5 March 2015 Characterization of porcine eyes based on autofluorescence lifetime imaging
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Proceedings Volume 9329, Multiphoton Microscopy in the Biomedical Sciences XV; 93290E (2015); doi: 10.1117/12.2077563
Event: SPIE BiOS, 2015, San Francisco, California, United States
Abstract
Multiphoton microscopy is a non-invasive imaging technique with ideal characteristics for biological applications. In this study, we propose to characterize three major structures of the porcine eye, the cornea, crystalline lens, and retina using two-photon excitation fluorescence lifetime imaging microscopy (2PE-FLIM).

Samples were imaged using a laser-scanning microscope, consisting of a broadband sub-15 femtosecond (fs) near-infrared laser. Signal detection was performed using a 16-channel photomultiplier tube (PMT) detector (PML-16PMT). Therefore, spectral analysis of the fluorescence lifetime data was possible. To ensure a correct spectral analysis of the autofluorescence lifetime data, the spectra of the individual endogenous fluorophores were acquired with the 16-channel PMT and with a spectrometer. All experiments were performed within 12h of the porcine eye enucleation.

We were able to image the cornea, crystalline lens, and retina at multiple depths. Discrimination of each structure based on their autofluorescence intensity and lifetimes was possible. Furthermore, discrimination between different layers of the same structure was also possible. To the best of our knowledge, this was the first time that 2PE-FLIM was used for porcine lens imaging and layer discrimination. With this study we further demonstrated the feasibility of 2PE-FLIM to image and differentiate three of the main components of the eye and its potential as an ophthalmologic technique.
© (2015) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Ana Batista, Hans Georg Breunig, Aisada Uchugonova, António Miguel Morgado, Karsten König, "Characterization of porcine eyes based on autofluorescence lifetime imaging", Proc. SPIE 9329, Multiphoton Microscopy in the Biomedical Sciences XV, 93290E (5 March 2015); doi: 10.1117/12.2077563; https://doi.org/10.1117/12.2077563
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KEYWORDS
Retina

Cornea

Eye

Fluorescence lifetime imaging

Second-harmonic generation

Luminescence

Collagen

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