23 February 2018 Second harmonic generation microscopy of the living human cornea
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Second Harmonic Generation (SHG) microscopy provides high-resolution structural imaging of the corneal stroma without the need of labelling techniques. This powerful tool has never been applied to living human eyes so far. Here, we present a new compact SHG microscope specifically developed to image the structural organization of the corneal lamellae in living healthy human volunteers. The research prototype incorporates a long-working distance dry objective that allows non-contact three-dimensional SHG imaging of the cornea. Safety assessment and effectiveness of the system were firstly tested in ex-vivo fresh eyes. The maximum average power of the used illumination laser was 20 mW, more than 10 times below the maximum permissible exposure (according to ANSI Z136.1-2000). The instrument was successfully employed to obtain non-contact and non-invasive SHG of the living human eye within well-established light safety limits. This represents the first recording of in vivo SHG images of the human cornea using a compact multiphoton microscope. This might become an important tool in Ophthalmology for early diagnosis and tracking ocular pathologies.
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Pablo Artal, Francisco Ávila, and Juan Bueno "Second harmonic generation microscopy of the living human cornea", Proc. SPIE 10498, Multiphoton Microscopy in the Biomedical Sciences XVIII, 1049810 (23 February 2018); doi: 10.1117/12.2290470; https://doi.org/10.1117/12.2290470

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