagen structure and nonlinear susceptibility: effects of heat, glycation, and enzymatic cleavage on second harmonic signal intensity," Lasers Surg. Med. 27, 329-335 (2000).
[CrossRef] [PubMed]

Sauviat, M.-P.

T. Boulesteix, A. M. Pena, N. Pagès, G. Godeau, M.-P. Sauviat, E. Beaurepaire and M. C. Schanne-Klein, "Micrometer scale ex vivo multiphoton imaging of unstained arterial wall structure," Cytometry 69A, 20-26 (2006).
[CrossRef]

Scanga, A.

Y. Yata, A. Scanga, A. Gillan, L. Yang, S. Reif, M. Breindl, D. A. Brenner and R. A. Rippe, "DNase I-hypersensitive sites enhance a1(I) collagen gene expression in Hepatic Stellate Cells," Hepatology 37, 267-276 (2003).
[CrossRef] [PubMed]

Schanne-Klein, M. C.

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

T. Boulesteix, A. M. Pena, N. Pagès, G. Godeau, M.-P. Sauviat, E. Beaurepaire and M. C. Schanne-Klein, "Micrometer scale ex vivo multiphoton imaging of unstained arterial wall structure," Cytometry 69A, 20-26 (2006).
[CrossRef]

Schanne-Klein, M.-C.

A.-M. Pena, A. Fabre, D. Débarre, J. Marchal-Somme, B. Crestani, J.-L. Martin, E. Beaurepaire and M.-C. Schanne-Klein, "Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy," Microsc. Res. Tech.in press (2007).
[CrossRef]

A.-M. Pena, T. Boulesteix, T. Dartigalongue and M.-C. Schanne-Klein, "Chiroptical effects in the second harmonic signal of collagens I and IV," J. Am. Chem. Soc. 127, 10314-10322 (2005).
[CrossRef] [PubMed]

Schenke-Layland, K.

K. Schenke-Layland, I. Riemann, U. A. Stock and K. König, "Imaging of cardiovascular structures using near-infrared femtosecond multiphoton laser scanning microscopy," J. Biomed. Opt. 10, 024017 (2005).
[CrossRef] [PubMed]

Seed, B.

E. Brown, T. Mckee, E. Ditomaso, 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]

Segall, J. E.

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

Singer, R. H.

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

Smith, M. C.

A. Fogo, J. A. Breyer, M. C. Smith, W. H. Cleveland, L. Agodoa, K. A. Kirk and R. Glassock, "Accuracy of the diagnosis of hypertensive nephrosclerosis in African Americans: a report from the African American Study of Kidney Disease (AASK) Trial. AASK pilot study investigators," Kidney Int. 51 244-252 (1997).
[CrossRef] [PubMed]

Sterenborg, H. J. C. M.

Stock, U. A.

K. Schenke-Layland, I. Riemann, U. A. Stock and K. König, "Imaging of cardiovascular structures using near-infrared femtosecond multiphoton laser scanning microscopy," J. Biomed. Opt. 10, 024017 (2005).
[CrossRef] [PubMed]

Stoller, P.

Suppato, W.

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

Taguchi, T.

M. S. Razzaque, M. Cheng, Y. Horita, M. Nishihara, T. Harada and T. Taguchi, "Immunohistochemical analysis of type III and IV collagens in tubulointerstitial damage in human benign nephrosclerosis," J. Int. Med. Res. 23, 480-486 (1995).
[PubMed]

Tan, H-Y.

W. Lo, S-W. Teng, H-Y. Tan, K. H. Kim, H-C. Chen, H-S. Lee, Y-F. Chen, P. T. C. So and C.-Y. Dong, "Intact corneal stroma visualization of GFP mouse revealed by multiphoton imaging," Microsc. Res. Tech. 69, 973-975 (2006)
[CrossRef] [PubMed]

Teng, S-W.

W. Lo, S-W. Teng, H-Y. Tan, K. H. Kim, H-C. Chen, H-S. Lee, Y-F. Chen, P. T. C. So and C.-Y. Dong, "Intact corneal stroma visualization of GFP mouse revealed by multiphoton imaging," Microsc. Res. Tech. 69, 973-975 (2006)
[CrossRef] [PubMed]

Terasaki, M.

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone and W. A. Mohler, "Three-dimensional high-resolution Second-Harmonic Generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

Tordjmann, T.

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

Tromberg, B. J.

A. Zoumi, X. Lu, G. S. Kassab and B. J. Tromberg, "Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy," Biophys. J. 87, 2778-2786 (2004).
[CrossRef] [PubMed]

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

A. K. Dunn, V. P. Wallace, M. Coleno, M. W. Berns and B. J. Tromberg, "Influence of optical properties on two-photon fluorescence imaging in turbid samples," Appl. Opt. 39, 1194-1201 (2000).
[CrossRef]

Wallace, V. P.

Wang, W.

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

Webb, W. W.

R. M. Williams, W. R. Zipfel and W. W. Webb, "Interpreting second-harmonic generation images of collagen fibrils," Biophys. J. 88, 1377-1386 (2005).
[CrossRef]

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]

White, J. G.

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

Whittaker, P.

P. Whittaker, R. A. Kloner, D. R. Boughner and J. G. Pickering, "Quantitative assessment of myocardial collagen with picrosirius red staining and circularly polarized light," Basic Res. Cardiol. 89, 397-410 (1994).
[CrossRef] [PubMed]

Williams, R. M.

R. M. Williams, W. R. Zipfel and W. W. Webb, "Interpreting second-harmonic generation images of collagen fibrils," Biophys. J. 88, 1377-1386 (2005).
[CrossRef]

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]

Wyckoff, J. B.

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

Yang, L.

Y. Yata, A. Scanga, A. Gillan, L. Yang, S. Reif, M. Breindl, D. A. Brenner and R. A. Rippe, "DNase I-hypersensitive sites enhance a1(I) collagen gene expression in Hepatic Stellate Cells," Hepatology 37, 267-276 (2003).
[CrossRef] [PubMed]

Yata, Y.

Y. Yata, A. Scanga, A. Gillan, L. Yang, S. Reif, M. Breindl, D. A. Brenner and R. A. Rippe, "DNase I-hypersensitive sites enhance a1(I) collagen gene expression in Hepatic Stellate Cells," Hepatology 37, 267-276 (2003).
[CrossRef] [PubMed]

Yeh, A.

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

Zavadil, J.

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

Zipfel, W. R.

R. M. Williams, W. R. Zipfel and W. W. Webb, "Interpreting second-harmonic generation images of collagen fibrils," Biophys. J. 88, 1377-1386 (2005).
[CrossRef]

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]

Zoumi, A.

A. Zoumi, X. Lu, G. S. Kassab and B. J. Tromberg, "Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy," Biophys. J. 87, 2778-2786 (2004).
[CrossRef] [PubMed]

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

Am. J. Kidney Dis.

K. A. Nath, "Tubulointerstitial changes as a major determinant in the progression of renal damage," Am. J. Kidney Dis. 20, 1-17 (1992)
[PubMed]

Appl. Opt.

Basic Res. Cardiol.

P. Whittaker, R. A. Kloner, D. R. Boughner and J. G. Pickering, "Quantitative assessment of myocardial collagen with picrosirius red staining and circularly polarized light," Basic Res. Cardiol. 89, 397-410 (1994).
[CrossRef] [PubMed]

Biophys. J.

R. M. Williams, W. R. Zipfel and W. W. Webb, "Interpreting second-harmonic generation images of collagen fibrils," Biophys. J. 88, 1377-1386 (2005).
[CrossRef]

P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone and W. A. Mohler, "Three-dimensional high-resolution Second-Harmonic Generation imaging of endogenous structural proteins in biological tissues," Biophys. J. 82, 493-508 (2002).
[CrossRef]

A. Zoumi, X. Lu, G. S. Kassab and B. J. Tromberg, "Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy," Biophys. J. 87, 2778-2786 (2004).
[CrossRef] [PubMed]

Cancer Res.

W. Wang, J. B. Wyckoff, V. C. Frohlich, Y. Oleynikov, S. Hüttelmaier, J. Zavadil, L. Cermak, E. P. Bottinger, R. H. Singer, J. G. White, J. E. Segall and J. S. Condeelis, "Single cell behavior in metastatic primary mammary tumors correlated with gene expressions patterns revealed by molecular profiling," Cancer Res. 62, 6278-6288 (2002).
[PubMed]

Cytometry

T. Boulesteix, A. M. Pena, N. Pagès, G. Godeau, M.-P. Sauviat, E. Beaurepaire and M. C. Schanne-Klein, "Micrometer scale ex vivo multiphoton imaging of unstained arterial wall structure," Cytometry 69A, 20-26 (2006).
[CrossRef]

Hepatology

Y. Yata, A. Scanga, A. Gillan, L. Yang, S. Reif, M. Breindl, D. A. Brenner and R. A. Rippe, "DNase I-hypersensitive sites enhance a1(I) collagen gene expression in Hepatic Stellate Cells," Hepatology 37, 267-276 (2003).
[CrossRef] [PubMed]

Histochem. J.

L. C. U. Junqueira, G. Bignolas and R. R. Brentani, "Picrosirius staining plus polarization microscopy, a specific method for collagen detection in tissue sections," Histochem. J. 11, 447-455 (1979).
[CrossRef] [PubMed]

J. Am. Chem. Soc.

A.-M. Pena, T. Boulesteix, T. Dartigalongue and M.-C. Schanne-Klein, "Chiroptical effects in the second harmonic signal of collagens I and IV," J. Am. Chem. Soc. 127, 10314-10322 (2005).
[CrossRef] [PubMed]

J. Biomed. Opt.

K. Schenke-Layland, I. Riemann, U. A. Stock and K. König, "Imaging of cardiovascular structures using near-infrared femtosecond multiphoton laser scanning microscopy," J. Biomed. Opt. 10, 024017 (2005).
[CrossRef] [PubMed]

K. König and I. Riemann, "High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution," J. Biomed. Opt. 8, 432-439 (2003).
[CrossRef] [PubMed]

J. Histochem. Cytochem.

K. L. Kusser and T. D. Randall, "Simultaneous detection of EGFP and cell surface markers by fluorescence Microscopy in Lymphoid Tissues," J. Histochem. Cytochem. 51, 5-14 (2003).
[CrossRef]

J. Int. Med. Res.

M. S. Razzaque, M. Cheng, Y. Horita, M. Nishihara, T. Harada and T. Taguchi, "Immunohistochemical analysis of type III and IV collagens in tubulointerstitial damage in human benign nephrosclerosis," J. Int. Med. Res. 23, 480-486 (1995).
[PubMed]

J. Neurosci. Methods

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

J. Pathol.

N. Buyukbabani and D. Droz, "Distribution of the extracellular matrix components in human glomerular lesions," J. Pathol. 172, 199-207 (1994).
[CrossRef] [PubMed]

J. Struct. Biol.

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

D. J. S. Hulmes, "Building collagen molecules, fibrils, and suprafibrillar structures," J. Struct. Biol. 137,2-10 (2002).
[CrossRef] [PubMed]

Kidney Int.

A. Fogo, J. A. Breyer, M. C. Smith, W. H. Cleveland, L. Agodoa, K. A. Kirk and R. Glassock, "Accuracy of the diagnosis of hypertensive nephrosclerosis in African Americans: a report from the African American Study of Kidney Disease (AASK) Trial. AASK pilot study investigators," Kidney Int. 51 244-252 (1997).
[CrossRef] [PubMed]

Lasers Surg. Med.

B. M. Kim, J. Eichler, K. M. Reiser, A. M. Rubenchik and L. B. Da Silva, "Collagen structure and nonlinear susceptibility: effects of heat, glycation, and enzymatic cleavage on second harmonic signal intensity," Lasers Surg. Med. 27, 329-335 (2000).
[CrossRef] [PubMed]

Microsc. Res. Tech.

W. Lo, S-W. Teng, H-Y. Tan, K. H. Kim, H-C. Chen, H-S. Lee, Y-F. Chen, P. T. C. So and C.-Y. Dong, "Intact corneal stroma visualization of GFP mouse revealed by multiphoton imaging," Microsc. Res. Tech. 69, 973-975 (2006)
[CrossRef] [PubMed]

A.-M. Pena, A. Fabre, D. Débarre, J. Marchal-Somme, B. Crestani, J.-L. Martin, E. Beaurepaire and M.-C. Schanne-Klein, "Three-dimensional investigation and scoring of extracellular matrix remodeling during lung fibrosis using multiphoton microscopy," Microsc. Res. Tech.in press (2007).
[CrossRef]

Nat. Med.

E. Brown, T. Mckee, E. Ditomaso, 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]

Nat. Methods

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

Opt. Commun.

J. Mertz and L. Moreaux, "Second-harmonic generation by focused excitation in inhomogeneously distributed scatterers," Opt. Commun. 196, 325-330 (2001).
[CrossRef]

Opt. Express

Opt. Lett.

Proc. Natl. Acad. Sci. USA

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

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

A.-M. Pena, M. Strupler, T. Boulesteix, G. Godeau and M.-C. Schanne-Klein, "Spectroscopic analysis of keratin endogenous signal for skin multiphoton microscopy," Opt. Express 13, 6268-6274; erratum: 13 6667 (2005).
[CrossRef] [PubMed]

Supplementary Material (3)

» Media 1: MOV (1327 KB)     
» Media 2: MOV (2277 KB)     
» Media 3: MOV (2219 KB)     

Cited By

OSA participates in CrossRef's Cited-By Linking service. Citing articles from OSA journals and other participating publishers are listed here.

Alert me when this article is cited.


Figures (6)

Fig. 1.
Fig. 1.

Comparison of SHG imaging to immunolabeling of collagens type I and type IV in serial frozen sections of murine renal tissue showing artery (A) and tubules (T). (a-c) 2PEF signals from FITC-coupled antibodies revealing (a) collagen I or (b) collagen IV, and (c) from endogenous chromophores such as elastin or cellular chromophores. (d-f) SHG signals in the adventitia. (g-h) Colocalization (white) of 2PEF (red) and SHG (green) signals showing that SHG arises from collagen I, not from collagen IV. (i) Merging of (c) and (f) showing the distribution of endogenous signals. Excitation: 25 mW at 800 nm, 20×0.9 NA objective, scale bar: 100 μm.

Fig. 2.
Fig. 2.

Comparison of histological staining and endogenous SHG in fibrotic kidney tissue: serial sections of mouse kidney (4 μm thick) visualized by (a-b) transmitted light microscopy, (c) polarized light microscopy, and (d,f) multiphoton microscopy (20 mW excitation at 860 nm, circular polarization, NA 0.9). G: glomerulus, A: artery, T: tubules, scale bar: 100 μm. (a) Masson’s trichrome staining; (b-c) Picrosirius red staining. (d) Multiphoton image showing SHG from collagen fibers (green) and endogenous 2PEF underlining the kidney morphology (red). (e) Spectra of these nonlinear endogenous signals (2PEF: red, and SHG: green), superimposed to the spectral transmittance of the SHG (blue) and 2PEF (pink) detection channels. (f) SHG image only (zoomed-in view in inset).

Fig. 3.
Fig. 3.

(a). (1.3 MB) Multimodal multiphoton z-stack of a fixed fibrotic transgenic mouse kidney: the endogenous 2PEF signals (red) reveal the morphology of the tissue, the SHG signals (green) map the distribution of fibrosis and the 2PEF signals from eGFP (white) evidence the collagen synthesis sites (860 nm excitation, 6 mW for the eGFP and 28 mW for the endogenous signals, 50 μm scale bar, 488×488×44 μm3, 13.4 s per image plane). [Media 1] (b) (2.3 MB) 3D reconstruction of SHG signal from collagen fibers in a fresh fibrotic mouse kidney (50 mW excitation at 860 nm, 477×406×58 μm3, 10.9 s per image plane). [Media 2]

Fig. 4.
Fig. 4.

Schematic of the image processing showing vignetting correction for 2PEF data, and vignetting correction, background correction and application of a threshold for SHG data. D stands for the mask image (pixel value=1 if significant SHG signal, 0 otherwise). S is the corrected SHG image where the pixels with no significant SHG signal are set to 0.

Fig. 5.
Fig. 5.

(2.2 MB) Corrected 2PEF/SHG image mapping 2.3×4.6 mm2 of an unlabeled fibrotic murine kidney with 0.8×0.8 μm2 pixel size (2967×5865 pixels scanned at 100 kHz, with 20×0.95-NA objective lens and 50 mW excitation at 860 nm, in a coronal fixed section). Intrinsic 2PEF signals (E700sp and GG455 filters, Chroma) arises from various cellular chromophores in the tubules and the collecting ducts, and SHG signals (E700sp and HQ430/20 filters, Chroma) map the distribution of fibrosis. The insets show the various distributions of collagen fibers scored in Table 1. [Media 3]

Fig. 6.
Fig. 6.

Morphological segmentation and SHG fibrosis scoring in the renal cortex of control versus treated mice. (a) Endogenous 2PEF image revealing the renal morphology and (b) SHG image evidencing collagen fibers, superimposed with the radial segmentation from the capsule (0 %) to the papilla (100 %) in the same unlabeled fibrotic kidney tissue as in Fig. 5(c) D score, expressed as a volume density of fibrosis, along the kidney cortex in control (light green) and Angiotensin II infused mice (dark green). Error bars correspond to standard errors of the mean.

Tables (2)

Tables Icon

Table 1. SHG scoring of areas enlarged in Fig. 5.

Tables Icon

Table 2. SHG scoring of control versus treated mice. Error bars correspond to standard errors of the mean.

Metrics