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Amit Paranjape, Sarah Peterson, Marc D Friedman, David Muller; Confocal Brillouin scanning microscope for estimation of corneal stiffness. Invest. Ophthalmol. Vis. Sci. 2014;55(13):3721.
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To develop a new non-contact confocal Brillouin Scanning Microscope for measurement of corneal stiffness.
A bench-type Brillouin confocal microscope was developed using a confocal scanning system. Fresh whole porcine globes were obtained <24 hours postmortem in saline on ice from Sioux-preme (Sioux City, IA). 8 Eyes were brought to 37°C and the epithelium debrided using a dull blade. Eyes were then soaked in a solution consisting of 0.1%RF and 0.85% saline for 20 minutes. Brillouin scattering microscopy measurements were performed on the 8 eyes. The corneas were divided into two groups and irradiated with doses of 5.4J/cm2 and 10.8J/cm2 with UVA light at 365nm with an irradiance of 30mW/cm2 pulse 1 sec on 1 second off. Brillouin microscopy was performed again and corneal stiffness values were calculated from the Brillouin shifts for pre and post cross-linking.
The new system was able to perform automated scanning. Corneal stiffness values pre and post crosslinking showed higher stiffness values after cross linking with increased stiffening with dose.
A new confocal Brillouin scanning microscopy system has been demonstrated. This system provides a measure of corneal stiffness pre and post cross-linking of riboflavin/UVA illuminated porcine eyes. Brillouin scanning microscopy may potentially serve as a useful non-contact tool for quantifying the biomechanical changes after cross linking in the cornea.
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