Reflective confocal laser scanning microscopy and nonlinear microscopy of cross-linked rabbit cornea

Alexander Krueger; Marina Hovakimyan; Diego F. Ramirez; Oliver Stachs; Rudolf F. Guthoff; Alexander Heisterkamp

doi:10.1117/12.831478

1 July 2009 Reflective confocal laser scanning microscopy and nonlinear microscopy of cross-linked rabbit cornea

Alexander Krueger, Marina Hovakimyan, Diego F. Ramirez, Oliver Stachs, Rudolf F. Guthoff, Alexander Heisterkamp

Author Affiliations +

Proceedings Volume 7367, Advanced Microscopy Techniques; 736719 (2009) https://doi.org/10.1117/12.831478
Event: European Conferences on Biomedical Optics, 2009, Munich, Germany

Abstract

Cross-linking of the cornea with application of Ribovlavin and UV-A light is an evolving clinical treatment of the eye disease keratoconus. Despite the positive clinical track record of corneal cross-linking, the complex wound healing process after the treatment is still under investigation. In this study an animal model was used to clarify the state of wound healing 5 weeks after treatment. Cross-linked rabbit corneae were imaged with reflective confocal laser scanning and nonlinear microscopy, namely second harmonic imaging microscopy (SHIM) and two-photon excited autofluorescence. First results show that the NAD(P) H-autofluorescence of the corneal keratocytes and their scattering signal still show a signature of the treatment five weeks after the cross-linking procedure. The SHIM signals show the structural morphology of the fibrous collagen sheets in the stroma of the cornea. SHIM detected in the forward direction differs substantially from backward SHIM, but no signature of treatment was found in both detection channels of the SHIM signal.

Citation Download Citation

Alexander Krueger, Marina Hovakimyan, Diego F. Ramirez, Oliver Stachs, Rudolf F. Guthoff, and Alexander Heisterkamp "Reflective confocal laser scanning microscopy and nonlinear microscopy of cross-linked rabbit cornea", Proc. SPIE 7367, Advanced Microscopy Techniques, 736719 (1 July 2009); https://doi.org/10.1117/12.831478

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KEYWORDS

Cornea

Second-harmonic generation

Confocal microscopy

Reflectivity

Microscopy

Collagen

Scanning helium ion microscopy

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