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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 3, Iss. 1 — Jan. 1, 2012
  • pp: 1–15

Optics InfoBase > Biomedical Optics Express > Volume 3 > Issue 1 > Page 1

In vivo structural imaging of the cornea by polarization-resolved second harmonic microscopy

Gaël Latour, Ivan Gusachenko, Laura Kowalczuk, Isabelle Lamarre, and Marie‑Claire Schanne-Klein  »View Author Affiliations


Biomedical Optics Express, Vol. 3, Issue 1, pp. 1-15 (2012)
http://dx.doi.org/10.1364/BOE.3.000001


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Abstract

The transparency and mechanical strength of the cornea are related to the highly organized three-dimensional distribution of collagen fibrils. It is of great interest to develop specific and contrasted in vivo imaging tools to probe these collagenous structures, which is not available yet. Second Harmonic Generation (SHG) microscopy is a unique tool to reveal fibrillar collagen within unstained tissues, but backward SHG images of cornea fail to reveal any spatial features due to the nanometric diameter of stromal collagen fibrils. To overcome this limitation, we performed polarization-resolved SHG imaging, which is highly sensitive to the sub-micrometer distribution of anisotropic structures. Using advanced data processing, we successfully retrieved the orientation of the collagenous fibrils at each depth of human corneas, even in backward SHG homogenous images. Quantitative information was also obtained about the submicrometer heterogeneities of the fibrillar collagen distribution by measuring the SHG anisotropy. All these results were consistent with numerical simulation of the polarization-resolved SHG response of cornea. Finally, we performed in vivo SHG imaging of rat corneas and achieved structural imaging of corneal stroma without any labeling. Epi-detected polarization-resolved SHG imaging should extend to other organs and become a new diagnosis tool for collagen remodeling.

© 2011 OSA

OCIS Codes
(120.5410) Instrumentation, measurement, and metrology : Polarimetry
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(170.4470) Medical optics and biotechnology : Ophthalmology
(190.4160) Nonlinear optics : Multiharmonic generation
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:
Ophthalmology Applications

History
Original Manuscript: November 7, 2011
Revised Manuscript: November 29, 2011
Manuscript Accepted: November 29, 2011
Published: December 1, 2011

Citation
Gaël Latour, Ivan Gusachenko, Laura Kowalczuk, Isabelle Lamarre, and Marie‑Claire Schanne-Klein, "In vivo structural imaging of the cornea by polarization-resolved second harmonic microscopy," Biomed. Opt. Express 3, 1-15 (2012)
http://www.opticsinfobase.org/boe/abstract.cfm?URI=boe-3-1-1


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References

  1. R. F. Guthoff, A. Zhivov, and O. Stachs, “In vivo confocal microscopy, an inner vision of the cornea - a major review,” Clin. Experiment. Ophthalmol.37(1), 100–117 (2009). [CrossRef] [PubMed]
  2. M. Gora, K. Karnowski, M. Szkulmowski, B. J. Kaluzny, R. Huber, A. Kowalczyk, and M. Wojtkowski, “Ultra high-speed swept source OCT imaging of the anterior segment of human eye at 200 kHz with adjustable imaging range,” Opt. Express17(17), 14880–14894 (2009). [CrossRef] [PubMed]
  3. G. Latour, G. Georges, L. S. Lamoine, C. Deumié, J. Conrath, and L. Hoffart, “Human graft cornea and laser incisions imaging with micrometer scale resolution full-field optical coherence tomography,” J. Biomed. Opt.15(5), 056006 (2010). [CrossRef] [PubMed]
  4. J. H. Krachmer, M. J. Mannis, and E. J. Holland, Cornea, 2nd ed. (Mosby, 2005).
  5. K. Plamann, F. Aptel, C. L. Arnold, A. Courjaud, C. Crotti, F. Deloison, F. Druon, P. Georges, M. Hanna, J.-M. Legeais, F. Morin, É. Mottay, V. Nuzzo, D. A. Peyrot, and M. Savoldelli, “Ultrashort pulse laser surgery of the cornea and the sclera,” J. Opt.12(8), 084002 (2010). [CrossRef]
  6. M. Hao, K. Flynn, C. Nien-Shy, B. E. Jester, M. Winkler, D. J. Brown, O. La Schiazza, J. Bille, and J. V. Jester, “In vivo non-linear optical (NLO) imaging in live rabbit eyes using the Heidelberg Two-Photon Laser Ophthalmoscope,” Exp. Eye Res.91(2), 308–314 (2010). [CrossRef] [PubMed]
  7. 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(1), 493–508 (2002). [CrossRef] [PubMed]
  8. 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. U.S.A.100(12), 7075–7080 (2003). [CrossRef] [PubMed]
  9. D. W. Piston, B. R. Masters, and W. W. Webb, “Three-dimensionally resolved NAD(P)H cellular metabolic redox imaging of the in situ cornea with two-photon excitation laser scanning microscopy,” J. Microsc.178(1), 20–27 (1995). [CrossRef] [PubMed]
  10. A. T. Yeh, N. Nassif, A. Zoumi, and B. J. Tromberg, “Selective corneal imaging using combined second-harmonic generation and two-photon excited fluorescence,” Opt. Lett.27(23), 2082–2084 (2002). [CrossRef] [PubMed]
  11. M. Han, G. Giese, and J. Bille, “Second harmonic generation imaging of collagen fibrils in cornea and sclera,” Opt. Express13(15), 5791–5797 (2005). [CrossRef] [PubMed]
  12. S.-W. Teng, H.-Y. Tan, J.-L. Peng, H.-H. Lin, K. H. Kim, W. Lo, Y. Sun, W.-C. Lin, S.-J. Lin, S.-H. Jee, P. T. C. So, and C.-Y. Dong, “Multiphoton autofluorescence and second-harmonic generation imaging of the ex vivo porcine eye,” Invest. Ophthalmol. Vis. Sci.47(3), 1216–1224 (2006). [CrossRef] [PubMed]
  13. N. Morishige, T. Nishida, and J. J. Jester, “Second harmonic generation for visualizing 3-dimensional structure of corneal collagen lamellae,” Cornea28(Suppl 1), S46–S53 (2009). [CrossRef]
  14. P. Matteini, F. Ratto, F. Rossi, R. Cicchi, C. Stringari, D. Kapsokalyvas, F. S. Pavone, and R. Pini, “Photothermally-induced disordered patterns of corneal collagen revealed by SHG imaging,” Opt. Express17(6), 4868–4878 (2009). [CrossRef] [PubMed]
  15. F. Aptel, N. Olivier, A. Deniset-Besseau, J.-M. Legeais, K. Plamann, M.-C. Schanne-Klein, and E. Beaurepaire, “Multimodal nonlinear imaging of the human cornea,” Invest. Ophthalmol. Vis. Sci.51(5), 2459–2465 (2010). [CrossRef] [PubMed]
  16. N. Olivier, F. Aptel, K. Plamann, M.-C. Schanne-Klein, and E. Beaurepaire, “Harmonic microscopy of isotropic and anisotropic microstructure of the human cornea,” Opt. Express18(5), 5028–5040 (2010). [CrossRef] [PubMed]
  17. J. M. Bueno, E. J. Gualda, A. Giakoumaki, P. Pérez-Merino, S. Marcos, and P. Artal, “Multiphoton microscopy of ex vivo corneas after collagen cross-linking,” Invest. Ophthalmol. Vis. Sci.52(8), 5325–5331 (2011). [CrossRef] [PubMed]
  18. M. Strupler, A.-M. Pena, M. Hernest, P.-L. Tharaux, J.-L. Martin, E. Beaurepaire, and M.-C. Schanne-Klein, “Second harmonic imaging and scoring of collagen in fibrotic tissues,” Opt. Express15(7), 4054–4065 (2007). [CrossRef] [PubMed]
  19. A. Deniset-Besseau, J. Duboisset, E. Benichou, F. Hache, P.-F. Brevet, and M.-C. Schanne-Klein, “Measurement of the second-order hyperpolarizability of the collagen triple helix and determination of its physical origin,” J. Phys. Chem. B113(40), 13437–13445 (2009). [CrossRef] [PubMed]
  20. R. Lacomb, O. Nadiarnykh, S. S. Townsend, and P. J. Campagnola, “Phase Matching considerations in Second Harmonic Generation from tissues: Effects on emission directionality, conversion efficiency and observed morphology,” Opt. Commun.281(7), 1823–1832 (2008). [CrossRef] [PubMed]
  21. M. Strupler and M.-C. Schanne-Klein, “Simulating second harmonic generation from tendon—do we see fibrils?” in Biomedical Optics, OSA Technical Digest (CD) (Optical Society of America, 2010), paper BTuD83.
  22. M. Rivard, M. Laliberté, A. Bertrand-Grenier, C. Harnagea, C. P. Pfeffer, M. Vallières, Y. St-Pierre, A. Pignolet, M. A. El Khakani, and F. Légaré, “The structural origin of second harmonic generation in fascia,” Biomed. Opt. Express2(1), 26–36 (2011). [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. Express15(19), 12286–12295 (2007). [CrossRef] [PubMed]
  24. P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, “Polarization-modulated second harmonic generation in collagen,” Biophys. J.82(6), 3330–3342 (2002). [CrossRef] [PubMed]
  25. O. Nadiarnykh and P. J. Campagnola, “Retention of polarization signatures in SHG microscopy of scattering tissues through optical clearing,” Opt. Express17(7), 5794–5806 (2009). [CrossRef] [PubMed]
  26. I. Gusachenko, G. Latour, and M.-C. Schanne-Klein, “Polarization-resolved Second Harmonic microscopy in anisotropic thick tissues,” Opt. Express18(18), 19339–19352 (2010). [CrossRef] [PubMed]
  27. S. Brasselet, D. Aït-Belkacem, A. Gasecka, F. Munhoz, S. Brustlein, and S. Brasselet, “Influence of birefringence on polarization resolved nonlinear microscopy and collagen SHG structural imaging,” Opt. Express18(14), 14859–14870 (2010). [CrossRef] [PubMed]
  28. S. Psilodimitrakopoulos, S. I. C. O. Santos, I. Amat-Roldan, A. K. N. Thayil, D. Artigas, and P. Loza-Alvarez, “In vivo, pixel-resolution mapping of thick filaments’ orientation in nonfibrilar muscle using polarization-sensitive second harmonic generation microscopy,” J. Biomed. Opt.14(1), 014001 (2009). [CrossRef] [PubMed]
  29. W.-L. Chen, T.-H. Li, P.-J. Su, C.-K. Chou, P. T. Fwu, S.-J. Lin, D. Kim, P. T. C. So, and C.-Y. Dong, “Second harmonic generation chi tensor microscopy for tissue imaging,” Appl. Phys. Lett.94(18), 183902 (2009). [CrossRef]
  30. V. Nucciotti, C. Stringari, L. Sacconi, F. Vanzi, L. Fusi, M. Linari, G. Piazzesi, V. Lombardi, and F. S. Pavone, “Probing myosin structural conformation in vivo by second-harmonic generation microscopy,” Proc. Natl. Acad. Sci. U.S.A.107(17), 7763–7768 (2010). [CrossRef] [PubMed]
  31. S. V. Plotnikov, A. C. Millard, P. J. Campagnola, and W. A. Mohler, “Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres,” Biophys. J.90(2), 693–703 (2006). [CrossRef] [PubMed]
  32. E. E. B. A. Directory, 18th ed. (European Eye Bank Association, 2010).
  33. P. Réfrégier, M. Roche, and S. Brasselet, “Precision analysis in polarization-resolved second harmonic generation microscopy,” Opt. Lett.36(11), 2149–2151 (2011). [CrossRef] [PubMed]
  34. J. C. Mansfield, C. P. Winlove, J. Moger, and S. J. Matcher, “Collagen fiber arrangement in normal and diseased cartilage studied by polarization sensitive nonlinear microscopy,” J. Biomed. Opt.13(4), 044020 (2008). [CrossRef] [PubMed]
  35. L. J. Bour, “Polarized light and the eye” in Visual Optics and Instrumentation, W. N. Charman, ed. (CRC Press,1991), Vol. 1, Chap. 13.
  36. L. Novotny and B. Hecht, Principles of Nano-Optics (Cambridge University Press, 2006).
  37. W. Radner, M. Zehetmayer, R. Aufreiter, and R. Mallinger, “Interlacing and cross-angle distribution of collagen lamellae in the human cornea,” Cornea17(5), 537–543 (1998). [CrossRef] [PubMed]
  38. S. Allgeier, A. Zhivov, F. Eberle, B. Koehler, S. Maier, G. Bretthauer, R. F. Guthoff, and O. Stachs, “Image reconstruction of the subbasal nerve plexus with in vivo confocal microscopy,” Invest. Ophthalmol. Vis. Sci.52(9), 5022–5028 (2011). [CrossRef] [PubMed]

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