Second harmonic generation microscopy of the living human cornea

Pablo Artal; Francisco Ávila; Juan Bueno

doi:10.1117/12.2290470

23 February 2018 Second harmonic generation microscopy of the living human cornea

Pablo Artal, Francisco Ávila, Juan Bueno

Proceedings Volume 10498, Multiphoton Microscopy in the Biomedical Sciences XVIII; 1049810 (2018) https://doi.org/10.1117/12.2290470
Event: SPIE BiOS, 2018, San Francisco, California, United States

Abstract

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.

Citation Download Citation

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); https://doi.org/10.1117/12.2290470

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