Adaptive Optics Optical Coherence Tomography Research Articles

Key Developments for Partial Coherence Biometry and Optical Coherence Tomography in the Human Eye Made in Vienna.

To describe key developments of optical biometry and optical coherence tomography (OCT) for ophthalmic applications made by one of the pioneering research groups. Partial coherence interferometry (PCI) as the basic ranging technology for modern optical biometry and for OCT was introduced for biomedical applications in the 1980s. Later, Fourier domain (FD) OCT was introduced and demonstrated to provide superior sensitivity as compared to time domain OCT. Further developments comprised ultrahigh-resolution OCT and deep-penetration OCT at wavelengths of approximately 1050 nm. Important functional extensions comprise Doppler OCT/OCT angiography, polarization-sensitive OCT, and adaptive optics OCT. High-precision PCI biometry has found extensive applications in cataract surgery and in research on intraocular lens design. Optical coherence tomography, especially in the second-generation variant of FD OCT, is now indispensable for ocular diagnostics in general and for retinal diagnostics in particular; 1050 nm OCT shows improved penetration into deeper layers like the choroid. The contributions of the Vienna research group helped to establish PCI biometry and FD OCT as the gold standards in their respective fields.

Modal content of living human cone photoreceptors.

Decades of experimental and theoretical investigations have established that photoreceptors capture light based on the principles of optical waveguiding. Yet considerable uncertainty remains, even for the most basic prediction as to whether photoreceptors support more than a single waveguide mode. To test for modal behavior in human cone photoreceptors in the near infrared, we took advantage of adaptive-optics optical coherence tomography (AO-OCT, λc = 785 nm) to noninvasively image in three dimensions the reflectance profile of cones. Modal content of reflections generated at the cone inner segment and outer segment junction (IS/OS) and cone outer segment tip (COST) was examined over a range of cone diameters in 1,802 cones from 0.6° to 10° retinal eccentricity. Second moment analysis in conjunction with theoretical predictions indicate cone IS and OS have optical properties consistent of waveguides, which depend on segment diameter and refractive index. Cone IS was found to support a single mode near the fovea (≤3°) and multiple modes further away (>4°). In contrast, no evidence of multiple modes was found in the cone OSs. The IS/OS and COST reflections share a common optical aperture, are most circular near the fovea, show no orientation preference, and are temporally stable. We tested mode predictions of a conventional step-index fiber model and found that in order to fit our AO-OCT results required a lower estimate of the IS refractive index and introduction of an IS focusing/tapering effect.

Adaptive optics optical coherence tomography at 1 MHz.

Image acquisition speed of optical coherence tomography (OCT) remains a fundamental barrier that limits its scientific and clinical utility. Here we demonstrate a novel multi-camera adaptive optics (AO-)OCT system for ophthalmologic use that operates at 1 million A-lines/s at a wavelength of 790 nm with 5.3 μm axial resolution in retinal tissue. Central to the spectral-domain design is a novel detection channel based on four high-speed spectrometers that receive light sequentially from a 1 × 4 optical switch assembly. Absence of moving parts enables ultra-fast (50ns) and precise switching with low insertion loss (-0.18 dB per channel). This manner of control makes use of all available light in the detection channel and avoids camera dead-time, both critical for imaging at high speeds. Additional benefit in signal-to-noise accrues from the larger numerical aperture afforded by the use of AO and yields retinal images of comparable dynamic range to that of clinical OCT. We validated system performance by a series of experiments that included imaging in both model and human eyes. We demonstrated the performance of our MHz AO-OCT system to capture detailed images of individual retinal nerve fiber bundles and cone photoreceptors. This is the fastest ophthalmic OCT system we know of in the 700 to 915 nm spectral band.

The cellular origins of the outer retinal bands in optical coherence tomography images.

To test the recently proposed hypothesis that the second outer retinal band, observed in clinical OCT images, originates from the inner segment ellipsoid, by measuring: (1) the thickness of this band within single cone photoreceptors, and (2) its respective distance from the putative external limiting membrane (band 1) and cone outer segment tips (band 3). Adaptive optics-optical coherence tomography images were acquired from four subjects without known retinal disease. Images were obtained at foveal (2°) and perifoveal (5°) locations. Cone photoreceptors (n = 9593) were identified and segmented in three dimensions using custom software. Features corresponding to bands 1, 2, and 3 were automatically identified. The thickness of band 2 was assessed in each cell by fitting the longitudinal reflectance profile of the band with a Gaussian function. Distances between bands 1 and 2, and between 2 and 3, respectively, were also measured in each cell. Two independent calibration techniques were employed to determine the depth scale (physical length per pixel) of the imaging system. When resolved within single cells, the thickness of band 2 is a factor of three to four times narrower than in corresponding clinical OCT images. The distribution of band 2 thickness across subjects and eccentricities had a modal value of 4.7 μm, with 48% of the cones falling between 4.1 and 5.2 μm. No significant differences were found between cells in the fovea and perifovea. The distance separating bands 1 and 2 was found to be larger than the distance between bands 2 and 3, across subjects and eccentricities, with a significantly larger difference at 5° than 2°. On the basis of these findings, we suggest that ascription of the outer retinal band 2 to the inner segment ellipsoid is unjustified, because the ellipsoid is both too thick and proximally located to produce the band.

Adaptive-optics optical coherence tomography processing using a graphics processing unit.

Graphics processing units are increasingly being used for scientific computing for their powerful parallel processing abilities, and moderate price compared to super computers and computing grids. In this paper we have used a general purpose graphics processing unit to process adaptive-optics optical coherence tomography (AOOCT) images in real time. Increasing the processing speed of AOOCT is an essential step in moving the super high resolution technology closer to clinical viability.

Adaptive optics optical coherence tomography with dynamic retinal tracking.

Adaptive optics optical coherence tomography (AO-OCT) is a highly sensitive and noninvasive method for three dimensional imaging of the microscopic retina. Like all in vivo retinal imaging techniques, however, it suffers the effects of involuntary eye movements that occur even under normal fixation. In this study we investigated dynamic retinal tracking to measure and correct eye motion at KHz rates for AO-OCT imaging. A customized retina tracking module was integrated into the sample arm of the 2nd-generation Indiana AO-OCT system and images were acquired on three subjects. Analyses were developed based on temporal amplitude and spatial power spectra in conjunction with strip-wise registration to independently measure AO-OCT tracking performance. After optimization of the tracker parameters, the system was found to correct eye movements up to 100 Hz and reduce residual motion to 10 µm root mean square. Between session precision was 33 µm. Performance was limited by tracker-generated noise at high temporal frequencies.

Improved visualization of outer retinal morphology with aberration cancelling reflective optical design for adaptive optics - optical coherence tomography

We present an aberration cancelling optical design for a reflective adaptive optics - optical coherence tomography (AO-OCT) retinal imaging system. The optical performance of this instrument is compared to our previous multimodal AO-OCT/AO-SLO retinal imaging system. The feasibility of new instrumentation for improved visualization of microscopic retinal structures is discussed. Examples of images acquired with this new AO-OCT instrument are presented.

Optical coherence tomography imaging in uveitis

Optical coherence tomography (OCT) is a non-contact noninvasive technique that allows in vivo imaging of the retina, choroid, optic nerve head, retinal nerve fiber layer, and the anterior structures of the eye. It was introduced into clinical practice two decades ago. Advances in OCT technology have been achieved by searching ultra-high-resolution OCT, adaptive optics OCT, eye-tracking OCT, and changes in signal detection technique from time-domain (TD) to spectral-domain (SD) detection. Today, SD OCT has become a part of routine uveitis practice. Apart from its diagnostic value in uveitis, OCT has enabled objective assessment of treatment response and provided predictive value for visual recovery and prognosis of uveitic entities. It is the standard diagnostic technique in the detection, monitoring of treatment, and determination of prognosis in uveitic macular edema as well as other inflammatory macular pathologies, including epiretinal membrane formation, vitreomacular traction, foveal atrophy, and lamellar/full-thickness macular holes. OCT has also shed light on the pathophysiology of several posterior uveitic entities. SD OCT has enabled visualization of four lines in the sensory retina which represent the external limiting membrane, the photoreceptor inner and outer segment junction, the photoreceptor outer segment and the retina pigment epithelium junction, and the retina pigment epithelium-choriocapillaris complex. Thus, we have gained substantial information about the pathologic and structural changes in uveitic conditions with primary or secondary outer retinal involvement. SD OCT has also provided invaluable information on the inner retinal and the vitreoretinal interface changes in uveitic conditions. With the introduction of enhanced depth imaging, visualization of the choroid and choriocapillaries has become possible. Therefore, OCT has become an indispensible ancillary test in the diagnosis and management of inflammatory diseases involving the retina and/or the choroid. As OCT technology continues to develop further it will provide new insights into the retinal and choroidal structure and the pathogenesis of posterior uveitic entities.

Multimodal Assessment of Microscopic Morphology and Retinal Function in Patients With Geographic Atrophy

To correlate retinal function and visual sensitivity with retinal morphology revealed by ultrahigh-resolution imaging with adaptive optics-optical coherence tomography (AO-OCT), on patients with geographic atrophy. Five eyes from five subjects were tested (four with geographic atrophy [66.3 ± 6.4 years, mean ± 1 SD] and one normal [61 years]). Photopic and scotopic multifocal electroretinograms (mfERGs) were recorded. Visual fields were assessed with microperimetry (mP) combined with a scanning laser ophthalmoscope for high-resolution confocal retinal fundus imaging. The eye tracker of the microperimeter identified the preferred retinal locus that was then used as a reference for precise targeting of areas for advanced retinal imaging. Images were obtained with purpose-built, in-house, ultrahigh resolution AO-OCT. Fundus autofluorescence (FAF) and color fundus (CF) photographs were also acquired. The AO-OCT imaging provided detailed cross-sectional structural representation of the retina. Up to 12 retinal layers were identified in the normal subject while many severe retinal abnormalities (i.e., calcified drusen, drusenoid pigment epithelium detachment, outer retinal tubulation) were identified in the retinae of the GA patients. The functional tests showed preservation of sensitivities, although somewhat compromised, at the border of the GA. The images provided here advance our knowledge of the morphology of retinal layers in GA patients. While there was a strong correlation between altered retinal structure and reduction in visual function, there were a number of examples in which the photoreceptor inner/outer segment (IS/OS) junctions lost reflectivity at the margins of GA, while visual function was still demonstrated. This was shown to be due to changes in photoreceptor orientation near the GA border.

Complex conjugate artifact-free adaptive optics optical coherence tomography of in vivo human optic nerve head

We acquired in vivo images of the human optic nerve head (ONH) using an adaptive optics-optical coherence tomography (AO-OCT) system. In order to improve imaging of the lamina cribrosa in the ONH with high lateral resolution and sensitivity, we implemented a complex conjugate artifact-free Fourier domain OCT (Fd-OCT) acquisition scheme with a reference arm-based phase shifting method. This allowed positioning of the lamina cribrosa structures near the zero path length difference where AO-OCT imaging achieves highest sensitivity. Implementation of our complex conjugate artifact removal (CCR) method required constant phase shifts between consecutive axial scans (A-scans), generated by continuous beam path-length changes from offsetting the pivot point of the scanning mirror placed in the reference arm. Fourier transform along the transverse axis and a filtering algorithm allowed reconstruction of CCR AO-OCT images. The suppression ratio of the mirror artifact was approximately 22 dB (at 18,000 A-scans per second acquisition speed) with a paperboard test target and an optimum phase-shift value. Finally, we reconstructed the three-dimensional structure of human ONH with enhanced depth range and sensitivity using CCR AO-OCT.

New developments in optical coherence tomography

AbstractPurpose Optical coherence tomography (OCT) has become a standard tool in imaging the retina, the optic nerve head and the anterior segment of the eye. In the recent years much effort was directed towards improving the acquisition speed and the resolution of the technique. In addition, functional extensions of the technique were presented.Methods Ultrahigh‐speed swept sources can be used to improve A‐scan rate in OCT, a technique also called optical frequency domain imaging. Increasing the bandwidth can improve longitudinal resolution in OCT and longer wavelengths in the near infrared improve penetration depth. Adaptive optics OCT improves transversal resolution. Functional extensions include measurement of blood flow and oxygenation.Results High‐speed imaging allows for three‐dimensional imaging of the posterior and anterior segment of the eye. Using adaptive optics volumetric cellular resolution imaging becomes possible. 1050 nm OCT allows for visualization of the sclera and therefore measurement of choroidal thickness. Functional OCT provides insight into retinal metabolism.Conclusion Improvements and extensions of OCT have been reported that will find its way into clinical routine. This is expected to result in improved understanding of retinal and optic nerve head disease.

Adaptive optics scanning ophthalmoscopy with annular pupils

Annular apodization of the illumination and/or imaging pupils of an adaptive optics scanning light ophthalmoscope (AOSLO) for improving transverse resolution was evaluated using three different normalized inner radii (0.26, 0.39 and 0.52). In vivo imaging of the human photoreceptor mosaic at 0.5 and 10° from fixation indicates that the use of an annular illumination pupil and a circular imaging pupil provides the most benefit of all configurations when using a one Airy disk diameter pinhole, in agreement with the paraxial confocal microscopy theory. Annular illumination pupils with 0.26 and 0.39 normalized inner radii performed best in terms of the narrowing of the autocorrelation central lobe (between 7 and 12%), and the increase in manual and automated photoreceptor counts (8 to 20% more cones and 11 to 29% more rods). It was observed that the use of annular pupils with large inner radii can result in multi-modal cone photoreceptor intensity profiles. The effect of the annular masks on the average photoreceptor intensity is consistent with the Stiles-Crawford effect (SCE). This indicates that combinations of images of the same photoreceptors with different apodization configurations and/or annular masks can be used to distinguish cones from rods, even when the former have complex multi-modal intensity profiles. In addition to narrowing the point spread function transversally, the use of annular apodizing masks also elongates it axially, a fact that can be used for extending the depth of focus of techniques such as adaptive optics optical coherence tomography (AOOCT). Finally, the positive results from this work suggest that annular pupil apodization could be used in refractive or catadioptric adaptive optics ophthalmoscopes to mitigate undesired back-reflections.

Performance of 97-elements ALPAO membrane magnetic deformable mirror in Adaptive Optics - Optical Coherence Tomography system for in vivo imaging of human retina

We present an evaluation of two different configurations of the ALPAO 97-actuator membrane magnetic deformable mirror for wavefront correction, using either 7 or 9 actuators across the eye pupil. These tests included monitoring of aberration correction for a human subject. This AO-sub system is part of the UC Davis high-resolution human retinal imaging adaptive optics - optical coherence tomography (AO-OCT) instrument. AO-OCT allows the three-dimensional (3D) visualization of different retinal structures in vivo with high volumetric resolution (3x3x3?m 3 ). Full Text: PDF

Outer retinal abnormalities associated with inner retinal pathology in nonglaucomatous and glaucomatous optic neuropathies

Inner and outer retinal morphology were quantified in vivo for 6 nonglaucomatous and 10 glaucomatous optic neuropathy patients. Custom, ultrahigh-resolution imaging modalities were used to evaluate segmented retinal layer thickness in 3D volumes (Fourier-domain optical coherence tomography), cone photoreceptor density (adaptive optics fundus camera), and the length of inner and outer segments of cone photoreceptors (adaptive optics-optical coherence tomography). Quantitative comparisons were made with age-matched controls, or by comparing affected and nonaffected retinal areas defined by changes in visual fields. The integrity of outer retinal layers on optical coherence tomography B-scans and density of cone photoreceptors were correlated with visual field sensitivity at corresponding retinal locations following reductions in inner retinal thickness. The photoreceptor outer segments were shorter and exhibited greater variability in retinal areas associated with visual field losses compared with normal or less affected areas of the same patient's visual field. These results demonstrate that nonglaucomatous and glaucomatous optic neuropathies are associated with outer retinal changes following long-term inner retinal pathology.

AO adapted to SD‐OCT

Abstract Purpose Optical coherence tomography (OCT) has emerged as a leading technique in ophthalmic imaging due to its capability to non‐invasively resolve tissue morphology with high sensitivity and high axial resolution. Despite increases in axial resolution, monochromatic ocular aberrations limited the transverse resolution for retinal imaging to ~20 μm, which is too large for visualization of cellular structures. Adaptive optics (AO) may be used to correct such aberrations, leading to an improvement in image contrast and lateral resolution. Methods A successful combination of ultra‐high speed (120,000 depth scans/s), ultra‐high resolution optical coherence tomography with adaptive optics and an achromatizing lens for compensation of monochromatic and longitudinal chromatic ocular aberrations, respectively, allows for non‐invasive volumetric imaging in normal and pathologic human retinas at cellular resolution. Results The capability of this imaging system is demonstrated through preliminary studies by probing cellular intraretinal structures that have not been accessible so far with in vivo, non‐invasive, label‐free imaging techniques, including pigment epithelial cells, micro‐vasculature of the choriocapillaris, single nerve fibre bundles and collagenous plates of the lamina cribrosa in the optic nerve head. In addition, the volumetric extent of cone loss in two colour‐blinds can be quantified for the first time. Conclusion AO OCT might provide opportunities to enhance the understanding of retinal pathogenesis and early diagnosis of retinal diseases. Commercial interest

Simultaneous high-resolution retinal imaging and high-penetration choroidal imaging by one-micrometer adaptive optics optical coherence tomography

Adaptive optics optical coherence tomography (AO-OCT) provides three-dimensional high-isotropic-resolution retinal images in vivo. We developed AO-OCT with a 1.03-mum probing beam and demonstrated high-penetration, high-resolution retinal imaging. Axial scans are acquired with a speed of 47,000 lines/s. AO closed loop is configured with a single deformable mirror. Seven eyes of 7 normal subjects were examined. Signal enhancement was found for all subjects. A rippled interface between nerve fiber layer and ganglion cell layer, boundary between ganglion cell layer and inner plexiform layer, and chorioscleral interface were identified. Simultaneous high-resolution and high-penetration choroidal imaging may be useful for microstructural investigation of photoreceptors and glaucomatous nerve-fiber abnormalities.

Impact of enhanced resolution, speed and penetration on three-dimensional retinal optical coherence tomography

: 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.

Volumetric retinal imaging with ultrahigh-resolution spectral-domain optical coherence tomography and adaptive optics using two broadband light sources

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.

Cellular resolution volumetric in vivo retinal imaging with adaptive optics–optical coherence tomography

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.

Ultrahigh-resolution optical coherence tomography with monochromatic and chromatic aberration correction.

We have developed an improved adaptive optics - optical coherence tomography (AO-OCT) system and evaluated its performance for in vivo imaging of normal and pathologic retina. The instrument provides unprecedented image quality at the retina with isotropic 3D resolution of 3.5 x 3.5 x 3.5 microm(3). Critical to the instrument's resolution is a customized achromatizing lens that corrects for the eye's longitudinal chromatic aberration and an ultra broadband light source (Delta lambda=112 nm lambda(0)= approximately 836 nm). The eye's transverse chromatic aberrations is modeled and predicted to be sufficiently small for the imaging conditions considered. The achromatizing lens was strategically placed at the light input of the AO-OCT sample arm. This location simplifies use of the achromatizing lens and allows straightforward implementation into existing OCT systems. Lateral resolution was achieved with an AO system that cascades two wavefront correctors, a large stroke bimorph deformable mirror (DM) and a micro-electromechanical system (MEMS) DM with a high number of actuators. This combination yielded diffraction-limited imaging in the eyes examined. An added benefit of the broadband light source is the reduction of speckle size in the axial dimension. Additionally, speckle contrast was reduced by averaging multiple B-scans of the same proximal patch of retina. The combination of improved micron-scale 3D resolution, and reduced speckle size and contrast were found to significantly improve visibility of microscopic structures in the retina.