Choroidal Haller's and Sattler's layer thickness measurement using 3-dimensional 1060-nm optical coherence tomography.

OBJECTIVES: To examine the feasibility of automatically segmented choroidal vessels in three-dimensional (3D) 1060-nmOCT by testing repeatability in healthy and AMD eyes and by mapping Haller's and Sattler's layer thickness in healthy eyes. METHODS: Fifty-five eyes (from 45 healthy subjects and 10 with non-neovascular age-related macular degeneration (AMD) subjects) were imaged by 3D-1060-nmOCT over a 36°x36° field of view. Haller's and Sattler's layer were automatically segmented, mapped and averaged across the Early Treatment Diabetic Retinopathy Study grid. For ten AMD eyes and ten healthy eyes, imaging was repeated within the same session and on another day. Outcomes were the repeatability agreement of Haller's and Sattler's layer thicknesses in healthy and AMD eyes, the validation with ICGA and the statistical analysis of the effect of age and axial eye length (AL) on both healthy choroidal sublayers. RESULTS: The coefficients of repeatability for Sattler's and Haller's layers were 35% and 21% in healthy eyes and 44% and 31% in AMD eyes, respectively. The mean±SD healthy central submacular field thickness for Sattler's and Haller's was 87±56 µm and 141±50 µm, respectively, with a significant relationship for AL (P<.001). CONCLUSIONS: Automated Sattler's and Haller's thickness segmentation generates rapid 3D measurements with a repeatability corresponding to reported manual segmentation. Sublayers in healthy eyes thinned significantly with increasing AL. In the presence of the thinned Sattler's layer in AMD, careful measurement interpretation is needed. Automatic choroidal vascular layer mapping may help to explain if pathological choroidal thinning affects medium and large choroidal vasculature in addition to choriocapillaris loss.

Non-invasive detection of early retinal neuronal degeneration by ultrahigh resolution optical coherence tomography.

Optical coherence tomography (OCT) has revolutionises the diagnosis of retinal disease based on the detection of microscopic rather than subcellular changes in retinal anatomy. However, currently the technique is limited to the detection of microscopic rather than subcellular changes in retinal anatomy. However, coherence based imaging is extremely sensitive to both changes in optical contrast and cellular events at the micrometer scale, and can generate subtle changes in the spectral content of the OCT image. Here we test the hypothesis that OCT image speckle (image texture) contains information regarding otherwise unresolvable features such as organelle changes arising in the early stages of neuronal degeneration. Using ultrahigh resolution (UHR) OCT imaging at 800 nm (spectral width 140 nm) we developed a robust method of OCT image analyses, based on spatial wavelet and texture-based parameterisation of the image speckle pattern. For the first time we show that this approach allows the non-invasive detection and quantification of early apoptotic changes in neurons within 30 min of neuronal trauma sufficient to result in apoptosis. We show a positive correlation between immunofluorescent labelling of mitochondria (a potential source of changes in cellular optical contrast) with changes in the texture of the OCT images of cultured neurons. Moreover, similar changes in optical contrast were also seen in the retinal ganglion cell- inner plexiform layer in retinal explants following optic nerve transection. The optical clarity of the explants was maintained throughout in the absence of histologically detectable change. Our data suggest that UHR OCT can be used for the non-invasive quantitative assessment of neuronal health, with a particular application to the assessment of early retinal disease.

Automated three-dimensional choroidal vessel segmentation of 3D 1060 nm OCT retinal data.

A fully automated, robust vessel segmentation algorithm has been developed for choroidal OCT, employing multiscale 3D edge filtering and projection of "probability cones" to determine the vessel "core", even in the tomograms with low signal-to-noise ratio (SNR). Based on the ideal vessel response after registration and multiscale filtering, with computed depth related SNR, the vessel core estimate is dilated to quantify the full vessel diameter. As a consequence, various statistics can be computed using the 3D choroidal vessel information, such as ratios of inner (smaller) to outer (larger) choroidal vessels or the absolute/relative volume of choroid vessels. Choroidal vessel quantification can be displayed in various forms, focused and averaged within a special region of interest, or analyzed as the function of image depth. In this way, the proposed algorithm enables unique visualization of choroidal watershed zones, as well as the vessel size reduction when investigating the choroid from the sclera towards the retinal pigment epithelium (RPE). To the best of our knowledge, this is the first time that an automatic choroidal vessel segmentation algorithm is successfully applied to 1060 nm 3D OCT of healthy and diseased eyes.

Choroidal thinning in diabetes type 1 detected by 3-dimensional 1060 nm optical coherence tomography.

PURPOSE: To map choroidal (ChT) and retinal thickness (RT) in patients with diabetes type 1 with and without maculopathy and retinopathy in order to compare them with healthy subjects using high speed 3-dimensional (3D) 1060 nm optical coherence tomography (OCT). METHODS: Thirty-three eyes from 33 diabetes type 1 subjects (23-57 years, 15 male) divided into groups of without pathology (NDR) and with pathology (DR; including microaneurysms, exudates, clinically significant macular-oedema and proliferative retinopathy) were compared with 20 healthy axial eye length and age-matched subjects (24-57 years, 9 male), imaged by high speed (60.000 A-scans/s) 3D 1060 nm OCT performed over 36° × 36° field of view. Ocular health status, disease duration, body mass index, haemoglobin-A1c, and blood pressure (bp) measurements were recorded. Subfoveal ChT, and 2D topographic maps between retinal pigment epithelium and the choroidal/scleral-interface, were automatically generated and statistically analyzed. RESULTS: Subfoveal ChT (mean ± SD, µm) for healthy eyes was 388 ± 109; significantly thicker than all diabetic groups, 291 ± 64 for NDR, and 303 ± 82 for DR (ANOVA P < 0.004, Tukey P = 0.01 for NDR and DR). Thinning did not relate to recorded factors (multi-regression analysis, P > 0.05). Compared with healthy eyes and the NDR, the averaged DR ChT-map demonstrated temporal thinning that extended superiorly and temporal-inferiorly (unpaired t-test, P < 0.05). Foveal RT and RT-maps showed no statistically significant difference between groups (mean SD, µm, healthy 212 ± 17, NDR 217 ± 15, DR 216 ± 27, ANOVA P > 0.05). CONCLUSIONS: ChT is decreased in diabetes type 1, independent of the absence of pathology and of diabetic disease duration. In eyes with pathology, 3D 1060 nm OCT averaged maps showed an extension of the thinning area matching retinal lesions and suggesting its involvement on onset or progression of disease.

Automated choroidal segmentation of 1060 nm OCT in healthy and pathologic eyes using a statistical model.

A two stage statistical model based on texture and shape for fully automatic choroidal segmentation of normal and pathologic eyes obtained by a 1060 nm optical coherence tomography (OCT) system is developed. A novel dynamic programming approach is implemented to determine location of the retinal pigment epithelium/ Bruch's membrane /choriocapillaris (RBC) boundary. The choroid-sclera interface (CSI) is segmented using a statistical model. The algorithm is robust even in presence of speckle noise, low signal (thick choroid), retinal pigment epithelium (RPE) detachments and atrophy, drusen, shadowing and other artifacts. Evaluation against a set of 871 manually segmented cross-sectional scans from 12 eyes achieves an average error rate of 13%, computed per tomogram as a ratio of incorrectly classified pixels and the total layer surface. For the first time a fully automatic choroidal segmentation algorithm is successfully applied to a wide range of clinical volumetric OCT data.

Mapping choroidal and retinal thickness variation in type 2 diabetes using three-dimensional 1060-nm optical coherence tomography.

PURPOSE: To map choroidal (ChT) and retinal thickness (RT) in healthy subjects and patients with diabetes with and without maculopathy using three dimensional 1060-nm optical coherence tomography (3D-1060nm-OCT). METHODS: Sixty-three eyes from 42 diabetic subjects (41-82 years of age; 11 females) grouped according to a custom scheme using Early Treatment Diabetic Retinopathy Study definitions for pathology within 1 disc-diameter of fovea (without pathology [NDR], microaneurysms [M1], exudates [M2], clinically significant macular edema [CSME]) and 16 eyes from 16 healthy age matched subjects (38-79 years of age; 11 females) were imaged by 3D-1060nm-OCT performed over a 36° × 36° field of view. Axial length, 45° fundus photographs, body mass index, plasma glucose, and blood pressure measurements were recorded. The ChT at the subfoveal location and ChT maps between RPE and the choroidal-scleral interface were generated and statistically analyzed. RESULTS: RT maps show thinning in the NDR group but an increase in thickness with increasing maculopathy in the temporal and central regions (unpaired t-test; P < 0.05). ChT mapping of all diabetic patients revealed central and inferior thinning compared to healthy eyes (unpaired t-test; P < 0.001). Subfoveal ChT (mean ± SD) for healthy eyes was 327 ± 74 µm, which was significantly thicker than all diabetic groups (214 ± 55 µm for NDR, 208 ± 49 µm for M1, 205 ± 54 µm for M2, and 211 ± 76 µm for CSME (ANOVA P < 0.001; Tukey P < 0.001). CONCLUSIONS: 3D-1060nm-OCT has shown that the central choroid is thinner in all type 2 diabetic eyes regardless of disease stage. The choroidal thinning may exceed the magnitude of possible choriocapillaris atrophy. In contrast to the conventional assessment of pathologic thickness change in several locations, thickness maps allow investigation of the choroid over the extent of affected areas.

Artefact reduction for cell migration visualization using spectral domain optical coherence tomography.

Visualization of cell migration during chemotaxis using spectral domain optical coherence tomography (OCT) requires non-standard processing techniques. Stripe artefacts and camera noise floor present in OCT data prevent detailed computer-assisted reconstruction and quantification of cell locomotion. Furthermore, imaging artefacts lead to unreliable results in automated texture based cell analysis. Here we characterize three pronounced artefacts that become visible when imaging sample structures with high dynamic range, e.g. cultured cells: (i) time-varying fixed-pattern noise; (ii) stripe artefacts generated by background estimation using tomogram averaging; (iii) image modulations due to spectral shaping. We evaluate techniques to minimize the above mentioned artefacts using an 800 nm optical coherence microscope. Effect of artefact reduction is shown exemplarily on two cell cultures, i.e. Dictyostelium on nitrocellulose substrate, and retinal ganglion cells (RGC-5) cultured on a glass coverslip. Retinal imaging also profits from the proposed processing techniques.

Robust segmentation of intraretinal layers in the normal human fovea using a novel statistical model based on texture and shape analysis.

A novel statistical model based on texture and shape for fully automatic intraretinal layer segmentation of normal retinal tomograms obtained by a commercial 800nm optical coherence tomography (OCT) system is developed. While existing algorithms often fail dramatically due to strong speckle noise, non-optimal imaging conditions, shadows and other artefacts, the novel algorithm's accuracy only slowly deteriorates when progressively increasing segmentation task difficulty. Evaluation against a large set of manual segmentations shows unprecedented robustness, even in the presence of additional strong speckle noise, with dynamic range tested down to 12dB, enabling segmentation of almost all intraretinal layers in cases previously inaccessible to the existing algorithms. For the first time, an error measure is computed from a large, representative manually segmented data set (466 B-scans from 17 eyes, segmented twice by different operators) and compared to the automatic segmentation with a difference of only 2.6% against the inter-observer variability.

Three-dimensional 1060-nm OCT: choroidal thickness maps in normal subjects and improved posterior segment visualization in cataract patients.

PURPOSE: To evaluate the performance and potential clinical role of three-dimensional (3D) 1060-nm OCT by generating choroidal thickness (ChT) maps in patients of different ages with different degrees of ametropia and axial lengths and to investigate the effect of cataract grade on OCT retinal imaging quality. METHODS: Axial lengths (ALs) and 45° fundus photographs were acquired from 64 eyes (34 healthy subjects, 19 to 80 years, ametropia +3 to -10 D). 3D 1060-nm OCT was performed over a 36° × 36° field of view with ∼7-µm axial resolution and up to 70 frames/s (512 A-scans/frame). ChT maps between retinal pigment epithelium and the choroidal-scleral interface, were generated and statistically analyzed. A further 30 eyes (19 subjects), with cataracts assessed with the LOCS III scale, were imaged with 3D 1060-nm OCT and 800-nm OCT, and visualization of the posterior segment was compared qualitatively. RESULTS: In 64 eyes, ChT maps displayed a thickness decrease with increasing AL. Subfoveal ChT was 315 ± 106 µm (mean ± SD), negatively correlated with AL (R(2) = -0.47, P < 0.001). Averaged ChT maps of eyes with AL < 23.39 mm showed an increased ChT in an area ∼1500 µm inferior, compared with subfoveal ChT. Eyes with AL > 24.5 mm showed a larger variation and a thicker ChT superiorly than inferiorly. Reduced signal strength in cataractous eyes was found in 65% of the 800-nm OCT images, but in only 10% of the 1060-nm OCT images. CONCLUSIONS: The imaging performance of 3D 1060-nm OCT is unique, producing maps that show the variation in ChT over the entire field of view, in relation to axial length. This imaging system has the potential of visualizing a novel clinical diagnostic biomarker. Compared with 800-nm OCT, it provides superior visualization of the posterior pole in cataractous eyes.

Wide-field optical coherence tomography of the choroid in vivo.

PURPOSE: To demonstrate high-speed, high axial resolution optical coherence tomography (OCT) at 1060 nm with penetration to the sclera. The clinical feasibility of dense, high-speed sampling for higher levels of detail at the macula and optic nerve head is explored with respect to motion artifacts. METHODS: A three-dimensional (3D) OCT system making use of a high-speed camera operating at 47,000 depth scans/s was developed. The 1010- to 1080-nm wavelength band leads to 6.7 microm effective axial resolution and enables the acquisition of retinal and choroidal 130 Megavoxel volumes of human subjects within 7 seconds. Motion artifacts were reduced by numeric postprocessing techniques. RESULTS: Drift motion artifacts could be suppressed within fields up to 38 degrees x 38 degrees (approximately 1 cm(2)) using acquisition speeds of up to 74 frames/s at 512 x 512 pixel/frame. This isotropic OCT sampling of the human retina in vivo allowed reconstruction of the retinal microvasculature solely on vessel reflectivity, without the use of contrast agents, and revealed three interconnected capillary meshworks. Simultaneously in the choroid, the structure of the choriocapillaris, Sattler's layer, and Haller's layer were differentiated, and the choroidal-scleral interface was clearly delineated in densely sampled narrow- and wide-angle scans (>38 degrees). At the optic nerve head, the 3D fine structure of the lamina cribrosa and the circle of Zinn-Haller were visualized. CONCLUSIONS: OCT almost centered within the 1060-nm water transmission window significantly profits from lower scattering and allows investigation of the retina and choroid at an unprecedented combination of penetration and high speed at high resolution and may provide superior clinical feasibility to commercial 800-nm devices.