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OBJECTIVE To compare image quality and acquisition time of corneal and retinal spectral domain optical coherence tomography (SD-OCT) under 3 different sedation-anesthesia conditions in horses. ANIMALS 6 middle-aged geldings free of ocular disease. PROCEDURES 1 randomly selected eye of each horse was evaluated via SD-OCT under the following 3 conditions: standing sedation without retrobulbar anesthetic block (RB), standing sedation with RB, and general anesthesia with RB. Five regions of interest were evaluated in the cornea (axial and 12, 3, 6, and 9 o’clock positions) and fundus (optic nerve head). Three diagnostic scans of predetermined quality were obtained per anatomical region. Image acquisition times and total scans per site were recorded. Corneal and retinal SD-OCT image quality was graded on a subjective scale from 0 (nondiagnostic) to 4 (excellent). RESULTS Mean values for the standing sedation without RB, standing sedation with RB, and general anesthesia conditions were 24, 23, and 17, respectively, for total cornea scan attempts; 23, 19, and 19 for total retina-scan attempts; 14.6, 13.2, and 9.2 minutes for total cornea scan time; 19.1, 9.2, and 13.0 for total retina scan time; 2.0, 2.3, and 2.5 for cornea grade; and 2.7, 2.9, and 2.5 for retina grade. CONCLUSIONS AND CLINICAL RELEVANCE The RB facilitated globe akinesia and improved the percentage of scans in frame and region of interest accuracy for retinal imaging via OCT in horses. Retrobulbar blocks improved clinical image acquisition while minimizing motion artifact. more...

Background: In ophthalmology, thickness and vessel density (VD) measurements for the 6 x 6 mm inner macular retinal area have received increasing attention in glaucomatous progression research. For this area, the Angiovue optical coherence tomography system introduced a 304 x 304 A/B scans function (classic Angio Retina scan) in 2014, and a 400 x 400 A/B scans function (high-definition [HD] Angio Retina scan) in 2017. These scan types cannot be used in combination for the software provided for progression analysis.Purpose: Since losing data for 3 years may negatively influence progression analysis, we investigated whether clinically significant differences exist between consecutive measurements acquired with these scan types on the same eyes.Methods: As a part of our noninterventional prospective glaucoma imaging study, primary-open-angle glaucoma patients (POAG group), and ocular hypertensive and healthy control participants (structurally undamaged group) were imagedusing both the classic and the HD Angio Retina scans, respectively, without changing the patients’ position. High-quality images were obtained on 12 POAG eyes of 12 consecutive POAG patients, and 10 healthy and ocular hypertensive eyes of 10 consecutive participants before the data collection had to be suspended due to the new coronavirus epidemic.Results: For Early Treatment Diabetic Retinopathy Study image area, the mean difference (classic minus HD value) was 0.02 ± 0.37 μm for inner retinal thickness (P = 0.869) and 0.33 ± 1.33 % (P = 0.452) for superficial capillary VD in the structurally normal group (between-methods difference: ≤ 0.8% of the respective normal value). In the POAG group, the corresponding figures were -0.07 ± 1.22 μm for inner retinal thickness (P = 0.854; between-methods difference: 0.6% of the normal value) and 1.12 ± 2.58 % for superficial capillary VD (P = 0.158; classic scan value minus HD scan value: 1.12 ± 2.58 %; 2.3% of the normal value).Conclusion: Our results suggest that combined use of thickness and VD values for structurally normal eyes and thickness values for POAG eyes derived from classic and HD scans, respectively, for progression analysis can be reasonable since the differences between the corresponding values are small. However, combining the corresponding VD parameters for POAG eyes is useful only when the follow-up time before the scan type change is long enough to counterbalance the effect of the change on the result. more...

Purpose: To evaluate the feasibility and safety of a coaxial dual-wavelength optical coherence tomography (OCT) device (marked as Hydra-OCT). Methods: Healthy participants without ocular pathology underwent retinal imaging using the Hydra-OCT allowing for simultaneous measurement of retinal scanning of 840 and 1,072 nm wavelength. Before and after measurement, best-corrected visual acuity and patients’ comfort were assessed. Representative OCT images from both wavelengths were compared by 5 independent graders using a subjective grading scheme. Results: A total of 30 eyes of 30 participants (8 females and 22 males) with a mean age of 26.5 years (range from 19 to 55 years) were included. Dual-wavelength image acquisition was made possible in each subject. The participant’s effort and comfort assessment using the Hydra-OCT imaging revealed an equivalent value as compared to the commercially available OCT machine. No adverse events were reported, and visual acuity was not altered by the Hydra-OCT. Imaging between the systems was comparable. Conclusions: This study provides evidence for the feasibility and safety of a coaxial dual-wavelength OCT imaging method under real-life conditions. The novel Hydra-OCT imaging device may offer additional insights into the pathology of retinal and choroidal diseases. more...

The accurate segmentation of retinal layers of the eye in a 3D optical coherence tomography (OCT) data provides relevant clinical information. This chapter introduces a three-dimensional (3D) segmentation approach that uses an adaptive patient-specific retinal atlas as well as an appearance model for 3D OCT data. To reconstruct that atlas of 3D retinal scan, we first segment the central area of the macula at which we can clearly identify the fovea. Markov Gibbs random field (MGRF) including intensity, shape, and spatial information of 12 layers of retina was all used to segment the selected area of retinal fovea. A set of coregistered OCT scans that were gathered from 200 different individuals were used to build a 2D shape prior. This shape prior was adapted in a following step to the first-order appearance and second-order spatial interaction MGRF model. After segmenting the center of the macula “foveal area,” the labels and appearances of the layers that have been segmented were used to have the adjacent slices segmented as well. The last step was then repeated recursively until the 3D OCT scan of the patient is segmented. This approach was tested on 30 individuals while some of them were normal and others were pathological, and then compared to a manually segmented ground truth and finally these results were verified by medical retina experts. Metrics such as dice similarity coefficient, agreement coefficient, and average deviation metrics were used to measure the performance of the proposed approach. Accomplished accuracy by the proposed approach shows promising results with noticeable advantages over the state-of-the-art 3D OCT approach. more...

The guest editorial provides an introduction to the Special Section on Advanced Retinal Imaging: Instrumentation, Methods, and Applications.

It has been recently demonstrated that structures corresponding to the cell bodies of highly transparent cells in the retinal ganglion cell layer could be visualized noninvasively in the living human eye by optical coherence tomography (OCT) via temporal averaging. Inspired by this development, we explored the application of volumetric temporal averaging in mice, which are important models for studying human retinal diseases and therapeutic interventions. A general framework of temporal speckle-averaging (TSA) of OCT and optical coherence tomography angiography (OCTA) is presented and applied to mouse retinal volumetric data. Based on the image analysis, the eyes of mice under anesthesia exhibit only minor motions, corresponding to lateral displacements of a few micrometers and rotations of a fraction of 1 deg. Moreover, due to reduced eye movements under anesthesia, there is a negligible amount of motion artifacts within the volumes that need to be corrected to achieve volume coregistration. In addition, the relatively good optical quality of the mouse ocular media allows for cellular-resolution imaging without adaptive optics (AO), greatly simplifying the experimental system, making the proposed framework feasible for large studies. The TSA OCT and TSA OCTA results provide rich information about new structures previously not visualized in living mice with non-AO-OCT. The mechanism of TSA relies on improving signal-to-noise ratio as well as efficient suppression of speckle contrast due to temporal decorrelation of the speckle patterns, enabling full utilization of the high volumetric resolution offered by OCT and OCTA. more...

Polarised near-infra-red light is reflected from the foveal area in a detectable bow-tie pattern of polarisation states, offering the opportunity for eye tracking. A coaxial optical transducer was developed, consisting of a laser diode, a polariser, a filter, and a photodetector. Several such transducers may be used to interrogate different spots on the retina, thus eliminating the requirement for scanning systems with moving parts. To test the signal quality obtainable, using just one transducer, a test subject was asked to fixate successively on twelve targets located on a circle around the transducer, to simulate the retina’s being interrogated by twelve sensors placed on a 30 diameter circle surrounding the projection of the fovea. The resulting signal is close to the “ideal” sine wave that would have been recorded from a propeller-type birefringence pattern from a human fovea. The transducer can be used in the detection of fixation for medical and other purposes. It does not require calibrati... more...

Early diagnosis of glaucoma, which is a leading cause for visual impairment, is critical for successful treatment. It has been shown that Imaging polarimetry has advantages in early detection of structural changes in the retina. Here, we theoretically and experimentally present a snapshot Mueller Matrix Polarimeter fundus camera, which has the potential to record the polarization-altering characteristics of retina with a single snapshot. It is made by incorporating polarization gratings into a fundus camera design. Complete Mueller Matrix data sets can be obtained by analyzing the polarization fringes projected onto the image plane. In this paper, we describe the experimental implementation of the snapshot retinal imaging Mueller matrix polarimeter (SRIMMP), highlight issues related to calibration, and provide preliminary images acquired from the camera. more...

Purpose To study a novel and fast optical coherence tomography (OCT) device for home-based monitoring in age-related macular degeneration (AMD) in a small sample yielding sparse OCT (spOCT) data and to compare the device to a commercially available reference device. Methods In this prospective study, both eyes of 31 participants with AMD were included. The subjects underwent scanning with an OCT prototype and a spectral-domain OCT to compare the accuracy of the central retinal thickness (CRT) measurements. Results Sixty-two eyes in 31 participants (21 females and 10 males) were included. The mean age was 79.6 years (age range, 69-92 years). The mean difference in the CRT measurements between the devices was 4.52 μm (SD ± 20.0 μm; range, -65.6 to 41.5 μm). The inter- and intrarater reliability coefficients of the OCT prototype were both >0.95. The laser power delivered was more...

Diabetic Retinopathy (DR) is a medical condition caused by fluctuating insulin level in the blood which causes vision loss in case of severity. Timely treatment of such risks requires identification of the first clinical symptoms like microaneurysms (MAs) and hemorrhages (HMAs). The presence of those symptoms are visible in the digital color photographs of the retina and appear as round dark red spots in the image. In this paper, two approaches in the detection of MAs and HMAs are proposed. First, the semi automated approach applies semi automated hessian-based candidate selection algorithm (SHCS) followed by thresholding to detect true MAs and HMAs. The automated approach applies automated hessian-based candidate selection algorithm (AHCS) followed by feature extraction and SVM classification that uses twenty images for training manually annotated by medical domain experts. Implementations of both the approaches have been tested on real world images from retinal scan. From the results, the detection rate of automated algorithm when compared with that of the semi automated algorithm has been found to be significantly lesser with a probability p more...

Optical coherence tomography (OCT) has become standard in retinal imaging at the pre-clinical and clinical level by allowing non-invasive, three-dimensional imaging of the tissue structure. However, OCT lacks specificity to contrast agents that could be used for in vivo molecular imaging. We have performed in vivo photothermal optical coherence tomography (PT-OCT) of targeted gold nanorods in the mouse retina after the mice were injected systemically with the contrast agent. To our knowledge, we are the first to perform PT-OCT in the eye and image targeted gold nanorods with this technology. As a model of age-related wet macular degeneration, lesions were induced by laser photocoagulation in each mouse retina (n=12 eyes). Untargeted and targeted (anti-mouse CD102 antibody, labeling neovasculature) gold nanorods (peak absorption λ=750nm) were injected intravenously by tail-vein injection five days after lesion induction, and imaged the same day with PT-OCT. Our instrument is a spectral domain OCT system (λ=860nm) with a Titanium:Sapphire laser (λ=750nm) added to the beam path using a 50:50 coupler to heat the gold nanorods. We acquired PT-OCT volumes of one lesion per mouse eye. There was a significant increase in photothermal intensity per unit area of the lesion in the targeted gold nanorods group versus the saline control group and the untargeted gold nanorods group. This experiment demonstrates the feasibility of PT-OCT to image the distribution of molecular contrast agents in the mouse retina, including in highly scattering lesions. In the future we will use this method to identify new biomarkers linked with retinal disease. more...

Adaptive optics (AO) is essential for achieving diffraction limited resolution in large numerical aperture (NA) in-vivo retinal imaging in small animals. Cellular-resolution in-vivo imaging of fluorescently labeled cells is highly desirable for studying pathophysiology in animal models of retina diseases in pre-clinical vision research. Currently, wavefront sensor-based (WFS-based) AO is widely used for retinal imaging and has demonstrated great success. However, the performance can be limited by several factors including common path errors, wavefront reconstruction errors and an ill-defined reference plane on the retina. Wavefront sensorless (WFS-less) AO has the advantage of avoiding these issues at the cost of algorithmic execution time. We have investigated WFS-less AO on a fluorescence scanning laser ophthalmoscopy (fSLO) system that was originally designed for WFS-based AO. The WFS-based AO uses a Shack-Hartmann WFS and a continuous surface deformable mirror in a closed-loop control system to measure and correct for aberrations induced by the mouse eye. The WFS-less AO performs an open-loop modal optimization with an image quality metric. After WFS-less AO aberration correction, the WFS was used as a control of the closed-loop WFS-less AO operation. We can easily switch between WFS-based and WFS-less control of the deformable mirror multiple times within an imaging session for the same mouse. This allows for a direct comparison between these two types of AO correction for fSLO. Our results demonstrate volumetric AO-fSLO imaging of mouse retinal cells labeled with GFP. Most significantly, we have analyzed and compared the aberration correction results for WFS-based and WFS-less AO imaging. more...

Adaptive optics has been successfully applied to cellular resolution imaging of the retina, enabling visualization of the characteristic mosaic patterns of the outer retina. Wavefront sensorless adaptive optics (WSAO) is a novel technique that facilitates high resolution ophthalmic imaging; it replaces the Hartmann-Shack Wavefront Sensor with an image-driven optimization algorithm and mitigates some the challenges encountered with sensor-based designs. However, WSAO generally requires longer time to perform aberrations correction than the conventional closed-loop adaptive optics. When used for in vivo retinal imaging applications, motion artifacts during the WSAO optimization process will affect the quality of the aberration correction. A faster converging optimization scheme needs to be developed to account for rapid temporal variation of the wavefront and continuously apply corrections. In this project, we investigate the Databased Online Nonlinear Extremum-seeker (DONE), a novel non-linear multivariate optimization algorithm in combination with in vivo human WSAO OCT imaging. We also report both hardware and software updates of our compact lens based WSAO 1060nm swept source OCT human retinal imaging system, including real time retinal layer segmentation and tracking (ILM and RPE), hysteresis correction for the multi-actuator adaptive lens, precise synchronization control for the 200kHz laser source, and a zoom lens unit for rapid switching of the field of view. Cross sectional images of the retinal layers and en face images of the cone photoreceptor mosaic acquired in vivo from research volunteers before and after WSAO optimization are presented. more...

A dual illumination system is proposed for cornea and retina imaging using spectral domain optical coherence tomography (SD-OCT). The system is designed to acquire cornea and retina imaging with dual illumination with limited optics and using a single spectrometer. The beam propagation for cornea and retina imaging in dual illumination enables to acquire the images of different segments. This approach will reduce the imaging time for separate corneal and retinal imaging. The in vivo imaging of both the cornea and retina of a health volunteer shows the feasibility of the system for clinical applications more...

Optical coherence tomography (OCT) retinal imaging contributes to understanding central nervous system (CNS) diseases because the eye is an anatomical “window to the brain” with direct optical access to nonmylenated retinal ganglion cells. However, many CNS diseases are associated with neuronal changes beyond the resolution of standard OCT retinal imaging systems. Though studies have shown the utility of scattering angle resolved (SAR) OCT for particle sizing and detecting disease states ex vivo, a compact SAR-OCT system for in vivo rodent retinal imaging has not previously been reported. We report a fiber-based SAR-OCT system (swept source at 1310 nm ± 65 nm, 100 kHz scan rate) for mouse retinal imaging with a partial glass window (center aperture) for angular discrimination of backscattered light. This design incorporates a dual-axis MEMS mirror conjugate to the ocular pupil plane and a high collection efficiency objective. A muring retina is imaged during euthanasia, and the proposed SAR-index is examined versus time. Results show a positive correlation between the SAR-index and the sub-cellular hypoxic response of neurons to isoflurane overdose during euthanasia. The proposed SAR-OCT design and image process technique offer a contrast mechanism able to detect sub-resolution neuronal changes for murine retinal imaging. more...

High resolution is important for sensitive detection of subtle distortions of retinal morphology at an early stage of eye diseases. We demonstrate virtually structured detection (VSD) as a feasible method to achieve in vivo super-resolution ophthalmoscopy. A line-scanning strategy was employed to achieve a super-resolution imaging speed up to 127 ?? frames / s with a frame size of 512 × 512 ?? pixels . The proof-of-concept experiment was performed on anesthetized frogs. VSD-based super-resolution images reveal individual photoreceptors and nerve fiber bundles unambiguously. Both image contrast and signal-to-noise ratio are significantly improved due to the VSD implementation. more...

Hand-held wide-field contact color fundus photography is currently the standard method to acquire diagnostic images of children during examination under anesthesia and in the neonatal intensive care unit. The recent development of portable non-contact hand-held OCT retinal imaging systems has proved that OCT is of tremendous help to complement fundus photography in the management of pediatric patients. Currently, there is no commercial or research system that combines color wide-field digital fundus and OCT imaging in a contact-fashion. The contact of the probe with the cornea has the advantages of reducing motion experienced by the photographer during the imaging and providing fundus and OCT images with wider field of view that includes the periphery of the retina. In this study we produce proof of concept for a contact-type hand-held unit for simultaneous color fundus and OCT live view of the retina of pediatric patients. The front piece of the hand-held unit consists of a contact ophthalmoscopy lens integrating a circular light guide that was recovered from a digital fundus camera for pediatric imaging. The custom-made rear piece consists of the optics to: 1) fold the visible aerial image of the fundus generated by the ophthalmoscopy lens on a miniaturized level board digital color camera; 2) conjugate the eye pupil to the galvanometric scanning mirrors of an OCT delivery system. Wide-field color fundus and OCT images were simultaneously obtained in an eye model and sequentially obtained on the eye of a conscious 25 year-old human subject with healthy retina. more...

We demonstrate a novel optical coherence tomography system specifically developed and validated for clinical imaging of retinoblastoma tumors in pediatric patients. The existing treatment options for this malignant tumor of the retina aim at reduction of tumor (re)growth risks, and vision preservation. The choice of optimal treatment strongly depends on skilled and detailed clinical assessment. Due to the limitations of the existing real-time diagnostic tools the patients at risk are periodically monitored with retinal imaging to confirm the absence of new tumor seedings. Three-dimensional visualization of tissue layer and microvasculature at improved axial and lateral resolution of interference-based OCT imaging provides sensitivity for detection of vital tumor tissue concurrent with local treatment. Our METC-approved system accommodates for the range of optical parameters of infants’ eyes, and uses the 1050nm wavelength to access the deeper choroid layers of retina. The prototype is designed for patients in supine position under general anesthesia, where ergonomic handheld module is connected to fiber-based optical setup via umbilical cord. The system conforms to clinical safety requirements, including fully isolated low-voltage electric circuit. Focusing is performed with a mechanically tunable lens, where resolution is 6 µm axially, and varies with focusing at 10-18µm laterally. We will present optical design, performance limitations, and results of the ongoing clinical study, including the increased OCT diagnostic sensitivity in three dimensions in comparison with the established clinical imaging modalities. We will discuss images of early, active, and treated tumors, as well as follow-up on patients after local and systemic treatments. more...

OCT has become a standard in retina imaging at the pre-clinical and clinical level by allowing non-invasive, three-dimensional imaging of the tissue structure. However, OCT lacks specificity to contrast agents that could be used for in vivo molecular imaging. We have performed in vivo photothermal optical coherence tomography (PTOCT) of gold nanorods in the mouse retina after the mice were injected intravenously with the contrast agent. To our knowledge, we are the first team to perform PTOCT in the eye. Four lesions were induced by laser photocoagulation in each mouse retina (n=6 mice) and gold nanorods (untargeted and targeted with anti-mouse CD102 antibody, which labels neovasculature, peak absorption λ=750nm) were injected intravenously by tail-vein injection five days later in four mice (two mice are controls). The mice were imaged with PTOCT the same day. Our instrument is a spectral domain OCT system (λ=860nm) with a Titanium:Sapphire laser (λ=750nm) added to the beam path using a 50:50 splitter to target the gold nanorods. We acquired PTOCT B-scans over one lesion per mouse eye. There was a significant increase in photothermal intensity at the center of the lesion in the gold nanorod group versus the control group. This experiment demonstrates the feasibility of PTOCT to image the distribution of contrast agents in the mouse retina. In the future we will use this method to optimize drug delivery to the retina in pre-clinical models. more...

Objective To perform in vivo analysis of retinal functional and structural parameters in healthy mouse eyes. Animal Studied Adult C57BL/6 male mice (n = 37). Procedures Retinal function was evaluated using pattern electroretinography (pERG) and the chromatic pupil light reflex (cPLR). Structural properties of the retina and nerve fiber layer (NFL) were evaluated using spectral-domain optical coherence tomography (SD-OCT). Results The average pERG amplitudes were found to be 11.2 ± 0.7 lV (P50-N95, mean ± SEM), with an implicit time for P50-N95 interval of 90.4 ± 5.4 ms. Total retinal thickness was 229.5 ± 1.7 lm (mean ± SEM) in the area centralis region. The thickness of the retinal nerve fiber layer (mean ± SEM) using a circular peripapillary retinal scan centered on the optic nerve was 46.7 ± 0.9 lm (temporal), 46.1 ± 0.9 lm (superior), 45.8 ± 0.9 lm (nasal), and 48.4 ± 1 lm (inferior). The baseline pupil diameter was 2.1 ± 0.05 mm in darkness, and 1.1 ± 0.05 and 0.56 ± 0.03 mm after stimulation with red (630 nm, luminance 200 kcd/m 2 ) or blue (480 nm, luminance 200 kcd/m 2 ) light illumination, respectively. more...

Rod-dominated transient retinal phototropism (TRP) has been observed in freshly isolated retinas, promising a noninvasive biomarker for objective assessment of retinal physiology. However, in vivo mapping of TRP is challenging due to its subcellular signal magnitude and fast time course. We report here a virtually structured detection-based super-resolution ophthalmoscope to achieve subcellular spatial resolution and millisecond temporal resolution for in vivo imaging of TRP. Spatiotemporal properties of in vivo TRP were characterized corresponding to variable light intensity stimuli, confirming that TRP is tightly correlated with early stages of phototransduction. more...

Due to the advancement of microoptoelectromechanical systems and microelectromechanical systems (MEMS) technologies, novel display architectures have emerged. One of the most successful and well-known examples is the Digital Micromirror Device from Texas Instruments, a 2-D array of bistable MEMS mirrors, which function as spatial light modulators for the projection display. This concept of employing an array of modulators is also seen in the grating light valve and the interferometric modulator display, where the modulation mechanism is based on optical diffraction and interference, respectively. Along with this trend comes the laser scanning display, which requires a single scanning device with a large scan angle and a high scan frequency. A special example in this category is the retinal scanning display, which is a head-up wearable module that laser-scans the image directly onto the retina. MEMS technologies are also found in other display-related research, such as stereoscopic (3-D) displays and plastic thin-film displays. more...

Optical coherence tomography (OCT) is a noninvasive imaging modality that has begun to find widespread use in retinal imaging for the detection of a variety of ocular diseases. In addition to structural changes in the form of altered retinal layer thicknesses, pathological conditions may also cause the formation of edema within the retina. In multiple sclerosis, for instance, the nerve fiber and ganglion cell layers are known to thin. Additionally, the formation of pseudocysts called microcystic macular edema (MME) have also been observed in the eyes of about 5% of MS patients, and its presence has been shown to be correlated with disease severity. Previously, we proposed separate algorithms for the segmentation of retinal layers and MME, but since MME mainly occurs within specific regions of the retina, a simultaneous approach is advantageous. In this work, we propose an automated globally optimal graph-theoretic approach that simultaneously segments the retinal layers and the MME in volumetric OCT scans. SD-OCT scans from one eye of 12 MS patients with known MME and 8 healthy controls were acquired and the pseudocysts manually traced. The overall precision and recall of the pseudocyst detection was found to be 86.0% and 79.5%, respectively. more...

Scanning the retinae of the human eyes with a laser beam is an approved diagnosis method in ophthalmology; moreover the retinal blood vessels form a biometric modality for identifying persons. Medical applied Scanning Laser Ophthalmoscopes (SLOs) usually contain galvanometric mirror systems to move the laser spot with a defined speed across the retina. Hence, the load of laser radiation is uniformly distributed and eye safety requirements can be easily complied. Micro machined mirrors also known as Micro Electro Mechanical Systems (MEMS) are interesting alternatives for designing retina scanning systems. In particular double-resonant MEMS are well suited for mass fabrication at low cost. However, their Lissajous-shaped scanning figure requires a particular analysis and specific measures to meet the requirements for a Class 1 laser device, i.e. eye-safe operation. The scanning laser spot causes a non-uniform pulsing radiation load hitting the retinal elements within the field of view (FoV). The relevant laser safety standards define a smallest considerable element for eye-related impacts to be a point source that is visible with an angle of maximum 1.5 mrad. For non-uniform pulsing expositions onto retinal elements the standard requires to consider all particular impacts, i.e. single pulses, pulse sequences in certain time intervals and cumulated laser radiation loads. As it may be expected, a Lissajous scanning figure causes the most critical radiation loads at its edges and borders. Depending on the applied power the laser has to be switched off here to avoid any retinal injury. more...

Adaptive optics (AO) is necessary to correct aberrations when imaging the mouse eye with high numerical aperture. In order to obtain cellular resolution, we have implemented wavefront sensorless adaptive optics for in vivo fluorescence imaging of mouse retina. Our approach includes a lens-based system and MEMS deformable mirror for aberration correction. The AO system was constructed with a reflectance channel for structural images and fluorescence channel for functional images. The structural imaging was used in real-time for navigation on the retina using landmarks such as blood vessels. We have also implemented a tunable liquid lens to select the retinal layer of interest at which to perform the optimization. At the desired location on the mouse retina, the optimization algorithm used the fluorescence image data to drive a modal hill-climbing algorithm using an intensity or sharpness image quality metric. The optimization requires ~30 seconds to complete a search up to the 20th Zernike mode. In this report, we have demonstrated the AO performance for high-resolution images of the capillaries in a fluorescence angiography. We have also made progress on an approach to AO with pupil segmentation as a possible sensorless technique suitable for small animal retinal imaging. Pupil segmentation AO was implemented on the same ophthalmic system and imaging performance was demonstrated on fluorescent beads with induced aberrations. more...

Observations of RPE disruption and autofluorescence (AF) photobleaching at light levels below the ANSI photochemical maximum permissible exposure (MPE) (Morgan et al., 2008) indicates a demand to modify future light safety standards to protect the retina from harm. To establish safe light exposures, we measured the visible light action spectrum for RPE disruption in an in vivo monkey model with fluorescence adaptive optics retinal imaging. Using this high resolution imaging modality can provide insight into the consequences of light on a cellular level and allow for longitudinal monitoring of retinal changes. The threshold retinal radiant exposures (RRE) for RPE disruption were determined for 4 wavelengths (460, 488, 544, and 594 nm). The anaesthetized macaque retina was exposed to a uniform 0.5° × 0.5° field of view (FOV). Imaging within a 2° × 2° FOV was performed before, immediately after and at 2 week intervals for 10 weeks. At each wavelength, multiple RREs were tested with 4 repetitions each to determine the threshold for RPE disruption. For qualitative analysis, RPE disruption is defined as any detectable change from the pre exposure condition in the cell mosaic in the exposed region relative to the corresponding mosaic in the immediately surrounding area. We have tested several metrics to evaluate the RPE images obtained before and after exposure. The measured action spectrum for photochemical RPE disruption has a shallower slope than the current ANSI photochemical MPE for the same conditions and suggests that longer wavelength light is more hazardous than other measurements would suggest. more...

It is well known that patient-specific ocular aberrations limit imaging resolution in the human retina. Previously, hardware adaptive optics (HAO) has been employed to measure and correct these aberrations to acquire high-resolution images of various retinal structures. While the resulting aberration-corrected images are of great clinical importance, clinical use of HAO has not been widespread due to the cost and complexity of these systems. We present a technique termed computational adaptive optics (CAO) for aberration correction in the living human retina without the use of hardware adaptive optics components. In CAO, complex interferometric data acquired using optical coherence tomography (OCT) is manipulated in post-processing to adjust the phase of the optical wavefront. In this way, the aberrated wavefront can be corrected. We summarize recent results in this technology for retinal imaging, including aberration-corrected imaging in multiple retinal layers and practical considerations such as phase stability and image optimization. more...

This paper presents the unsupervised retinal vessels segmentation method, SEVERE (SEgmenting VEssels in REtina images), which is based on the direction map of retina scan images assigning each pixel one out of twelve discrete directions. SEVERE works on the green channel of RGB retina scan images. It does not require any pre-processing phase and all the computations are done exclusively on the direction map. SEVERE has been checked on publicly available datasets producing qualitatively satisfactory results and outperforming other existing methods in terms of quantitative performance evaluation parameters, such as accuracy and sensitivity. (C) 2015 Elsevier B.V. All rights reserved. more...

Copyright © 2016 SPIE. Optical coherence tomography (OCT) is a noninvasive method for retinal imaging. In this work, we present an in vivo human retinal microvascular network measurement by an intensity-based Doppler variance (IBDV) based on sweptsource OCT. In addition, an automatic three-dimensional (3-D) segmentation method was used for segmenting intraretinal layers. The microvascular networks were divided into six layers by visualizing of each individual layer with enhanced imaging contrast. This method has potential for earlier diagnosis and precise monitoring in retinal vascular diseases. more...

We present a parallel optical coherence tomography(OCT) system using a complementary metaloxide-semiconductor(CMOS) smart array detector. This paper includes descriptions of the CMOS smart array detector, and the experimental results of scattering surface targets including a wet tissue on an Air Force target, and ex vivo gold fish retina where the boundary between the vitreous humor and the ganglion cell layer could be identified in the acquired images. After three-dimensional volume rendering, several other layers between the ganglion cell layer and the choroid layer could be observed even though the contrast between the layers was weak. Control of the current version of the CMOS smart array detector requires significant time for maintenance. Although the current CMOS smart array detector needs improvement, the design is a promising ophthalmologic instrument and can provide fast retinal scanning and prompt display of the acquired images. more...

Scanning laser ophthalmoscopy (SLO) and optical coherence tomography (OCT) provide complementary views of the retina, with the former collecting fluorescence data with good lateral but relatively low-axial resolution, and the latter collecting label-free backscattering data with comparable lateral but much higher axial resolution. To take maximal advantage of the information of both modalities in mouse retinal imaging, we have constructed a compact, four-channel, wide-field (∼50 deg) system that simultaneously acquires and automatically coregisters three channels of confocal SLO and Fourier domain OCT data. The scanner control system allows “zoomed” imaging of a region of interest identified in a wide-field image, providing efficient digital sampling and localization of cellular resolution features in longitudinal imaging of individual mice. The SLO is equipped with a “flip-in” spectrometer that enables spectral “fingerprinting” of fluorochromes. Segmentation of retina layers and en face display facilitate spatial comparison of OCT data with SLO fluorescence patterns. We demonstrate that the system can be used to image an individual retinal ganglion cell over many months, to simultaneously image microglia and Muller glia expressing different fluorochromes, to characterize the distinctive spatial distributions and clearance times of circulating fluorochromes with different molecular sizes, and to produce unequivocal images of the heretofore uncharacterized mouse choroidal vasculature. more...

High-resolution optical coherence tomography (OCT) retinal imaging is important to noninvasively visualize the various retinal structures to aid in better understanding of the pathogenesis of vision-robbing diseases. However, conventional OCT systems have a trade-off between lateral resolution and depth-of-focus. In this report, we present the development of a focus-stacking OCT system with automatic focus optimization for high-resolution, extended-focal-range clinical retinal imaging by incorporating a variable-focus liquid lens into the sample arm optics. Retinal layer tracking and selection was performed using a graphics processing unit accelerated processing platform for focus optimization, providing real-time layer-specific en face visualization. After optimization, multiple volumes focused at different depths were acquired, registered, and stitched together to yield a single, high-resolution focus-stacked dataset. Using this system, we show high-resolution images of the retina and optic nerve head, from which we extracted clinically relevant parameters such as the nerve fiber layer thickness and lamina cribrosa microarchitecture. more...

Animal models play an important role for understanding the pathophysiology of glaucoma and age-related macular degeneration. With these models, longitudinal studies can be performed and therefore there is need for non-invasive evaluation of disease progress. For that purpose optical coherence tomography (OCT) can be used. Since tissues with different polarization properties are important in these diseases, polarization sensitive OCT (PS-OCT) could be a valuable tool in preclinical research. In this work a high resolution PS-OCT (HR-PS-OCT) system was used in-vivo for rodent retinal imaging. A super luminescent diode with a bandwidth of 100 nm was used as a light source that yielded an axial resolution of 5.1 μm in air (3.8 μm in tissue). The A-scan rate was 83 kHz, a whole 3D dataset was acquired in a few seconds (1536x1024x200 pixels in 3.5 s) which reduced motion artifacts. Rats (Sprague-Dawley, Long-Evans and Brown Norway) as well as mice (C57BL/6) were imaged. High resolution reflectivity images showed all retinal layers in all animals. From acquired data also phase retardation, fast axis orientation and degree of polarization uniformity (DOPU) images were calculated. On phase retardation images sclera was identified as birefringent and retinal pigment epithelium (RPE) and choroid as depolarizing tissues. Our results demonstrate the suitability of the system for high speed/resolution imaging in follow up studies on rodents. more...

Rodent models are indispensable in studying various retinal diseases. Noninvasive, high-resolution retinal imaging of rodent models is highly desired for longitudinally investigating the pathogenesis and therapeutic strategies. However, due to severe aberrations, the retinal image quality in rodents can be much worse than that in humans. We numerically and experimentally investigated the influence of chromatic aberration and optical illumination bandwidth on retinal imaging. We confirmed that the rat retinal image quality decreased with increasing illumination bandwidth. We achieved the retinal image resolution of 10 μm using a 19 nm illumination bandwidth centered at 580 nm in a home-built fundus camera. Furthermore, we observed higher chromatic aberration in albino rat eyes than in pigmented rat eyes. This study provides a design guide for high-resolution fundus camera for rodents. Our method is also beneficial to dispersion compensation in multiwavelength retinal imaging applications. more...

Intrinsic optical signal (IOS) imaging promises to be a noninvasive method for high-resolution examination of retinal physiology, which can advance the study and diagnosis of eye diseases. While specialized optical instruments are desirable for functional IOS imaging of retinal physiology, in depth understanding of multiple IOS sources in the complex retinal neural network is essential for optimizing instrument designs. We provide a brief overview of IOS studies and relationships in rod outer segment suspensions, isolated retinas, and intact eyes. Recent developments of line-scan confocal and functional optical coherence tomography (OCT) instruments have allowed in vivo IOS mapping of photoreceptor physiology. Further improvements of the line-scan confocal and functional OCT systems may provide a feasible solution to pursue functional IOS mapping of human photoreceptors. Some interesting IOSs have already been detected in inner retinal layers, but better development of the IOS instruments and software algorithms is required to achieve optimal physiological assessment of inner retinal neurons. more...

A compact, non-invasive multi-modal system has been developed for in vivo mouse retina imaging. It is configured for simultaneously detecting green and red fluorescent protein signals with scanning laser ophthalmoscopy (SLO) back-scattered light from the SLO illumination beam, and depth information about different retinal layers by means of Optical Coherence Tomography (OCT). Simultaneous assessment of retinal characteristics with different modalities can provide a wealth of information about the structural and functional changes in the retinal neural tissue and chorio-retinal vasculature in vivo . Additionally, simultaneous acquisition of multiple channels facilitates analysis of the data of different modalities by automatic temporal and structural co-registration. As an example of the instrument’s performance we imaged the retina of a mouse with constitutive expression of GFP in microglia cells (Cx3 cr1 GFP/+ ), and which also expressed the red fluorescent protein mCherry in Muller glial cells by means of adeno-associated virus delivery (AAV2) of an mCherry cDNA driven by the GFAP (glial fibrillary acid protein) promoter. more...

Many applications could benefit from miniaturized systems to scan blood vessels behind the retina in the human eye, so called „retina scanning“. This reaches from access control to sophisticated security applications and medical devices. High volume systems for consumer applications require low cost and a user friendly operation. For example this includes no need for removal of glasses and self-adjustment, in turn guidance of focus and point of attraction by simultaneous projection for the user. A new system has been designed based on the well-known resonantly driven 2-d scanner mirror of Fraunhofer IPMS. A combined NIR and VIS laser system illuminates the eye through an eye piece designed for an operating distance allowing the use of glasses and granting sufficient field of view. This usability feature was considered to be more important than highest miniaturization. The modulated VIS laser facilitates the projection of an image directly onto the retina. The backscattered light from the continuous NIR laser contains the information of the blood vessels and is detected by a highly sensitive photo diode. A demonstrational setup has been realized including readout and driving electronics. The laser power was adjusted to an eye-secure level. Additional security features were integrated. Test measurements revealed promising results. In a first demonstration application the detection of biometric pattern of the blood vessels was evaluated for issues authentication in. more...

© 2015 SPIE. We describe the design and performance of a recently implemented retinal imaging system for the human eye that combines adaptive optics (AO) with spectral domain optical coherence tomography (OCT) and scanning laser ophthalmoscopy (SLO). The AO-OCT-SLO system simultaneously acquires SLO frames and OCT B-scans at 60 Hz with an OCT volume acquisition scan rate of 0.24 Hz. The SLO images are used to correct for eye motion during the registration of OCT B-scans. Key optical design considerations are discussed including: minimizing system aberrations through the use of off-axis relay telescopes; choice of telescope magnification based on pupil plane requirements and restrictions; and the use of dichroic beam splitters to separate and re-combine OCT and SLO beams around the nonshared horizontal scanning mirrors. We include an analysis of closed-loop AO correction on a model eye and compare these findings with system performance in vivo. The 2D and 3D OCT scans included in this work demonstrate the ability of this system to laterally and axially resolve individual cone photoreceptors, while the corresponding SLO images show the en face mosaics at the photoreceptor layer showing rods and cones. Images from both healthy and diseased retina are presented. more...

We present a multiscale sensorless adaptive optics (SAO) OCT system capable of imaging retinal structure and vasculature with various fields-of-view (FOV) and resolutions. Using a single deformable mirror and exploiting the polarization properties of light, the SAO-OCT-A was implemented in a compact and easy to operate system. With the ability to adjust the beam diameter at the pupil, retinal imaging was demonstrated at two different numerical apertures with the same system. The general morphological structure and retinal vasculature could be observed with a few tens of micrometer-scale lateral resolution with conventional OCT and OCT-A scanning protocols with a 1.7-mm-diameter beam incident at the pupil and a large FOV (15 deg× 15 deg). Changing the system to a higher numerical aperture with a 5.0-mm-diameter beam incident at the pupil and the SAO aberration correction, the FOV was reduced to 3 deg× 3 deg for fine detailed imaging of morphological structure and microvasculature such as the photoreceptor mosaic and capillaries. Multiscale functional SAO-OCT imaging was performed on four healthy subjects, demonstrating its functionality and potential for clinical utility. more...

With the miniaturization of scanning mirrors and the emergence of wearable health monitoring, an intriguing step is to investigate the potential of a laser scanning ophthalmoscope (LSO) for retinal imaging with wearable glasses. In addition to providing morphological information of the retina, such as vasculature, LSO images could also be used to provide information on general health conditions. A compact eyeglass with LSO capability would give access, on demand, to retinal parameters without disturbing the subject's activity. One of the main challenges in this field is the creation of a device that does not interrupt the user's field of view. We report, to our knowledge, the first see-through ophthalmoscope. The system is analyzed with three-dimensional simulations and tested in a proof-of-concept setup with the same key parameters of a wearable device. Finally, image quality is analyzed by acquiring images of an ex-vivo human eye sample. (C) The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI. more...

We present a 3-path optical system for studying the retinal movement of jumping spiders: a visible OLED virtual reality system presents stimulus, while NIR illumination and imaging systems observe retinal movement. more...

A compact, high-speed line scanning quasi-confocal ophthalmoscope (LSO) for retina imaging is presented in this paper. By using a line beam to illuminate the retina, meanwhile a linear array sensor is used for imaging the retina, the LSO system significantly reduces the size, complexity, and cost comparing to a conventional confocal scanning laser ophthalmoscope (CSLO). With only one moving scanner to provide raster scanning of the line beam of the retina, the imaging frequency achieves 160 Hz and the lateral resolution is nearly 10 μm for 1024×330 pixels imaging mode. Preliminary experiments are performed for imaging the macula, the optic nerve head and other targets, providing high resolution and high speed videos of human retina. more...

There is an increasing need for reliable authentication for a number of applications such as e commerce. Common authentication methods based on ownership (ID card) or knowledge factors (password, PIN) are often prone to manipulations and may therefore be not safe enough. Various inherence factor based methods like fingerprint, retinal pattern or voice identifications are considered more secure. Retina scanning in particular offers both low false rejection rate (FRR) and low false acceptance rate (FAR) with about one in a million. Images of the retina with its characteristic pattern of blood vessels can be made with either a fundus camera or laser scanning methods. The present work describes the optical design of a new compact retina laser scanner which is based on MEMS (Micro Electric Mechanical System) technology. The use of a dual axis micro scanning mirror for laser beam deflection enables a more compact and robust design compared to classical systems. The scanner exhibits a full field of view of 10° which corresponds to an area of 4 mm2 on the retinal surface surrounding the optical disc. The system works in the near infrared and is designed for use under ambient light conditions, which implies a pupil diameter of 1.5 mm. Furthermore it features a long eye relief of 30 mm so that it can be conveniently used by persons wearing glasses. The optical design requirements and the optical performance are discussed in terms of spot diagrams and ray fan plots. more...

In regenerative medicine, the zebrafish is a prominent animal model for studying degeneration and regeneration processes, e.g. of photoreceptor cells in the retina. By means of optical coherence tomography (OCT), these studies can be conducted over weeks using the same individual and hence reducing the variability of the results. To allow an improvement of zebrafish retinal OCT imaging by suitable optics, we developed a zebrafish eye model using geometrical data obtained by in vivo dispersion encoded full range OCT as well as a dispersion comprising gradient index (GRIN) lens model based on refractive index data found in the literature. Using non-sequential ray tracing, the focal length of the spherical GRIN lens (diameter of 0.96 mm) was determined to be 1.22 mm at 800 nm wavelength giving a Matheissen’s ratio (ratio of focal length to radius of the lens) of 2.54, which fits well into the range between 2.19 and 2.82, found for various fish lenses. Additionally, a mean refractive index of 1.64 at 800 nm could be retrieved for the lens to yield the same focal position as found for the GRIN condition. With the aid of the zebrafish eye model, the optics of the OCT scanner head were adjusted to provide high-resolution retinal images with a field of view of 30° x 30°. The introduced model therefore provides the basis for improved retinal imaging with OCT and can be further used to study the image formation within the zebrafish eye. more...

When using scanning laser ophthalmoscopy to produce images of the eye fundus, maximum permissible exposure (MPE) limits must be considered. These limits are set out in international standards such as the National Standards Institute ANSI Z136.1 Safe Use of Lasers (USA) and BS EN 60825-1: 1994 (UK) and corresponding Euro norms but these documents do not explicitly consider the case of scanned beams. Our study aims to show how MPE values can be calculated for the specific case of retinal scanning by taking into account an array of parameters, such as wavelength, exposure duration, type of scanning, line rate and field size, and how each set of initial parameters results in MPE values that correspond to thermal or photochemical damage to the retina. more...

The Virtual Retinal Display (VRD) is a visual display that scans modulated laser light on to the retina of the viewer's eye to create an image. Maximum permissible exposures (MPE) have been calculated for the VRD in both normal viewing and possible failure modes. The MPE power levels are compared to the measured power that enters the eye while viewing images with the VRD. The power levels indicate that the VRD is safe in both normal operating mode and in failure modes. more...

The microvasculature network of retina plays an important role in the study and diagnosis of retinal diseases (age-related macular degeneration and diabetic retinopathy for example). Although it is possible to noninvasively acquire high-resolution retinal images with modern retinal imaging technologies, non-uniform illumination, the low contrast of thin vessels and the background noises all make it difficult for diagnosis. In this paper, we introduce a novel retinal vessel extraction algorithm based on gradient vector flow and matched filtering to segment retinal vessels with different likelihood. Firstly, we use isotropic Gaussian kernel and adaptive histogram equalization to smooth and enhance the retinal images respectively. Secondly, a multi-scale matched filtering method is adopted to extract the retinal vessels. Then, the gradient vector flow algorithm is introduced to locate the edge of the retinal vessels. Finally, we combine the results of matched filtering method and gradient vector flow algorithm to extract the vessels at different likelihood levels. The experiments demonstrate that our algorithm is efficient and the intensities of vessel images exactly represent the likelihood of the vessels. more...

Retinal imaging with optical coherence tomography (OCT) has rapidly advanced in ophthalmic applications with the broad availability of Fourier domain (FD) technology in commercial systems. The high sensitivity afforded by FD-OCT has enabled imaging of the choroid, a layer of blood vessels serving the outer retina. Improved visualization of the choroid and the choroid-sclera boundary has been investigated using techniques such as enhanced depth imaging (EDI), and also with OCT systems operating in the 1060-nm wavelength range. We report on a comparison of imaging the macular choroid with commercial and prototype OCT systems, and present automated 3D segmentation of the choroid-scleral layer using a graph cut algorithm. The thickness of the choroid is an important measurement to investigate for possible correlation with severity, or possibly early diagnosis, of diseases such as age-related macular degeneration. more...

Small animal models of retinal diseases are important to vision research, and noninvasive high resolution in vivo rodent retinal imaging is becoming an increasingly important tool used in this field. We present a custom Fourier domain optical coherence tomography (FD-OCT) instrument for high resolution imaging of mouse retina. In order to overcome aberrations in the mouse eye, we incorporated a commercial adaptive optics system into the sample arm of the refractive FD-OCT system. Additionally, a commercially available refraction canceling lens was used to reduce lower order aberrations and specular back-reflection from the cornea. Performance of the adaptive optics (AO) system for correcting residual wavefront aberration in the mice eyes is presented. Results of AO FD-OCT images of mouse retina acquired in vivo with and without AO correction are shown as well. more...