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PURPOSE: Use projection-resolved OCT angiography to investigate the autoregulatory response in the 3 parafoveal retinal plexuses under hyperoxia. DESIGN: Prospective cohort study. PARTICIPANTS: Nine eyes from 9 healthy participants. METHODS: One eye from each participant was scanned using a commercial spectral-domain OCT system. Two repeated macular scans (3 × 3 mm2) were acquired at baseline and during oxygen breathing. The split-spectrum amplitude-decorrelation algorithm was used to detect blood flow. The projection-resolved algorithm was used to suppress projection artifacts and resolve blood flow in 3 distinct parafoveal plexuses. The Wilcoxon signed-rank test was used to compare baseline and hyperoxic parameters. The coefficient of variation, intraclass correlation coefficient, and pooled standard deviation were used to assess the reliability of OCT angiography measurements. MAIN OUTCOME MEASURES: Flow index and vessel density were calculated from the en face angiograms of each of the 3 plexuses, as well as from the all-plexus inner retinal slab. RESULTS: Hyperoxia induced significant reduction in the flow index (-11%) and vessel density (-7.8%) of only the deep capillary plexus (P < 0.001) and in the flow index of the all-plexus slab (P = 0.015). The flow index also decreased in the intermediate capillary plexus and the superficial vascular complex, but these changes were small and not statistically significant. The projection-resolved OCT angiography showed good within-session baseline repeatability (coefficient of variation, 0.8%-5.2%; intraclass correlation coefficient, 0.93-0.98) in all parameters. Relatively large between-day response reproducibility was observed (pooled standard deviation, 1.7%-9.4%). CONCLUSIONS: Projection-resolved OCT angiography was able to show that the retinal autoregulatory response to hyperoxia affects only the deep capillary plexus, but not the intermediate capillary plexus or superficial vascular complex.

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AIM: To assess total retinal blood flow (TRBF) in diabetic retinopathy (DR) using multiplane en face Doppler optical coherence tomography (OCT). METHODS: A 70 kHz spectral-domain OCT system scanned a 2×2 mm area centred at the optic disc of the eyes with DR and healthy participants. The multiplane en face Doppler OCT algorithm generated a three-dimensional volumetric data set consisting of 195 en face planes. The TRBF was calculated from the maximum flow values of each branching retinal vein at an optimised en face plane. DR severity was graded according to the international clinical classification system. The generalised linear model method was used to compare flow values between DR groups and the control group. RESULTS: A total of 71 eyes from 71 participants were included. Ten eyes were excluded due to poor image quality. The within-visit repeatability of scans was 4.1% (coefficient of variation). There was no significant difference in the TRBF between the healthy (46.7±10.2 µL/min) and mild/moderate non-proliferative DR (44.9±12.6 µL/min) groups. The TRBF in severe non-proliferative DR (39.1±12.6 µL/min) and proliferative DR (28.9±8.85 µL/min) groups were significantly lower (p=0.04 and p<0.0001, respectively) than that of the healthy group. TRBF was correlated with DR disease severity (p<0.0001, linear trend test). CONCLUSION: The novel multiplane en face Doppler OCT method provided reliable measurements of TRBF in DR eyes. This may be a useful tool in understanding the pathophysiology of DR.

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BACKGROUND: Motion artifacts degrade the quality of optical coherence tomography angiography (OCTA). Orthogonal registration can eliminate the majority of these artifacts, but some artifacts persist in most clinical images. We evaluate an automated registration algorithm with selective merging and filtering to remove remaining artifacts and improve the quality of images. METHODS: A 70 kHz commercial spectral domain OCT was used to obtain 3 mm × 3 mm OCTA in 10 healthy, 5 age-related macular degeneration (AMD), and 31 diabetic retinopathy (DR) participants. Projection artifacts were removed and images were segmented into 3 inner retinal plexuses. Amplitude thresholding identified lines containing a residual artifact and correlation between neighboring lines identified distorted stripes. Then the angiograms were registered and the lines selectively merged. A vesselness filter was applied to the resulting images. The images were evaluated for signal-to-noise ratio (SNR), image entropy, vessel connectivity and vessel density. RESULTS: Registration and selective merging (RSM) algorithm improved the SNR (P<0.02) compared to orthogonal registration alone. RSM with vesselness filter increased the image entropy (P<10-8) and reduced inter-subject variability (standard error ≤3%, n=10) in healthy eyes. The method improved vessel details and connectivity in OCTA of healthy, DR and neovascular AMD eyes. CONCLUSIONS: This automated registration method eliminates residual motion artifacts and enhances the visualization of vessels in OCTA.

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A microfluidic chip with microchannels ranging from 8 to 96 µm was used to mimic blood vessels down to the capillary level. Blood flow within the microfluidic channels was analyzed with split-spectrum amplitude-decorrelation angiography (SSADA)-based optical coherence tomography (OCT) angiography. It was found that the SSADA decorrelation value was related to both blood flow speed and channel width. SSADA could differentiate nonflowing blood inside the microfluidic channels from static paper. The SSADA decorrelation value was approximately linear with blood flow velocity up to a threshold Vsat of 5.83±1.33 mm/s (mean±standard deviation over the range of channel widths). Beyond this threshold, it approached a saturation value Dsat. Dsat was higher for wider channels, and approached a maximum value Dsm as the channel width became much larger than the beam focal spot diameter. These results indicate that decorrelation values (flow signal) in capillary networks would be proportional to both flow velocity and vessel caliber but would be capped at a saturation value in larger blood vessels. These findings are useful for interpretation and quantification of clinical OCT angiography results.

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A phase gradient angiography (PGA) method is proposed for optical coherence tomography (OCT). This method allows the use of phase information to map the microvasculature in tissue without the correction of bulk motion and laser trigger jitter induced phase artifacts. PGA can also be combined with the amplitude/intensity to improve the performance. Split-spectrum technique can further increase the signal to noise ratio by more than two times. In-vivo imaging of human retinal circulation is shown with a 70 kHz, 840 nm spectral domain OCT system and a 200 kHz, 1050 nm swept source OCT system. Four different OCT angiography methods are compared. The best performance was achieved with split-spectrum amplitude and phase-gradient angiography.

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We propose a novel automated volumetric segmentation method to detect and quantify retinal fluid on optical coherence tomography (OCT). The fuzzy level set method was introduced for identifying the boundaries of fluid filled regions on B-scans (x and y-axes) and C-scans (z-axis). The boundaries identified from three types of scans were combined to generate a comprehensive volumetric segmentation of retinal fluid. Then, artefactual fluid regions were removed using morphological characteristics and by identifying vascular shadowing with OCT angiography obtained from the same scan. The accuracy of retinal fluid detection and quantification was evaluated on 10 eyes with diabetic macular edema. Automated segmentation had good agreement with manual segmentation qualitatively and quantitatively. The fluid map can be integrated with OCT angiogram for intuitive clinical evaluation.

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We propose an innovative registration method to correct motion artifacts for wide-field optical coherence tomography angiography (OCTA) acquired by ultrahigh-speed swept-source OCT (>200 kHz A-scan rate). Considering that the number of A-scans along the fast axis is much higher than the number of positions along slow axis in the wide-field OCTA scan, a non-orthogonal scheme is introduced. Two en face angiograms in the vertical priority (2 y-fast) are divided into microsaccade-free parallel strips. A gross registration based on large vessels and a fine registration based on small vessels are sequentially applied to register parallel strips into a composite image. This technique is extended to automatically montage individual registered, motion-free angiograms into an ultrawide-field view.

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PURPOSE: To use multiplane en face Doppler optical coherence tomography (OCT) to measure the change in total retinal blood flow (TRBF) in response to hyperoxia. METHODS: One eye of each healthy human participant (n = 8) was scanned with a commercial high-speed (70-kHz) spectral OCT system. Three repeated scans were captured at baseline and after 10 minutes of oxygen (hyperoxia) by open nasal mask. The procedure was performed twice on day 1 and once more on day 2. Blood flow of each vein was estimated using Doppler OCT at an optimized en face plane. The TRBF was summed from all veins at the optic disc. The TRBF hyperoxic response was calculated as the TRBF percent change from baseline. RESULTS: Participants experienced a 23.6% ± 10.7% (mean ± standard deviation [SD]) decrease (P < 0.001, paired t-test) in TRBF during hyperoxia. The within-day repeatability of baseline TRBF was 4.1% and the between-day reproducibility was 10.9% coefficient of variation (CV). Between-grader reproducibility was 3.9% CV. The repeatability and reproducibility (pooled SD) of hyperoxic response were 6.1% and 6.4%, respectively. CONCLUSIONS: The multiplane en face Doppler OCT algorithm was able to detect, in all participants, a decreased TRBF in response to hyperoxia. The response magnitude for each participant varied among repeated trials, and the averaging of multiple trials was helpful in establishing the individual response. This technique shows good potential for the clinical investigation of vascular autoregulation.

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This article provides an overview of advanced image processing for three dimensional (3D) optical coherence tomographic (OCT) angiography of macular diseases, including age-related macular degeneration (AMD) and diabetic retinopathy (DR). A fast automated retinal layers segmentation algorithm using directional graph search was introduced to separates 3D flow data into different layers in the presence of pathologies. Intelligent manual correction methods are also systematically addressed which can be done rapidly on a single frame and then automatically propagated to full 3D volume with accuracy better than 1 pixel. Methods to visualize and analyze the abnormalities including retinal and choroidal neovascularization, retinal ischemia, and macular edema were presented to facilitate the clinical use of OCT angiography.

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A custom-built dynamic-focus swept-source optical coherence tomography (SS-OCT) system with a central wavelength of 1310 nm was used to image the anterior eye from the cornea to the lens. An electrically tunable lens was utilized to dynamically control the positions of focusing planes over the imaging range of 10 mm. The B-scan images were acquired consecutively at the same position but with different focus settings. The B-scan images were then registered and averaged after filtering the out-of-focus regions using a Gaussian window. By fusing images obtained at different depth focus locations, high-resolution and high signal-strength images were obtained over the entire imaging depth. In vivo imaging of human anterior segment was demonstrated. The performance of the system was compared with two commercial OCT systems. The human eye ciliary body was better visualized with the dynamic-focusing SS-OCT system than using the commercial 840 and 1310 nm OCT systems. The sulcus-to-sulcus distance was measured, and the result agreed with that acquired with ultrasound biomicroscopy

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Retinal blood supply is tightly regulated under a variety of hemodynamic considerations in order to satisfy a high metabolic need and maintain both vessel structure and function. Simulation of the human eye can induce hemodynamics alterations, and attempt to assess the vascular reactivity response has been well documented in the scientific literature. Advancements in noninvasive imaging technologies have led to the characterization of magnitude and time course in retinal blood flow response to stimuli. This allowed for a better understanding of the mechanism in which blood flow is regulated, as well as identifying functional impairments in the diseased eye. Clinically, the ability to detect retinal blood flow reactivity during stimulation of the eye offers potential for the detection, differentiation, and diagnosis of diseases.

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IMPORTANCE: Vascular factors may have important roles in the pathophysiology of glaucoma. A practical method for the clinical evaluation of ocular perfusion is needed to improve glaucoma management. OBJECTIVE: To detect peripapillary retinal perfusion in glaucomatous eyes compared with normal eyes using optical coherence tomography (OCT) angiography. DESIGN, SETTING, AND PARTICIPANTS: Prospective observational study performed from July 24, 2013, to April 17, 2014. Participants were recruited and tested at Casey Eye Institute, Oregon Health & Science University. In total, 12 glaucomatous eyes and 12 age-matched normal eyes were analyzed. The optic disc region was imaged twice using a 3 × 3-mm scan by a 70-kHz, 840-nm-wavelength spectral OCT system. The split-spectrum amplitude-decorrelation angiography algorithm was used. Peripapillary flow index was calculated as the mean decorrelation value in the peripapillary region, defined as a 700-µm-wide elliptical annulus around the disc. Peripapillary vessel density was the percentage area occupied by vessels. The data statistical analysis was performed from October 30, 2013, to May 30, 2014. MAIN OUTCOMES AND MEASURES: Variability was assessed by the coefficient of variation. The Mann-Whitney test was used to compare the 2 groups of eyes. Correlations between vascular and visual field variables were assessed by linear regression analysis. RESULTS: In 12 normal eyes, a dense microvascular network around the disc was visible on OCT angiography. In 12 glaucomatous eyes, this network was visibly attenuated globally and focally. In normal eyes, between-visit reproducibilities of peripapillary flow index and peripapillary vessel density were 4.3% and 2.7% of the coefficient of variation, respectively, while the population variabilities of peripapillary flow index and peripapillary vessel density were 8.2% and 3.0% of the coefficient of variation, respectively. Peripapillary flow index and peripapillary vessel density in glaucomatous eyes were lower than those in normal eyes (P < .001 for both). Peripapillary flow index (Pearson r = -0.808) and peripapillary vessel density (Pearson r = -0.835) were highly correlated with visual field pattern standard deviation in glaucomatous eyes (P = .001 for both). The areas under the receiver operating characteristic curve for normal vs glaucomatous eyes were 0.892 for peripapillary flow index and 0.938 for peripapillary vessel density. CONCLUSIONS AND RELEVANCE: Using OCT angiography, reduced peripapillary retinal perfusion in glaucomatous eyes can be visualized as focal defects and quantified as peripapillary flow index and peripapillary vessel density, with high repeatability and reproducibility. Quantitative OCT angiography may have value in future studies to determine its potential usefulness in glaucoma evaluation.

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An automated algorithm was developed for total retinal blood flow (TRBF) using 70-kHz spectral optical coherence tomography (OCT). The OCT was calibrated for the transformation from Doppler shift to speed based on a flow phantom. The TRBF scan pattern contained five repeated volume scans (2 x 2 mm) obtained in 3 s and centered on central retinal vessels in the optic disc. The TRBF was calculated using an en face Doppler technique. For each retinal vein, blood flow was measured at an optimal plane where the calculated flow was maximized. The TRBF was calculated by summing flow in all veins. The algorithm tracked vascular branching so that either root or branch veins are summed, but never both. The TRBF in five repeated volumes were averaged to reduce variation due to cardiac cycle pulsation. Finally, the TRBF was corrected for eye length variation. Twelve healthy eyes and 12 glaucomatous eyes were enrolled to test the algorithm. The TRBF was 45.4 ± 6.7 µl/min for healthy control and 34.7 ± 7.6 µl/min for glaucomatous participants (p-value = 0.01). The intravisit repeatability was 8.6% for healthy controls and 8.4% for glaucoma participants. The proposed automated method provided repeatable TRBF measurement.

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PURPOSE: To measure the change in peripapillary retinal blood flow in response to hyperoxia by using optical coherence tomography (OCT) angiography. METHODS: One eye of each healthy human participants (six) was scanned with a commercial high-speed (70 kHz) spectral OCT. Scans were captured twice after 10-minute exposures to normal breathing (baseline) and hyperoxia. Blood flow was detected by the split-spectrum amplitude-decorrelation angiography (SSADA) algorithm. Peripapillary retinal blood flow index and vessel density were calculated from en face maximum projections of the retinal layers. The experiment was performed on 2 separate days for each participant. Coefficient of variation (CV) was used to measure within-day repeatability and between-day reproducibility. Paired t-tests were used to compare means of baseline and hyperoxic peripapillary retinal blood flow. RESULTS: A decrease of 8.87% ± 3.09% (mean ± standard deviation) in flow index and 2.61% ± 1.50% in vessel density was observed under hyperoxia. The within-day repeatability CV of baseline measurements was 5.75% for flow index and 1.67% for vessel density. The between-day reproducibility CV for baseline flow index and vessel density was 11.1% and 1.14%, respectively. The between-day reproducibility of the hyperoxic response was 3.71% and 1.67% for flow index and vessel density, respectively. CONCLUSIONS: Optical coherence tomography angiography with SSADA was able to detect a decrease in peripapillary retinal blood flow in response to hyperoxia. The response was larger than the variability of baseline measurements. The magnitude of an individual's hyperoxic response was highly variable between days. Thus, reliable assessment may require averaging multiple measurements.

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We propose methods to align interferograms affected by trigger jitter to a reference interferogram based on the information (amplitude/phase) at a fixed-pattern noise location to reduce residual fixed-pattern noise and improve the phase stability of swept source optical coherence tomography (SS-OCT) systems. One proposed method achieved this by introducing a wavenumber shift (k-shift) in the interferograms of interest and searching for the k-shift that minimized the fixed-pattern noise amplitude. The other method calculated the relative k-shift using the phase information at the residual fixed-pattern noise location. Repeating this wavenumber alignment procedure for all A-lines of interest produced fixed-pattern noise free and phase stable OCT images. A system incorporating these correction routines was used for human retina OCT and Doppler OCT imaging. The results from the two methods were compared, and it was found that the intensity-based method provided better results.

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We demonstrate the proof of concept of a novel Fourier-domain optical coherence tomography contrast mechanism using gold nanorod contrast agents and a spectral fractionation processing technique. The methodology detects the spectral shift of the backscattered light from the nanorods by comparing the ratio between the short and long wavelength halves of the optical coherence tomography signal intensity. Spectral fractionation further divides the halves into sub-bands to improve spectral contrast and suppress speckle noise. Herein, we show that this technique can detect gold nanorods in intralipid tissue phantoms. Furthermore, cellular labeling by gold nanorods was demonstrated using retinal pigment epithelial cells in vitro.

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