Retinal Layer Segmentation Research Articles

HyFormer: a hybrid transformer-CNN architecture for retinal OCT image segmentation

Optical coherence tomography (OCT) has become the leading imaging technique in diagnosing and treatment planning for retinal diseases. Retinal OCT image segmentation involves extracting lesions and/or tissue structures to aid in the decisions of ophthalmologists, and multi-class segmentation is commonly needed. As the target regions often spread widely inside the retina, and the intensities and locations of different categories can be close, good segmentation networks must possess both global modeling capabilities and the ability to capture fine details. To address the challenge in capturing both global and local features simultaneously, we propose HyFormer, an efficient, lightweight, and robust hybrid network architecture. The proposed architecture features parallel Transformer and convolutional encoders for independent feature capture. A multi-scale gated attention block and a group positional embedding block are introduced within the Transformer encoder to enhance feature extraction. Feature integration is achieved in the decoder composed of the proposed three-path fusion modules. A class activation map-based cross-entropy loss function is also proposed to improve segmentation results. Evaluations are performed on a private dataset with myopic traction maculopathy lesions and the public AROI dataset for retinal layer and lesion segmentation with age-related degeneration. The results demonstrate HyFormer's superior segmentation performance and robustness compared to existing methods, showing promise for accurate and efficient OCT image segmentation. .

GCN-Enhanced Spatial-Spectral Dual-Encoder Network for Simultaneous Segmentation of Retinal Layers and Fluid in OCT Images

Changes in retinal thickness and the morphology of fluid provide valuable diagnostic information for Macular Edema (ME). Optical Coherence Tomography (OCT) is a pivotal diagnostic tool for ME, and the accurate segmentation of OCT retinal images aids ophthalmologists in monitoring disease progression. However, the unpredictable nature of ME’s occurrence poses significant challenges for existing methods in achieving precise segmentation of both retinal layers and fluids simultaneously. In this study, a novel U-shaped encoder-decoder architecture named GD-Net, is introduced for simultaneously segmenting retinal layers of diverse thicknesses and fluids. Specifically, a novel Fast Fourier Encoder is integrated within the encoder branch to capture spectral domain features overlooked by the spatial encoder. Additionally, Multi-scale Graph Convolution module is inserted between the encoder and decoder branches to leverage retinal layer topology for spatial reasoning. GD-Net is further optimized using a joint loss function, which includes a weighted version of region segmentation loss and contour loss, effectively mitigating the issues of blurred fluid edges and internal holes. The method was validated on three publicly available datasets: DUKE DME, Peripapillary OCT, and RETOUCH, achieving overall mean Dice score of 0.839, 0.826, and 0.872, respectively, outperforming the other state-of-the-art techniques. GD-Net demonstrated consistent high-precision segmentation on B-scans from different devices, proving its robustness and surpassing other methods in terms of visual quality, thus offering substantial support for clinical ophthalmologists in diagnosing ME. The code and weights are publicly available at: https://github.com/DBook111/GD-Net.

HDB-Net: hierarchical dual-branch network for retinal layer segmentation in diseased OCT images.

Optical coherence tomography (OCT) retinal layer segmentation is a critical procedure of the modern ophthalmic process, which can be used for diagnosis and treatment of diseases such as diabetic macular edema (DME) and multiple sclerosis (MS). Due to the difficulties of low OCT image quality, highly similar retinal interlayer morphology, and the uncertain presence, shape and size of lesions, the existing algorithms do not perform well. In this work, we design an HDB-Net network for retinal layer segmentation in diseased OCT images, which solves this problem by combining global and detailed features. First, the proposed network uses a Swin transformer and Res50 as a parallel backbone network, combined with the pyramid structure in UperNet, to extract global context and aggregate multi-scale information from images. Secondly, a feature aggregation module (FAM) is designed to extract global context information from the Swin transformer and local feature information from ResNet by introducing mixed attention mechanism. Finally, the boundary awareness and feature enhancement module (BA-FEM) is used to extract the retinal layer boundary information and topological order from the low-resolution features of the shallow layer. Our approach has been validated on two public datasets, and Dice scores were 87.61% and 92.44, respectively, both outperforming other state-of-the-art technologies.

Evaluation of thickness of individual macular retinal layers in diabetic eyes from optical coherence tomography

The effect of diabetes mellitus (DM) on individual retinal layers remains incompletely understood. We evaluated the intra-retinal layer thickness alterations in 71 DM eyes with no diabetic retinopathy (DR), 90 with mild DR, and 63 with moderate DR without macular edema, using spectral-domain optical coherence tomography (SD-OCT) and the Iowa Reference Algorithm for automated retinal layer segmentation. The average thickness of 10 intra-retinal layers was then corrected for ocular magnification using axial length measurements, and pairwise comparisons were made using multivariable linear regression models adjusted for gender and race. In DM no DR eyes, significant thinning was evident in the ganglion cell layer (GCL; p < 0.001), inner nuclear layer (INL; p = 0.001), and retinal pigment epithelium (RPE; p = 0.014) compared to normal eyes. Additionally, mild DR eyes exhibited a thinner inner plexiform layer (IPL; p = 0.008) than DM no DR eyes. Conversely, moderate DR eyes displayed thickening in the INL, outer nuclear layer, IPL, and retinal nerve fiber layer (all p ≤ 0.002), with notably worse vision. These findings highlight distinctive patterns: early diabetic eyes experience thinning in specific retinal layers, while moderate DR eyes exhibit thickening of certain layers and slightly compromised visual acuity, despite the absence of macular edema. Understanding these structural changes is crucial for comprehending diabetic eye complications.

Enhancing OCT patch-based segmentation with improved GAN data augmentation and semi-supervised learning

For optimum performance, deep learning methods, such as those applied for retinal and choroidal layer segmentation in optical coherence tomography (OCT) images, require sufficiently large and diverse labelled datasets for training. However, the acquisition and labelling of such data can be difficult or infeasible due to privacy reasons (particularly in the medical domain), accessing patient images such as those with specific pathologies, and the cost and time investment to annotate large volumes of data by clinical experts. Data augmentation is one solution to address this issue, either using simple variations and transformations of the images (e.g. flips, brightness) or using synthetic data from sophisticated generative methods such as generative adversarial networks (GANs). Semi-supervised learning (SSL) is another technique which aims to utilise unlabelled data to enhance the performance of deep learning methods and is beneficial where significant amounts of data may be available but are not labelled. In this study, we aim to enhance patch-based OCT retinal and choroidal layer segmentation with both GAN-based data augmentation and SSL. In particular, we employ a conditional StyleGAN2 to generate synthetic patches for data augmentation and a similar unconditional GAN for pre-training the patch classifier to perform SSL. In doing so, we propose a new patch classifier architecture based on the discriminator architecture to improve performance, in addition to the SSL benefit. Compared to previous methods, the proposed data augmentation approach provides an improved data augmentation performance for patch classification with its effectiveness widespread, particularly in the case of low data, across three different OCT datasets encompassing a range of scanning parameters, noise levels, pathology and participant variability. The method provides some subsequent improvements in boundary delineation which is of high importance from a clinical perspective. Additionally, the proposed SSL approach boosts classification performance and boundary delineation performance in some cases which provides further usefulness in the case of low data. The proposed methods can be utilised to enhance OCT segmentation methods, which may be of considerable benefit for both clinicians and researchers.

Boundary-Repairing Dual-Path Network for Retinal Layer Segmentation in OCT Image with Pigment Epithelial Detachment.

Automatic retinal layer segmentation in optical coherence tomography (OCT) images is crucial for the diagnosis of ocular diseases. Currently, automatic retinal layer segmentation works well with normal OCT images. However, pigment epithelial detachment (PED) dramatically alters the retinal structure, causing blurred boundaries and partial disappearance of the Bruch's Membrane (BM), thus posing challenges to the segmentation. To tackle these problems, we propose a novel dual-path U-shaped network for simultaneous layer segmentation and boundary regression. This network first designs a feature interaction fusion (FIF) module to strengthen the boundary shape constraints in the layer path. To address the challenge posed by partial BM disappearance and boundary-blurring, we propose a layer boundary repair (LBR) module. This module aims to use contrastive loss to enhance the confidence of blurred boundary regions and refine the segmentation of layer boundaries through the re-prediction head. In addition, we introduce a novel bilateral threshold distance map (BTDM) designed for the boundary path. The BTDM serves to emphasize information within boundary regions. This map, combined with the updated probability map, culminates in topology-guaranteed segmentation results achieved through a topology correction (TC) module. We investigated the proposed network on two severely deformed datasets (i.e., OCTA-500 and Aier-PED) and one slightly deformed dataset (i.e., DUKE). The proposed method achieves an average Dice score of 94.26% on the OCTA-500 dataset, which was 1.5% higher than BAU-Net and outperformed other methods. In the DUKE and Aier-PED datasets, the proposed method achieved average Dice scores of 91.65% and 95.75%, respectively.

Hybrid deep learning and optimal graph search method for optical coherence tomography layer segmentation in diseases affecting the optic nerve.

Accurate segmentation of retinal layers in optical coherence tomography (OCT) images is critical for assessing diseases that affect the optic nerve, but existing automated algorithms often fail when pathology causes irregular layer topology, such as extreme thinning of the ganglion cell-inner plexiform layer (GCIPL). Deep LOGISMOS, a hybrid approach that combines the strengths of deep learning and 3D graph search to overcome their limitations, was developed to improve the accuracy, robustness and generalizability of retinal layer segmentation. The method was trained on 124 OCT volumes from both eyes of 31 non-arteritic anterior ischemic optic neuropathy (NAION) patients and tested on three cross-sectional datasets with available reference tracings: Test-NAION (40 volumes from both eyes of 20 NAION subjects), Test-G (29 volumes from 29 glaucoma subjects/eyes), and Test-JHU (35 volumes from 21 multiple sclerosis and 14 control subjects/eyes) and one longitudinal dataset without reference tracings: Test-G-L (155 volumes from 15 glaucoma patients/eyes). In the three test datasets with reference tracings (Test-NAION, Test-G, and Test-JHU), Deep LOGISMOS achieved very high Dice similarity coefficients (%) on GCIPL: 89.97±3.59, 90.63±2.56, and 94.06±1.76, respectively. In the same context, Deep LOGISMOS outperformed the Iowa reference algorithms by improving the Dice score by 17.5, 5.4, and 7.5, and also surpassed the deep learning framework nnU-Net with improvements of 4.4, 3.7, and 1.0. For the 15 severe glaucoma eyes with marked GCIPL thinning (Test-G-L), it demonstrated reliable regional GCIPL thickness measurement over five years. The proposed Deep LOGISMOS approach has potential to enhance precise quantification of retinal structures, aiding diagnosis and treatment management of optic nerve diseases.

Human versus Artificial Intelligence: Validation of a Deep Learning Model for Retinal Layer and Fluid Segmentation in Optical Coherence Tomography Images from Patients with Age-Related Macular Degeneration.

Artificial intelligence (AI) models have received considerable attention in recent years for their ability to identify optical coherence tomography (OCT) biomarkers with clinical diagnostic potential and predict disease progression. This study aims to externally validate a deep learning (DL) algorithm by comparing its segmentation of retinal layers and fluid with a gold-standard method for manually adjusting the automatic segmentation of the Heidelberg Spectralis HRA + OCT software Version 6.16.8.0. A total of sixty OCT images of healthy subjects and patients with intermediate and exudative age-related macular degeneration (AMD) were included. A quantitative analysis of the retinal thickness and fluid area was performed, and the discrepancy between these methods was investigated. The results showed a moderate-to-strong correlation between the metrics extracted by both software types, in all the groups, and an overall near-perfect area overlap was observed, except for in the inner segment ellipsoid (ISE) layer. The DL system detected a significant difference in the outer retinal thickness across disease stages and accurately identified fluid in exudative cases. In more diseased eyes, there was significantly more disagreement between these methods. This DL system appears to be a reliable method for accessing important OCT biomarkers in AMD. However, further accuracy testing should be conducted to confirm its validity in real-world settings to ultimately aid ophthalmologists in OCT imaging management and guide timely treatment approaches.

Long-term impact of low-energy femtosecond laser and manual cataract surgery on macular layer thickness: A prospective randomized study.

To evaluate change in retinal layers 18 months after femtosecond laser-assisted cataract surgery (LCS) and manual cataract surgery (MCS) in a representative age-related cataract population using artificial intelligence (AI)-based automated retinal layer segmentation. This was a prospective, randomized and intraindividual-controlled study including 60 patients at the Medical University of Vienna, Austria. Bilateral same-day LCS and MCS were performed in a randomized sequence. To provide insight into the development of cystoid macular oedema (CME), retinal layer thickness was measured pre-operatively and up to 18 months post-operatively in the central 1 mm, 3 mm and 6 mm. Fifty-six patients completed all follow-up visits. LCS compared to MCS did not impact any of the investigated retinal layers at any follow-up visit (p > 0.05). For the central 1 mm, a significant increase in total retinal thickness (TRT) was seen after 1 week followed by an elevated plateau thereafter. For the 3 mm and 6 mm, TRT increased only after 3 weeks and 6 weeks and decreased again until 18 months. TRT remained significantly increased compared to pre-operative thickness (p < 0.001). Visual acuity remained unaffected by the macular thickening and no case of CME was observed. Inner nuclear layer (INL) and outer nuclear layer (ONL) were the main causative layers for the total TRT increase. Photoreceptors (PR) declined 1 week after surgery but regained pre-operative values 18 months after surgery. Low-energy femtosecond laser pre-treatment did not influence thickness of the retinal layers in any topographic zone compared to manual high fluidic phacoemulsification. TRT did not return to pre-operative values 18 months after surgery. The causative layers for subclinical development of CME were successfully identified.

Assessment of area and structural irregularity of retinal layers in diabetic retinopathy using machine learning and image processing techniques

Diabetes retinopathy prevention necessitates early detection, monitoring, and treatment. Non-invasive optical coherence tomography (OCT) shows structural changes in the retinal layer. OCT image evaluation necessitates retinal layer segmentation. The ability of our automated retinal layer segmentation to distinguish between normal, non-proliferative (NPDR), and proliferative diabetic retinopathy (PDR) was investigated in this study using quantifiable biomarkers such as retina layer smoothness index (SI) and area (S) in horizontal and vertical OCT images for each zone (fovea, superior, inferior, nasal, and temporal). This research includes 84 eyes from 57 individuals. The study shows a significant difference in the Area (S) of inner nuclear layer (INL) and outer nuclear layer (ONL) in the horizontal foveal zone across the three groups (p < 0.001). In the horizontal scan, there is a significant difference in the smoothness index (SI) of the inner plexiform layer (IPL) and the upper border of the outer plexiform layer (OPL) among three groups (p < 0.05). There is also a significant difference in the area (S) of the OPL in the foveal zone among the three groups (p = 0.003). The area (S) of the INL in the foveal region of horizontal slabs performed best for distinguishing diabetic patients (NPDR and PDR) from normal individuals, with an accuracy of 87.6%. The smoothness index (SI) of IPL in the nasal zone of horizontal foveal slabs was the most accurate at 97.2% in distinguishing PDR from NPDR. The smoothness index of the top border of the OPL in the nasal zone of horizontal slabs was 84.1% accurate in distinguishing NPDR from PDR. Smoothness index of IPL in the temporal zone of horizontal slabs was 89.8% accurate in identifying NPDR from PDR patients. In conclusion, optical coherence tomography can assess the smoothness index and irregularity of the inner and outer plexiform layers, particularly in the nasal and temporal regions of horizontal foveal slabs, to distinguish non-proliferative from proliferative diabetic retinopathy. The evolution of diabetic retinopathy throughout severity levels and its effects on retinal layer irregularity need more study.

Deep Learning-Based Retinal Layer Segmentation in Optical Coherence Tomography Scans of Patients with Inherited Retinal Diseases.

In optical coherence tomography (OCT) scans of patients with inherited retinal diseases (IRDs), the measurement of the thickness of the outer nuclear layer (ONL) has been well established as a surrogate marker for photoreceptor preservation. Current automatic segmentation tools fail in OCT segmentation in IRDs, and manual segmentation is time-consuming. Patients with IRD and an available OCT scan were screened for the present study. Additionally, OCT scans of patients without retinal disease were included to provide training data for artificial intelligence (AI). We trained a U-net-based model on healthy patients and applied a domain adaption technique to the IRD patients' scans. We established an AI-based image segmentation algorithm that reliably segments the ONL in OCT scans of IRD patients. In a test dataset, the dice score of the algorithm was 98.7%. Furthermore, we generated thickness maps of the full retinal thickness and the ONL layer for each patient. Accurate segmentation of anatomical layers on OCT scans plays a crucial role for predictive models linking retinal structure to visual function. Our algorithm for segmentation of OCT images could provide the basis for further studies on IRDs.

Self-attention CNN for retinal layer segmentation in OCT.

The structure of the retinal layers provides valuable diagnostic information for many ophthalmic diseases. Optical coherence tomography (OCT) obtains cross-sectional images of the retina, which reveals information about the retinal layers. The U-net based approaches are prominent in retinal layering methods, which are usually beneficial to local characteristics but not good at obtaining long-distance dependence for contextual information. Furthermore, the morphology of retinal layers with the disease is more complex, which brings more significant challenges to the task of retinal layer segmentation. We propose a U-shaped network combining an encoder-decoder architecture and self-attention mechanisms. In response to the characteristics of retinal OCT cross-sectional images, a self-attentive module in the vertical direction is added to the bottom of the U-shaped network, and an attention mechanism is also added in skip connection and up-sampling to enhance essential features. In this method, the transformer's self-attentive mechanism obtains the global field of perception, thus providing the missing context information for convolutions, and the convolutional neural network also efficiently extracts local features, compensating the local details the transformer ignores. The experiment results showed that our method is accurate and better than other methods for segmentation of the retinal layers, with the average Dice scores of 0.871 and 0.820, respectively, on two public retinal OCT image datasets. To perform the layer segmentation of retinal OCT image better, the proposed method incorporates the transformer's self-attention mechanism in a U-shaped network, which is helpful for ophthalmic disease diagnosis.

Radiation effects on retinal layers revealed by OCT, OCT-A, and perimetry as a function of dose and time from treatment

Optical coherence tomography (OCT) has become a key method for diagnosing and staging radiation retinopathy, based mainly on the presence of fluid in the central macula. A robust retinal layer segmentation method is required for identification of the specific layers involved in radiation-induced pathology in individual eyes over time, in order to determine damage driven by radiation injury to the microvessels and to the inner retinal neurons. Here, we utilized OCT, OCT-angiography, visual field testing, and patient-specific dosimetry models to analyze abnormal retinal layer thickening and thinning relative to microvessel density, visual function, radiation dose, and time from radiotherapy in a cross-sectional cohort of uveal melanoma patients treated with 125I-plaque brachytherapy. Within the first 24 months of radiotherapy, we show differential thickening and thinning of the two inner retinal layers, suggestive of microvessel leakage and neurodegeneration, mostly favoring thickening. Four out of 13 eyes showed decreased inner retinal capillary density associated with a corresponding normal inner retinal thickness, indicating early microvascular pathology. Two eyes showed the opposite: significant inner retinal layer thinning and normal capillary density, indicating early neuronal damage preceding a decrease in capillary density. At later time points, inner retinal thinning becomes the dominant pathology and correlates significantly with decreased vascularity, vision loss, and dose to the optic nerve. Stable multiple retinal layer segmentation provided by 3D graph-based methods aids in assessing the microvascular and neuronal response to radiation, information needed to target therapeutics for radiation retinopathy and vision loss.

Retinal Layer Segmentation in OCT Images With Boundary Regression and Feature Polarization.

The geometry of retinal layers is an important imaging feature for the diagnosis of some ophthalmic diseases. In recent years, retinal layer segmentation methods for optical coherence tomography (OCT) images have emerged one after another, and huge progress has been achieved. However, challenges due to interference factors such as noise, blurring, fundus effusion, and tissue artifacts remain in existing methods, primarily manifesting as intra-layer false positives and inter-layer boundary deviation. To solve these problems, we propose a method called Tightly combined Cross-Convolution and Transformer with Boundary regression and feature Polarization (TCCT-BP). This method uses a hybrid architecture of CNN and lightweight Transformer to improve the perception of retinal layers. In addition, a feature grouping and sampling method and the corresponding polarization loss function are designed to maximize the differentiation of the feature vectors of different retinal layers, and a boundary regression loss function is devised to constrain the retinal boundary distribution for a better fit to the ground truth. Extensive experiments on four benchmark datasets demonstrate that the proposed method achieves state-of-the-art performance in dealing with problems of false positives and boundary distortion. The proposed method ranked first in the OCT Layer Segmentation task of GOALS challenge held by MICCAI 2022. The source code is available at https://www.github.com/tyb311/TCCT.

Early inner plexiform layer thinning and retinal nerve fiber layer thickening in excitotoxic retinal injury using deep learning-assisted optical coherence tomography

Excitotoxicity from the impairment of glutamate uptake constitutes an important mechanism in neurodegenerative diseases such as Alzheimer’s, multiple sclerosis, and Parkinson's disease. Within the eye, excitotoxicity is thought to play a critical role in retinal ganglion cell death in glaucoma, diabetic retinopathy, retinal ischemia, and optic nerve injury, yet how excitotoxic injury impacts different retinal layers is not well understood. Here, we investigated the longitudinal effects of N-methyl-D-aspartate (NMDA)-induced excitotoxic retinal injury in a rat model using deep learning-assisted retinal layer thickness estimation. Before and after unilateral intravitreal NMDA injection in nine adult Long Evans rats, spectral-domain optical coherence tomography (OCT) was used to acquire volumetric retinal images in both eyes over 4 weeks. Ten retinal layers were automatically segmented from the OCT data using our deep learning-based algorithm. Retinal degeneration was evaluated using layer-specific retinal thickness changes at each time point (before, and at 3, 7, and 28 days after NMDA injection). Within the inner retina, our OCT results showed that retinal thinning occurred first in the inner plexiform layer at 3 days after NMDA injection, followed by the inner nuclear layer at 7 days post-injury. In contrast, the retinal nerve fiber layer exhibited an initial thickening 3 days after NMDA injection, followed by normalization and thinning up to 4 weeks post-injury. Our results demonstrated the pathological cascades of NMDA-induced neurotoxicity across different layers of the retina. The early inner plexiform layer thinning suggests early dendritic shrinkage, whereas the initial retinal nerve fiber layer thickening before subsequent normalization and thinning indicates early inflammation before axonal loss and cell death. These findings implicate the inner plexiform layer as an early imaging biomarker of excitotoxic retinal degeneration, whereas caution is warranted when interpreting the ganglion cell complex combining retinal nerve fiber layer, ganglion cell layer, and inner plexiform layer thicknesses in conventional OCT measures. Deep learning-assisted retinal layer segmentation and longitudinal OCT monitoring can help evaluate the different phases of retinal layer damage upon excitotoxicity.

Diverse Data Generation for Retinal Layer Segmentation With Potential Structure Modeling.

Accurate retinal layer segmentation on optical coherence tomography (OCT) images is hampered by the challenges of collecting OCT images with diverse pathological characterization and balanced distribution. Current generative models can produce high-realistic images and corresponding labels without quantitative limitations by fitting distributions of real collected data. Nevertheless, the diversity of their generated data is still limited due to the inherent imbalance of training data. To address these issues, we propose an image-label pair generation framework that generates diverse and balanced potential data from imbalanced real samples. Specifically, the framework first generates diverse layer masks, and then generates plausible OCT images corresponding to these layer masks using two customized diffusion probabilistic models respectively. To learn from imbalanced data and facilitate balanced generation, we introduce pathological-related conditions to guide the generation processes. To enhance the diversity of the generated image-label pairs, we propose a potential structure modeling technique that transfers the knowledge of diverse sub-structures from lowly- or non-pathological samples to highly pathological samples. We conducted extensive experiments on two public datasets for retinal layer segmentation. Firstly, our method generates OCT images with higher image quality and diversity compared to other generative methods. Furthermore, based on the extensive training with the generated OCT images, downstream retinal layer segmentation tasks demonstrate improved results. The code is publicly available at: https://github.com/nicetomeetu21/GenPSM.

Interactive Deep Learning-Based Retinal OCT Layer Segmentation Refinement by Regressing Translation Maps

Interactive Deep Learning-Based Retinal OCT Layer Segmentation Refinement by Regressing Translation Maps

Self-Guided Adversarial Network for Domain Adaptive Retinal Layer Segmentation

Self-Guided Adversarial Network for Domain Adaptive Retinal Layer Segmentation

Lightweight Retinal Layer Segmentation With Global Reasoning

Lightweight Retinal Layer Segmentation With Global Reasoning

Evaluation of Retinal Layers in Individuals With Pseudoexfoliation Syndrome and Ocular Hypertension.

This study investigated the retinal segmental thicknesses in individuals with pseudoexfoliation syndrome and ocular hypertension. Maximum thinning was found at 6 mm inferior to the inner plexiform layer. This layer is very important for the early diagnosis of glaucoma. To analyze the thickness of the peripapillary retinal nerve fiber layer and 8 macular layers using optical coherence tomography in eyes with ocular hypertension (OHT) and pseudoexfoliation syndrome (PXS) and healthy eyes and to evaluate between-group differences. In a prospective study, the macular segmentation of retinal layers in 120 eyes of 120 participants was performed automatically using current Heidelberg Spectralis optical coherence tomography software, which provides measurements for 8 retinal layers. Thickness maps divided into nine subfields (ie, 1, 3, and 6mm) were extracted from the software for each retinal layer and compared between groups. The thinnest macular layers appeared in the ocular hypertensive PXS, normotensive PXS, and OHT groups in that order. In the inner retinal layers (macular retinal nerve fiber layer, ganglion cell layer, and inner plexiform layer), statistically significant differences emerged between the PXS and control groups in all quadrants of the 3 and 6mm areas. No significant difference between the OHT group and control group appeared except in the 6mm temporal quadrant of the inner plexiform layer (IPL). Receiving operating characteristic analysis revealed quadrants with high area-under-the-curve values at 3 and 6mm in macular segments in all 3 groups compared with the control group. In macular segment analysis, the inner retinal layers showed the most thinning in patients with ocular hypertensive PXS. According to receiving operating characteristic curve analysis, examinations performed 6mm inferior to the IPL, as the quadrant with the highest area under the curve in all 3 groups, are critical for the early diagnosis of glaucoma.