Granzyme B degrades extracellular matrix and promotes inflammation and choroidal neovascularization.

Age-related macular degeneration (AMD) is a common retinal neurodegenerative disease among the elderly. Neovascular AMD (nAMD), a leading cause of AMD-related blindness, involves choroidal neovascularization (CNV), which can be suppressed by anti-angiogenic treatments. However, current CNV treatments do not work in all nAMD patients. Here we investigate a novel target for AMD. Granzyme B (GzmB) is a serine protease that promotes aging, chronic inflammation and vascular permeability through the degradation of the extracellular matrix (ECM) and tight junctions. Extracellular GzmB is increased in retina pigment epithelium (RPE) and mast cells in the choroid of the healthy aging outer retina. It is further increased in donor eyes exhibiting features of nAMD and CNV. Here, we show in RPE-choroidal explant cultures that exogenous GzmB degrades the RPE-choroid ECM, promotes retinal/choroidal inflammation and angiogenesis while diminishing anti-angiogenic factor, thrombospondin-1 (TSP-1). The pharmacological inhibition of either GzmB or mast-cell degranulation significantly reduces choroidal angiogenesis. In line with our in vitro data, GzmB-deficiency reduces the extent of laser-induced CNV lesions and the age-related deterioration of electroretinogram (ERG) responses in mice. These findings suggest that targeting GzmB, a serine protease with no known endogenous inhibitors, may be a potential novel therapeutic approach to suppress CNV in nAMD.

In vivo multi-contrast depth-resolved choroidal imaging of a mouse using polarization-diversity optical coherence tomography.

The results of depth-resolved multi-contrast in vivo mouse choroidal imaging using a polarization-diversity optical coherence tomography (PD-OCT) system are presented. A selectively chosen depth of focus that was fine-tuned with a sensorless adaptive optics technique and a simple segmentation based on the degree of polarization uniformity signal visualizes the detailed features of a mouse choroid from the OCT angiography images. A comprehensive image analysis of the choroid revealed the distinctive pathological characteristics of the laser-induced choroidal neovascularization mouse.

Image-based cross-calibration method for multiple spectrometer-based OCT.

A fast and practical computational cross-calibration of multiple spectrometers is described. A signal correlation matrix (CM) can be constructed from paired B-scans in a multiple-spectrometer optical coherence tomography (OCT), where the wavelength-corresponding pixels are indicated by high cross correlation. The CM can be used to either guide the physical alignment of spectrometers or to numerically match the spectra in the post-process. The performance is comparable to the previously reported optimization approach, as demonstrated by the mirror tests, qualitative comparison of OCT and optical coherence tomography angiography (OCTA) images, and quantitative comparison of image metrics.

Phase-corrected buffer averaging for enhanced OCT angiography using FDML laser.

Megahertz-rate optical coherence tomography angiography (OCTA) is highly anticipated as an ultrafast imaging tool in clinical settings. However, shot-noise-limited sensitivity is inevitably reduced in high-speed imaging systems. In this Letter, we present a coherent buffer averaging technique for use with a Fourier-domain mode-locked (FDML) laser to improve OCTA contrast at 1060 nm MHz-rate retinal imaging. Full characterization of spectral variations among the FDML buffers and a numerical correction method are also presented, with the results demonstrating a 10-fold increase in the phase alignment among buffers. Coherent buffer averaging provided better OCTA contrast than the conventional multi-frame averaging approach with a faster acquisition time.

Visible light sensorless adaptive optics for retinal structure and fluorescence imaging.

Optical coherence tomography (OCT) has emerged as a powerful imaging instrument and technology in biomedicine. OCT imaging is predominantly performed using wavelengths in the near infrared; however, visible light (VIS) has been recently employed in OCT systems with encouraging results for high-resolution retinal imaging. Using a broadband supercontinuum VIS source, we present a sensorless adaptive optics (SAO) multimodal imaging system driven by VIS-OCT for volumetric retinal structural imaging, followed by the acquisition of fluorescence emission. The coherence-gated, depth-resolved VIS-OCT images used for image-guided SAO aberration correction enable high-resolution structural and fluorescence imaging.

Prominin-1 null Xenopus laevis develop subretinal drusenoid-like deposits, cone-rod dystrophy, and RPE atrophy.

Mutations in the PROMININ-1 (PROM1) gene are associated with inherited, non-syndromic vision loss. Here, we used CRISPR/Cas9 to induce truncating prom1-null mutations in Xenopus laevis to create a disease model. We then tracked progression of retinal degeneration in these animals from the ages of 6 weeks to 3 years old. We found that retinal degeneration caused by prom1-null is age-dependent and likely involves death or damage to the retinal pigment epithelium (RPE) that precedes photoreceptor degeneration. As prom1-null frogs age, they develop large cellular debris deposits in the subretinal space and outer segment layer which resemble subretinal drusenoid deposits (SDD) in their location, histology, and representation in color fundus photography and optical coherence tomography (OCT). In older frogs, these SDD-like deposits accumulate in size and number, and they are present before retinal degeneration occurs. Evidence for an RPE origin of these deposits includes infiltration of pigment granules into the deposits, thinning of RPE as measured by OCT, and RPE disorganization as measured by histology and OCT. The appearance and accumulation of SDD-like deposits and RPE thinning and disorganization in our animal model suggests an underlying disease mechanism for prom1-null mediated blindness of death and dysfunction of the RPE preceding photoreceptor degeneration, instead of direct effects upon photoreceptor outer segment morphogenesis, as was previously hypothesized.

Generalized 3D registration algorithm for enhancing retinal optical coherence tomography images.

Significance: Optical coherence tomography (OCT) has emerged as the standard of care for diagnosing and monitoring the treatment of various ocular disorders due to its noninvasive nature and in vivo volumetric acquisition capability. Despite its widespread applications in ophthalmology, motion artifacts remain a challenge in OCT imaging, adversely impacting image quality. While several multivolume registration algorithms have been developed to address this issue, they are often designed to cater to one specific OCT system or acquisition protocol. Aim: We aim to generate an OCT volume free of motion artifacts using a system-agnostic registration algorithm that is independent of system specifications or protocol. Approach: We developed a B-scan registration algorithm that removes motion and corrects for both translational eye movements and rotational angle differences between volumes. Tests were carried out on various datasets obtained from two different types of custom-built OCT systems and one commercially available system to determine the reliability of the proposed algorithm. Additionally, different system specifications were used, with variations in axial resolution, lateral resolution, signal-to-noise ratio, and real-time motion tracking. The accuracy of this method has further been evaluated through mean squared error (MSE) and multiscale structural similarity index measure (MS-SSIM). Results: The results demonstrate improvements in the overall contrast of the images, facilitating detailed visualization of retinal vasculatures in both superficial and deep vasculature plexus. Finer features of the inner and outer retina, such as photoreceptors and other pathology-specific features, are discernible after multivolume registration and averaging. Quantitative analyses affirm that increasing the number of averaged registered volumes will decrease MSE and increase MS-SSIM as compared to the reference volume. Conclusions: The multivolume registered data obtained from this algorithm offers significantly improved visualization of the retinal microvascular network as well as retinal morphological features. Furthermore, we have validated that the versatility of our methodology extends beyond specific OCT modalities, thereby enhancing the clinical utility of OCT for the diagnosis and monitoring of ocular pathologies.

Early inner plexiform layer thinning and retinal nerve fiber layer thickening in excitotoxic retinal injury using deep learning-assisted optical coherence tomography.

Excitotoxicity from the impairment of glutamate uptake constitutes an important mechanism in neurodegenerative diseases such as Alzheimer's, multiple sclerosis, and Parkinson's disease. Within the eye, excitotoxicity is thought to play a critical role in retinal ganglion cell death in glaucoma, diabetic retinopathy, retinal ischemia, and optic nerve injury, yet how excitotoxic injury impacts different retinal layers is not well understood. Here, we investigated the longitudinal effects of N-methyl-D-aspartate (NMDA)-induced excitotoxic retinal injury in a rat model using deep learning-assisted retinal layer thickness estimation. Before and after unilateral intravitreal NMDA injection in nine adult Long Evans rats, spectral-domain optical coherence tomography (OCT) was used to acquire volumetric retinal images in both eyes over 4 weeks. Ten retinal layers were automatically segmented from the OCT data using our deep learning-based algorithm. Retinal degeneration was evaluated using layer-specific retinal thickness changes at each time point (before, and at 3, 7, and 28 days after NMDA injection). Within the inner retina, our OCT results showed that retinal thinning occurred first in the inner plexiform layer at 3 days after NMDA injection, followed by the inner nuclear layer at 7 days post-injury. In contrast, the retinal nerve fiber layer exhibited an initial thickening 3 days after NMDA injection, followed by normalization and thinning up to 4 weeks post-injury. Our results demonstrated the pathological cascades of NMDA-induced neurotoxicity across different layers of the retina. The early inner plexiform layer thinning suggests early dendritic shrinkage, whereas the initial retinal nerve fiber layer thickening before subsequent normalization and thinning indicates early inflammation before axonal loss and cell death. These findings implicate the inner plexiform layer as an early imaging biomarker of excitotoxic retinal degeneration, whereas caution is warranted when interpreting the ganglion cell complex combining retinal nerve fiber layer, ganglion cell layer, and inner plexiform layer thicknesses in conventional OCT measures. Deep learning-assisted retinal layer segmentation and longitudinal OCT monitoring can help evaluate the different phases of retinal layer damage upon excitotoxicity.

Advanced multi-contrast Jones matrix optical coherence tomography for Doppler and polarization sensitive imaging.

An advanced version of Jones matrix optical coherence tomography (JMT) is demonstrated for Doppler and polarization sensitive imaging of the posterior eye. JMT is capable of providing localized flow tomography by Doppler detection and investigating the birefringence property of tissue through a three-dimensional (3-D) Jones matrix measurement. Owing to an incident polarization multiplexing scheme based on passive optical components, this system is stable, safe in a clinical environment, and cost effective. Since the properties of this version of JMT provide intrinsic compensation for system imperfection, the system is easy to calibrate. Compared with the previous version of JMT, this advanced JMT achieves a sufficiently long depth measurement range for clinical cases of posterior eye disease. Furthermore, a fine spectral shift compensation method based on the cross-correlation of calibration signals was devised for stabilizing the phase of OCT, which enables a high sensitivity Doppler OCT measurement. In addition, a new theory of JMT which integrates the Jones matrix measurement, Doppler measurement, and scattering measurement is presented. This theory enables a sensitivity-enhanced scattering OCT and high-sensitivity Doppler OCT. These new features enable the application of this system to clinical cases. A healthy subject and a geographic atrophy patient were measured in vivo, and simultaneous imaging of choroidal vasculature and birefringence structures are demonstrated.

Polarization-Diversity Optical Coherence Tomography Assessment of Choroidal Nevi.

Purpose: The purpose of this study was to demonstrate the utility of polarization-diversity optical coherence tomography (PD-OCT), a noninvasive imaging technique with melanin-specific contrast, in the quantitative and qualitative assessment of choroidal nevi. Methods: Nevi were imaged with a custom-built 55-degree field-of-view (FOV) 400 kHz PD-OCT system. Imaging features on PD-OCT were compared to those on fundus photography, auto-fluorescence, ultrasound, and non-PD-OCT images. Lesions were manually segmented for size measurement and metrics for objective assessment of melanin distributions were calculated, including degree of polarization uniformity (DOPU), attenuation coefficient, and melanin occupancy rate (MOR). Results: We imaged 17 patients (mean age = 69.5 years, range = 37-90) with 11 pigmented, 3 non-pigmented, and 3 mixed pigmentation nevi. Nevi with full margin acquisition had an average longest basal diameter of 5.1 mm (range = 2.99-8.72 mm) and average height of 0.72 mm (range = 0.37 mm-2.09 mm). PD-OCT provided clear contrast of choroidal melanin content, distribution, and delineation of nevus margins for melanotic nevi. Pigmented nevi were found to have lower DOPU, higher attenuation coefficient, and higher MOR than non-pigmented lesions. Melanin content on PD-OCT was consistent with pigmentation on fundus in 15 of 17 nevi (88%). Conclusions: PD-OCT allows objective assessment of choroidal nevi melanin content and distribution. In addition, melanin-specific contrast by PD-OCT enables clear nevus margin delineation and may improve serial growth surveillance. Further investigation is needed to determine the clinical significance and prognostic value of melanin characterization by PD-OCT in the evaluation of choroidal nevi.

Segmentation-guided domain adaptation and data harmonization of multi-device retinal optical coherence tomography using cycle-consistent generative adversarial networks.

BACKGROUND: Medical images such as Optical Coherence Tomography (OCT) images acquired from different devices may show significantly different intensity profiles. An automatic segmentation model trained on images from one device may perform poorly when applied to images acquired using another device, resulting in a lack of generalizability. This study addresses this issue using domain adaptation methods improved by Cycle-Consistent Generative Adversarial Networks (CycleGAN), especially when the ground-truth labels are only available in the source domain. METHODS: A two-stage pipeline is proposed to generate segmentation in the target domain. The first stage involves the training of a state-of-the-art segmentation model in the source domain. The second stage aims to adapt the images from the target domain to the source domain. The adapted target domain images are segmented using the model in the first stage. Ablation tests were performed with integration of different loss functions, and the statistical significance of these models is reported. Both the segmentation performance and the adapted image quality metrics were evaluated. RESULTS: Regarding the segmentation Dice score, the proposed model ssppg achieves a significant improvement of 46.24% compared to without adaptation and reaches 87.4% of the upper limit of the segmentation performance. Furthermore, image quality metrics, including FID and KID scores, indicate that adapted images with better segmentation also have better image qualities. CONCLUSION: The proposed method demonstrates the effectiveness of segmentation-driven domain adaptation in retinal imaging processing. It reduces the labor cost of manual labeling, incorporates prior anatomic information to regulate and guide domain adaptation, and provides insights into improving segmentation qualities in image domains without labels.

High-penetration swept source Doppler optical coherence angiography by fully numerical phase stabilization.

A high-penetration swept-source optical coherence tomography (HP-SS-OCT) system based on a 1-µm short cavity laser is developed. Doppler OCT processing is applied, along with a custom-made numerical phase stabilization algorithm; this process does not require additional calibration hardware. Thus, our phase stabilization method is simple and can be employed in a variety of SS-OCT systems. The bidirectional blood flow and vasculature in the deep choroid was successfully imaged via two Doppler modes that use different time intervals for Doppler processing. En face projection image of squared power of Doppler shift is compared to ICGA, and the utility of our method is verified.

Numerical calibration method for a multiple spectrometer-based OCT system.

The present paper introduces a numerical calibration method for the easy and practical implementation of multiple spectrometer-based spectral-domain optical coherence tomography (SD-OCT) systems. To address the limitations of the traditional hardware-based spectrometer alignment across more than one spectrometer, we applied a numerical spectral calibration algorithm where the pixels corresponding to the same wavelength in each unit are identified through spatial- and frequency-domain interferometric signatures of a mirror sample. The utility of dual spectrometer-based SD-OCT imaging is demonstrated through in vivo retinal imaging at two different operation modes with high-speed and dual balanced acquisitions, respectively, in which the spectral alignment is critical to achieve improved retinal image data without any artifacts caused by misalignment of the spectrometers.

ADAPTIVE OPTICS OPTICAL COHERENCE TOMOGRAPHY IN A CASE OF ACUTE ZONAL OCCULT OUTER RETINOPATHY.

PURPOSE: To report a case of acute zonal occult outer retinopathy in which adaptive optics (AO) facilitated visualization of abnormal photoreceptors previously thought to be in an area of normal retina on conventional optical coherence tomography (OCT). METHODS: Case report. RESULTS: A 51-year-old woman presents with 11-month history of photopsias and scotoma in the temporal visual field of her left eye. Ocular imaging including fluorescein angiography, fundus autofluorescence and OCT suggested the diagnosis of acute zonal occult outer retinopathy in the left eye. Adaptive optics optical coherence tomography (AO-OCT) revealed photoreceptor abnormalities not previously identified in conventional OCT, in areas apparently normal on multimodal imaging. On enface and cross-sectional AO-OCT, round and evenly spaced hyperreflectivity corresponding to normal cone mosaic (Pattern 1) was adjacent to unevenly and disrupted cone hyperreflectivity (Pattern 2) and areas with hyporeflectivity or no cone reflectivity (Pattern 3). Cross-sectional AO-OCT of Patterns 2 and 3 also revealed attenuation of ellipsoid zone with loss of interdigitation zone. CONCLUSION: Adaptive optics OCT documented cone photoreceptors in finer details than conventional OCT and revealed early changes in a patient with acute zonal occult outer retinopathy, in an area of the retina thought to be normal on conventional multimodal imaging. These findings may provide important insight into pathogenesis and progression of the disease.

Corneal imaging with blue-light optical coherence microscopy.

Corneal imaging is important for the diagnostic and therapeutic evaluation of many eye diseases. Optical coherence tomography (OCT) is extensively used in ocular imaging due to its non-invasive and high-resolution volumetric imaging characteristics. Optical coherence microscopy (OCM) is a technical variation of OCT that can image the cornea with cellular resolution. Here, we demonstrate a blue-light OCM as a low-cost and easily reproducible system to visualize corneal cellular structures such as epithelial cells, endothelial cells, keratocytes, and collagen bundles within stromal lamellae. Our blue-light OCM system achieved an axial resolution of 12 µm in tissue over a 1.2 mm imaging depth, and a lateral resolution of 1.6 µm over a field of view of 750 µm × 750 µm.

Clinical explainable differential diagnosis of polypoidal choroidal vasculopathy and age-related macular degeneration using deep learning.

BACKGROUND: This study aims to achieve an automatic differential diagnosis between two types of retinal pathologies with similar pathological features - Polypoidal choroidal vasculopathy (PCV) and wet age-related macular degeneration (AMD) from volumetric optical coherence tomography (OCT) images, and identify clinically-relevant pathological features, using an explainable deep-learning-based framework. METHODS: This is a retrospective study with data from a cross-sectional cohort. The OCT volume of 73 eyes from 59 patients was included in this study. Disease differentiation was achieved through single-B-scan-based classification followed by a volumetric probability prediction aggregation step. We compared different labeling strategies with and without identifying pathological B-scans within each OCT volume. Clinical interpretability was achieved through normalized aggregation of B-scan-based saliency maps followed by maximum-intensity-projection onto the en face plane. We derived the PCV score from the proposed differential diagnosis framework with different labeling strategies. The en face projection of saliency map was validated with the pathologies identified in Indocyanine green angiography (ICGA). RESULTS: Model trained with both labeling strategies achieved similar level differentiation power (>90%), with good correspondence between pathological features detected from the projected en face saliency map and ICGA. CONCLUSIONS: This study demonstrated the potential clinical application of non-invasive differential diagnosis using AI-driven OCT-based analysis, with minimal requirement of labeling efforts, along with clinical explainability achieved through automatically detected disease-related pathologies.

Multimodal analysis of pearls and pearl treatments by using optical coherence tomography and fluorescence spectroscopy.

We present an integrated optical system that consists of optical coherence tomography (OCT) and laser-induced fluorescence (LIF) spectroscopy for multimodal analysis of pearls and pearl treatments. The OCT source and the LIF excitation beams were aligned together to illuminate the same spot of a pearl fixed on the sample stage that was under rotation. As a result, both OCT images and LIF spectra of the pearls were detected at the same time and also at the same place. For OCT, a 1310 nm-centered swept laser source was used. For LIF, a 405 nm laser diode was used and a lensed multimode fiber was utilized as a fluorescence probe. The tomographic investigation on the internal structure of a pearl allowed us to evaluate and categorize the pearl nondestructively as was previously reported. In addition, the measurements of fluorescence spectrum and its decaying rate helped to determine the species of mother oyster. The proposed multimodal analysis made it possible to classify the pearls and also to disclose the treatments made on the pearls.

Optimizing 3D retinal vasculature imaging in diabetic retinopathy using registration and averaging of OCT-A.

High resolution visualization of optical coherence tomography (OCT) and OCT angiography (OCT-A) data is required to fully take advantage of the imaging modality's three-dimensional nature. However, artifacts induced by patient motion often degrade OCT-A data quality. This is especially true for patients with deteriorated focal vision, such as those with diabetic retinopathy (DR). We propose a novel methodology for software-based OCT-A motion correction achieved through serial acquisition, volumetric registration, and averaging. Motion artifacts are removed via a multi-step 3D registration process, and visibility is significantly enhanced through volumetric averaging. We demonstrate that this method permits clear 3D visualization of retinal pathologies and their surrounding features, 3D visualization of inner retinal capillary connections, as well as reliable visualization of the choriocapillaris layer.

Effect of optical coherence tomography and angiography sampling rate towards diabetic retinopathy severity classification.

Optical coherence tomography (OCT) and OCT angiography (OCT-A) may benefit the screening of diabetic retinopathy (DR). This study investigated the effect of laterally subsampling OCT/OCT-A en face scans by up to a factor of 8 when using deep neural networks for automated referable DR classification. There was no significant difference in the classification performance across all evaluation metrics when subsampling up to a factor of 3, and only minimal differences up to a factor of 8. Our findings suggest that OCT/OCT-A can reduce the number of samples (and hence the acquisition time) for a volume for a given field of view on the retina that is acquired for rDR classification.