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Purpose: The purpose of this tutorial is to provide visual scientists with various approaches for comparing two or more groups of data using parametric statistical tests, which require that the distribution of data within each group is normal (Gaussian). Non-parametric tests are used for inference when the sample data are not normally distributed or the sample is too small to assess its true distribution., Methods: Methods are reviewed using retinal thickness, as measured by optical coherence tomography (OCT), as an example for comparing two or more group means. The following parametric statistical approaches are presented for different situations: two-sample t-test, Analysis of Variance (ANOVA), paired t-test, and the analysis of repeated measures data using a linear mixed-effects model approach., Results: Analyzing differences between means using various approaches is demonstrated, and follow-up procedures to analyze pairwise differences between means when there are more than two comparison groups are discussed. The assumption of equal variance between groups and methods to test for equal variances are examined. Examples of repeated measures analysis for right and left eyes on subjects, across spatial segments within the same eye (e.g. quadrants of each retina), and over time are given., Conclusions: This tutorial outlines parametric inference tests for comparing means of two or more groups and discusses how to interpret the output from statistical software packages. Critical assumptions made by the tests and ways of checking these assumptions are discussed. Efficient study designs increase the likelihood of detecting differences between groups if such differences exist. Situations commonly encountered by vision scientists involve repeated measures from the same subject over time, measurements on both right and left eyes from the same subject, and measurements from different locations within the same eye. Repeated measurements are usually correlated, and the statistical analysis needs to account for the correlation. Doing this the right way helps to ensure rigor so that the results can be repeated and validated.

Introduction: Non-invasive multimodal imaging, including optical coherence tomography angiography (OCTA), has demonstrated high sensitivity and specificity for the management of retinal diseases. Since the availability of OCTA in 2015, we have developed a policy of "as little fluorescein angiography as possible". In this study, we describe the rate of OCTA and fluorescein angiography (FA) performed and their indications., Methods: Retrospective descriptive single-centre study. Chart review of patients examined for retinal disorders by one medical retina specialist between January 2015 and June 2018., Results: 3487 patients were examined for a retinal disease. The charts of 651 of these patients (1170 eyes) were randomly selected for analysis (study group). The mean age in the study group was 72 years (60% female). Overall, OCTA and FA were performed in 49.4% and 6.5% of the studied eyes respectively. After the first year, the rate of FA decreased to 2%. The main indications for OCTA were age-related macular degeneration (AMD): 50%, central serous chorioretinopathy (CSC): 12% and retinal vascular occlusion: 10%. The most frequent indications for FA were AMD: 49%, diabetic retinopathy (DR): 18% and CSC: 14%. FA was performed in 16%, 11% and 10% of eyes diagnosed with DR, CSC and AMD respectively (P<0.05)., Conclusion: In routine practice, OCTA was performed in half of the eyes examined for a retinal disorder. The overall rate of FA was 6.5% and dropped to 2% after the first year of OCTA use. DR was the main disorder still requiring FA., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Purpose: To determine whether ultra-widefield (UWF) retinal imaging changes the staging or management of sickle cell retinopathy compared with clinical examination., Methods: Prospective, observational study including patients with sickle cell disease. All patients underwent dilated fundus examination by a fellowship-trained retina specialist, as well as UWF fundus photography (FF) and fluorescein angiography (FA). Sickle retinopathy stage and treatment recommendation per eye were determined after clinical examination, UWF-FF, and UWF-FA, respectively, and differences in retinopathy stage and treatment recommendation were compared., Results: A total of 70 eyes from 35 patients (17 women, 48.6%), mean age 30.4 years, were included. Sickle genotypes included 26 patients with sickle SS (74.3%), 7 SC (20.0%), and 2 β(+)thalassemia (5.7%). Based on examination, most eyes (42/70; 60.0%) had no visible retinopathy. Based on UWF-FF, about half of the eyes were found to be Goldberg Stage 2 or above (36/70; 51.4%). Based on UWF-FA, nearly all eyes were Goldberg Stage 2 or above (63/70; 90%). However, clinical examination reliably detected neovascularization, and in no case did the addition of UWF imaging change management relative to examination alone., Conclusion: Ultra-widefield imaging detects a higher stage of sickle cell retinopathy compared with clinical examination alone, but these differences may not be clinically significant.

Background and Objective: Accurate segmentation of the intra-retinal tissue layers in Optical Coherence Tomography (OCT) images plays an important role in the diagnosis and treatment of ocular diseases such as Age-Related Macular Degeneration (AMD) and Diabetic Macular Edema (DME). The existing energy minimization based methods employ multiple, manually handcrafted cost terms and often fail in the presence of pathologies. In this work, we eliminate the need to handcraft the energy by learning it from training images in an end-to-end manner. Our method can be easily adapted to pathologies by re-training it on an appropriate dataset., Methods: We propose a Conditional Random Field (CRF) framework for the joint multi-layer segmentation of OCT B-scans. The appearance of each retinal layer and boundary is modeled by two convolutional filter banks and the shape priors are modeled using Gaussian distributions. The total CRF energy is linearly parameterized to allow a joint, end-to-end training by employing the Structured Support Vector Machine formulation., Results: The proposed method outperformed three benchmark algorithms on four public datasets. The NORMAL-1 and NORMAL-2 datasets contain healthy OCT B-scans while the AMD-1 and DME-1 dataset contain B-scans of AMD and DME cases respectively. The proposed method achieved an average unsigned boundary localization error (U-BLE) of 1.52 pixels on NORMAL-1, 1.11 pixels on NORMAL-2 and 2.04 pixels on the combined NORMAL-1 and DME-1 dataset across the eight layer boundaries, outperforming the three benchmark methods in each case. The Dice coefficient was 0.87 on NORMAL-1, 0.89 on NORMAL-2 and 0.84 on the combined NORMAL-1 and DME-1 dataset across the seven retinal layers. On the combined NORMAL-1 and AMD-1 dataset, we achieved an average U-BLE of 1.86 pixels on the ILM, inner and outer RPE boundaries and a Dice of 0.98 for the ILM-RPE in region and 0.81 for the RPE layer., Conclusion: We have proposed a supervised CRF based method to jointly segment multiple tissue layers in OCT images. It can aid the ophthalmologists in the quantitative analysis of structural changes in the retinal tissue layers for clinical practice and large-scale clinical studies., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The purpose of this study was to identify the normative values of ganglion cell-inner plexiform layer (GCIPL) thickness in healthy Korean children using swept-source optical coherence tomography (SS-OCT) and to investigate the correlations of age, refractive error, axial length (AL), retinal nerve fiber layer (RNFL) thickness and cup-to-disc (C/D) ratio with GCIPL thickness. Children aged between 3 and 17 who had visited our pediatric ophthalmology clinic were enrolled. Each subject underwent full ophthalmic examinations including RNFL thickness, C/D ratio and GCIPL thickness measurement by SS-OCT as well as AL measurement by partial-coherence interferometry. A total of 254 eyes of 127 children were included. The mean average GCIPL thickness was 71.5 ± 5.35 μm; the thickest sector was the superonasal and the thinnest the inferior. According to multivariate regression analysis, average GCIPL thickness was significantly associated with spherical equivalent and RNFL thickness (P < 0.0001 for both): the higher the myopia or the thinner the RNFL thickness, the thinner the GCIPL thickness. In conclusion, this study provides an SS-OCT-based pediatric normative database of GCIPL thickness that can serve as a reference for early detection and follow-up of glaucoma and optic nerve diseases in children.

Spectral-domain Optical coherence tomography (OCT) has shown remarkable utility in the study of retinal disease and has helped to characterize the fovea in Parkinson disease (PD) patients. We developed a detailed mathematical model based on raw OCT data to allow differentiation of foveae of PD patients from healthy controls. Of the various models we tested, a difference of a Gaussian and a polynomial was found to have "the best fit". Decision was based on mathematical evaluation of the fit of the model to the data of 45 control eyes versus 50 PD eyes. We compared the model parameters in the two groups using receiver-operating characteristics (ROC). A single parameter discriminated 70 % of PD eyes from controls, while using seven of the eight parameters of the model allowed 76 % to be discriminated. The future clinical utility of mathematical modeling in study of diffuse neurodegenerative conditions that also affect the fovea is discussed.

Purpose: To evaluate a simplified method to measure peripapillary choroidal thickness using commercially available, three-dimensional optical coherence tomography (3D-OCT)., Methods: 3D-OCT images of normal eyes were consecutively obtained from the 3D-OCT database of Korea University Medical Center On the peripapillary images for retinal nerve fiber layer (RNFL) analysis, choroidal thickness was measured by adjusting the segmentation line for the retinal pigment epithelium to the chorioscleral junction using the modification tool built into the 3D-OCT image viewer program. Variations of choroidal thickness at 12 sectors of the peripapillary area were evaluated., Results: We were able to measure the peripapillary choroidal thickness in 40 eyes of our 40 participants, who had a mean age of 41.2 years (range, 15 to 84 years). Choroidal thickness measurements had strong inter-observer correlation at each sector (r = 0.901 to 0.991, p < 0.001). The mean choroidal thickness was 191 ± 62 µm. Choroidal thickness was greatest at the temporal quadrant (mean ± SD, 210 ± 78 µm), followed by the superior (202 ± 66 µm), nasal (187 ± 64 µm), and inferior quadrants (152 ± 59 µm)., Conclusions: The measurement of choroidal thickness on peripapillary circle scan images for RNFL analysis using the 3D-OCT viewing program was highly reliable and efficient.

We present a novel, high-speed, polarization-sensitive, optical coherence tomography set-up for retinal imaging operating at a central wavelength of 1060 nm which was tested for in vivo imaging in healthy human volunteers. We use the system in combination with a Fourier domain mode locked laser with active spectral shaping which enables the use of forward and backward sweep in order to double the imaging speed without a buffering stage. With this approach and with a custom designed data acquisition system, we show polarization-sensitive imaging with an A-scan rate of 350 kHz. The acquired three-dimensional data sets of healthy human volunteers show different polarization characteristics in the eye, such as depolarization in the retinal pigment epithelium and birefringence in retinal nerve fiber layer and sclera. The increased speed allows imaging of large volumes with reduced motion artifacts. Moreover, averaging several two-dimensional frames allows the generation of high-definition B-scans without the use of an eye-tracking system. The increased penetration depth of the system, which is caused by the longer probing beam wavelength, is beneficial for imaging choroidal and scleral structures and allows automated segmentation of these layers based on their polarization characteristics.

ABSTRACT. Image enhancement of retinal structures, in optical coherence tomography (OCT) scans through denoising, has the potential to aid in the diagnosis of several eye diseases. In this paper, a locally adaptive denoising algorithm using double-density dual-tree complex wavelet transform, a combination of the double-density wavelet transform and the dual-tree complex wavelet transform, is applied to reduce speckle noise in OCT images of the retina. The algorithm overcomes the limitations of commonly used multiple frame averaging technique, namely the limited number of frames that can be recorded due to eye movements, by providing a comparable image quality in significantly less acquisition time equal to an order of magnitude less time compared to the averaging method. In addition, improvements of image quality metrics and 5 dB increase in the signal-to-noise ratio are attained.

The review provides a concise explanation of principles of operation of different optical coherence tomography methods. A comparative analysis of their advantages and disadvantages is presented in relation to specific applications. The review will assist the reader in making an educated choice on the most suitable optical coherence tomography method to be used in a particular application., (© 2012 The Authors Journal of Microscopy © 2012 Royal Microscopical Society.)

Aims: To determine the retinal nerve fibre layer (RNFL) thickness at which visual field (VF) damage becomes detectable and associated with structural loss., Methods: In a prospective cross-sectional study, 72 healthy and 40 glaucoma subjects (one eye per subject) recruited from an academic institution had VF examinations and spectral domain optical coherence tomography (SD-OCT) optic disc cube scans (Humphrey field analyser and Cirrus HD-OCT, respectively). Comparison of global mean and sectoral RNFL thicknesses with VF threshold values showed a plateau of threshold values at high RNFL thicknesses and a sharp decrease at lower RNFL thicknesses. A 'broken stick' statistical model was fitted to global and sectoral data to estimate the RNFL thickness 'tipping point' where the VF threshold values become associated with the structural measurements. The slope for the association between structure and function was computed for data above and below the tipping point., Results: The mean RNFL thickness threshold for VF loss was 75.3 μm (95% CI: 68.9 to 81.8), reflecting a 17.3% RNFL thickness loss from age-matched normative value. Above the tipping point, the slope for RNFL thickness and threshold value was 0.03 dB/μm (CI: -0.02 to 0.08) and below the tipping point, it was 0.28 dB/μm (CI: 0.18 to 0.38); the difference between the slopes was statistically significant (p<0.001). A similar pattern was observed for quadrant and clock-hour analysis., Conclusions: Substantial structural loss (∼17%) appears to be necessary for functional loss to be detectable using the current testing methods.

A robust multiscale stereo matching algorithm is proposed to find reliable correspondences between low contrast and weakly textured retinal image pairs with radiometric differences. Existing algorithms designed to deal with piecewise planar surfaces with distinct features and Lambertian reflectance do not apply in applications such as 3D reconstruction of medical images including stereo retinal images. In this paper, robust pixel feature vectors are formulated to extract discriminative features in the presence of noise in scale space, through which the response of low-frequency mechanisms alter and interact with the response of high-frequency mechanisms. The deep structures of the scene are represented with the evolution of disparity estimates in scale space, which distributes the matching ambiguity along the scale dimension to obtain globally coherent reconstructions. The performance is verified both qualitatively by face validity and quantitatively on our collection of stereo fundus image sets with ground truth, which have been made publicly available as an extension of standard test images for performance evaluation.

Objective: Thiazolidinediones (TZDs) are insulin-sensitizing agents that are associated with peripheral edema and have been reported to be associated with diabetic macular edema (DME). We hypothesized that TZDs produce subclinical increases in retinal thickness that may be detected by optical coherence tomography (OCT) but are not seen on routine dilated funduscopic examination., Research Design and Methods: We used OCT to screen for subclinical DME in a cross-sectional study of patients with type 2 diabetes; 29 patients were taking TZDs and 58 were not taking TZDs. We analyzed data using multiple linear regression analysis to investigate associations of retinal thickness with clinical characteristics., Results: There was no significant difference between the central subfield retinal thickness in the non-TZD group (206.4 ± 28.0 microns; n = 59) and TZD group (204.1 ± 26.1 microns; n = 29) (p = .72) nor were there significant differences in any other retinal subfield. There was no significant correlation of retinal thickness with laboratory results studies--peripheral edema, gender, age, duration of diabetes, individual, or combinations of medications. Retinal thickness differences between regions displayed normal anatomical variation. However, ethnic differences were found in which African-Americans had thinner retinas in all regions than Caucasians regardless of whether or not they used TZDs., Conclusions: These data suggest that TZDs do not cause subclinical DME in a demographically diverse patient population with diabetes. The established normal ranges for macular thickness may require adjustment based on ethnicity., (© 2011 Diabetes Technology Society.)

Polarization sensitive OCT has recently been shown to provide tissue specific contrast, enabling direct identification of retinal layers based on the intrinsic properties of their interaction with light. However, the capabilities of displaying and analyzing 3D datasets in scientific publications were rather limited. Within the framework of the Interactive Science Publishing project, we present new ways of displaying and analyzing 3D sets of various polarization parameters recorded in healthy and diseased human retinas. These datasets can be interactively explored by the reader. Furthermore, we provide data of the 3D distribution of backscattered Stokes vectors to allow the reader to develop and test their own data processing algorithms.

The purpose of this study was to investigate the effect of image quality on retinal nerve fiber layer (RNFL) and retinal thickness measurements obtained using three commercially available spectral domain-optical coherence tomographers (SD-OCT). Subjectively determined good, medium and poor quality images were obtained from four healthy and one glaucoma suspect eyes. RNFL and retinal thickness measurements were compared as a function of image quality. Results indicate that when image quality is within the range specified as acceptable by SD-OCT manufacturers, RNFL and retinal thickness measurements are comparable.

Ultrahigh-resolution adaptive optics-optical coherence tomography (UHR-AO-OCT) instrumentation allowing monochromatic and chromatic aberration correction was used for volumetric in vivo retinal imaging of various retinal structures including the macula and optic nerve head (ONH). Novel visualization methods that simplify AO-OCT data viewing are presented, and include co-registration of AO-OCT volumes with fundus photography and stitching of multiple AO-OCT sub-volumes to create a large field of view (FOV) high-resolution volume. Additionally, we explored the utility of Interactive Science Publishing by linking all presented AO-OCT datasets with the OSA ISP software.

Ultrabroadband sources, such as multiplexed superluminescent diodes (SLDs) and femtosecond lasers, have been successfully employed in adaptive optics optical coherence tomography (AO-OCT) systems for ultrahigh resolution retinal imaging. The large cost differential of these sources, however, motivates the need for a performance comparison. Here, we compare the performance of a Femtolasers Integral Ti:Sapphire laser and a Superlum BroadLighter T840, using the same AO-OCT system and the same subject. In addition, we investigate the capability of our instrument equipped with the Integral to capture volume images of the fovea and adjacent regions on a second subject using the AO to control focus in the retina and custom and freeware image registration software to reduce eye motion artifacts. Monochromatic ocular aberrations were corrected with a woofer-tweeter AO system. Coherence lengths of the Integral and BroadLighter were measured in vivo at 3.2 microm and 3.3 microm, respectively. The difference in dynamic range was 5 dB, close to the expected variability of the experiment. Individual cone photoreceptors, retinal capillaries and nerve fiber bundles were distinguished in all three dimensions with both sources. The acquired retinal volumes are provided for viewing in OSA ISP, allowing the reader to data mine at the microscope level.

We demonstrate and compare two image processing methods for visualization and analysis of three-dimensional optical coherence tomography (OCT) data acquired in eyes with different retinal pathologies. A method of retinal layer segmentation based on a multiple intensity thresholding algorithm was implemented in order to generate simultaneously outer retinal topography maps and reflectivity maps. We compare the applicability of the two methods to the diagnosis of retinal diseases and their progression. The data presented in this contribution were acquired with a high speed (25,000 A-scans/s), high resolution (4.5 microm) spectral OCT prototype instrument operating in the ophthalmology clinic.

Ultrahigh-resolution (UHR) spectral-domain optical coherence tomography (SD-OCT) and speckle noise-reduced SD-OCT instruments were compared with regard to their ability to visualize retinal microstructures and detect micropathologies in the same series of eyes in this hospital-based study. Both the instruments identically visualized normal retinal structures, except for the retinal ganglion cell layer, which was better delineated by the speckle noise-reduced SD-OCT instrument. Retinal pigment epithelium (RPE) and Bruch's membrane were better delineated by UHR SD-OCT. Retinal and sub-RPE pathologies were also identically visualized by both the instruments. Layer differentiation for locating each pathology was better visualized by speckle noise-reduced SD-OCT.

Fast T-scanning (transverse scanning, en-face) was used to build B-scan or C-scan optical coherence tomography (OCT) images of the retina. Several unique signature patterns of en-face (coronal) are reviewed in conjunction with associated confocal images of the fundus and B-scan OCT images. Benefits in combining T-scan OCT with confocal imaging to generate pairs of OCT and confocal images similar to those generated by scanning laser ophthalmoscopy (SLO) are discussed in comparison with the spectral OCT systems. The multichannel potential of the OCT/SLO system is demonstrated with the addition of a third hardware channel which acquires and generates indocyanine green (ICG) fluorescence images. The OCT, confocal SLO and ICG fluorescence images are simultaneously presented in a two or a three screen format. A fourth channel which displays a live mix of frames of the ICG sequence superimposed on the corresponding coronal OCT slices for immediate multidimensional comparison, is also included. OSA ISP software is employed to illustrate the synergy between the simultaneously provided perspectives. This synergy promotes interpretation of information by enhancing diagnostic comparisons and facilitates internal correction of movement artifacts within C-scan and B-scan OCT images using information provided by the SLO channel.

This study examines the ability of RTVue, Cirrus and Spectralis OCT Spectral domain-optical coherence tomographs (SD-OCT) to detect localized retinal nerve fiber layer defects in glaucomatous eyes. In this observational case series, four glaucoma patients (8 eyes) were selected from the University of California, San Diego Shiley Eye Center and the Diagnostic Innovations in Glaucoma Study (DIGS) based on the presence of documented localized RNFL defects in at least one eye confirmed by masked stereophotograph assessment. One RTVue 3D Disc scan, one RTVue NHM4 scan, one Cirrus Optic Disk Cube 200x200 scan and one Spectralis scan centered on the optic disc (15x15 scan angle, 768 A-scans x 73 B-scans) were obtained on all undilated eyes within a single session. Results were compared with those obtained from stereophotographs. In 6 eyes the presence of localized RNFL defects was detected by stereophotography. In general, by qualitatively evaluating the retinal thickness maps generated, all SD-OCT instruments examined were able to confirm the presence of localized glaucomatous structural damage seen on stereophotographs. This study confirms SD-OCT is a promising technology for glaucoma detection as it may assist clinicians identify the presence of localized glaucomatous structural damage.

: Recent substantial developments in light source and detector technology have initiated a paradigm shift in retinal optical coherence tomography (OCT) performance. Broad bandwidth light sources in the 800 nm and 1060 nm wavelength region enable axial OCT resolutions of 2-3 mum and 5-7 mum, respectively. Novel high speed silicon based CMOS cameras at 800 nm and InGaAs based CCD cameras in combination with frequency domain OCT technology enable data acquisition speeds of up to 47,000 A-scans/s at 1060 nm and up to 312,500 A-scans/s at 800 nm. Combining ultrahigh axial resolution, ultrahigh speed OCT at 800 nm with pancorrected adaptive optics allows volumetric in vivo cellular resolution retinal imaging. Commercially available three-dimensional (3D) retinal OCT at 800 nm (20,000 A-scans/s, 6 mum axial resolution) is compared to ultrahigh speed 3D retinal imaging at 800 nm (160,000 A-scans/s, 2-3 mum axial resolution), high speed 3D choroidal imaging at 1060 nm (47,000 Ascan/ second, 6-7 mum axial resolution) and cellular resolution retinal imaging at 800 nm using adaptive optics OCT at 160,000 A-scans/second with isotropic resolution of ~2 mum. Analysis of the performance of these four imaging modalities applied in normal and pathologic eyes focusing on motion artifact free volumetric retinal imaging and revealing novel, complementary morphological information due to enhanced resolution, speed and penetration is presented.

To diagnose glaucoma and other diseases of the retinal ganglion cell/ optic nerve, the thickness of the retinal nerve fiber layer (RNFL) is routinely measured with optical coherence tomography. Until recently, these OCT measurements were made almost exclusively with a time domain OCT (tdOCT) machine from a single manufacturer. Recently, a number of OCT machines, based upon an improved frequency domain OCT technology (fdOCT), have appeared. We compared measurements made using a new fdOCT machine to those from the older tdOCT machine. The results were comparable. More importantly, we learned that the key factor determining whether results from different machines will be comparable is the algorithm used to segment RNFL thickness, not the type of OCT.

Optical coherence tomography (OCT) is a new noninvasive high-resolution method that measures the retinal nerve fiber layer (RNFL) thickness. An overview of the use in optic neuritis (ON) and multiple sclerosis (MS) is presented. Literature survey of PubMed was carried out. RNFL thickness in eyes of healthy control subjects was 102.9-111.11 microm, in eyes affected by ON 59.79-85 microm, and in fellow eyes 82.73-99.8 microm. All studies found a significant reduction in RNFL in eyes affected by ON compared with fellow eyes and eyes of healthy controls. Two out of three studies found a significant reduction in RNFL in fellow eyes compared with control eyes. RNFL thickness correlated with visual acuity, visual field, low-contrast letter acuity, contrast sensitivity, and color vision. Correlations were also found with the optic nerve area evaluated by magnetic resonance imaging, neurologic impairment score, and increasing disease duration. One of two studies found a significant correlation with amplitudes of visual evoked potentials, neither correlated with latencies. OCT is a promising new tool for evaluating atrophy in patients with ON and MS.