KEYWORDS: Depolarization, Signal to noise ratio, Eye, Retina, Tissues, Polarization, In vivo imaging, Optical calibration, Calibration, Image segmentation
SignificanceA data-based calibration method with enhanced depolarization contrast in polarization-sensitive optical coherence tomography (PS-OCT) was developed and demonstrated effective for detecting melanin content in the eye.AimWe aim to mitigate the dependence between the measured depolarization metric and the intensity signal-to-noise ratio (SNR) for improved visualization of depolarizing tissues, especially in low SNR regions, and to demonstrate the enhanced depolarization contrast to evaluate melanin presence.ApproachA function for calibrating the depolarization metric was experimentally derived from the young albino guinea pig, assuming depolarization free in the retina. A longitudinal study of guinea pigs (9 weeks) was conducted to assess the accumulation of melanin during early eye growth. Furthermore, the melanin content of the sub-macular choroid was compared in eyes with light and dark irides involving 14 human subjects in early middle adulthood.ResultsWe observed an increase in the improved depolarization contrast, which indicates potential melanin accumulation in the early eye development with age in the pigmented guinea pig eyes. We found a significant difference in melanin content between human eyes with light and dark colors.ConclusionsOur proposed calibration method enhanced the visualization of depolarizing structures in PS-OCT, which can be generalized to all kinds of polarization-sensitive imaging and can potentially monitor melanin in healthy and pathological eyes.
Previous studies in optoretinography often rely on AO-OCT systems to resolve individual cells or use intensity-based image processing to extract the dynamics of the retinal layer as an ensemble. For non-AO point-scan OCT, investigating tissue dynamics from obscure speckle patterns while preserving the spatial heterogeneity of signals remains challenging. Here, we developed phase-restoring subpixel image registration and unsupervised machine learning algorithms to accurately extract spatially-resolved OCT phase signals from the outer retina in rodents. In addition to observing light-evoked deformation of the photoreceptors outer segments, we discovered an optical signature of the retinal pigment epithelium (RPE) response to visual stimuli.
In-vivo imaging of the light-evoked responses of retinal cells in rodents can provide valuable insights into the correlation between optoretinography (ORG) signals and retinal degeneration. However, interpreting outer retina dynamics in rodents is challenging due to the limited resolution of optical coherence tomography, which often results in the superposition of outer retinal layers, such as the rod outer segment (ROS), retinal pigment epithelium (RPE), and Bruch’s membrane, within speckle patterns. Here, we present an automated, unbiased approach for extracting spatially-resolved outer retinal dynamics from complicated speckle patterns. Using this approach, we revealed the light-evoked dynamics of both ROS and RPE in rodents.
Small animals, such as rodents, are attractive options for investigating the intrinsic process of retinal degeneration. In this study, we used phase-sensitive optical coherence tomography to explore the comprehensive dynamics of rats’ outer retinas in response to visual stimuli. By calculating the temporal phase difference between different outer retinal bands, we revealed highly reproducible retinal dynamics, on the order of tens of nanometers, related to different parts of the outer retina. Our approach may pave the way for preclinical optoretinography study in small animals, facilitating clinical translations for the early detection of neurodegenerative diseases.
Recently, there has been vast interest in probing photoreceptor dynamics using optical coherence tomography (OCT). Most successful demonstrations implemented adaptive optics or digital adaptive optics to resolve individual cones or rods in human subjects. Here we use phase information to trace the photoreceptor response in rodents using an ultrahigh-resolution, phase-sensitive, spectral-domain OCT. Brown Norway rats (six to 14 weeks) were sedated using a ketamine and xylazine cocktail. Repeated scans were registered by a phase-restoring subpixel motion correction algorithm to isolate the bulk motion, and two hyperreflective bands (inner segment/outer segment junction – IS/OS; outer segment tip + retinal pigment epithelium + Bruch's membrane) were segmented automatically. As a result, two types of nanoscale signals (biphasic Type-I and monophasic Type-II) were detected with a clear separation in depth. We tested the repeatability, scotopic stimulus strength dependency, and photopic background intensity dependency. Besides, we demonstrated enface mapping of the ORG signals in a wide field of 20°, analogous to the multifocal electroretinogram but with a much higher resolution, revealing the spatial distribution of the outer retina function. This method could be extended to study animal models with photoreceptor degeneration and clinical studies to investigate early photoreceptor dysfunction with high spatiotemporal resolution.
KEYWORDS: Image registration, Optical coherence tomography, Image segmentation, Visualization, Tissues, Speckle pattern, Signal detection, Motion models, In vivo imaging, Image processing algorithms and systems
Phase-sensitive OCT can be used for imaging the photoreceptor deformations in response to the light stimulus or optoretinography (ORG). Here, we propose a phase-restoring subpixel image registration method and an automated signal extraction algorithm for optoretinography using phase-sensitive OCTs. We validated these methods in simulations, phantom experiments, and in-vivo optoretinogram imaging. Our image registration method yields better amplitude stability and higher phase accuracy compared with conventional approaches, and we found two types of signals (one monophasic and the other biphasic) simultaneously in rodent ORG imaging. These results can be beneficial to the ongoing preclinical/clinical ORG studies.
KEYWORDS: Optical coherence tomography, In vivo imaging, Adaptive optics, Signal detection, Optical signal processing, Human subjects, Head, Animal model studies
In recent years, there have been vast interests in probing photoreceptor dynamics using optical coherence tomography (OCT). Most of the successful demonstrations implemented adaptive optics or digital adaptive optics to resolve individual cones or rods in human subjects. Here we use phase information to trace the photoreceptor response in rodents using an ultrahigh-resolution, phase-sensitive, spectral-domain OCT. As a result, two types of nanoscale signals (monophasic and biphasic) were detected with a clear separation in depth. The monophasic signal is less susceptible to stimulus intensity and saturated from a 3% breach rate.
Polarization-sensitive OCT (PS-OCT) derives image contrast from tissue birefringence. Here, we introduced triple-input polarization sensitive optical coherence tomography (TRIPS-OCT), a new polarimetric modulation and reconstruction strategy for depth-resolved tomographic birefringence imaging in-vivo. We modulated the polarization states between three repeated frames and enabled the reconstruction of the Mueller matrix at each location within the triple-measured frames. We demonstrated a 2-fold reduction of the birefringence noise floor compared to the conventional dual-input reconstruction method, and a 3-fold reduction of the measurement error of optic axis orientation in retinal imaging with the compensation of corneal retardance and diattenuation.
Here, we demonstrate the visual evoked blood flow change in trilaminar retinal vasculature in rodents using OCT capillary velocimetry. A custom-built spectral-domain optical coherence tomography operates on 800 nm region performed all the imaging. In-vivo measurements were conducted on dark-adapted Brown Norway rats. The retinal blood flow velocity was calculated using a modified dynamic light scattering (DLS) – OCT method. As a result, the blood flow velocity in arterioles, venules, and capillaries in the retinal vasculature can be resolved. Flicker stimulation triggered heterogeneous responses – reduced flow velocity in large vessels vs. increased flow velocity in capillaries.
Phase-sensitive optical coherence tomography (OCT) enables label-free imaging of structural dynamics with nanoscopic sensitivity. However, the inevitable bulk tissue motions degrade the signal stability and introduce extra phase error. To suppress the motion-induced phase error, we propose a phase-restoring subpixel motion correction method for post-hoc motion correction in Fourier domain OCT, which enables translational shifts of complex-valued OCT images by arbitrary distance. Phantom and in-vivo rodent optoretinogram imaging experiments were conducted to demonstrate the advantages of the proposed method over conventional pixel-level method and the Fourier transform based method.
We investigate the influence of the OCT system resolution on high-quality en face corneal endothelial cell images in vivo, to allow for quantitative analysis of cell density. We vary the lateral resolution of the ultrahigh-resolution (UHR) OCT system (centered at 850 nm) by using different objectives, and the axial resolution by windowing the source spectrum. We are able to obtain a high-quality en face corneal endothelial cell map in vivo using UHR OCT for the first time. Quantitative analysis result of cell density from in vivo en face corneal endothelial cell map agrees with previously reported data.
Choriocapillaris is a unique vascular plexus located posterior to the retinal pigment epithelium. In the recent years, there is an increasing interest to investigate choriocapillaris alteration and progression of eye diseases and aging, using the optical coherence tomography angiography (OCTA). However, standardized algorithm for analysis has not been developed. Herein, we present non-invasive, in-vivo, high-resolution images of the non-human primates’ choriocapillaris using OCTA. Images were acquired with a prototype swept-source OCTA (SS-OCTA) system with 100kHz A-scan/s rate, over regions of 3×3 mm2 and 12×12 mm2. The non-perfusion area, also called flow voids, were segmented with an intensity damped, illuminance-compensated algorithm. The optimized quantification of the choriocapillaris flow voids may have applications in a wide array of eye diseases including age-related macular degeneration (AMD) and visualization of choriocapillaris in animal models could aid future studies on choroid involvement in models of eye disease.
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