Ganglion cell-inner plexiform layer measurements derived from widefield compared to montaged 9-field optical coherence tomography.

CLINICAL RELEVANCE: With equivalent inner retinal thickness measurements compared to a more conventional composite optical coherence tomography (OCT) protocol, Widefield optical coherence tomography (WF-OCT) is a clinically viable, time-saving option facilitating detection of ocular pathologies within the central 55° of the retina. PURPOSE: To compare ganglion cell-inner plexiform layer (GCIPL) thicknesses obtained using a single WF-OCT scan and standard composite OCT scans acquired in 9 fields of gaze (9F-OCT). METHODS: Thirteen healthy participants underwent WF-OCT and 9F-OCT using the Spectralis OCT. The GCIPL was automatically segmented with a manual review for 9F-OCT and was manually segmented for WF-OCT. After registration, differences in GCIPL thicknesses were compared using 95% confidence intervals computed from one-sample t-tests and Bland-Altman analyses. Location-specific differences in B-scan tilt were analysed using Spearman correlations and linear regression models. To determine whether B-scan tilt influences GCIPL measurements, regression models of tilt versus differences between perpendicular and axial GCIPL thickness were applied. RESULTS: While scattered locations demonstrated significant GCIPL thickness differences between WF-OCT and 9F-OCT, most differences did not exceed the axial pixel resolution of the instrument of 3.87 µm. Bland-Altman analyses indicated no notable bias using WF-OCT. Moderate correlations indicating significant location-specific differences in B-scan tilt were observed for temporal, central and inferior B-scans (r = -0.62 to 0.72), with linear regression models predicting a maximum difference in the tilt of 4.65°. The quadratic regression model indicated that at tilts greater than 27.3°, perpendicular GCIPL measurements become increasingly thin relative to axial measurements. CONCLUSIONS: GCIPL thicknesses and B-scan tilts from WF-OCT are comparable to 9F-OCT, indicating that WF-OCT can be applied clinically to obtain valid inner retinal OCT measurements over 55° of the central retina with relative ease. However, for peripheral locations, B-scan tilt may need to be considered when measuring GCIPL thicknesses.

Custom extraction of macular ganglion cell-inner plexiform layer thickness more precisely co-localizes structural measurements with visual fields test grids.

We aimed to evaluate methods of extracting optical coherence tomography (OCT)-derived macular ganglion cell-inner plexiform layer (GCIPL) thickness measurements over retinal locations corresponding to standard visual field (VF) test grids. A custom algorithm was developed to automatically extract GCIPL thickness measurements from locations corresponding to Humphrey Field Analyser 10-2 and 30-2 test grids over Goldmann II, III and V stimulus sizes from a healthy cohort of 478 participants. Differences between GCIPL thickness measurements based on VF test grids (VF-based paradigms) and the 8 × 8 grid, as per instrument review software, were analyzed, as were impacts of fovea to optic disc tilt and areas over which GCIPL thickness measurements were extracted. Significant differences between the VF-based paradigms and the 8 × 8 grid were observed at up to 55% of locations across the macula, with the greatest deviations at the fovea (median 25.5 µm, 95% CI 25.24-25.72 µm, P < .0001). While significant correlations with fovea to optic disc tilt were noted at up to 33% of locations distributed 6°-8° from the foveal center, there were no marked differences in GCIPL thickness measurements between VF-based paradigms using different stimulus sizes. As such, standard high-density OCT measurement paradigms do not adequately reflect GCIPL measurements at retinal locations tested with standard VF patterns, with the central macular region contributing most to the observed differences and with further correction required for fovea to optic disc tilt. Spatial direction of GCIPL thickness measurements will improve future comparisons of structure and function, thereby improving methods designed to detect pathology affecting the inner retina.

Macula Ganglion Cell Thickness Changes Display Location-Specific Variation Patterns in Intermediate Age-Related Macular Degeneration.

Purpose: The purpose of this study was to examine changes in the ganglion cell layer (GCL) of individuals with intermediate age-related macular degeneration (AMD) using grid-wise analysis for macular optical coherence tomography (OCT) volume scans. We also aim to validate the use of age-correction functions for GCL thickness in diseased eyes. Methods: OCT macular cube scans covering 30° × 25° were acquired using Spectralis spectral-domain OCT for 87 eyes with intermediate AMD, 77 age-matched normal eyes, and 254 non-age-matched normal eyes. The thickness of the ganglion cell layer (GCL) was defined after segmentation at 60 locations across an 8 × 8 grid centered on the fovea, where each grid location covered 0.74 mm2 (approximately 3° × 3°) within the macula. Each GCL location of normal eyes (n = 77) were assigned to a specific iso-ganglion cell density cluster in the macula, based on patterns of age-related GCL thickness loss. Analyses were then performed comparing AMD GCL grid-wise data against corresponding spatial clusters, and significant AMD GCL thickness changes were denoted as values outside the 95% distribution limits. Results: Analysis of GCL thickness changes revealed significant differences between spatial clusters, with thinning toward the fovea, and thickening toward the peripheral macula. The direction of GCL thickness changes in AMD were associated more so with thickening than thinning in all analyses. Results were corroborated by the application of GCL thickness age-correction functions. Conclusions: GCL thickness changed significantly and nonuniformly within the macula of intermediate AMD eyes. Further characterization of these changes is critical to improve diagnoses and monitoring of GCL-altering pathologies.

Development of a Spatial Model of Age-Related Change in the Macular Ganglion Cell Layer to Predict Function From Structural Changes.

PURPOSE: To develop location-specific models of normal, age-related changes in the macular ganglion cell layer (GCL) from optical coherence tomography (OCT). Using these OCT-derived models, we predicted visual field (VF) sensitivities and compared these results to actual VF sensitivities. DESIGN: Retrospective cohort study. METHODS: Single eyes of 254 normal participants were retrospectively enrolled from the Centre for Eye Health (Sydney, Australia). Macular GCL measurements were obtained using Spectralis OCT. Cluster algorithms were performed to identify spatial patterns demonstrating similar age-related change. Quadratic and linear regression models were subsequently used to characterize age-related GCL decline. Forty participants underwent additional testing with Humphrey VFs, and 95% prediction intervals were calculated to measure the predictive ability of structure-function models incorporating cluster-based pooling, age correction, and consideration of spatial summation. RESULTS: Quadratic GCL regression models provided a superior fit (P value <.0001-.0066), establishing that GCL decline commences in the late 30s across the macula. The equivalent linear rates of GCL decline showed eccentricity-dependent variation (0.13 µm/yr centrally vs 0.06 µm/yr peripherally); however, average, normalized GCL loss per year was consistent across the 64 macular measurement locations at 0.26%. The 95% prediction intervals describing predicted VF sensitivities were significantly narrower across all cluster-based structure-function models (3.79-4.99 dB) compared with models without clustering applied (5.66-6.73 dB, P < .0001). CONCLUSIONS: Combining spatial clustering with age-correction based on regression models allowed the development of robust models describing GCL changes with age. The resultant superior predictive ability of VF sensitivity from ganglion cell measurements may be applied to future models of disease development to improve detection of early macular GCL pathology.

Consistency of Structure-Function Correlation Between Spatially Scaled Visual Field Stimuli and In Vivo OCT Ganglion Cell Counts.

Purpose: To investigate the effect of stimulus size and disease status on the structure-function relationship within the central retina, we correlated the differential light sensitivity (DLS) with Goldmann stimulus size I to V (GI-V) and optical coherence tomography (OCT) derived in vivo ganglion cell count per stimulus area (GCc) within the macular area in normal subjects and patients with early glaucoma. Methods: Humphrey Field Analyzer 10-2 visual field data with GI through V and Spectralis OCT macular ganglion cell layer (GCL) thickness measurements were collected from normal and early glaucoma cohorts including 25 subjects each. GCc was calculated from GCL thickness data and correlated with DLSs for different stimulus sizes. Results: Correlation coefficients attained with smaller stimulus size were higher compared to larger stimulus sizes in both normal (GI-GII: R2 = 0.41-0.43, GIII-GV: R2 = 0.16-0.41) and diseased cohorts (GI-GII: R2 = 0.33-0.41, GIII-GV: R2 = 0.19-0.36). Quadratic regression curves for combined GI to V data demonstrated high correlation (R2= 0.82-0.90) and differed less than 1 dB of visual sensitivity within the GCc range between cohorts. The established structure-function relationship was compatible with a histologically derived model correlation spanning the range predicted by stimulus sizes GI to GIII. Conclusions: Stimulus sizes within critical spatial summation area (GI-II) improved structure-function correlations in the central visual field. The structure-function relationship was identical in both normal and diseased cohort when GI to GV data were combined. Congruency of GI and GII structure-function correlation with those previously derived with GIII from more peripheral locations further suggests that the structure-function relationship is governed by the number of ganglion cell per stimulus area.

Pattern Recognition Analysis of Age-Related Retinal Ganglion Cell Signatures in the Human Eye.

Purpose: To characterize macular ganglion cell layer (GCL) changes with age and provide a framework to assess changes in ocular disease. This study used data clustering to analyze macular GCL patterns from optical coherence tomography (OCT) in a large cohort of subjects without ocular disease. Methods: Single eyes of 201 patients evaluated at the Centre for Eye Health (Sydney, Australia) were retrospectively enrolled (age range, 20-85); 8 × 8 grid locations obtained from Spectralis OCT macular scans were analyzed with unsupervised classification into statistically separable classes sharing common GCL thickness and change with age. The resulting classes and gridwise data were fitted with linear and segmented linear regression curves. Additionally, normalized data were analyzed to determine regression as a percentage. Accuracy of each model was examined through comparison of predicted 50-year-old equivalent macular GCL thickness for the entire cohort to a true 50-year-old reference cohort. Results: Pattern recognition clustered GCL thickness across the macula into five to eight spatially concentric classes. F-test demonstrated segmented linear regression to be the most appropriate model for macular GCL change. The pattern recognition-derived and normalized model revealed less difference between the predicted macular GCL thickness and the reference cohort (average ± SD 0.19 ± 0.92 and -0.30 ± 0.61 µm) than a gridwise model (average ± SD 0.62 ± 1.43 µm). Conclusions: Pattern recognition successfully identified statistically separable macular areas that undergo a segmented linear reduction with age. This regression model better predicted macular GCL thickness. The various unique spatial patterns revealed by pattern recognition combined with core GCL thickness data provide a framework to analyze GCL loss in ocular disease.

Repeatability of Heidelberg Retinal Tomography 3 and effect of alignment algorithm on glaucoma suspects.

BACKGROUND: The Heidelberg Retina Tomograph (HRT) is a commonly-used clinical instrument for glaucoma diagnosis; however, the repeatability of the two most commonly used analysis tools, Moorfield regression analysis (MRA) and 'glaucoma probability score' (GPS) is not known and could have significant implications for patients at risk or suspected of developing glaucoma. Thus, the intra-visit repeatability of the HRT3 (an objective measure of instrument-induced variability) was investigated in a glaucoma suspect cohort. METHODS: Two repeat 15° × 15° optic nerve head scans were taken from 164 eyes of 84 patients using the HRT (HRT3, software version 3) during a single visit. The variability of global and sectoral rim area, rim volume and GPS were analysed with and without image alignment using SPSS 22.0. Repeatability was evaluated as absolute difference between the two measurements. RESULTS: Repeatability of the global rim area, rim volume and GPS were high with and without image registration. Variability increased by a small, yet significant amount without image alignment (p < 0.001 to p = 0.002). This increase was more prominent for sectoral analysis with the exception of the rim area and volume in the temporal sector (p = 0.034 to p < 0.001). Increase in the variability of the GPS was also significant (p < 0.001). CONCLUSION: HRT3 demonstrated high short-term repeatability, which significantly improved with image registration for all global measurements. Sectoral analysis demonstrated higher repeatability with image registration for some of the sectors in rim area and volume analysis and all sectors for the GPS. Therefore, depending upon the assessed parameters, image registration may play a significant role in the interpretation of results in glaucoma suspects.

Influence of education and diagnostic modes on glaucoma assessment by optometrists.

PURPOSE: To evaluate the influence of different clinical examination techniques, including optic nerve head (ONH) photography, visual field tests, and adjunct imaging on the diagnosis of glaucoma by Australian and New Zealand optometrists. The effect of a short-term, didactic teaching module on these is also explored. METHODS: Clinical data of 30 patients previously seen at the Centre for Eye Health was collected and compiled into glaucoma diagnostic assessment modules. Each of six modules contained different combinations of clinical examination results and required a classification of the cases as normal, suspicious or glaucoma. A cohort of 54 Australian and New Zealand optometrists were recruited for the study and allocated into two cohorts. The intervention group completed a glaucoma training course prior to the assessment while the control group completed the assessment without additional training. Diagnostic accuracy was compared between modules and optometrist groups. RESULTS: High false negative rates were observed with ONH photography, which were drastically reduced with the addition of visual field, albeit at the cost of increased false positive rates. Addition of adjunct imaging techniques partially compensated for the increase in the false positive rate from the visual field, but had limited effect on false negative rate. Educational intervention resulted in larger improvement in the diagnostic ability when multiple imaging modalities were provided. CONCLUSION: The study highlighted the importance of combining both structural and functional assessments in glaucoma. Current imaging technology demonstrated limited usefulness for event diagnosis due to the persistent difficulties of defining structural and functional loss in glaucoma, thus highlighting the need for new glaucoma assessment techniques. Short-term didactic teaching programs may only result in limited improvement of glaucoma diagnostic ability in optometrists, and hence, it may need to be combined with long-term and/or non-didactic training components to obtain a greater effect.

Cirrus HD-OCT short-term repeatability of clinical retinal nerve fiber layer measurements.

PURPOSE: The detection of changes in the retinal nerve fiber layer (RNFL) as measured by optical coherence tomography (OCT) is crucial in glaucoma diagnosis and management. We investigated the short-term repeatability of peripapillary RNFL measurements in a commercially available spectral domain OCT focusing on a broad clinical spectrum of patients. METHODS: Two consecutive peripapillary RNFL measurements were taken on 227 eyes with Cirrus HD-OCT (Carl Zeiss Meditec, Version 6.5 software) using the optic disc 200 × 200 protocol. Repeatability was assessed as Bland-Altman limits of agreement and intraclass coefficients (ICCs). RESULTS: Limits of agreement showed the greatest variability in the superior RNFL quadrant at ±7.5 µm and the least variability in the temporal quadrant at ±5.2 µm. The short-term repeatability for the average RNFL thickness resulted in an ICC of 0.98 and variability of 3.81 µm. Individual quadrants were similar, excepting the nasal RNFL quadrant with an ICC of 0.94. Inferior and temporal quadrants were the most repeatable with a variability of 2 to 3% instrument error. CONCLUSIONS: Cirrus HD-OCT has excellent short-term repeatability for peripapillary RNFL measurements in a mixed patient cohort. Retinal nerve fiber layer measurements are less reliable in the nasal RNFL quadrant. As other quadrants are used in glaucoma diagnosis, the detection of glaucomatous progression would be reliable.