Research Papers

Imaging corneal pathology in a transgenic mouse model using nonlinear microscopy

[+] Author Affiliations
Julia G. Lyubovitsky

University of California, Beckman Laser Institute, Laser Microbeam and Medical Program, Irvine, California 92612

Joel A. Spencer

University of California, Departments of Medicine and Biological Chemistry, Irvine, California 92697

Tatiana B. Krasieva

University of California, Beckman Laser Institute, Laser Microbeam and Medical Program, Irvine, California 92612

Bogi Andersen

University of California, Departments of Medicine and Biological Chemistry, Irvine, California 92697

Bruce J. Tromberg

University of California, Beckman Laser Institute, Laser Microbeam and Medical Program, Irvine, California 92612

J. Biomed. Opt. 11(1), 014013 (January 31, 2006). doi:10.1117/1.2163254
History: Received May 25, 2005; Accepted September 01, 2005; Published January 31, 2006
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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.

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© 2006 Society of Photo-Optical Instrumentation Engineers

Citation

Julia G. Lyubovitsky ; Joel A. Spencer ; Tatiana B. Krasieva ; Bogi Andersen and Bruce J. Tromberg
"Imaging corneal pathology in a transgenic mouse model using nonlinear microscopy", J. Biomed. Opt. 11(1), 014013 (January 31, 2006). ; http://dx.doi.org/10.1117/1.2163254


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