Ultrahigh-resolution Spectral Domain Optical Coherence Research Articles

Microstructural changes of photoreceptor layers detected by ultrahigh-resolution SD-OCT in patients with autosomal recessive bestrophinopathy

PurposeTo determine the changes in the microstructures of the photoreceptors in patients with autosomal recessive bestrophinopathy (ARB) by ultrahigh-resolution spectral-domain optical coherence tomography (UHR-SD-OCT). MethodsFive eyes of 4 patients with ARB were studied. Cross-sectional images of the fovea were recorded by the UHR-SD-OCT system with a depth resolution of <2.0 μm. ResultsThe UHR-SD-OCT images revealed changes in the outer retinal structures that were dependent on the severity of the photoreceptor atrophy. There was an increase in the reflectivity and appearance of small hyperreflective dots (HRDs) in the outer segments, followed by an irregularity and decrease in the length of the outer segments, then a disruption of the ellipsoid zone (EZ) band, and appearance of large HRDs corresponding to the segmented ellipsoids. Finally, there was a disappearance of the large HRDs followed by a localized thinning of the outer nuclear layer and appearance of hyperreflective foci above the region of the disrupted EZ. ConclusionsUHR-SD-OCT can record images that show detailed changes of the microstructures of the photoreceptors at different stages of ARB. These observations should help in determining the mechanisms involved in retinal pathology and should provide important information on the effectiveness of treatments.

Detailed analyses of microstructure of photoreceptor layer at different severities of occult macular dystrophy by ultrahigh-resolution SD-OCT

PurposeTo analyze the microstructures of the photoreceptor layer in detail in eyes with occult macular dystrophy (OMD, Miyake's disease) by ultrahigh-resolution spectral-domain optical coherence tomography (UHR-SD-OCT). ObservationsTwenty-eight normal subjects and 5 patients with OMD of different severities were studied. Cross-sectional images through the fovea were recorded with a UHR-SD-OCT system with a depth resolution of <2.0 μm. In patients with OMD, the UHR-SD-OCT images revealed abnormal photoreceptor microstructures which were not detected in the conventional SD-OCT images. The UHR-SD-OCT images showed that the interdigitation zone (IZ) was not present and the outer segments were hyperreflective with hyperreflective dots (HRDs) aligned like string of pearls during the earlier stages. There was a disruption of the ellipsoid zone (EZ) which appeared as clusters of larger HRDs, and these HRDs became less apparent with increasing time. The outer segments became hyporeflective and rod IZ became apparent with longer duration of the disease process. Conclusions and ImportanceThe UHR-SD-OCT images show detailed characteristics of the photoreceptor microstructures of different severities during the progression of OMD. These detailed observations will help in understanding the mechanisms involved in the retinal pathology and should provide important information for their treatments.

High-Speed, Ultrahigh-Resolution Spectral-Domain OCT with Extended Imaging Range Using Reference Arm Length Matching.

PurposeTo develop high-speed, extended-range, ultrahigh-resolution spectral-domain optical coherence tomography (UHR SD-OCT) and demonstrate scan protocols for clinical retinal imaging.MethodsA UHR SD-OCT operating at 840-nm with 150-nm bandwidths was developed. The axial imaging range was extended by dynamically matching reference arm length to the retinal contour during acquisition. Two scan protocols were demonstrated for imaging healthy participants and patients with dry age-related macular degeneration. A high-definition raster protocol with intra–B-scan reference arm length matching (ReALM) was used for high-quality cross-sectional imaging. A cube volume scan using horizontal and vertical rasters with inter–B-scan ReALM and software motion correction was used for en face and cross-sectional imaging. Linear OCT signal display enhanced visualization of outer retinal features.ResultsUHR SD-OCT was demonstrated at 128- and 250-kHz A-scan rates with 2.7 µm axial resolution and a 1.2-mm, 6-dB imaging range in the eye. Dynamic ReALM was used to maintain the retina within the 6-dB imaging range over wider fields of view. Outer retinal features, including the rod and cone interdigitation zones, retinal pigment epithelium, and Bruch's membrane were visualized and alterations observed in age-related macular degeneration eyes.ConclusionsTechnological advances and dynamic ReALM improve the imaging performance and clinical usability of UHR SD-OCT.Translational RelevanceThese advances should simplify clinical imaging workflow, reduce imaging session times, and improve yield of high quality images. Improved visualization of photoreceptors, retinal pigment epithelium, and Bruch's membrane may facilitate diagnosis and monitoring of age-related macular degeneration and other retinal diseases.

New clinical ultrahigh-resolution SD-OCT using A-scan matching algorithm.

A new clinical ultrahigh-resolution spectral domain optical coherence tomography (UHR-SD-OCT) system using an original averaging technique named "A-scan matching algorithm" was developed. The aim of this study was to determine whether our new UHR-SD-OCT system can obtain clearer sectional images of the retina than conventional standard resolution SD-OCT systems (SR-SD-OCT). We recorded horizontal B-scan images of 42 normal eyes using our new UHR-SD-OCT device (Bi-μ, Kowa) and a conventional SR-SD-OCT (Spectralis, Heidelberg). To evaluate the clarity of the interdigitation zone (IZ) subjectively, the integrity of IZ was divided into three types by two raters. To evaluate the clarity of the IZ objectively, a peak height score (PHS) was calculated at five different points of the macula using the longitudinal reflectivity profile. The mean (± SD) of the subjective visibility score of the IZ in the UHR-SD-OCT images was 2.64 ± 0.54 which was significantly higher than the 2.46 ± 0.46 in the SR-SD-OCT images (P = 0.02). The PHS was also significantly higher for the UHR-SD-OCT than for the SR-SD-OCT images at all five locations (all P < 0.01). The results indicate that the newly developed clinical UHR-SD-OCT instrument using the A-scan matching algorithm can obtain clearer images of the IZ, and they suggest that this device should be clinically useful in detecting finer structural abnormalities of the outer retina.

Measurement of the Retention Time of Different Ophthalmic Formulations with Ultrahigh-Resolution Optical Coherence Tomography

ABSTRACTPurpose/aim of the study: The purpose of this study was to measure the pre-corneal retention time of two marketed formulations (eye drops and eye gel) of a steroid–antibiotic fixed combination (FC) containing 0.1% dexamethasone and 0.3% netilmicin.Materials and methods: Pre-corneal retention time was evaluated in 16 healthy subjects using an ultrahigh-resolution anterior segment spectral domain optical coherence tomography (OCT). All subjects randomly received both formulations of the FC (Netildex, SIFI, Italy). Central tear film thickness (CTFT) was measured before instillation (time 0) and then after 1, 10, 20, 30, 40 50, 60 and 120 min. The pre-corneal retention time was calculated by plotting CTFT as a function of time. Differences between time points and groups were analyzed by Student’s t-test.Results: CTFT increased significantly after the instillation of the eye gel formulation (p < 0.001). CTFT reached its maximum value 1 min after instillation and returned to baseline after 60 min. No effect on CTFT was observed after the instillation of eye drops. The difference between the two formulations was statistically significant at time 1 min (p < 0.0001), 10 min (p < 0.001) and 20 min (p < 0.01).Conclusions: The FC formulated as eye gel was retained on the ocular surface longer than the corresponding eye drop solution. Consequently, the use of the eye gel might extend the interval between instillations and decrease the frequency of administration.

Full-range ultrahigh-resolution spectral-domain optical coherence tomography in 1.7 µm wavelength region for deep-penetration and high-resolution imaging of turbid tissues

For the first time, we developed a full-range ultrahigh-resolution (UHR) spectral-domain optical coherence tomography (SD-OCT) technique working in the 1.7 µm wavelength region. This technique allowed high-resolution, deep-tissue imaging. By using a supercontinuum source operating at a wavelength of 1.7 µm, an axial resolution of 3.6 µm in a tissue specimen was achieved. To enhance the imaging depth of UHR-SD-OCT, we performed full-range OCT imaging based on a phase modulation method. We demonstrated the three-dimensional (3D) imaging of a mouse brain with the developed system, and specific structures in the mouse brain were clearly visualized at depths up to 1.7 mm.

Myocardial imaging using ultrahigh-resolution spectral domain optical coherence tomography.

We present an ultrahigh-resolution spectral domain optical coherence tomography (OCT) system in 800 nm with a low-noise supercontinuum source (SC) optimized for myocardial imaging. The system was demonstrated to have an axial resolution of 2.72 μm with a large imaging depth of 1.78 mm and a 6-dB falloff range of 0.89 mm. The lateral resolution (5.52 μm) was compromised to enhance the image penetration required for myocardial imaging. The noise of the SC source was analyzed extensively and an imaging protocol was proposed for SC-based OCT imaging with appreciable contrast. Three-dimensional datasets were acquired ex vivo on the endocardium side of tissue specimens from different chambers of fresh human and swine hearts. With the increased resolution and contrast, features such as elastic fibers, Purkinje fibers, and collagen fiber bundles were observed. The correlation between the structural information revealed in the OCT images and tissue pathology was discussed as well.

Visual abilities in two raptors with different ecology.

Differences in visual capabilities are known to reflect differences in foraging behaviour even among closely related species. Among birds, the foraging of diurnal raptors is assumed to be guided mainly by vision but their foraging tactics include both scavenging upon immobile prey and the aerial pursuit of highly mobile prey. We studied how visual capabilities differ between two diurnal raptor species of similar size: Harris's hawks, Parabuteo unicinctus, which take mobile prey, and black kites, Milvus migrans, which are primarily carrion eaters. We measured visual acuity, foveal characteristics and visual fields in both species. Visual acuity was determined using a behavioural training technique; foveal characteristics were determined using ultra-high resolution spectral-domain optical coherence tomography (OCT); and visual field parameters were determined using an ophthalmoscopic reflex technique. We found that these two raptors differ in their visual capacities. Harris's hawks have a visual acuity slightly higher than that of black kites. Among the five Harris's hawks tested, individuals with higher estimated visual acuity made more horizontal head movements before making a decision. This may reflect an increase in the use of monocular vision. Harris's hawks have two foveas (one central and one temporal), while black kites have only one central fovea and a temporal area. Black kites have a wider visual field than Harris's hawks. This may facilitate the detection of conspecifics when they are scavenging. These differences in the visual capabilities of these two raptors may reflect differences in the perceptual demands of their foraging behaviours.

High-speed ultrahigh-resolution spectral domain optical coherence tomography using high-power supercontinuum at 0.8 µm wavelength

We demonstrated high-speed ultrahigh-resolution (UHR) optical coherence tomography (OCT) in the 800 nm wavelength region. A high-power coherent supercontinuum (SC) and a high-speed line scan camera were used to construct a spectral domain OCT. The axial resolution was 3.1 µm in air and 2.3 µm in tissue. The dependence of sensitivity on the SC power and A-scan rate was examined. For the A-scan rate of 70 kHz, the sensitivity of 104 dB was achieved for the SC power higher than 60 mW. High-speed in vivo UHR-OCT imaging was demonstrated for zebrafish embryo and swimming medaka.

Ultrahigh-Resolution Spectral Domain Optical Coherence Tomography Based on a Linear-Wavenumber Spectrometer

In this study we demonstrate ultrahigh-resolution spectral domain optical coherence tomography (UHR SD-OCT) with a linear-wavenumber (k) spectrometer, to accelerate signal processing and to display two-dimensional (2-D) images in real time. First, we performed a numerical simulation to find the optimal parameters for the linear-k spectrometer to achieve ultrahigh axial resolution, such as the number of grooves in a grating, the material for a dispersive prism, and the rotational angle between the grating and the dispersive prism. We found that a grating with 1200 grooves and an F2 equilateral prism at a rotational angle of <TEX>$26.07^{\circ}$</TEX>, in combination with a lens of focal length 85.1 mm, are suitable for UHR SD-OCT with the imaging depth range (limited by spectrometer resolution) set at 2.0 mm. As guided by the simulation results, we constructed the linear-k spectrometer needed to implement a UHR SD-OCT. The actual imaging depth range was measured to be approximately 2.1 mm, and axial resolution of <TEX>$3.8{\mu}m$</TEX> in air was achieved, corresponding to <TEX>$2</TEX><TEX>.8{\mu}m$</TEX> in tissue (n = 1.35). The sensitivity was -91 dB with -10 dB roll-off at 1.5 mm depth. We demonstrated a 128.2 fps acquisition rate for OCT images with 800 lines/frame, by taking advantage of NVIDIA's compute unified device architecture (CUDA) technology, which allowed for real-time signal processing compatible with the speed of the spectrometer's data acquisition.

Ultrahigh-resolution optical coherence tomography and its application in inspection of industrial materials

Since many industrial materials have micro or submicro structures on the surface or subsurface, utrahigh-resolution is required in the inspection of these materials. Ultrahigh-resolution optical coherence tomography uses broadband light sources to achieve axial image resolutions on the scale of a few microns. We have been investigating an ultrahigh-resolution spectral-domain optical coherence tomography (SD-OCT) system using supercontinuum sources (SC) in free space. The effective SC spectrum has a full width at half maximum of 230 nm centered around 665 nm, and the imaging setup has an ultrahigh axial resolution of 0.9 μm in air, and a lateral resolution of 3.9 μm, with the system measurement range being 0.6 mm in axial direction. At a 50 μm axial position, the sensitivity can be 63 dB with 28600 axial scans per second at 2048 pixels per axial scan. Images of polystyrene microspheres solution with an average diameter of 5 μm and different sizes of industrial abrasive papers are presented to illustrate the performance of the system.

Measurement of Tear Film Thickness Using Ultrahigh-Resolution Optical Coherence Tomography

To visualize the precorneal tear film with ultrahigh-resolution spectral domain optical coherence tomography, enabling quantification of tear film thickness in healthy subjects. A custom-built spectral domain optical coherence tomography system comprising a broadband titanium:sapphire laser operating at 800 nm and a high-speed charge coupled device (CCD) camera with a read-out rate of 47 kHz was used for measurement of precorneal tear film thickness. The system provides a theoretical axial resolution of 1.2 μm in tissue. The signal-to-noise ratio close to the zero delay was measured with 94 dB. A total of 26 healthy subjects were included in this study. Measurement was started immediately after blinking and averaged over a period of 1 second. In a subset of eight healthy subjects, the reproducibility of the approach was studied by measuring the tear film thickness every 10 minutes over 1 hour. The average central tear film thickness of the measured population was 4.79 ± 0.88 μm. Reproducibility was very high, with an intraclass correlation coefficient of 0.97. A breakup of the tear film was observed in one subject after 14 seconds. Our data indicate that the human precorneal tear film can be measured with excellent reproducibility using ultrahigh-resolution optical coherence tomography. This technique may be a valuable tool in the management of dry eye syndrome.

Ultrahigh resolution spectral-domain optical coherence tomography at 1.3 μm using a broadband superluminescent diode light source

We present an ultrahigh resolution spectral-domain optical coherence tomography imaging system using a broadband superluminescent diode light source emitting at a center wavelength of 1.3μm. The light source consists of two spectrally shifted superluminescent diodes that are coupled together into a single mode fiber. The effective emission power spectrum has a full width at half maximum of 200nm and the source output power is 10mW. The imaging system has an axial resolution of 3.9μm in air (<3.0μm in biological tissue), and a lateral resolution of 6.5μm. The sensitivity and the maximum line rate are 95dB and 46kHz, respectively. Images of an infrared viewing card and a cornea from human eye suffering from glaucoma showing Schlemm's canal are presented to illustrate the performance of the system.

Topographic Thickness of Bowman's Layer Determined by Ultra-High Resolution Spectral Domain–Optical Coherence Tomography

To characterize the thickness profile of the corneal epithelium and the Bowman's layer across the horizontal meridian. Forty-four eyes of 22 healthy subjects were investigated in this study. Ultra-high resolution anterior segment spectral domain-optical coherence tomography (SD-OCT) was used to assess the topographic thickness of the epithelium and the Bowman's layer across the cornea. Thicknesses at five locations, including the center, midperiphery, and periphery close to the limbus, on both the nasal and the temporal sides along the horizontal meridian, were analyzed. Mean epithelial thickness at the central cornea was 52.5 ± 2.4 μm. It increased gradually from the center to the periphery (P < 0.001). There was no significant difference between the nasal side and the temporal side for epithelial thickness. The central Bowman's layer thickness was 17.7 ± 1.6 μm, and it remained constant from the center to the midperiphery (P > 0.05). However, thicknesses at the nasal and temporal periphery, 20.0 ± 1.9 μm and 19.8 ± 2.2 μm, respectively, were significantly greater than the central and midperipheral thicknesses (P < 0.001). Nasal and temporal thicknesses were similar on either side of the center. The epithelium and the Bowman's layer were not evenly distributed across the horizontal meridian of the cornea. SD-OCT provided useful information about topographic thickness of the different corneal layers in vivo.

Structural Grading of Foveal Hypoplasia Using Spectral-Domain Optical Coherence Tomography: A Predictor of Visual Acuity?

To characterize and grade the spectrum of foveal hypoplasia based on different stages of arrested development of the fovea. Grading was performed using morphologic findings obtained by ultra high-resolution spectral-domain optical coherence tomography. Best-corrected visual acuity (BCVA) was calculated for different grades. Observational case series. Sixty-nine patients with foveal hypoplasia (albinism, n = 34; PAX6 mutations, n = 10; isolated cases, n = 14; achromatopsia, n = 11) and 65 control subjects were examined. A 7×7-mm retinal area was sampled using a 3-dimensional scanning protocol (743×75, A scans×B scans) with ultra high-resolution spectral-domain optical coherence tomography (SOCT Copernicus HR; 3-μm axial resolution). Gross morphologic abnormalities were documented. B-scans at the fovea were segmented using a longitudinal reflectivity profile. Logarithm of the minimum angle of resolution BCVA was obtained. Grading was based on presence or absence of foveal pit and widening of the outer nuclear layer (ONL) and outer segment (OS) at the fovea. Quantitative measurements were obtained for comparing atypical foveal hypoplasia in achromatopsia. Best-corrected visual acuity was compared with the grade of foveal hypoplasia. Four grades of foveal hypoplasia were distinguished: grade 1, shallow foveal pit, presence of ONL widening, presence of OS lengthening; grade 2, grade 1 but absence of foveal pit; grade 3, grade 2 but absence of OS lengthening; grade 4, grade 3 but absence of ONL widening. There was significant difference in visual acuity (VA) associated with each grade (P<0.0001). Grade 1 was associated with the best VA (median VA, 0.2), whereas grades 2, 3, and 4 were associated with progressively poorer VA with a median VA of 0.44, 0.60, and 0.78, respectively. The atypical features seen with foveal hypoplasia associated with achromatopsia were characterized by decreased retinal and ONL thickness and deeper foveal depth. A structural grading system for foveal hypoplasia was developed based on the stage at which foveal development was arrested, which helps to provide a prognostic indicator for VA and is applicable in a range of disorders associated with foveal hypoplasia. Atypical foveal hypoplasia in achromatopsia shows different characteristics. The author(s) have no proprietary or commercial interest in any materials discussed in this article.

In vivo volumetric imaging of chicken retina with ultrahigh-resolution spectral domain optical coherence tomography

The chicken retina is an established animal model for myopia and light-associated growth studies. It has a unique morphology: it is afoveate and avascular; oxygen and nutrition to the inner retina is delivered by a vascular tissue (pecten) that protrudes into the vitreous. Here we present, to the best of our knowledge, the first in vivo, volumetric high-resolution images of the chicken retina. Images were acquired with an ultrahigh-resolution optical coherence tomography (UHROCT) system with 3.5 µm axial resolution in the retina, at the rate of 47,000 A-scans/s. Spatial variations in the thickness of the nerve fiber and ganglion cell layers were mapped by segmenting and measuring the layer thickness with a semi-automatic segmentation algorithm. Volumetric visualization of the morphology and morphometric analysis of the chicken retina could aid significantly studies with chicken retinal models of ophthalmic diseases.

High-Resolution In Vivo Imaging in Achromatopsia

To characterize the retinal changes in patients with achromatopsia using an ultrahigh-resolution (UHR) spectral-domain optical coherence tomography (OCT) to examine how human achromatopsia corresponds to its animal model. Comparative case series. Ultrahigh-resolution OCT (Copernicus; OPTOPOL Technology S.A., Zawiercie, Poland; 3-μm axial resolution) was used to obtain scans from 13 patients (26 eyes) with achromatopsia and from 20 controls (40 eyes). A 3-dimensional scan program (743×75; A×B scan) sampling a 7×7-mm retinal area centered at the fovea was used to obtain tomograms of the fovea. Individual B-scans at the fovea were exported and analyzed using ImageJ (Wayne Rasband, National Institute of Health) for reflectance profiles and morphologic abnormalities. Gross morphologic changes in OCT were characterized. Specifically, inner segment and outer segment (IS/OS) junction and cone outer segment tip (COST) disruption was noted. Using the reflectance profiles, foveal depth, thickness of the outer nuclear layer (ONL), and retinal thickness (RT) were measured. A characteristic so-called punched out hyporeflective zone (HRZ) was noted in 7 of 13 patients; this was age-dependent (P = 0.001). The area of the HRZ was asymmetric with the nasal area being significantly greater than the temporal area (P = 0.002). In all patients, there was disruption of the IS/OS junction at the foveal or parafoveal regions, or both. Five of 13 patients also had a disrupted COST reflectivity. There was significant (P = 1.1×10(-6)) ONL thinning in the achromats compared with controls, which was age-dependent (P = 0.0002). Foveal maldevelopment was seen in 9 of 13 patients. The achromats also had a significantly reduced foveal depth (P = 7.7×10(-6)) and RT (P = 1.46×10(-9)) compared with controls. A range of signs in achromatopsia are described that can be detected using UHR OCT. The IS/OS junction and COST reflectivity disruption and presence of HRZ and ONL thinning are signs of cone photoreceptor degeneration. The latter 2 are age-dependent, which suggests that achromatopsia is a progressive disorder. In addition, foveal maldevelopment is described; this represents a fetal developmental defect linked to cone photoreceptor degeneration.

Retinal Structure of Birds of Prey Revealed by Ultra-High Resolution Spectral-Domain Optical Coherence Tomography

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

Retinal tumor imaging and volume quantification in mouse model using spectral-domain optical coherence tomography

We have successfully imaged the retinal tumor in a mouse model using an ultra-high resolution spectral-domain optical coherence tomography (SD-OCT) designed for small animal retinal imaging. For segmentation of the tumor boundaries and calculation of the tumor volume, we developed a novel segmentation algorithm. The algorithm is based on parametric deformable models (active contours) and is driven by machine learning-based region classification, namely a Conditional Random Field. With this algorithm we are able to obtain the tumor boundaries automatically, while the user can specify additional constraints (points on the boundary) to correct the segmentation result, if needed. The system and algorithm were successfully applied to studies on retinal tumor progression and monitoring treatment effects quantitatively in a mouse model of retinoblastoma.

Ultrahigh-resolution versus speckle noise-reduction in spectral-domain optical coherence tomography

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.