A real-time line-field optical coherence tomography (LF-OCT) system is demonstrated with image acquisition rates of up to 5000 B-frames or 2.5 million A-lines per second for 500 A-lines per B-frame. The system uses a high-speed low-cost camera to achieve continuous data transfer rates required for real-time imaging, allowing the evaluation of future applications in clinical or intraoperative environments. The light source is an 840 nm super-luminescent diode. Leveraging parallel computing with GPU and high speed CoaXPress data transfer interface, we were able to acquire, process, and display OCT data with low latency. The studied system uses anamorphic beam shaping in the detector arm, optimizing the field of view and sensitivity for imaging biological tissue at cellular resolution. The lateral and axial resolution measured in air were 1.7 µm and 6.3 µm, respectively. Experimental results demonstrate real-time inspection of the trabecular meshwork and Schlemm’s canal on ex vivo corneoscleral wedges and real-time imaging of endothelial cells of human subjects in vivo .
This study presents a 3D in vitro cell culture model, meticulously 3D printed to replicate the conventional aqueous outflow pathway anatomical structure, facilitating the study of trabecular meshwork (TM) cellular responses under glaucomatous conditions. Glaucoma affects TM cell functionality, leading to extracellular matrix (ECM) stiffening, enhanced cell-ECM adhesion, and obstructed aqueous humor outflow. Our model, reconstructed from polyacrylamide gel with elastic moduli of 1.5 and 21.7 kPa, is based on serial block-face scanning electron microscopy images of the outflow pathway. It allows for quantifying 3D, depth-dependent, dynamic traction forces exerted by both normal and glaucomatous TM cells within an active fluid-structure interaction (FSI) environment. In our experimental design, we designed two scenarios: a control group with TM cells observed over 20 hours without flow (static setting), focusing on intrinsic cellular contractile forces, and a second scenario incorporating active FSI to evaluate its impact on traction forces (dynamic setting). Our observations revealed that active FSI results in higher traction forces (normal: 1.83-fold and glaucoma: 2.24-fold) and shear strains (normal: 1.81-fold and glaucoma: 2.41-fold), with stiffer substrates amplifying this effect. Glaucomatous cells consistently exhibited larger forces than normal cells. Increasing gel stiffness led to enhanced stress fiber formation in TM cells, particularly in glaucomatous cells. Exposure to active FSI dramatically altered actin organization in both normal and glaucomatous TM cells, particularly affecting cortical actin stress fiber arrangement. This model while preliminary offers a new method in understanding TM cell biomechanics and ECM stiffening in glaucoma, highlighting the importance of FSI in these processes.Statement of Significance: This pioneering project presents an advanced 3D in vitro model, meticulously replicating the human trabecular meshwork's anatomy for glaucoma research. It enables precise quantification of cellular forces in a dynamic fluid-structure interaction, a leap forward from existing 2D models. This advancement promises significant insights into trabecular meshwork cell biomechanics and the stiffening of the extracellular matrix in glaucoma, offering potential pathways for innovative treatments. This research is positioned at the forefront of ocular disease study, with implications that extend to broader biomedical applications.
Corneal disease is the fifth leading cause of global blindless. Optical coherence tomography (OCT) for anterior imaging is extensively used 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 visible-light OCM as a low-cost and easily reproducible system to visualize various corneal cellular structures such as epithelial cells, endothelial cells, keratocytes, and collagen bundles within stromal lamellae. The visible-light OCM was also used to study pressure changes in anterior segment human donor eyes. The 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 up to 750 μm × 750 μm.
Corneal diseases are the fifth leading cause of visual loss globally. Current clinical imaging instruments such as In Vivo Confocal Microscopy (IVCM) offer high lateral resolution to observe cellular structures but lack large field of view and volumetric imaging capability and require high operator skill to focus and align. To overcome these limitations, we have developed blue (450 nm) and green (510 nm) light Optical Coherence Microscopy (OCM) to image cellular structures. Imaging was demonstrated in ex vivo samples including human donor eyes. The OCM systems were based on a spectral-domain optical coherence tomography engine and achieved 750 μm × 750 µm field of view, 1.2 mm imaging depth, and 1.6 μm lateral resolution. Epithelial cells, endothelial cells, and keratocytes of ex vivo rabbit cornea were visualized. Additionally, collagen fibers were observed in stromal lamellae with striated patterns. En face and cross-sectional images of trabecular meshwork and Schlemm’s canal in a donor human eye wedge were observed at various trans-meshwork pressures controlled by cannulation of the canal. Microbes such as filamentous fungi and bacteria were observed.
Purpose: To assess the repeatability and accuracy of corneal astigmatism measurement with a spectral-domain optical coherence tomography (OCT) system (Avanti, Optovue) and compare them with Scheimpflug imaging (Pentacam HR, Oculus) and swept-source optical biometry (IOLMaster 700, Carl Zeiss Meditec AG). Setting: Casey Eye Institute, Oregon Health & Science University, Portland, Oregon. Design: Prospective cross-sectional observational study. Methods: 60 pseudophakic eyes with monofocal nontoric intraocular lens that previously had refractive surgery were analyzed. To assess accuracy, simulated keratometry (SimK) and net corneal astigmatism, obtained from each device, were compared with subjective manifest refraction astigmatism. Repeatability for corneal astigmatism was assessed for OCT and Pentacam HR by the coefficient of repeatability from 3 repeated measures. Results: Compared with manifest refraction, SimK readings produced with-the-rule astigmatic bias that was reduced for net astigmatism for the 3 devices. Except for OCT net astigmatism, all instruments significantly overestimated the magnitude of the astigmatism (linear mixed-effects model [LMM], P < .05). OCT net astigmatism showed the highest accuracy for manifest astigmatism prediction with the smaller 95% confidence ellipse for the mean difference vector. OCT net mean absolute difference was 0.57 diopters (D), significantly smaller than that of the other modalities (LMM, P < .05). Net corneal astigmatism measured with OCT showed the best repeatability (coefficient of repeatability = 0.29 D). Conclusions: OCT has the capability to measure net corneal astigmatism with higher precision and accuracy than Pentacam HR Scheimpflug imaging and IOLMaster 700 swept-source optical biometry in postrefractive subjects.
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.
PURPOSE:To demonstrate a noncontact device to enrich oxygen concentration during corneal cross-linking (CXL).METHODS:An oxygen delivery device was tested in a laboratory mock-up. The device comprises a clear polycarbonate tube of 14 cm in length and 1.58 cm inner diameter. Compressed oxygen gas is delivered to the tube from a side opening. The oximeter was attached to a sampling tube 3 mm above the apex of a scleral lens that simulates the cornea. The lens was mounted on a mannequin face. During each experimental run, the oximeter reading was recorded manually every 30 seconds for 4.5 minutes after the flow regulator was opened to the preset flow rate. Three flow rates of 0.25, 0.50, and 1 L/min were tested with all three cornea-tube distances of 8, 10, and 14 mm.RESULTS:The baseline oxygen concentration was 20.9%. The oxygen concentration reached plateau levels after 2 to 3.5 minutes. Oxygen measurements were averaged over the three time points in the plateau phase between 3.5 and 4.5 minutes. Atmospheric oxygen concentration above the simulated cornea was found to be strongly dependent on the oxygen flow rate up to 1 L/min. At the 1 L/min flow rate, 99% concentration was achieved at 8 to 10 mm of cornea-tube distances, and dropped to 90% at 14 mm.CONCLUSIONS:Atmospheric oxygen concentration can be boosted to more than 90% using a noncontact device. This could potentially improve the effectiveness of accelerated CXL by boosting oxygen transport more than fourfold. [J Refract Surg. 2020;36(9):613-616.].
Objective: To detect non-exudative choroidal neovascularization (CNV) in age-related macular degeneration with optical coherence tomography angiography (OCTA) and determine risk of developing exudative CNV compared to eyes without non-exudative CNV. Design: Prospective longitudinal observational study Participants: Consecutive patients with drusen and pigmentary changes in the study eye and exudative neovascular AMD in the fellow eye. Methods: Study participants underwent spectral domain OCTA (AngioVue, Optovue, Inc), clinical exam, and structural OCT at baseline and six-month intervals for two years. OCTA images were exported for custom processing to remove projection artifact and calculate CNV vessel area. Main Outcome: Rate of developing exudation in eyes with and without non-exudative CNV as detected by OCTA on regular follow-up. Results: Sixty-three prospective study participants were followed every 6 months and 48 completed the 2-year study. Mean age was 78 years and 60.3% were female. On the baseline visit, 5 eyes (7.9%) were found to have non-exudative CNV by OCTA and 3 of them developed exudation. Over the 2 years of follow-up, 5 more eyes developed non-exudative CNV on a follow-Corresponding up visit and all of them developed exudation. Overall, 8 of the 10 eyes with non-exudative CNV developed exudation with a mean time of 8 months and a mean CNV area growth rate of 20%/ month (exponential model, p=0.014). Initiation of anti-angiogenic treatment halted their growth. In comparison, exudation occurred in only 6 of the 53 eyes in which no non-exudative CNV were detected. Cox proportional hazard analysis showed that having a non-exudative CNV detected was associated with 18.1-fold increase in the rate of subsequently developing exudation (P<0.0001). Conclusions: Non-exudative CNVs are frequently detected by OCTA in the fellow eyes of exudative CNV. These lesions carry a high risk of developing exudation within the first year after detection and could benefit from close monitoring. The high risk of progression may justify prophylactic treatment; further studies are needed.
Purpose: To detect nonexudative choroidal neovascularization (CNV) in age-related macular degeneration (AMD) with OCT angiography (OCTA) and determine the risk of exudative CNV developing compared with eyes without nonexudative CNV. Design: Prospective, longitudinal, observational study. Participants: Consecutive patients with drusen and pigmentary changes in the study eye and exudative neovascular AMD in the fellow eye. Methods: In this prospective observational study, participants underwent spectral-domain OCTA (AngioVue; Optovue, Inc, Fremont, CA), clinical examination, and structural OCT at baseline and 6-month intervals for 2 years. OCT angiography images were exported for custom processing to remove projection artifact and calculate CNV vessel area. Main Outcome Measures: Rate of developing exudation in eyes with and without nonexudative CNV as detected by OCTA on regular follow-up. Results: Sixty-three study participants were followed up every 6 months and 48 completed the 2-year study. Mean age was 78 years and 60.3% were female. On the baseline visit, 5 eyes (7.9%) were found to have nonexudative CNV by OCTA, and 3 of them demonstrated exudation. Of 58 eyes with a normal OCTA on baseline visit, 5 eyes developed nonexudative CNV during a follow-up visit. All 5 of these nonexudative CNV went on to develop exudation in subsequent visits. Overall, 8 of the 10 eyes with nonexudative CNV developed exudation with a mean time of 8 months and mean CNV area growth rate of 20% per month (P = 0.014, exponential model). Initiation of antiangiogenic treatment halted their growth. In comparison, exudation occurred in only 6 of the 53 eyes (11%) that lacked a precursor nonexudative CNV. Cox proportional hazard analysis showed that having nonexudative CNV detected was associated with an 18.1-fold increase in the rate of exudation subsequently developing (P < 0.0001). Conclusions: Nonexudative CNV frequently is detected by OCTA in the fellow eyes of those with exudative CNV. These lesions carry a high risk of exudation developing within the first year after detection and could benefit from close monitoring. The high risk of progression may justify prophylactic treatment; further studies are needed. (C) 2019 by the American Academy of Ophthalmology