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Related Experiment Videos

Imaging corneal pathology in a transgenic mouse model using nonlinear microscopy.

Julia G Lyubovitsky1, Joel A Spencer, Tatiana B Krasieva

  • 1University of California, Beckman Laser Institute, Laser Microbeam and Medical Program, Irvine, California 92612, USA. jlyubovi@uci.edu

Journal of Biomedical Optics
|March 11, 2006
PubMed
Summary

Nonlinear optical microscopy (NLOM) reveals corneal abnormalities in transgenic mice with blocked Clim gene function. These changes include epithelial dysplasia, stromal collagen irregularities, and endothelial compromise, impacting corneal structure and metabolism.

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Area of Science:

  • Ophthalmology
  • Biomedical Engineering
  • Molecular Biology

Background:

  • Gene alterations can lead to significant structural and metabolic changes in tissues.
  • Nonlinear optical microscopy (NLOM) offers advanced imaging capabilities for biological tissues.

Purpose of the Study:

  • To investigate the sensitivity of NLOM to detect corneal perturbations caused by genetic modification.
  • To correlate gene expression changes with structural and metabolic alterations in the cornea.

Main Methods:

  • Utilized a transgenic mouse model with partially blocked Clim gene function in the corneal epithelium.
  • Employed intrinsic reflectance NLOM to image corneal structure, including epithelium, stroma, and endothelium.
  • Applied ratiometric redox fluorometry (two-photon excited cellular fluorescence of NAD(P)H and FAD) to assess mitochondrial energy metabolism.

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  • Confirmed NLOM findings with histological sections and biochemical data.

    • Main Results:

    • NLOM detected epithelial dysplasia, irregular collagen arrays in the stroma, and a compromised posterior endothelium in transgenic corneas.
    • Collagen alterations were linked to increased synthesis and deposition of collagen III during corneal remodeling.
    • Ratiometric redox fluorometry revealed higher metabolic activity in the corneal endothelium compared to the epithelium.
    • Combined TPF and SHG imaging elucidated the relationship between gene alternation and structural changes.

    Conclusions:

    • NLOM is sensitive to genetic perturbations affecting corneal structure and cellular metabolism.
    • Clim gene modification induces significant corneal abnormalities, including stromal and endothelial damage.
    • The combination of NLOM and metabolic imaging provides a powerful tool for studying gene-environment interactions in corneal diseases.