Smoothness assessment of corneal stromal surfaces.

Purpose: To assess the accuracy of the scanning electron microscopy (SEM) and present alternative approaches to quantify surface roughness based on numerical analysis.

Setting: Department of Ophthalmology, Maisonneuve-Rosemont Hospital, University of Montreal, Montreal, Quebec, Canada.

Design: Experimental study.

Methods: Lamellar stromal cuts were performed on human corneas using a femtosecond laser or a microkeratome. The photodisrupted stromal surfaces were processed for SEM, and images were acquired at ×1000 magnification. First, images were evaluated by independent observers. Second, images were analyzed based on first-order and second-order statistics of gray-level intensities. Third, 3-dimensional (3-D) surface reconstructions were generated from pairs of SEM images acquired at 2 angles.

Results: Results show that traditional assessment of roughness based on evaluating SEM images by independent observers can be replaced by computer-image texture analysis; an algorithm was developed to avoid subjective and time-consuming observations. The 3-D reconstructions allowed additional characterization of surface properties that was not possible with SEM images alone. Significant fluctuations in surface height were lost, although they could be retrieved using 3-D reconstructions.

Conclusions: Image texture analysis allowed objective and repeatable assessment of stromal surface roughness; however, full assessments of surface-height fluctuations required 3-D reconstruction. These complementary methodologies offer a more comprehensive assessment of corneal surface roughness in clinical applications.