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Purpose: Although the frequency in which practitioners are fitting scleral
contact lenses is increasing, the recommendation for proper tear layer depth
(thickness) varies amongst experts. The main goal of this paper is to clinically
verify the effect of varying tear layer depths on induced corneal edema during
lens wear.
Methods: Ten subjects with healthy eyes were fitted with scleral lenses on their
right eye. Each of them was fit with two different lenses: one with an apical
clearance of 200 μm and another with an apical clearance of 600 μm. They wore
the lenses for 8 hours on two different days, with at least a one week wash-out
period. Lenses were applied at 8 a.m. on each of the testing days. Pachymetry
measurements were taken one day prior to lens wear at 4 p.m., on the day of
wear prior to lens application, and after removal of the lenses at 4 p.m.
Measurements were collected using both the Pentacam® HR Corneal
Tomographer, as well as the Visante Anterior Segment Optical Coherence
Tomographer (OCT). The apical clearance was measured using the
Visante OCT at two intervals during the test day: immediately after application of
the lens and immediately prior to the removal of the lens.
Results: In this study, there was found to be no significant difference in corneal
edematous response during lens wear between the two test groups. The study
shows that the eyes with the lenses have a statistically significantly thicker
cornea compared to the non-lens-wearing eye after wearing either lens for 8
hours, lying within clinically and physiologically acceptable limits.
Conclusion: Our clinical results do not correlate with current theoretical
calculations, which predict a greater amount of corneal swelling with increasing
tear layer thickness. It has to be evaluated if the effect on corneal edema
changes with longer wearing periods, larger samples or other influences.
Key words: scleral (contact) lens, corneal edema, pachymetry, tear layer
thickness, vaulting, apical clearance
Pharmaceutical agents or drugs often have a pronounced impact on protein-protein interactions in cells, and in particular, cell membranes. Changes of molecular conformations as well as of intermolecular interactions may affect dipole-dipole interaction between chromophoric groups, which can be proven by measuring the Förster resonance energy transfer (FRET). If these chromophores are located within or in close proximity to the plasma membrane, they are excited preferentially by an evanescent electromagnetic wave upon total internal reflection (TIR) of an incident laser beam. For the TIR-FRET screening of larger cell collectives, we performed three separate steps: (1) setting up of a membrane associated test system for probing the interaction between the epidermal growth factor receptor (EGFR) and the growth factor receptor-bound protein 2; (2) use of the Epac-SH188 sensor for quantitative evaluation under the microscope; and (3) application of a TIR fluorescence reader to probe the interaction of GFP with Nile Red. In the first two steps, we measured FRET from cyan (CFP) to yellow fluorescent protein (YFP) by spectral analysis and fluorescence lifetime imaging (FLIM) upon illumination of whole cells (epi-illumination) as well as selective illumination of their plasma membranes by TIR. In particular, TIR excitation permitted FRET measurements with high sensitivity and low background. The Epac sensor showed a more rapid response to pharmaceutical agents, e.g., Forskolin or the A2B adenosine receptor agonist NECA, in close proximity to the plasma membrane compared to the cytosol. Finally, FRET from a membrane associated GFP to Nile Red was used to test a multi-well TIR fluorescence reader with simultaneous detection of a larger number of samples.