September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
In vivo linear and nonlinear imaging of corneal structures on BALB/c and streptozotocin-diabetic Thy1-YFP mice
Author Affiliations & Notes
  • Oliver Stachs
    Department of Ophthalmology, University of Rostock, Rostock, Germany
  • Tobias Ehmke
    Laser Zentrum Hannover e.V., Hannover, Germany
  • Janine Leckelt
    Institute of Medical Biochemistry and Molecular Biology, University of Rostock, Rostock, Germany
  • Maria Reichard
    Department of Ophthalmology, University of Rostock, Rostock, Germany
  • Heike Weiss
    Institute of Medical Biochemistry and Molecular Biology, University of Rostock, Rostock, Germany
  • Marine Hovakimyan
    Institute for Biomedical Engineering, University of Rostock, Rostock, Germany
  • Alexander Heisterkamp
    Institute of Quantum Optics, Leibniz University Hannover, Hannover, Germany
  • Simone Baltrusch
    Institute of Medical Biochemistry and Molecular Biology, University of Rostock, Rostock, Germany
  • Footnotes
    Commercial Relationships   Oliver Stachs, None; Tobias Ehmke, None; Janine Leckelt, None; Maria Reichard, None; Heike Weiss, None; Marine Hovakimyan, None; Alexander Heisterkamp, None; Simone Baltrusch, None
  • Footnotes
    Support  German Diabetes Association (DDG); German Federal Ministry of Education and Research (REMEDIS)
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 2190. doi:
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      Oliver Stachs, Tobias Ehmke, Janine Leckelt, Maria Reichard, Heike Weiss, Marine Hovakimyan, Alexander Heisterkamp, Simone Baltrusch; In vivo linear and nonlinear imaging of corneal structures on BALB/c and streptozotocin-diabetic Thy1-YFP mice. Invest. Ophthalmol. Vis. Sci. 2016;57(12):2190.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Two-photon microscopy (TPM) allows high contrast imaging at a subcellular resolution scale. In this work, TPM was applied to visualize corneal structures in two mouse models (BALB/c and B6.Cg-Tg(Thy1-YFP)16Jrs/J) in vivo. In particular, the transgenic Thy1-YFP mice expressing the yellow fluorescent protein (YFP) in all motor and sensory neurons had been used for investigating the nerve fiber density in healthy and streptozotocin(STZ)-diabetic mice.

Methods : We present nonlinear microscopy (modified MPM200, Thorlabs, New Jersey, USA) as a complementary technique for visualizing corneal structures in in vivo mouse models (BALB/c and Thy1-YFP). In particular, TPM was used in addition to CLSM (HRT II + RCM, Heidelberg Engineering, Heidelberg, Germany) to visualize and quantify the corneal nerve fiber density in STZ diabetic Thy1-YFP mice in comparison to controls.

Results : In a first set of experiments, corneas of age-matched healthy BALB/c mice had been imaged in order to demonstrate the applicability of both techniques in vivo. Furthermore STZ-treated mice developed a significant increase of blood glucose concentrations (20.24 ± 1.42 mmol/l (diabetic) versus 7.42 ± 0.56 mmol/l (healthy)) and were classified as diabetic. The corneal nerve fiber density was compared between STZ-diabetic and control Thy1-YFP mice via CLSM. STZ-diabetic Thy1-YFP mice exhibited a significantly lower subbasal nerve fiber density, when compared to controls (healthy 20.84 ± 0.81 mm/mm2; STZ-diabetic 16.32 ± 0.72 mm/mm2). To evaluate the subbasal nerve plexus in the same animals by a complementary imaging technique to CLSM, TPM was established in healthy and STZ-diabetic Thy1-YFP mouse. Although higher absolute nerve fiber density values were observed when compared to CLSM, also the TPM analyses revealed a significant decrease of nerve fibers in STZ-diabetic mice compared to controls (healthy: 30.2 ± 0.70 mm/mm2; STZ-diabetic: 21.15 ± 1.49 mm/mm2). Correlation of individual nerve fiber density from Thy1-YFP mice revealed a significant correlation between both methods.

Conclusions : Thus, the present study provides further evidence of corneal confocal microscopy as a promising technique for non-invasive detection of diabetic neuropathy. Information derived from these experiments may become clinically relevant and help to develop new drugs for treatment of diabetic neuropathy.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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