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Journal of Biomedical Optics • new

High-resolution, label-free two-photon imaging of diseased human corneas
Author(s): Ana Batista; Hans Georg Breunig; Aisada König; Andreas Schindele; Tobias Hager; Berthold Seitz; Karsten König
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Paper Abstract

The diagnosis of corneal diseases may be improved by monitoring the metabolism of cells and the structural organization of the stroma using two-photon imaging (TPI). We used TPI to assess the differences between nonpathological (NP) human corneas and corneas diagnosed with either keratoconus, Acanthamoeba keratitis, or stromal corneal scars. Images were acquired using a custom-built five-dimensional laser-scanning microscope with a broadband sub-15 femtosecond near-infrared pulsed excitation laser and a 16-channel photomultiplier tube detector in combination with a time-correlated single photon counting module. Morphological alterations of epithelial cells were observed for all pathologies. Moreover, diseased corneas showed alterations to the cells’ metabolism that were revealed using the NAD(P)H free to protein-bound ratios. The mean autofluorescence lifetime of the stroma and the organization of the collagen fibers were also significantly altered due to the pathologies. We demonstrate that TPI can be used to distinguish between NP and diseased human corneas, based not only on alterations of the cells’ morphology, which can also be evaluated using current clinical devices, but on additional morphological and functional features such as the organization of the stroma and the cells’ metabolism. Therefore, TPI could become an efficient tool for diagnosing corneal diseases and better understanding the biological processes of the diseases.

Paper Details

Date Published: 2 March 2018
PDF: 8 pages
J. Biomed. Opt. 23(3) 036002 doi: 10.1117/1.JBO.23.3.036002
Published in: Journal of Biomedical Optics Volume 23, Issue 3
Show Author Affiliations
Ana Batista, Univ. des Saarlandes (Germany)
JenLab GmbH (Germany)
Hans Georg Breunig, Univ. des Saarlandes (Germany)
JenLab GmbH (Germany)
Aisada König, Univ. des Saarlandes (Germany)
JenLab GmbH (Germany)
Andreas Schindele, JenLab GmbH (Germany)
Tobias Hager, Univ. des Saarlandes (Germany)
Berthold Seitz, Universitätsklinikum des Saarlandes (Germany)
Univ. des Saarlandes (Germany)
Karsten König, JenLab GmbH (Germany)
Univ. des Saarlandes (Germany)

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