Imaging of retinal vasculature using adaptive optics SLO/OCT.
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ABSTRACT:
SUBMITTER: Felberer F
PROVIDER: S-EPMC4399679 | biostudies-other | 2015 Apr
REPOSITORIES: biostudies-other
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Project description:We describe the details of a multimodal retinal imaging system which combines adaptive optics (AO) with an integrated scanning light ophthalmoscopy (SLO) and optical coherence tomography (OCT) imaging system. The OCT subsystem consisted of a swept-source, Fourier-domain mode-locked (FDML) laser, with a very high A-scan rate (1.6 MHz), whose beam was raster scanned on the retina by two scanners-one resonant scanner and one galvanometer. The high sweep rate of the FDML permitted the SLO and OCT to utilize the same scanners for in vivo retinal imaging and, unlike existing multimodal systems, concurrently acquired SLO frames and OCT volumes with approximate en face correspondence at a rate of 6 Hz. The AO provided diffraction-limited cellular resolution for both imaging channels.
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Project description:Adaptive optics (AO) is an insightful tool that has been increasingly applied to existing imaging systems for viewing the retina at a cellular level. By correcting for individual optical aberrations, AO offers an improvement in transverse resolution from 10-15 μm to ~2 μm, enabling assessment of individual retinal cell types. One of the settings in which its utility has been recognised is that of the inherited retinal diseases (IRDs), the genetic and clinical heterogeneity of which warrants better cellular characterisation. In this review, we provide a summary of the basic principles of AO, its integration into multiple retinal imaging modalities and its clinical applications, focusing primarily on IRDs. Furthermore, we present a comprehensive summary of AO-based cellular findings in IRDs according to their associated disease-causing genes.
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Project description:Adaptive optics scanning laser ophthalmoscopy (AO-SLO) has recently been used to achieve exquisite subcellular resolution imaging of the mouse retina. Wavefront sensing-based AO typically restricts the field of view to a few degrees of visual angle. As a consequence the relationship between AO-SLO data and larger scale retinal structures and cellular patterns can be difficult to assess. The retinal vasculature affords a large-scale 3D map on which cells and structures can be located during in vivo imaging. Phase-variance OCT (pv-OCT) can efficiently image the vasculature with near-infrared light in a label-free manner, allowing 3D vascular reconstruction with high precision. We combined widefield pv-OCT and SLO imaging with AO-SLO reflection and fluorescence imaging to localize two types of fluorescent cells within the retinal layers: GFP-expressing microglia, the resident macrophages of the retina, and GFP-expressing cone photoreceptor cells. We describe in detail a reflective afocal AO-SLO retinal imaging system designed for high resolution retinal imaging in mice. The optical performance of this instrument is compared to other state-of-the-art AO-based mouse retinal imaging systems. The spatial and temporal resolution of the new AO instrumentation was characterized with angiography of retinal capillaries, including blood-flow velocity analysis. Depth-resolved AO-SLO fluorescent images of microglia and cone photoreceptors are visualized in parallel with 469 nm and 663 nm reflectance images of the microvasculature and other structures. Additional applications of the new instrumentation are discussed.
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Project description:Multiphoton microscopy enables imaging deep into scattering tissues. The efficient generation of non-linear optical effects is related to both the pulse duration (typically on the order of femtoseconds) and the size of the focused spot. Aberrations introduced by refractive index inhomogeneity in the sample distort the wavefront and enlarge the focal spot, which reduces the multiphoton signal. Traditional approaches to adaptive optics wavefront correction are not effective in thick or multi-layered scattering media. In this report, we present sensorless adaptive optics (SAO) using low-coherence interferometric detection of the excitation light for depth-resolved aberration correction of two-photon excited fluorescence (TPEF) in biological tissue. We demonstrate coherence-gated SAO TPEF using a transmissive multi-actuator adaptive lens for in vivo imaging in a mouse retina. This configuration has significant potential for reducing the laser power required for adaptive optics multiphoton imaging, and for facilitating integration with existing systems.
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Project description:Adaptive optics (AO) is a technology used to improve the performance of optical systems by reducing the effects of optical aberrations. The direct visualization of the photoreceptor cells, capillaries and nerve fiber bundles represents the major benefit of adding AO to retinal imaging. Adaptive optics is opening a new frontier for clinical research in ophthalmology, providing new information on the early pathological changes of the retinal microstructures in various retinal diseases. We have reviewed AO technology for retinal imaging, providing information on the core components of an AO retinal camera. The most commonly used wavefront sensing and correcting elements are discussed. Furthermore, we discuss current applications of AO imaging to a population of healthy adults and to the most frequent causes of blindness, including diabetic retinopathy, age-related macular degeneration and glaucoma. We conclude our work with a discussion on future clinical prospects for AO retinal imaging.
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Project description:PurposeUltrahigh-resolution optical coherence tomography (UHR-OCT) with adaptive optics (AO) provides micrometer-scale 3D resolution that is attractive for imaging the retinal microvasculature. Such imaging may be useful for early detection of pathologic changes as in diabetic retinopathy. Here the authors investigate this potential for detecting individual capillaries in healthy subjects.MethodsUHR-AO-OCT volumes centered on the fovea were acquired from seven subjects (age range, 25-61 years) with three preselected with no foveal avascular zone (FAZ). Images were compared with entoptic diagrams using the capillaries at the rim of the FAZ. Methods of comparison were testing for the presence of a FAZ, noting distinct features in the capillary pattern, and measuring the size of the FAZ. Additional analysis included measurements of capillary diameter and depth range with retinal eccentricity.ResultsUHR-AO-OCT results are consistent with entoptic observations for all three methods of comparison. FAZ diameters measured by UHR-AO-OCT and entoptic imaging are strongly correlated (R(2) = 0.86). Average capillary diameter near the FAZ rim is 5.1 (4.6) ± 1.4 μm, with the value in parentheses accounting for axial image blur. This is consistent with histology (average, ~4.7 μm). Depth range of the capillaries increases monotonically with eccentricity (0°-1.25°) and is larger and more variable for subjects without FAZ.ConclusionsUHR-AO-OCT permits observation of many of the capillaries proximal to the FAZ, including those of average size based on published histology. This supports the view that the vast majority of capillaries in the retina are likely detectable with UHR-AO-OCT.
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Project description:Early detection of axonal tissue loss in retinal nerve fiber layer (RNFL) is critical for effective treatment and management of diseases such as glaucoma. This study aims to evaluate the capability of ultrahigh-resolution optical coherence tomography with adaptive optics (UHR-AO-OCT) for imaging the RNFL axonal bundles (RNFBs) with 3×3×3μm(3) resolution in the eye. We used a research-grade UHR-AO-OCT system to acquire 3°×3° volumes in four normal subjects and one subject with an arcuate retinal nerve fiber layer defect (n=5; 29-62years). Cross section (B-scans) and en face (C-scan) slices extracted from the volumes were used to assess visibility and size distribution of individual RNFBs. In one subject, we reimaged the same RNFBs twice over a 7month interval and compared bundle width and thickness between the two imaging sessions. Lastly we compared images of an arcuate RNFL defect acquired with UHR-AO-OCT and commercial OCT (Heidelberg Spectralis). Individual RNFBs were distinguishable in all subjects at 3° retinal eccentricity in both cross-sectional and en face views (width: 30-50μm, thickness: 10-15μm). At 6° retinal eccentricity, RNFBs were distinguishable in three of the five subjects in both views (width: 30-45μm, thickness: 20-40μm). Width and thickness RNFB measurements taken 7months apart were strongly correlated (p<0.0005). Mean difference and standard deviation of the differences between the two measurement sessions were -0.1±4.0μm (width) and 0.3±1.5μm (thickness). UHR-AO-OCT outperformed commercial OCT in terms of clarity of the microscopic retina. To our knowledge, these are the first measurements of RNFB cross section reported in the living human eye.
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Project description:PurposeTo describe the feasibility of peripheral OCT imaging in retinal diseases using a novel full-field device.MethodsA total of 134 consecutive eyes were referred and imaged on the Optos Silverstone swept-source OCT (SS-OCT) (Optos PLC; Dunfermline, UK). Scanning laser ophthalmoscope (SLO) images and the associated SS-OCT images were obtained in the posterior pole, mid-periphery or far periphery based on the nature of the referral and on new areas of interest observed in the optomap images at the time of imaging.ResultsA total of 134 eyes (96 patients) were enrolled in the study. One hundred and twenty-five eyes (91 patients) with 38 retinal pathologies were prospectively assessed and 9 eyes (5 patients) were excluded due to incomplete image acquisition. The average age of the subjects was 54 years (range 21-92 years). Thirty-nine out of 125 eyes (31%) had macular pathologies. Eighty-six out of 125 eyes (69%) had peripheral only pathologies, an area which cannot be visualized by standard OCT devices with a 50 degree field-of-view.ConclusionsThe ability to capture peripheral pathologies using an integrated SLO-UWF imaging with full-field swept-source provided high-grade anatomical insight that confirmed the medical and surgical management in a majority of cases. Its use in the mid- and far periphery provides a holistic clinical picture, which can potentially aid in the understanding of various retinal pathologies.
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