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Journal of Biomedical Optics

Imaging corneal pathology in a transgenic mouse model using nonlinear microscopy
Author(s): Julia G. Lyubovitsky; Joel A. Spencer; Tatiana B. Krasieva; Bogi Andersen; Bruce Jason Tromberg
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Paper Abstract

A transgenic mouse model with a Clim [co-factor of LIM (a combination of first letters of Lin-11 (C. elegans), ISL1 (rat), and Mec-3 (C. elegans) gene names) domain proteins] gene partially blocked in the epithelial compartment of its tissues is used to establish the sensitivity of intrinsic reflectance nonlinear optical microscopy (NLOM) to stromal and cellular perturbations in the cornea. Our results indicate dysplasia in the squamous epithelium, irregular collagen arrays in the stroma, and a compromised posterior endothelium in the corneas of these mice. As suggested by biochemical data, the collagen alterations are likely due to collagen III synthesis and deposition during healing and remodeling of transgenic mice corneal stromas. All of the topographic features seen in NLOM images of normal and aberrant corneas are confirmed by coregistration with histological sections. In this work, we also use ratiometric redox fluorometry based on two-photon excited cellular fluorescence from reduced nicotinamide adenine dinucleotide (NAD)(P)H and oxidized flavin adenine dinucleotide (FAD) to study mitocondrial energy metabolism. Employing this method, we detect higher metabolic activity in the endothelial layer of cornea compared to an epithelial layer located further away from the metabolites. The combination of two-photon excited fluorescence (TPF) with second harmonic generation (SHG) signals allows imaging to aid in understanding the relationship between alternation of specific genes and structural changes in cells and extracellular matrix.

Paper Details

Date Published: 1 January 2006
PDF: 6 pages
J. Biomed. Opt. 11(1) 014013 doi: 10.1117/1.2163254
Published in: Journal of Biomedical Optics Volume 11, Issue 1
Show Author Affiliations
Julia G. Lyubovitsky, Univ. of California/Irvine (United States)
Joel A. Spencer, Univ. of California/Irvine (United States)
Tatiana B. Krasieva, Univ. of California/Irvine (United States)
Bogi Andersen, Univ. of California/Irvine (United States)
Bruce Jason Tromberg, Univ. of California/Irvine (United States)

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