NDLI: Two-Photon Fluorescence Microscopy for Determination of the Riboflavin Concentration in the Anterior Corneal Stroma When Using the Dresden Protocol.

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Two-Photon Fluorescence Microscopy for Determination of the Riboflavin Concentration in the Anterior Corneal Stroma When Using the Dresden Protocol.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Stachs, Oliver Fischinger, Isaak Seiler, Theo G. Ehmke, Tobias Zapp, Daniel Heisterkamp, Alexander Seiler, Theo
Description	Country affiliation: Switzerland Author Affiliation: Seiler TG ( Klinik und Poliklinik für Augenheilkunde, Technische Universität München, Munich, Germany 2Institut für Refraktive und Ophthalmo-Chirurgie (IROC), Zurich, Switzerland.); Ehmke T ( Laser Zentrum Hannover e.V., Hanover, Germany.); Fischinger I ( Klinik und Poliklinik für Augenheilkunde, Technische Universität München, Munich, Germany 2Institut für Refraktive und Ophthalmo-Chirurgie (IROC), Zurich, Switzerland.); Zapp D ( Klinik und Poliklinik für Augenheilkunde, Technische Universität München, Munich, Germany.); Stachs O ( Department of Ophthalmology, Rostock University Medical Center, Rostock, Germany.); Seiler T ( Institut für Refraktive und Ophthalmo-Chirurgie (IROC), Zurich, Switzerland.); Heisterkamp A ( Laser Zentrum Hannover e.V., Hanover, Germany 5Institut für Quantenoptik, Leibniz Universität Hannover, Hanover, Germany.)
Abstract	PURPOSE: To determine the riboflavin concentration gradient in the anterior corneal stroma when using the Dresden protocol with different dextran solutions. METHODS: Three different groups of porcine corneas, five each, were compared regarding the riboflavin concentration in the anterior stroma. Before all experiments, stable hydration conditions were established for the corresponding solution. All groups were treated with 0.1% riboflavin in different dextran solutions (15%, 16%, 20%). After imbibition, two-photon microscopy was used to determine fluorescence intensity. For signal attenuation and concentration determination corneas were saturated and measured a second time by two-photon microscopy. Additionally, the distribution was calculated mathematically and compared to the empiric results. RESULTS: Riboflavin concentration is decreasing with depth for all dextran solutions. A nearly constant concentration could be determined over the first 75 µm. Analysis of the fit functions leads to diffusion coefficients of D = 2.97 × 10-7 cm2/s for the 15% dextran solution, D = 2.34 × 10-7 cm2/s for the 16% dextran solution, and D = 1.28 × 10-7 cm2/s for the 20% dextran solution. The riboflavin gradients of the 20% dextran group were statistically significantly different from 15% dextran starting at a depth of 220 µm and deeper (P = 0.047). The 16% dextran group differed statistically at a depth of 250 µm and deeper (P = 0.047). These results show a significant difference to those published previously. CONCLUSIONS: With correct settings two-photon microscopy is a precise way to determine the concentration of riboflavin in cornea. The measured gradient is excellently fit by a Gaussian distribution, which comes out as a solution of Fick's second law.
ISSN	01460404
e-ISSN	15525783
Journal	Investigative Opthalmology & Visual Science
Issue Number	11
Volume Number	56
Language	English
Publisher	Association for Research in Vision and Ophthalmology
Publisher Date	2015-10-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Anterior Eye Segment Metabolism Corneal Stroma Microscopy, Fluorescence, Multiphoton Photosensitizing Agents Pharmacokinetics Riboflavin Animals Dextrans Ophthalmic Solutions Swine Tissue Distribution Discipline Ophthalmology
Content Type	Text
Resource Type	Article
Subject	Ophthalmology Sensory Systems Cellular and Molecular Neuroscience

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