Temporal change of retinal nerve fiber layer reflectance speckle in normal and hypertensive retinas.

This study investigated temporal change of retinal nerve fiber layer (RNFL) reflectance speckle in retinas with ocular hypertensive (OHT) damage and in control retinas from untreated eyes. Experimental OHT damage to rat retinas was induced by laser photocoagulation of the trabecular meshwork. A series of 660 nm reflectance images was collected from isolated retinas at 10-sec intervals. Areas containing speckled texture were selected on nerve fiber bundles. Correlation coefficients between images with different imaging delays were calculated and plotted as a function of delay. To evaluate the temporal change of speckles, decay of correlation coefficients with time was fitted with an exponential function characterized by a time constant τ. Reflectance per unit thickness (σ) of the areas was also measured and low σ was used as a surrogate of OHT damage. Speckle phenomena occurred in the control RNFL and the RNFL with reduced σ. In the control retinas, τ and σ were nearly constant along bundles but differed significantly among bundles in the same retinas. Among the control retinas, σ was similar, whereas τ varied significantly. In the retinas with OHT damage (low σ) τ could be within, greater or lower than the range in controls. The parameters τ and σ provide independent assessment of the RNFL with OHT damage. Measurements of temporal change of RNFL reflectance speckle may offer a method for detecting functional abnormality of the RNFL.

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.

Cytoskeletal Alteration and Change of Retinal Nerve Fiber Layer Birefringence in Hypertensive Retina.

PURPOSE: Glaucoma damages the retinal nerve fiber layer (RNFL). Both RNFL thickness and retardance can be used to assess the damage, but birefringence, the ratio of retardance to thickness, is a property of the tissue itself. This study investigated the relationship between axonal cytoskeleton and RNFL birefringence in retinas with hypertensive damage. MATERIALS AND METHODS: High intraocular pressure (IOP) was induced unilaterally in rat eyes. RNFL retardance in isolated retinas was measured. Cytostructural organization and bundle thickness were evaluated by confocal imaging of immunohistochemical staining of the cytoskeletal components: microtubules (MTs), F-actin, and neurofilaments. Bundles with different appearances of MT stain were studied, and their birefringence was calculated at different radii from the optic nerve head (ONH) center. RESULTS: Forty bundles in eight normal retinas and 37 bundles in 10 treated retinas were examined. In normal retinas, the stain of axonal cytoskeleton was approximately uniform within bundles, and RNFL birefringence did not change along bundles. In treated retinas, elevation of IOP caused non-uniform alteration of axonal cytoskeleton across the retina, and distortion of axonal MTs was associated with decreased birefringence. The study further demonstrated that change of RNFL birefringence profiles along bundles can imply altered axonal cytoskeleton, suggesting that ultrastructural change of the RNFL can be inferred from clinical measurements of RNFL birefringence. The study also demonstrated that measuring RNFL birefringence profiles along bundles, instead of at a single location, may provide a more sensitive way to detect axonal ultrastructural change. CONCLUSIONS: Measurement of RNFL birefringence along bundles can provide estimation of cytoskeleton alteration and sensitive detection of glaucomatous damage.

Retinal nerve fiber layer reflectometry must consider directional reflectance.

Recent studies reveal that measurements of retinal nerve fiber layer (RNFL) reflectance provide more sensitive detection of glaucomatous damage than RNFL thickness, but most do not consider directional reflectance of the RNFL, an important source of variability. This study quantitatively compared RNFL directional reflectance, represented by an angular spread function (ASF), measured at different scattering angles, different wavelengths and different distances from the optic nerve head (ONH) and for bundles with different thicknesses (T). An ASF was characterized by its amplitude (A) and width (W). Internal reflectance of a bundle was expressed as A/T. The study found that A varied significantly with scattering angle and wavelength and that A/T was different among bundles but constant along the same bundle, indicating that the internal structure of axons may vary among bundles but does not change with distance. This study also found that W was larger near the ONH and at longer wavelengths, but did not depend on scattering angle or T. Because a 4.3° change in incident angle can change reflected intensity by a factor of 2.7, accounting for directional reflectance should improve the accuracy and reproducibility of RNFL reflectance measurements.

Reflectance speckle of retinal nerve fiber layer reveals axonal activity.

PURPOSE: This study investigated the retinal nerve fiber layer (RNFL) reflectance speckle and tested the hypothesis that temporal change of RNFL speckle reveals axonal dynamic activity. METHODS: RNFL reflectance speckle of isolated rat retinas was studied with monochromatic illumination. A series of reflectance images was collected every 5 seconds for approximately 15 minutes. Correlation coefficients (CC) of selected areas between a reference and subsequent images were calculated and plotted as a function of the time intervals between images. An exponential function fit to the time course was used to evaluate temporal change of speckle pattern. To relate temporal change of speckle to axonal activity, in vitro living retina perfused at a normal (34°C) and a lower (24°C) temperature, paraformaldehyde-fixed retina, and retina treated with microtubule depolymerization were used. RESULTS: RNFL reflectance was not uniform; rather nerve fiber bundles had a speckled texture that changed with time. In normally perfused retina, the time constant of the CC change was 0.56 ± 0.26 minutes. In retinas treated with lower temperature and microtubule depolymerization, the time constants increased by two to four times, indicating that the speckle pattern changed more slowly. The speckled texture in fixed retina was stationary. CONCLUSIONS: Fixation stops axonal activity; treatments with either lower temperature or microtubule depolymerization are known to decrease axonal transport. The results obtained in this study suggest that temporal change of RNFL speckle reveals structural change due to axonal activity. Assessment of RNFL reflectance speckle may offer a new means of evaluating axonal function.

The shape of the ganglion cell plus inner plexiform layers of the normal human macula.

PURPOSE: To use surfaces generated by two-dimensional penalized splines (2D P-splines) to characterize the shape of the macular ganglion cell plus inner plexiform layers (GCL+IPL) in a group of normal humans. METHODS: Macular images of the right eyes of 23 normal subjects ranging in age from 18 to 75 years were obtained with spectral-domain optical coherence tomography (SD-OCT). The thickness of GCL+IPL was determined by manual segmentation, areas with blood vessels were removed, and the resulting maps were fit by smooth surfaces in polar coordinates centered on the fovea. RESULTS: Smooth surfaces based on 2D P-splines could precisely represent GCL+IPL thickness data, with errors comparable to the axial resolution of the SD-OCT instrument. Metrics were developed for the size, shape, and slope of the edge of the foveal depression and size and shape of the surrounding macular ridge. The slope of the foveal edge was negatively correlated with foveal size (r = -0.60). The size of the macular ridge was positively correlated with foveal size (r = 0.75), with a slope near unity (0.90 ± 0.18). The centroids of the foveal edge and macular ridge clustered near the foveal center. The foveal edge and macular ridge were well fit by ellipses. The mean GCL+IPL thickness formed an elliptical annulus elongated by approximately 30% in the horizontal direction. CONCLUSIONS: The methods developed here provide precise characterization of retinal layers for the study of glaucoma, foveal development, and other applications.

Wavelength-dependent change of retinal nerve fiber layer reflectance in glaucomatous retinas.

PURPOSE: Retinal nerve fiber layer (RNFL) reflectance is often used in optical methods for RNFL assessment in clinical diagnosis of glaucoma, yet little is known about the reflectance property of the RNFL under the development of glaucoma. This study measured the changes in RNFL reflectance spectra that occurred in retinal nerve fiber bundles with different degrees of glaucomatous damage. METHODS: A rat model of glaucoma with laser photocoagulation of trabecular meshwork was used. Reflectance of the RNFL in an isolated retina was measured at wavelengths of 400-830 nm. Cytostructural distribution of the bundles measured optically was evaluated by confocal imaging of immunohistochemistry staining of cytoskeletal components, F-actin, microtubules, and neurofilaments. RNFL reflectance spectra were studied in bundles with normal-looking appearance, early F-actin distortion, and apparent damage of all cytoskeletal components. Changes of RNFL reflectance spectra were studied at different radii (0.22, 0.33, and 0.44 mm) from the optic nerve head (ONH). RESULTS: Bundles in 30 control retinas and 41 glaucomatous retinas were examined. In normal retinas, reflectance spectra were similar along the same bundles. In glaucomatous retinas, reflectance spectra changed along bundles with the spectra becoming flatter as bundle areas approached the ONH. CONCLUSIONS: Elevation of intraocular pressure (IOP) causes nonuniform changes in RNFL reflectance across wavelengths. Changes of reflectance spectra occur early in bundles near the ONH and prior to apparent cytoskeletal distortion. Using the ratio of RNFL reflectance measured at different wavelengths can provide early and sensitive detection of glaucomatous damage.

Variance reduction in a dataset of normal macular ganglion cell plus inner plexiform layer thickness maps with application to glaucoma diagnosis.

PURPOSE: To examine the similarities and differences in the shape of the macular ganglion cell plus inner plexiform layers (GCL+IPL) in a healthy human population, and seek methods to reduce population variance and improve discriminating power. METHODS: Macular images of the right eyes of 23 healthy subjects were obtained with spectral domain optical coherence tomography. The thickness of GCL+IPL was determined by manual segmentation, areas with blood vessels were removed, and the resulting maps were fit by smooth surfaces in polar coordinates centered on the fovea. RESULTS: The mean GCL+IPL thickness formed a horizontal elliptical annulus. The variance increased toward the center and was highest near the foveal edge. Individual maps differed in foveal size and overall GCL+IPL thickness. Foveal size correction by radially shifting individual maps to the same foveal size as the mean map reduced perifoveal variance. Thickness alignment by shifting individual maps axially, then radially, to match the mean map reduced overall variance. These transformations had very little effect on the population mean. CONCLUSIONS: Simple transformations of individual GCL+IPL thickness maps to a canonical form can considerably reduce the population variance in a sample of normal eyes, likely improving the ability to discriminate abnormal maps. The transformations considered here preserve the local geometry of the thickness maps. When used on a patient's map, they can produce a deviation map that provides a meaningful measurement of the size of local thickness deviations and allows estimation of the number of ganglion cells lost in a glaucomatous defect.

Dual-band spectral-domain optical coherence tomography for in vivo imaging the spectral contrasts of the retinal nerve fiber layer.

The ultimate goal of the study is to provide an imaging tool to detect the earliest signs of glaucoma before clinically visible damage occurs to the retinal nerve fiber layer (RNFL). Studies have shown that the optical reflectance of the damaged RNFL at short wavelength (<560 nm) is reduced much more than that at long wavelength, which provides spectral contrast for imaging the earliest damage to the RNFL. To image the spectral contrast we built a dual-band spectral-domain optical coherence tomography (SD-OCT) centered at 808 nm (NIR) and 415 nm (VIS). The light at the two bands was provided by the fundamental and frequency-doubled outputs of a broadband Ti:Sapphire laser. The depth resolution of the NIR and VIS OCT systems are 4.7 µm and 12.2 µm in the air, respectively. The system was applied to imaging the rat retina in vivo. Significantly different appearances between the OCT cross sectional images at the two bands were observed. The ratio of the light reflected from the RNFL over that reflected from the entire retina at the two bands were quantitatively compared. The experimental results showed that the dual-band OCT system is feasible for imaging the spectral contrasts of the RNFL.

Reflectance decreases before thickness changes in the retinal nerve fiber layer in glaucomatous retinas.

PURPOSE: Glaucoma damages the retinal never fiber layer (RNFL). RNFL thickness, measured with optical coherence tomography (OCT), is often used in clinical assessment of the damage. In this study the relation between the RNFL reflectance and thickness at early stages of glaucoma was investigated. METHODS: A rat model of glaucoma was used that involved laser photocoagulation of the trabecular meshwork. The reflectance of the RNFL in an isolated retina was measured, followed by immunohistochemical staining of the axonal cytoskeleton. RNFL thickness was measured by confocal fluorescence imaging. RNFL reflectance was calculated for bundle areas located at radii of 0.22, 0.33, and 0.44 mm from the optic nerve head (ONH) center. Linear regression was used to study the relation between reflectance and thickness. For glaucomatous eyes, only those bundles with no apparent structural damage were used. RESULTS: Bundles in 11 control retinas and 10 treated retinas were examined. Bundle thickness of both groups at each radius was similar (P = 0.89). The reflectance of the bundles at radii of 0.33 and 0.44 mm was found to be similar in both control and treated retinas (P > 0.5). However, the reflectance of the bundles at the 0.22-mm radius decreased significantly in the treated group (P = 0.005). CONCLUSIONS: Elevation of intraocular pressure causes decrease in RNFL reflectance for bundles near the ONH. Change in RNFL reflectance precedes thinning of the RNFL. The results suggest that a decrease in RNFL reflectance near the ONH is an early sign of glaucomatous damage.

Registration of OCT fundus images with color fundus photographs based on blood vessel ridges.

This paper proposes an algorithm to register OCT fundus images (OFIs) with color fundus photographs (CFPs). This makes it possible to correlate retinal features across the different imaging modalities. Blood vessel ridges are taken as features for registration. A specially defined distance, incorporating information of normal direction of blood vessel ridge pixels, is designed to calculate the distance between each pair of pixels to be matched in the pair image. Based on this distance a similarity function between the pair image is defined. Brute force search is used for a coarse registration and then an Iterative Closest Point (ICP) algorithm for a more accurate registration. The registration algorithm was tested on a sample set containing images of both normal eyes and eyes with pathologies. Three transformation models (similarity, affine and quadratic models) were tested on all image pairs respectively. The experimental results showed that the registration algorithm worked well. The average root mean square errors for the affine model are 31 µm (normal) and 59 µm (eyes with disease). The proposed algorithm can be easily adapted to registration for other modality retinal images.

Revealing Henle's fiber layer using spectral domain optical coherence tomography.

PURPOSE: Spectral domain optical coherence tomography (SD-OCT) uses infrared light to visualize the reflectivity of structures of differing optical properties within the retina. Despite their presence on histologic studies, traditionally acquired SD-OCT images are unable to delineate the axons of photoreceptor nuclei, Henle's fiber layer (HFL). The authors present a new method to reliably identify HFL by varying the entry position of the SD-OCT beam through the pupil. METHODS: Fifteen eyes from 11 subjects with normal vision were prospectively imaged using 1 of 2 commercial SD-OCT systems. For each eye, the entry position of the SD-OCT beam through the pupil was varied horizontally and vertically. The reflectivity of outer retinal layers was measured as a function of beam position, and thicknesses were recorded. RESULTS: The reflectivity of HFL was directionally dependent and increased with eccentricity on the side of the fovea opposite the entry position. When HFL was included in the measurement, the thickness of the outer nuclear layer (ONL) of central horizontal B-scans increased by an average of 52% in three subjects quantified. Four cases of pathology, in which alterations to the normal macular geometry affected HFL intensity, were identified. CONCLUSIONS: The authors demonstrated a novel method to distinguish HFL from true ONL. An accurate measurement of the ONL is critical to clinical studies measuring photoreceptor layer thickness using any SD-OCT system. Recognition of the optical properties of HFL can explain reflectivity changes imaged in this layer in association with macular pathology.

Retinal structure of birds of prey revealed by ultra-high resolution spectral-domain optical coherence tomography.

PURPOSE: To reveal three-dimensional (3-D) information about the retinal structures of birds of prey in vivo. METHODS: An ultra-high resolution spectral-domain optical coherence tomography (SD-OCT) system was built for in vivo imaging of retinas of birds of prey. The calibrated imaging depth and axial resolution of the system were 3.1 mm and 2.8 µm (in tissue), respectively. 3-D segmentation was performed for calculation of the retinal nerve fiber layer (RNFL) map. RESULTS: High-resolution OCT images were obtained of the retinas of four species of birds of prey: two diurnal hawks (Buteo platypterus and Buteo brachyurus) and two nocturnal owls (Bubo virginianus and Strix varia). These images showed the detailed retinal anatomy, including the retinal layers and the structure of the deep and shallow foveae. The calculated thickness map showed the RNFL distribution. Traumatic injury to one bird's retina was also successfully imaged. CONCLUSIONS: Ultra-high resolution SD-OCT provides unprecedented high-quality 2-D and 3-D in vivo visualization of the retinal structures of birds of prey. SD-OCT is a powerful imaging tool for vision research in birds of prey.

Characteristics of optic nerve head drusen on optical coherence tomography images.

BACKGROUND AND OBJECTIVE: To describe the characteristics of optic nerve head drusen in optical coherence tomography (OCT) images. PATIENTS AND METHODS: Cross-sectional images of the optic nerve were obtained in seven patients with optic nerve head drusen with Stratus and spectral-domain OCT (Carl Zeiss Meditec, Dublin, CA). These were compared to optic disc photographs, autofluorescence, and echography images. For comparison, these tests were performed on four patients with papilledema and three patients with small optic discs. RESULTS: Optic nerve head drusen typically elevated the disc surface and appeared as an optically empty cavity, sometimes with a perceptible reflection from the posterior surface. The disc surface was also elevated in cases of papilledema, but had a strong anterior reflectance behind which there was no visible structure. The surface of the small optic nerves was slightly elevated, but with less anterior reflectance. CONCLUSION: Optic nerves with drusen showed features in these OCT images that were distinct from cases of papilledema or small optic discs.

Altered F-actin distribution in retinal nerve fiber layer of a rat model of glaucoma.

Glaucoma damages the retinal nerve fiber layer (RNFL). The purpose of this study was to investigate the distribution in RNFL of axonal F-actin, a cytoskeletal component, under the development of glaucoma. Intraocular hypertension was induced in a rat model by translimbal laser photocoagulation of the trabecular meshwork. The retinas of control and treated eyes were obtained after different exposures to elevated IOP. Nerve fiber bundles were identified by fluorescent phalloidin staining of F-actin. Nuclei of cell bodies were identified by DAPI fluorescent counterstain. F-actin distribution in whole-mounted retinas was examined by confocal microscopy. En face and cross-sectional images of RNFL were collected around the optic nerve head (ONH). F-actin in normal RNFL was intensely and uniformly stained. In glaucomatous retina, F-actin staining was not uniform within bundles and total loss of F-actin staining was found in severely damaged areas. Altered F-actin often occurred near the ONH in bundles that appeared normal more peripherally. Both alteration and total loss of F-actin were found most often in dorsal retina. In normal RNFL, F-actin is rich and approximately uniformly distributed within nerve fiber bundles. Elevated IOP changes F-actin distribution in RNFL. Topographic features of F-actin alteration suggest that F-actin near the ONH is more sensitive to glaucomatous damage. The alteration pattern also suggests an ONH location for the glaucomatous insult in this rat model.

Spectral domain optical coherence tomographic imaging of geographic atrophy.

BACKGROUND AND OBJECTIVE: To compare images of geographic atrophy (GA) obtained using spectral domain optical coherence tomography (SD-OCT) with images obtained using fundus autofluorescence (FAF). PATIENTS AND METHODS: Five eyes from patients with dry AMD were imaged using SD-OCT and FAF, and the size and shape of the GA were compared. RESULTS: GA appears bright on SD-OCT compared with the surrounding areas with an intact retinal pigment epithelium because of increased reflectivity from the underlying choroid. SD-OCT and FAF both identified GA reproducibly, and measurement of the area of GA is comparable between the two methods with a mean difference of 2.7% of the total area. CONCLUSION: SD-OCT can identify and quantitate areas of GA. The size and shape of these areas correlate well to the areas of GA seen on autofluorescence images; however, SD-OCT imaging also provides important cross-sectional anatomic information.

Spectral domain optical coherence tomography for proliferative diabetic retinopathy with subhyaloid hemorrhage.

A prototype 6-microm axial resolution spectral domain optical coherence tomography (SD-OCT) device was used to image the retina of a patient with uncontrolled diabetes mellitus who had proliferative diabetic retinopathy with subhyaloid hemorrhage. A raster scan pattern with 128 B-scans covering a 6 X 6 X 2-mm volume of the retina was obtained. SD-OCT showed the presence of blood localized between the internal limiting membrane and the posterior hyaloid face and allowed visualization of the cross sectional retinal architecture and the vitreoretinal interface at different horizontal levels that could be registered with the color fundus photograph. SD-OCT provided useful information about the relationship of the hemorrhage to the posterior hyaloid and the retina.

Calibration of fundus images using spectral domain optical coherence tomography.

BACKGROUND AND OBJECTIVE: Measurements performed on fundus images using current software are not accurate. Accurate measurements can be obtained only by calibrating a fundus camera using measurements between fixed retinal landmarks, such as the dimensions of the optic nerve, or by relying on a calibrated model eye provided by a reading center. However, calibrated spectral domain OCT (SD-OCT) could offer a convenient alternative method for the calibration of any fundus image. PATIENTS AND METHODS: The ability to measure exact distances on SD-OCT fundus images was tested by measuring the distance between the center of the fovea and the optic nerve. Calibrated SD-OCT scans measuring 6 X 6 X 2 mm centered on the fovea and the optic nerve were analyzed in 50 healthy right eyes. The foveal center was identified using cross-sectional SD-OCT images, and the center of the optic nerve was identified manually. The SD-OCT scans were registered to each other, and the distances between the center of the optic nerve and fovea were calculated. The overlay of these SD-OCT fundus images on photographic fundus images was performed. RESULTS: Any image of the fundus could be calibrated by overlaying the SD-OCT fundus image, and the measurements were consistent with previously defined calibration methods. The mean distance between the center of the fovea and the center of the optic nerve was 4.32 +/-0.32 mm. The line from the center of the optic nerve to the foveal center had a mean declination of 7.67 +/- 3.88 degrees. Mean horizontal displacement and vertical displacement were 4.27 +/- 0.29 mm and 0.58 +/- 0.29 mm, respectively. CONCLUSIONS: The overlay of the SD-OCT fundus image provides a convenient method for calibrating any image of the fundus. This approach should provide a uniform standard when comparing images from different devices and from different reading centers.

Macular thickness measurements in normal eyes using spectral domain optical coherence tomography.

BACKGROUND AND OBJECTIVE: Knowledge of the macular thickness in a normal population is important for the evaluation of pathological macular change. The purpose of this study was to define and measure macular thickness in normal eyes using spectral domain optical coherence tomography (OCT). PATIENTS AND METHODS: Fifty eyes from 50 normal subjects (29 men and 21 women, aged 22 to 68 years) were scanned with a prototype Cirrus HD-OCT system (5 microm axial resolution) (Carl Zeiss Meditec, Inc.). The proprietary Cirrus segmentation algorithm was used to produce retinal thickness maps, which were then averaged over 9 regions defined by a circular target centered at the true fovea location. The macular thickness of 13 subjects scanned with both HD-OCT and StratusOCT were compared. RESULTS: After centering the fovea, the mean and standard deviation values for retinal thickness measurements were calculated point wise and averaged on standard regions. For patients scanned with both systems, the thickness measurements from HD-OCT were approximately 50 microm larger than those from StratusOCT. The difference between the two measurements decreased somewhat with eccentricity. CONCLUSION: Using HD-OCT, it is possible to acquire retinal data sets containing an unprecedented number of data points. Furthermore, it is possible to use OCT fundus images to evaluate the scan quality and to center the measurement at the fovea. These advantages, together with good automated segmentation, can produce more accurate retinal thickness measurements. Incorporation of the photoreceptor layer in the measurements is anatomically meaningful and may be significant in evaluating various retinal pathologies and visual acuity outcomes.

Corneal birefringence mapped by scanning laser polarimetry.

Corneal birefringence affects polarization-sensitive optical measurements of the eye. Recent literature supports the idea that corneal birefringence is biaxial, although with some disagreement among reports and without considering corneas with very low values of central retardance. This study measured corneal retardation in eyes with a wide range of central corneal retardance by means of scanning laser polarimetry (GDx-VCC, Carl Zeiss Meditec, Inc.), which computes the retardance and slow axis of the cornea from images of the bow tie pattern formed by the radial birefringence of the macula. Measurements were obtained at many points on the cornea by translating the instrument. Data were compared to calculations of the retardation produced by a curved biaxial material between two spherical surfaces. Most corneas showed one or two small areas of zero retardance where the refracted ray within the cornea aligned with an optical axis of the material. The retardation patterns in these corneas could be mimicked, but not accurately described, by the biaxial model. Two corneas with large areas of low retardance more closely resembled a uniaxial model. We conclude that the cornea, in general, behaves as a biaxial material with its fastest axis perpendicular to its surface. Some locations in a few corneas can be uniaxial with the optical axis perpendicular to the surface. Importantly, corneal birefringence varies greatly among people and, within a single cornea, significantly with position.