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H.–Y. Tan, S.–W. Teng, J.–L. Peng, H.–H. Lin, H.–Y. Wu, W. Lo, Y. Sun, W.–C. Lin, S.–J. Lin, C.–Y. Dong; Multiphoton Imaging of Porcine Eyes . Invest. Ophthalmol. Vis. Sci. 2005;46(13):1081.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To demonstrate the feasibility of using multiphoton microscopy in imaging ocular surface. Methods: The porcine eyes used for imaging were immersed in PBS buffer for viewing. The home–built multiphoton microscope used for this study was constructed from a commercial upright microscope (Nikon E800). Using a 40x, NA 0.8 water immersion objective (Fluor), the 780 nm photons from a titanium sapphire laser were used to induced multiphoton autofluorescence and second–harmonic generation(SHG) signals from different positions and depths of porcine ocular surface. Results:Without extrinsic fluorescent molecules, we were able to image the ocular surface. The cornea and limbal epithelial cells (autofluorescence), as well as the well–organized corneal stromal collagen fibers (SHG) are visible . The imaging results were compatible with the histological morphology. Conclusions: We demonstrate an excellent imaging of porcine ocular surface using multiphoton induced fluorescence and SHG signals, without the introduction of extrinsic fluorescent molecules. With additional developements, multiphoton microscopy may in future be applicated for in vivo investigation of ophthalmologic pathologies.
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