This study presents an exploratory approach to performing optical coherence tomographic angiography (OCTA) using motion contrast between repeated volumetric scans captured at 200 ms intervals, as opposed to conventional OCTA which measure motion contrast between repeat B-scans captured at shorter intervals. The proposed inter-volumetric OCTA (IV-OCTA) is made possible through an advanced image registration algorithm based on Mattes mutual information metric, and it is implemented on an ultra-high-resolution spectral domain 840 nm OCT system with a 250 KHz A-scan rate. IV-OCTA demonstrates a high detection sensitivity for microvessels with slow blood flow and reduces bulk motion artifacts using the embedded volume registration algorithm. Meanwhile, averaging repeated volumes can substantially reduce the speckle noise for clearer structural imaging compared to the averaged repeated B-scans.
KEYWORDS: Optical coherence tomography, Imaging systems, Cornea, Signal processing, Retina, Cameras, Signal detection, Real time imaging, In vivo imaging, Sensors
A line-field optical coherence tomography (LFOCT) application is implemented for real-time in vivo corneal and retinal imaging. In contrast to other described systems of LFOCT that use single-shot high-speed cameras, we describe the first results utilizing a camera with continuous high-speed data transfer and display. The system is based on a previously published design using a center wavelength of 840nm and a bandwidth of 50nm. The system’s B-frame and en-face display speed reaches up to 5000 frames per second corresponding to 2,500,000 A-lines. A visible light camera is used to detect the interferometric signal to reduce costs and improve optomechanical integration. Balancing the sensitivity vs. acquisition speed allows continuous high data transfer and processing rates and simplifies the implementation as a bedside system. Higher frame rates are important for scan positioning on non-compliant subjects such as infants and children.
Retinal capillary blood flow speeds quantification may provide biomarkers for retinal diseases. While OCTA visualizes the retinal vasculature, it provides limited information about the blood flow speeds. We present an OCTA-based method for measuring quantitative surrogate markers for blood flow speeds in retinal capillaries using a temporal autocorrelation decay model, by acquiring multiple OCT B-scan repeats and compiling OCTA measurements both spatially and temporally. A 600 kHz swept light source enabled short and multiple interscan times with fine A-scan spacing. We show blood flow speed differences among retinal vascular plexuses in healthy eyes and alterations in eyes with diabetic retinopathy.
KEYWORDS: Signal processing, Optical coherence tomography, Beam diameter, Light sources and illumination, Imaging systems, High speed cameras, Medical device development
A real-time high-speed line-field optical coherence tomography (LFOCT) configuration at 5,000 B-frames or 2,500,000 A-lines per second for clinical applications is described. The development of the LFOCT technology has experienced increasing interest despite the challenge of achieving sufficient sensitivity and resolution due to crosstalk. Publications of research systems usually apply expensive infrared cameras to improve image quality. The costs, complexity, and lack of real-time data transfer of dedicated highspeed IR cameras make it challenging to employ LFOCT for a more comprehensive application range. We demonstrate using a comparatively low-cost, high-speed camera for real-time data transfer and image display in conjunction with a light source at a center wavelength of 840 nm and a bandwidth of 50 nm. Although the camera and available sample power limits the sensitivity (75 dB), we can demonstrate imaging on a human fingertip (in vivo) and the cornea of a rabbit eye. Further development of the system will focus on compensating for the camera's limited sensitivity and improving resolution to target clinical in vivo imaging such as ophthalmic applications.
We demonstrate the first 1050nm MEMS-eVCSEL co-packaged with a wideband amplifier to achieve over 70nm wavelength tuning at over 30mW of output power and SMSR greater than 40dB. Ophthalmic Optical Coherence Tomography Angiography (OCTA) images acquired at 800kHz A-scan rates showcase the telecom grade 14pin butterfly co-package as a path to low cost swept source OCT engines. Device design employs a strain-compensated InGaAs/GaAsP gain region disposed on a wideband fully oxidized GaAs/AlxOy back mirror capable of tuning ranges beyond 100nm. It has been suggested the wideband fully oxidized GaAs/AlxOy back mirror may pose risk to device lifetime reliability. However, over 9000hrs of lifetime testing validates reliability and projects device lifetimes exceed 20,000hrs under continuous use.
Oxygen saturation (sO2) of RBCs in capillaries can indirectly assess local tissue oxygenation and metabolic function. For example, the altered retinal oxygenation in diabetic retinopathy and local hypoxia during tumor development in cancer are reflected by abnormal sO2 of local capillary networks. However, it is far from clear whether accurate label-free optical oximetry (i.e. measuring hemoglobin sO2) is feasible from dispersed red blood cells (RBCs) at the single-capillary level. The sO2-dependent hemoglobin absorption contrast present in optical scattering signal is complicated by geometry-dependent scattering from RBCs. Here we provide a theoretical model to calculate the backscattering spectra of single RBCs based on the first-order Born approximation, considering the orientation, size variation, and deformation of RBCs. We show that the oscillatory spectral behavior of RBC geometries is smoothed by variations in cell size and orientation, resulting in clear sO2-dependent spectral contrast. In addition, this spectral contrast persists with different deformations of RBCs, allowing the sO2 of individual RBCs in capillaries to be characterized. The theoretical model is verified by Mie theory and experiments using visible light optical coherence tomography (vis-OCT). Thus, this study shows for the first time the feasibility of, and provides a theoretical model for, label-free optical oximetry at the single-capillary level by backscattering-based imaging modalities, challenging the popular view that such measurements are impossible at the single-capillary level. This is promising for in vivo backscattering-based optical oximetry at the single-capillary level, to measure local capillary sO2 for early diagnosis, progression monitoring, and treatment evaluation of diabetic retinopathy and cancer.
Monitoring cortical hemodynamic response after ischemic stroke (IS) is essential for understanding the pathophysiological mechanisms behind IS-induced neuron loss. Functional optical coherence tomography (OCT) is an emerging technology that can fulfill the requirement, providing label-free, high-resolution 3D images of cerebral hemodynamics.
Unfortunately, strong tissue scattering pose a significant challenge for existing OCT oximetry techniques, as they either ignore the effect or compensate it numerically. Here we developed a novel dual-depth sampling and normalization strategy using visible-light OCT (vis-OCT) angiograms that can provide robust and precise sO2 estimations within cerebral circulation. The related theoretical formulation were established, and its implication and limitations were discussed.
We monitored mouse cortical hemodynamics using the newly-developed method. Focal ischemic stroke was induced through photothrombosis. The analysis on pre- and post-IS vis-OCT images revealed both vascular morphology and oxygenation altered substantially after the occlusion. First, the ischemic core could be clearly identified as angiographic intensity fell below the detection limit. In addition, vessel dilation presented universally in the penumbra region. Notably for pial arteriles, the percentage of increase demonstrated inverse relationship with their pre-occlusion, pre-dilation dimeter.
Vis-OCT oxygenation maps on intact cortex revealed spatial sO2 variations within pial vessels. Specifically, sO2 in arterioles decreased as it bifurcated and plunged into deeper tissue. Similarly, venous sO2 was higher in the larger, more superficial pial brunches. However, such difference was no longer appreciable after photothrombosis. Averaged arteriole sO2 dropped to 64% – 67% in the penumbra region.
Oxygen saturation (sO2) of red blood cells (RBCs) in capillaries can indirectly assess local tissue oxygenation and metabolic function. For example, the altered retinal oxygenation in diabetic retinopathy and local hypoxia during tumor development in cancer are reflected by abnormal sO2 of local capillary networks. However, it is far from clear whether accurate label-free optical oximetry (i.e., measuring hemoglobin sO2) is feasible from dispersed RBCs at the single capillary level. The sO2-dependent hemoglobin absorption contrast present in optical scattering signal is complicated by geometry-dependent scattering from RBCs. We present a numerical study of backscattering spectra from single RBCs based on the first-order Born approximation, considering practical factors: RBC orientations, size variation, and deformations. We show that the oscillatory spectral behavior of RBC geometries is smoothed by variations in cell size and orientation, resulting in clear sO2-dependent spectral contrast. In addition, this spectral contrast persists with different mean cellular hemoglobin content and different deformations of RBCs. This study shows for the first time the feasibility of, and provides a theoretical model for, label-free optical oximetry at the single capillary level using backscattering-based imaging modalities, challenging the popular view that such measurements are impossible at the single capillary level.
We developed a simultaneous visible-light (Vis) and near-infrared (NIR) dual-band optical coherence tomography (OCT) system using a single supercontinuum laser source. The goal was to benchmark our newly developed Vis-OCT against the well-developed NIR-OCT. The Vis-OCT subsystem operated at 91 nm full-width-at-half-maximum (FWHM) bandwidth centered at 566 nm; the NIR-OCT subsystem operated at 93 nm FWHM bandwidth centered at 841 nm. The axial resolutions were 1.8 and 4.4 μm in air for the Vis- and NIR-OCT subsystems, respectively. We compared the respective performances, including anatomical imaging, angiography, absolute retinal blood flow measurements, and spectroscopic analysis for retinal blood oxygen saturation (sO2), between the two subsystems in rodents in vivo. While demonstrating minor discrepancies related to operation wavelengths, both subsystems showed comparable performances in the first three tests. However, we were only able to retrieve sO2 using the Vis-OCT subsystem.
The oxygen-dependent absorption of hemoglobin provides the fundamental contrast for all label-free techniques measuring blood oxygenation. When hemoglobin is packaged into red blood cells (RBCs), the structure of the cells creates light scattering which also depends on the absorption based on the Kramers-Kronig relationship. Thus a proper characterization of the optical behaviors of blood has been a key to any accurate measurement of blood oxygenation, particularly at the capillary level where RBCs are dispersed individually in contrast to a densely packed whole blood. Here we provided a theoretical model under Born Approximation to characterize the oxygen dependent backscattering spectroscopic contrast from single RBCs. Using this theoretical model, we conducted simulations on both oxygenated and deoxygenated single RBCs with different sizes for standard and possible deformed cell geometries in blood flow, all which suggested similar backscattering spectroscopic contrast and were confirmed by Mie Theory and experiments using visible Optical Coherence Tomography (visOCT). As long as the cell size satisfies Gaussian distribution with a coefficient variance (C.V.) large enough, there is clear absorption contrast between the backscattering spectra of oxygenated and deoxygenated single RBCs calculated by this model, so oxygen saturation can then be characterized. Thus, this theoretical model can be extended to extract absorption features of other scattering particles as long as they satisfy Born Approximation.
The lack of capability to quantify oxygen metabolism noninvasively impedes both fundamental investigation and clinical diagnosis of a wide spectrum of diseases including all the major blinding diseases such as age-related macular degeneration, diabetic retinopathy, and glaucoma. Using visible light optical coherence tomography (vis-OCT), we demonstrated accurate and robust measurement of retinal oxygen metabolic rate (rMRO2) noninvasively in rat eyes. The rMRO2 was calculated by concurrent measurement of blood flow and blood oxygen saturation (sO2). Blood flow was calculated by the principle of Doppler optical coherence tomography, where the phase shift between two closely spaced A-lines measures the axial velocity. The distinct optical absorption spectra of oxy- and deoxy-hemoglobin provided the contrast for sO2 measurement, combined with the spectroscopic analysis of vis-OCT signal within the blood vessels. We continuously monitored the regulatory response of oxygen consumption to a progressive hypoxic challenge. We found that both oxygen delivery, and rMRO2 increased from the highly regulated retinal circulation (RC) under hypoxia, by 0.28±0.08 μL/min (p<0.001), and 0.20±0.04 μL/min (p<0.001) per 100 mmHg systemic pO2 reduction, respectively. The increased oxygen extraction compensated for the deficient oxygen supply from the poorly regulated choroidal circulation (CC).
We explored, both numerically and experimentally, whether OCT can be a good candidate to accurately measure retinal oxygen metabolism. We first used statistical methods to numerically simulate photon transport in the retina to mimic OCT working under different spectral ranges. Then we analyze accuracy of OCT oximetry subject to parameter variations such as vessel size, pigmentation, and oxygenation. We further developed an experimental OCT system based on the spectral range identified by our simulation work. We applied the newly developed OCT to measure both retinal hemoglobin oxygen saturation (sO2) and retinal retinal flow. After obtaining the retinal sO2 and blood velocity, we further measured retinal vessel diameter and calculated the retinal oxygen metabolism rate (MRO2). To test the capability of our OCT, we imaged wild-type Long-Evans rats ventilated with both normal air and air mixtures with various oxygen concentrations.
Our simulation suggested that OCT working within visible spectral range is able to provide accurate measurement of retinal MRO2 using inverse Fourier transform spectral reconstruction. We called this newly developed technology vis-OCT, and showed that vis-OCT was able to measure the sO2 value in every single major retinal vessel around the optical disk as well as in micro retinal vessels. When breathing normal air, the averaged sO2 in arterial and venous blood in Long-Evans rats was measured to be 95% and 72%, respectively. When we challenge the rats using air mixtures with different oxygen concentrations, vis-OCT measurement followed analytical models of retinal oxygen diffusion and pulse oximeter well.
Visible light optical coherence tomography (vis-OCT) is intrinsically capable of optical determination of blood oxygen saturation (sO2). Thanks to its 3D sectioning ability, confounding factors that plaque multi-wavelength fundus photography can be avoided. We further supplemented it with motion-enhanced angiography (vis-OCTA), which allowed us to resolve retinal micro vessels without losing spectral information. As a result, spectroscopic vis-OCTA can extract microvascular sO2 which are generally inaccessible. Here we extend the theoretical formulation of vis-OCTA oximetry to include optical attenuation, scattering and motion contrast. The model allows robust estimation of sO2, while also promising reduction of illuminating power to 1/3 of current value of ~1 mW. To demonstrate the capability of our approach, we performed oxygen challenge while taking vis-OCTA measurements on rat ocular circulation in vivo. We supplied the experiment animal with the following gas mixture: normal air, 5% CO2 air, pure O2 and 10% O2 air. For each inhalation gas, the OCTA measurements were compared with peripheral capillary sO2 (spO2) provided by a pulse oximeter. The retinal artery sO2 measurements corresponded well with spO2 reading as expected (R2 = 0.87). We found that both retinal and choroidal circulation sO2 moderately increased when we supplied 5% CO2 air. 100% O2 inhalation significantly increased both artery and vein oxygenation. On the contrary, 10% O2 air could deplete the oxygen reservoir in the circulation and lead to low sO2 readings.
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