Share Email Print

Proceedings Paper

Two-photon autofluorescence lifetime and SHG imaging of healthy and diseased human corneas
Author(s): Ana Batista; Hans Georg Breunig; Aisada Uchugonova; Berthold Seitz M.D.; António Miguel Morgado; Karsten König
Format Member Price Non-Member Price
PDF $17.00 $21.00

Paper Abstract

Corneal function can be drastically affected by several degenerations and dystrophies, leading to blindness. Early diagnosis of corneal disease is of major importance and it may be accomplished by monitoring changes of the metabolic state and structural organization, the first detectable pathological signs, by two-photon excitation autofluorescence lifetime and second-harmonic generation imaging. In this study, we propose to use these imaging techniques to differentiate between healthy and pathological corneas. Images were acquired using a laser-scanning microscope with a broadband sub-15 femtosecond near-infrared pulsed laser and a 16-channel photomultiplier tube detector for signal collection. This setup allows the simultaneous excitation of metabolic co-factors and to identify them based on their fluorescence spectra. We were able to discriminate between healthy and pathological corneas using two-photon excitation autofluorescence lifetime and second-harmonic generation imaging from corneal epithelium and stroma. Furthermore, differences between different pathologies were observed. Alterations in the metabolic state of corneal epithelial cells were observed using the autofluorescence lifetime of the metabolic co-factors. In the corneal stroma, we observed not only alterations in the collagen fibril structural organization but also alterations in the autofluorescence lifetime. Further tests are required as the number of pathological samples must be increased. In the future, we intend to establish a correlation between the metabolic and structural changes and the disease stage. This can be a step forward in achieving early diagnosis.

Paper Details

Date Published: 20 March 2015
PDF: 8 pages
Proc. SPIE 9307, Ophthalmic Technologies XXV, 93071Q (20 March 2015); doi: 10.1117/12.2077569
Show Author Affiliations
Ana Batista, IBILI, Univ. de Coimbra (Portugal)
Univ. des Saarlandes (Germany)
Hans Georg Breunig, Univ. des Saarlandes (Germany)
JenLab GmbH (Germany)
Aisada Uchugonova, Univ. des Saarlandes (Germany)
Berthold Seitz M.D., Univ. des Saarlandes (Germany)
António Miguel Morgado, IBILI, Univ. de Coimbra (Portugal)
Karsten König, Univ. des Saarlandes (Germany)
JenLab GmbH (Germany)

Published in SPIE Proceedings Vol. 9307:
Ophthalmic Technologies XXV
Fabrice Manns; Per G. Söderberg; Arthur Ho, Editor(s)

© SPIE. Terms of Use
Back to Top
Sign in to read the full article
Create a free SPIE account to get access to
premium articles and original research
Forgot your username?