26 February 2010 Nonlinear 3D microscopy of ex vivo corneas
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A multiphoton microscope has been developed to investigate the sources of nonlinear fluorescence (TPEF) and second harmonic generation (SHG) in non-stained samples of ex-vivo corneas. Stacks of images from different depths are recorded to reconstruct high-resolution 3D (volume) images of the cornea. The corneal epithelium and endothelium provide significant TPEF signal, while the only source of SHG is the stroma. Within the stroma, the keratocytes can also be visualized. Volumetric 3D images of the cornea combining TPEF and SHG signals are useful to characterize the organization of the corneal collagen and to describe the distribution of keratocytes. These images will help to better understand how different pathologies modify the corneal structure and to control the changes produced by surgical or healing processes.
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Juan M. Bueno, Juan M. Bueno, Emilio J. Gualda, Emilio J. Gualda, Pablo Artal, Pablo Artal, } "Nonlinear 3D microscopy of ex vivo corneas", Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 756937 (26 February 2010); doi: 10.1117/12.840720; https://doi.org/10.1117/12.840720

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