Reflectance Spectrum and Birefringence of the Retinal Nerve Fiber Layer With Hypertensive Damage of Axonal Cytoskeleton.

Purpose: Glaucoma damages the retinal nerve fiber layer (RNFL). This study used precise multimodal image registration to investigate the changes of the RNFL reflectance spectrum and birefringence in nerve fiber bundles with different degrees of axonal damage. Methods: The reflectance spectrum of individual nerve fiber bundles in normal rats and rats with experimental glaucoma was measured from 400 to 830 nm and their birefringence was measured at 500 nm. Optical measurements of the same bundles were made at different distances from the optic nerve head (ONH). After the optical measurements, the axonal cytoskeleton of the RNFL was evaluated by confocal microscopy to assess the severity of cytoskeletal change. Results: For normal bundles, the shape of the RNFL reflectance spectrum and the value of RNFL birefringence did not change along bundles. In treated retinas, damage to the cytoskeleton varied within and across retinas. The damage in retinal sectors was subjectively graded from normal-looking to severe. Change of spectral shape occurred near the ONH in all sectors studied. This change became more prominent and occurred farther from the ONH with increased damage severity. In contrast, RNFL birefringence did not show change in normal-looking sectors, but decreased in sectors with mild and moderate damage. The birefringence of severely damaged sectors was either within or below the normal range. Conclusions: Varying degrees of cytoskeletal damage affect the RNFL reflectance spectrum and birefringence differently, supporting differences in the ultrastructural basis for the two optical properties. Both properties, however, may provide a means to detect disease and to estimate ultrastructural damage of the RNFL in glaucoma.