Retinal Nerve Fiber Layer Assessment: Area versus Thickness Measurements from Elliptical Scans Centered on the Optic Nerve

Abstract

An evaluation of the retinal nerve fiber layer (RNFL) provides important information on the health of the optic nerve. Standard measurements of the RNFL consider only thickness, but an accurate assessment should also consider axial length, size of the optic nerve head (ONH), blood vessel contribution, and distance of the scan from the ONH margin. In addition, although most primate ONHs are elliptical, the circular scan centered on the ONH is the mainstay in both clinical and research analyses. The purpose of this study was to evaluate thickness and area measures of RNFL cross sections when axial length and ONH shape are included. Circular, raster, and radial scans of left eye optic nerves were acquired from 40 normal rhesus monkeys (Macaca mulatta) using spectral domain optical coherence tomography. The disc margin was identified by manually selecting the RPE/Bruch's membrane opening and ONH border tissue. With a pixel-to-micrometer conversion computed from a three-surface schematic eye, RNFL scans were interpolated at 300 to 600 μm (50-μm increments) from the edge of the ONH. The thickness and area of the RNFL at each distance were obtained by custom programs. Blood vessels in the RNFL were selected and removed from the overall RNFL measures. The average RNFL thickness decreased systematically from 149 ± 12.0 μm for scans 300 μm from the disc margin to 113 ± 7.2 μm at an eccentricity of 600 μm (P < 0.05). In contrast, the cross-sectional areas of the RNFL did not vary with scan location from the disc margin (0.85 ± 0.07 mm(2) at 300 μm compared with 0.86 ± 0.06 mm(2) at 600 μm). Blood vessels accounted for 9.3% of total RNFL thickness or area, but varied with retinal location. On average, 17.6% of the superior and 14.2% of the inferior RNFL was vascular, whereas blood vessels accounted for only 2.3% of areas of the temporal and nasal RNFL regions. In nonhuman primates, with appropriate transverse scaling and ONH shape analysis, the cross-sectional area of the RNFL is independent of scan distance, up to 600 μm from the rim margin, indicating that the axonal composition changes little over this range. The results suggest that, with incorporation of transverse scaling, the RNFL cross-sectional area, rather than RNFL thickness, provides an accurate assessment of the retinal ganglion cell axonal content within the eye.