Effects of Low-Level Light Therapy on Resting-State Connectivity Following Moderate Traumatic Brain Injury: Secondary Analyses of a Double-blinded Placebo-controlled Study.

Background Low-level light therapy (LLLT) has been shown to modulate recovery in patients with traumatic brain injury (TBI). However, the impact of LLLT on the functional connectivity of the brain when at rest has not been well studied. Purpose To use functional MRI to assess the effect of LLLT on whole-brain resting-state functional connectivity (RSFC) in patients with moderate TBI at acute (within 1 week), subacute (2-3 weeks), and late-subacute (3 months) recovery phases. Materials and Methods This is a secondary analysis of a prospective single-site double-blinded sham-controlled study conducted in patients presenting to the emergency department with moderate TBI from November 2015 to July 2019. Participants were randomized for LLLT and sham treatment. The primary outcome of the study was to assess structural connectivity, and RSFC was collected as the secondary outcome. MRI was used to measure RSFC in 82 brain regions in participants during the three recovery phases. Healthy individuals who did not receive treatment were imaged at a single time point to provide control values. The Pearson correlation coefficient was estimated to assess the connectivity strength for each brain region pair, and estimates of the differences in Fisher z-transformed correlation coefficients (hereafter, z differences) were compared between recovery phases and treatment groups using a linear mixed-effects regression model. These analyses were repeated for all brain region pairs. False discovery rate (FDR)-adjusted P values were computed to account for multiple comparisons. Quantile mixed-effects models were constructed to quantify the association between the Rivermead Postconcussion Symptoms Questionnaire (RPQ) score, recovery phase, and treatment group. Results RSFC was evaluated in 17 LLLT-treated participants (median age, 50 years [IQR, 25-67 years]; nine female), 21 sham-treated participants (median age, 50 years [IQR, 43-59 years]; 11 female), and 23 healthy control participants (median age, 42 years [IQR, 32-54 years]; 13 male). Seven brain region pairs exhibited a greater change in connectivity in LLLT-treated participants than in sham-treated participants between the acute and subacute phases (range of z differences, 0.37 [95% CI: 0.20, 0.53] to 0.45 [95% CI: 0.24, 0.67]; FDR-adjusted P value range, .010-.047). Thirteen different brain region pairs showed an increase in connectivity in sham-treated participants between the subacute and late-subacute phases (range of z differences, 0.17 [95% CI: 0.09, 0.25] to 0.26 [95% CI: 0.14, 0.39]; FDR-adjusted P value range, .020-.047). There was no evidence of a difference in clinical outcomes between LLLT-treated and sham-treated participants (range of differences in medians, -3.54 [95% CI: -12.65, 5.57] to -0.59 [95% CI: -7.31, 8.49]; P value range, .44-.99), as measured according to RPQ scores. Conclusion Despite the small sample size, the change in RSFC from the acute to subacute phases of recovery was greater in LLLT-treated than sham-treated participants, suggesting that acute-phase LLLT may have an impact on resting-state neuronal circuits in the early recovery phase of moderate TBI. ClinicalTrials.gov Identifier: NCT02233413 © RSNA, 2024 Supplemental material is available for this article.

Experimental and theoretical model of microvascular network remodeling and blood flow redistribution following minimally invasive microvessel laser ablation.

Overly dense microvascular networks are treated by selective reduction of vascular elements. Inappropriate manipulation of microvessels could result in loss of host tissue function or a worsening of the clinical problem. Here, experimental, and computational models were developed to induce blood flow changes via selective artery and vein laser ablation and study the compensatory collateral flow redistribution and vessel diameter remodeling. The microvasculature was imaged non-invasively by bright-field and multi-photon laser microscopy, and optical coherence tomography pre-ablation and up to 30 days post-ablation. A theoretical model of network remodeling was developed to compute blood flow and intravascular pressure and identify vessels most susceptible to changes in flow direction. The skin microvascular remodeling patterns were consistent among the five specimens studied. Significant remodeling occurred at various time points, beginning as early as days 1-3 and continuing beyond day 20. The remodeling patterns included collateral development, venous and arterial reopening, and both outward and inward remodeling, with variations in the time frames for each mouse. In a representative specimen, immediately post-ablation, the average artery and vein diameters increased by 14% and 23%, respectively. At day 20 post-ablation, the maximum increases in arterial and venous diameters were 2.5× and 3.3×, respectively. By day 30, the average artery diameter remained 11% increased whereas the vein diameters returned to near pre-ablation values. Some arteries regenerated across the ablation sites via endothelial cell migration, while veins either reconnected or rerouted flow around the ablation site, likely depending on local pressure driving forces. In the intact network, the theoretical model predicts that the vessels that act as collaterals after flow disruption are those most sensitive to distant changes in pressure. The model results correlate with the post-ablation microvascular remodeling patterns.

Needle guidance with Doppler-tracked polarization-sensitive optical coherence tomography.

Significance: Optical coherence tomography (OCT) can be integrated into needle probes to provide real-time navigational guidance. However, unscanned implementations, which are the simplest to build, often struggle to discriminate the relevant tissues. Aim: We explore the use of polarization-sensitive (PS) methods as a means to enhance signal interpretability within unscanned coherence tomography probes. Approach: Broadband light from a laser centered at 1310 nm was sent through a fiber that was embedded into a needle. The polarization signal from OCT fringes was combined with Doppler-based tracking to create visualizations of the birefringence properties of the tissue. Experiments were performed in (i) well-understood structured tissues (salmon and shrimp) and (ii) ex vivo porcine spine. The porcine experiments were selected to illustrate an epidural guidance use case. Results: In the porcine spine, unscanned and Doppler-tracked PS OCT imaging data successfully identified the skin, subcutaneous tissue, ligament, and epidural spaces during needle insertion. Conclusions: PS imaging within a needle probe improves signal interpretability relative to structural OCT methods and may advance the clinical utility of unscanned OCT needle probes in a variety of applications.

Needle guidance with Doppler-tracked polarization-sensitive optical coherence tomography.

We demonstrate that a simple, unscanned polarization-sensitive optical coherence tomography needle probe can be used to perform layer identification in biological tissues. Broadband light from a laser centered at 1310 nm was sent through a fiber that was embedded into a needle, and analysis of the polarization state of the returning light after interference coupled with Doppler-based tracking allowed the calculation of phase retardation and optic axis orientation at each needle location. Proof-of-concept phase retardation mapping was shown in Atlantic salmon tissue, while axis orientation mapping was demonstrated in white shrimp tissue. The needle probe was then tested on the ex vivo porcine spine, where mock epidural procedures were performed. Our imaging results demonstrate that unscanned, Doppler-tracked polarization-sensitive optical coherence tomography imaging successfully identified the skin, subcutaneous tissue, and ligament layers, before successfully reaching the target of the epidural space. The addition of polarization-sensitive imaging into the bore of a needle probe therefore allows layer identification at deeper locations in the tissue.

Development of polarization-sensitive optical coherence tomography imaging platform and metrics to quantify electrostimulation-induced peripheral nerve injury in vivo in a small animal model.

Significance: Neuromodulation devices are rapidly evolving for the treatment of neurological diseases and conditions. Injury from implantation or long-term use without obvious functional losses is often only detectable through terminal histology. New technologies are needed that assess the peripheral nervous system (PNS) under normal and diseased or injured conditions. Aim: We aim to demonstrate an imaging and stimulation platform that can elucidate the biological mechanisms and impacts of neurostimulation in the PNS and apply it to the sciatic nerve to extract imaging metrics indicating electrical overstimulation. Approach: A sciatic nerve injury model in a 15-rat cohort was observed using a newly developed imaging and stimulation platform that can detect electrical overstimulation effects with polarization-sensitive optical coherence tomography. The sciatic nerve was electrically stimulated using a custom-developed nerve holder with embedded electrodes for 1 h, followed by a 1-h recovery period, delivered at above-threshold Shannon model k -values in experimental groups: sham control (SC, n = 5 , 0.0 mA / 0 Hz ), stimulation level 1 (SL1, n = 5 , 3.4 mA / 50 Hz , and k = 2.57 ), and stimulation level 2 (SL2, n = 5 , 6.8 mA / 100 Hz , and k = 3.17 ). Results: The stimulation and imaging system successfully captured study data across the cohort. When compared to a SC after a 1-week recovery, the fascicle closest to the stimulation lead showed an average change of + 4 % / - 309 % (SL1/SL2) in phase retardation and - 79 % / - 148 % in optical attenuation relative to SC. Analysis of immunohistochemistry (IHC) shows a + 1 % / - 36 % difference in myelin pixel counts and - 13 % / + 29 % difference in axon pixel counts, and an overall increase in cell nuclei pixel count of + 20 % / + 35 % . These metrics were consistent with IHC and hematoxylin/eosin tissue section analysis. Conclusions: The poststimulation changes observed in our study are manifestations of nerve injury and repair, specifically degeneration and angiogenesis. Optical imaging metrics quantify these processes and may help evaluate the safety and efficacy of neuromodulation devices.

Feature issue introduction: ultrafast optical imaging.

This feature issue of Optics Express collects 20 articles that report the most recent progress of ultrafast optical imaging. This review provides a summary of these articles that cover the spectrum of ultrafast optical imaging, from new technologies to applications.

Improved in vivo optical coherence tomography imaging of animal peripheral nerves using a prism nerve holder.

Significance: Modern optical volumetric imaging modalities, such as optical coherence tomography (OCT), provide enormous information about the structure, function, and physiology of living tissue. Although optical imaging achieves lateral resolution on the order of the wavelength of light used, and OCT achieves axial resolution on a similar micron scale, tissue optical properties, particularly high scattering and absorption, limit light penetration to only a few millimeters. In addition, in vivo imaging modalities are susceptible to significant motion artifacts due to cardiac and respiratory function. These effects limit access to artifact-free optical measurements during peripheral neurosurgery to only a portion of the exposed nerve without further modification to the procedure. Aim: We aim to improve in vivo OCT imaging during peripheral neurosurgery in small and large animals by increasing the amount of visualized nerve volume as well as suppressing motion of the imaged area. Approach: We designed a nerve holder with embedded mirror prisms for peripheral nerve volumetric imaging as well as a specific beam steering strategy to acquire prism and direct view volumes in one session with minimal motion artifacts. Results: The axially imaged volumes from mirror prisms increased the OCT signal intensity by > 22 dB over a 1.25-mm imaging depth in tissue-mimicking phantoms. We then demonstrated the new imaging capabilities in visualizing peripheral nerves from direct and side views in living rats and minipigs using a polarization-sensitive OCT system. Prism views have shown nerve fascicles and vasculature from the bottom half of the imaged nerve which was not visible in direct view. Conclusions: We demonstrated improved OCT imaging during neurosurgery in small and large animals by combining the use of a prism nerve holder with a specifically designed beam scanning protocol. Our strategy can be applied to existing OCT imaging systems with minimal hardware modification, increasing the nerve tissue volume visualized. Enhanced imaging depth techniques may lead to a greater adoption of structural and functional optical biomarkers in preclinical and clinical medicine.

Wide-Field Three-Dimensional Depth-Invariant Cellular-Resolution Imaging of the Human Retina.

Three-dimensional (3D) cellular-resolution imaging of the living human retina over a large field of view will bring a great impact in clinical ophthalmology, potentially finding new biomarkers for early diagnosis and improving the pathophysiological understanding of ocular diseases. While hardware-based and computational adaptive optics (AO) optical coherence tomography (OCT) have been developed to achieve cellular-resolution retinal imaging, these approaches support limited 3D imaging fields, and their high cost and intrinsic hardware complexity limit their practical utility. Here, this work demonstrates 3D depth-invariant cellular-resolution imaging of the living human retina over a 3 × 3 mm field of view using the first intrinsically phase-stable multi-MHz retinal swept-source OCT and novel computational defocus and aberration correction methods. Single-acquisition imaging of photoreceptor cells, retinal nerve fiber layer, and retinal capillaries is presented across unprecedented imaging fields. By providing wide-field 3D cellular-resolution imaging in the human retina using a standard point-scan architecture routinely used in the clinic, this platform proposes a strategy for expanded utilization of high-resolution retinal imaging in both research and clinical settings.

Experimental and Theoretical Model of Single Vessel Minimally Invasive Micro-Laser Ablation: Inducing Microvascular Network Remodeling and Blood Flow Redistribution Without Compromising Host Tissue Function.

Overly dense microvascular networks are treated by selective reduction of vascular elements. Inappropriate manipulation of microvessels could result in loss of host tissue function or a worsening of the clinical problem. Here, experimental, and computational models were developed to induce blood flow changes via selective artery and vein laser ablation and study the compensatory collateral flow redistribution and vessel diameter remodeling. The microvasculature was imaged non-invasively by bright-field and multi-photon laser microscopy, and Optical Coherence Tomography pre-ablation and up to 30 days post-ablation. A theoretical model of network remodeling was developed to compute blood flow and intravascular pressure and identify vessels most susceptible to changes in flow direction. The skin microvascular remodeling patterns were consistent among the five specimens studied. Significant remodeling occurred at various time points, beginning as early as days 1-3 and continuing beyond day 20. The remodeling patterns included collateral development, venous and arterial reopening, and both outward and inward remodeling, with variations in the time frames for each mouse. In a representative specimen, immediately post-ablation, the average artery and vein diameters increased by 14% and 23%, respectively. At day 20 post-ablation, the maximum increases in arterial and venous diameters were 2.5x and 3.3x, respectively. By day 30, the average artery diameter remained 11% increased whereas the vein diameters returned to near pre-ablation values. Some arteries regenerated across the ablation sites via endothelial cell migration, while veins either reconnected or rerouted flow around the ablation site, likely depending on local pressure driving forces. In the intact network, the theoretical model predicts that the vessels that act as collaterals after flow disruption are those most sensitive to distant changes in pressure. The model results match the post-ablation microvascular remodeling patterns.

Using the dynamic forward scattering signal for optical coherence tomography based blood flow quantification.

To our knowledge, all existing optical coherence tomography approaches for quantifying blood flow, whether Doppler-based or decorrelation-based, analyze light that is back-scattered by moving red blood cells (RBCs). This work investigates the potential advantages of basing these measurements on light that is forward-scattered by RBCs, i.e., by looking at the signals back-scattered from below the vessel. We show experimentally that flowmetry based on forward-scattering is insensitive to vessel orientation for vessels that are approximately orthogonal to the imaging beam. We further provide proof-of-principle demonstrations of dynamic forward-scattering (DFS) flowmetry in human retinal and choroidal vessels.

Local vasoregulative interventions impact drug concentrations in the skin after topical laser-assisted delivery.

INTRODUCTION: The ability of ablative fractional lasers (AFL) to enhance topical drug uptake is well established. After AFL delivery, however, drug clearance by local vasculature is poorly understood. Modifications in vascular clearance may enhance AFL-assisted drug concentrations and prolong drug dwell time in the skin. Aiming to assess the role and modifiability of vascular clearance after AFL-assisted delivery, this study examined the impact of vasoregulative interventions on AFL-assisted 5-fluorouracil (5-FU) concentrations in in vivo skin. METHODS: 5-FU uptake was assessed in intact and AFL-exposed skin in a live pig model. After fractional CO2 laser exposure (15 mJ/microbeam, 5% density), vasoregulative intervention using topical brimonidine cream, epinephrine solution, or pulsed dye laser (PDL) was performed in designated treatment areas, followed by a single 5% 5-FU cream application. At 0, 1, 4, 48, and 72 h, 5-FU concentrations were measured in 500 and 1500 µm skin layers by mass spectrometry (n = 6). A supplemental assessment of blood flow following AFL ± vasoregulation was performed using optical coherence tomography (OCT) in a human volunteer. RESULTS: Compared to intact skin, AFL facilitated a prompt peak in 5-FU delivery that remained elevated up to 4 hours (1500 µm: 1.5 vs. 31.8 ng/ml [1 hour, p = 0.002]; 5.3 vs. 14.5 ng/ml [4 hours, p = 0.039]). However, AFL's impact was transient, with 5-FU concentrations comparable to intact skin at later time points. Overall, vasoregulative intervention with brimonidine or PDL led to significantly higher peak 5-FU concentrations, prolonging the drug's dwell time in the skin versus AFL delivery alone. As such, brimonidine and PDL led to twofold higher 5-FU concentrations than AFL alone in both skin layers by 1 hour (e.g., 500 µm: 107 ng/ml [brimonidine]; 96.9 ng/ml [PDL], 46.6 ng/ml [AFL alone], p ≤ 0.024), and remained significantly elevated at 4 hours (p ≤ 0.024). A similar pattern was observed for epinephrine, although trends remained nonsignificant (p ≥ 0.09). Prolonged 5-FU delivery was provided by PDL, resulting in sustained drug deposition compared to AFL alone at both 48 and 72 hours in the superficial skin layer (p ≤ 0.024). Supporting drug delivery findings, OCT revealed that increases in local blood flow after AFL were mitigated in test areas also exposed to PDL, brimonidine, or epinephrine, with PDL providing the greatest, sustained reduction in flow over 48 hours. CONCLUSION: Vasoregulative intervention in conjunction with AFL-assisted delivery enhances and prolongs 5-FU deposition in in vivo skin.

RF properties of circular-ranging OCT signals.

Circular-ranging optical coherence tomography (CR-OCT) systems that use a time-stepped frequency comb source generate interference fringe signals that are more complex than those of a conventional swept-source OCT system. Here, we define a common terminology for describing these signals, and we develop a mathematical framework that relates the radio-frequency (RF) properties of these fringe signals to the parameters of the frequency comb source. With this framework, we highlight non-intuitive mechanisms whereby the design of the frequency comb source can affect imaging performance. We show, for example, that amplitude-pulsed time-stepped frequency comb sources have a sensitivity advantage over constant power time-stepped frequency comb sources. More broadly, this framework and associated terminology provide a foundation on which to design and optimize time-stepped frequency comb sources and systems.

Relationship between axial resolution and signal-to-noise ratio in optical coherence tomography.

In optical coherence tomography (OCT), axial resolution and signal-to-noise ratio (SNR) are typically viewed as uncoupled parameters. We show that this is true only for mirror-like surfaces and that in diffuse scattering samples such as biological tissues there is an inherent coupling between axial resolution and measurement SNR. We explain the origin of this coupling and demonstrate that it can be used to achieve increased imaging penetration depth at the expense of resolution. Finally, we argue that this coupling should be considered during OCT system design processes that seek to balance the competing needs of resolution, sensitivity, and system/source complexity.

Robust and easy-to-operate stretched-pulse mode-locked wavelength-swept laser with an all-polarization-maintaining fiber cavity for 10 MHz A-line rate optical coherence tomography.

We demonstrate robust and easy-to-operate stretched-pulse mode-locked laser (SPML) architectures using all-polarization-maintaining fiber laser cavities. Because of the polarization-maintaining construction, the laser performance is unaffected by mechanical perturbation on the cavity fibers. The lasers automatically initiate linear-in-wavenumber sweeps across 100 nm centered at 1290 nm with a 10 MHz repetition rate. OCT imaging with a sensitivity of 98 dB and a single-sided 6 dB coherence length of 2.5 mm is demonstrated. OCT angiography of a mouse brain that visualized three-dimensional cerebral microvasculature over a field of 1.5mm×1.5mm (398 A-lines × 380 B-scans) at a rate of 5.26 volumes per second is also presented. The robust all-PMF SPML lasers are a turnkey, high-performance source for ultrahigh-speed OCT imaging.

Solid stress impairs lymphocyte infiltration into lymph-node metastases.

Strong and durable anticancer immune responses are associated with the generation of activated cancer-specific T cells in the draining lymph nodes. However, cancer cells can colonize lymph nodes and drive tumour progression. Here, we show that lymphocytes fail to penetrate metastatic lesions in lymph nodes. In tissue from patients with breast, colon, and head and neck cancers, as well as in mice with spontaneously developing breast-cancer lymph-node metastases, we found that lymphocyte exclusion from nodal lesions is associated with the presence of solid stress caused by lesion growth, that solid stress induces reductions in the number of functional high endothelial venules in the nodes, and that relieving solid stress in the mice increased the presence of lymphocytes in lymph-node lesions by about 15-fold. Solid-stress-mediated impairment of lymphocyte infiltration into lymph-node metastases suggests a therapeutic route for overcoming T-cell exclusion during immunotherapy.

Multi-beam OCT imaging based on an integrated, free-space interferometer.

While it is a common practice to increase the speed of swept-source optical coherence tomography (OCT) systems by using a high-speed source, this approach may not always be optimal. Parallelization in the form of multiple imaging beams is an alternative approach, but scalable and low-loss multi-beam OCT architectures are needed to capitalize on its advantages. In this study, we demonstrate an eight-beam OCT system using an interferometer architecture comprising planar lightwave circuits (PLC) splitters, V-groove assemblies (VGA), and optical ribbon fibers. We achieved an excess loss and heterodyne efficiency on each channel that was close to that of single-beam systems. In vivo structural imaging of a human finger and OCT angiography imaging of a mouse ear was performed to demonstrate the imaging performance of the system. This work provides further evidence supporting multi-beam architectures as a viable strategy for increasing OCT imaging speed.

Stepped frequency comb generation based on electro-optic phase-code mode-locking for moderate-speed circular-ranging OCT.

Circular-ranging (CR) optical coherence tomography (OCT) uses frequency comb sources to improve long-range imaging. While the initial development of CR-OCT focused on extremely high-speed imaging (i.e., operation at A-line rates of several to tens of MHz), there are many applications and imaging strategies for which more moderate speeds are preferred. However, we lack suitable frequency comb sources to enable moderate speed CR-OCT imaging. Here, we describe a novel phase-code mode-locking (PCML) laser architecture that can be operated from the kilohertz to megahertz range, while also offering novel features such as dynamic re-configurability and simplified linear-in-time frequency stepping. We demonstrate a prototype CR-OCT system with a PCML laser and present imaging results at A-line rates from 176 kHz to 3.52 MHz with coherence-length limited imaging depths as high as 170 mm.

Artifact Rates for 2D Retinal Nerve Fiber Layer Thickness Versus 3D Neuroretinal Rim Thickness Using Spectral-Domain Optical Coherence Tomography.

Purpose: To compare the rates of clinically significant artifacts for two-dimensional peripapillary retinal nerve fiber layer (RNFL) thickness versus three-dimensional (3D) neuroretinal rim thickness using spectral-domain optical coherence tomography (SD-OCT). Methods: Only one eye per patient was used for analysis of 120 glaucoma patients and 114 normal patients. For RNFL scans and optic nerve scans, 15 artifact types were calculated per B-scan and per eye. Neuroretinal rim tissue was quantified by the minimum distance band (MDB). Global MDB neuroretinal rim thicknesses were calculated before and after manual deletion of B-scans with artifacts and subsequent automated interpolation. A clinically significant artifact was defined as one requiring manual correction or repeat scanning. Results: Among glaucomatous eyes, artifact rates per B-scan were significantly more common in RNFL scans (61.7%, 74 of 120) compared to B-scans in neuroretinal rim volume scans (20.9%, 1423 of 6820) (95% confidence interval [CI], 31.6-50.0; P < 0.0001). For clinically significant artifact rates per eye, optic nerve scans had significantly fewer artifacts (15.8% of glaucomatous eyes, 13.2% of normal eyes) compared to RNFL scans (61.7% of glaucomatous eyes, 25.4% of normal eyes) (glaucoma group: 95% CI, 34.1-57.5, P < 0.0001; normal group: 95% CI, 1.3-23.3, P = 0.03). Conclusions: Compared to the most commonly used RNFL thickness scans, optic nerve volume scans less frequently require manual correction or repeat scanning to obtain accurate measurements. Translational Relevance: This paper illustrates the potential for 3D OCT algorithms to improve in vivo imaging in glaucoma.

Artifact Rates for 2D Retinal Nerve Fiber Layer Thickness Versus 3D Retinal Nerve Fiber Layer Volume.

Purpose: To compare artifact rates in two-dimensional (2D) versus three-dimensional (3D) retinal nerve fiber layer (RNFL) scans using Spectralis optical coherence tomography (OCT). Methods: Thirteen artifact types in 2D and 3D RNFL scans were identified in 106 glaucomatous eyes and 95 normal eyes. Artifact rates were calculated per B-scan and per eye. In 3D volume scans, artifacts were counted only for the 97 B-scans used to calculate RNFL parameters for the 2.5-3.5-mm annulus. 3D RNFL measurements were calculated twice, once before and again after deletion of B-scans with artifacts and subsequent automated interpolation. Results: For 2D scans, artifacts were present in 58.5% of B-scans (62 of 106) in glaucomatous eyes. For 3D scans, a mean of 35.4% of B-scans (34.3 of 97 B-scans per volume scan) contained an artifact in 106 glaucomatous eyes. For 3D data of glaucoma patients, mean global RNFL thickness values were similar before and after interpolation (77.0 ± 11.6 µm vs. 75.1 ± 11.2 µm, respectively; P = 0.23). Fewer clinically significant artifacts were noted in 3D RNFL scans, where only 7.5% of glaucomatous eyes (8 of 106) and 0% of normal eyes (0 of 95) had artifacts, compared to 2D RNFL scans, where 58.5% of glaucomatous eyes (62 of 106) and 14.7% of normal eyes (14 of 95) had artifacts. Conclusions: Compared to 2D RNFL scans, 3D RNFL volume scans less often require manual correction to obtain accurate measurements. Translational Relevance: 3D RNFL volume scans have fewer clinically significant artifacts compared to 2D RNFL thickness scans.

9.4 MHz A-line rate optical coherence tomography at 1300 nm using a wavelength-swept laser based on stretched-pulse active mode-locking.

In optical coherence tomography (OCT), high-speed systems based at 1300 nm are among the most broadly used. Here, we present 9.4 MHz A-line rate OCT system at 1300 nm. A wavelength-swept laser based on stretched-pulse active mode locking (SPML) provides a continuous and linear-in-wavenumber sweep from 1240 nm to 1340 nm, and the OCT system using this light source provides a sensitivity of 98 dB and a single-sided 6-dB roll-off depth of 2.5 mm. We present new capabilities of the 9.4 MHz SPML-OCT system in three microscopy applications. First, we demonstrate high quality OCTA imaging at a rate of 1.3 volumes/s. Second, by utilizing its inherent phase stable characteristics, we present wide dynamic range en face Doppler OCT imaging with multiple time intervals ranging from 0.25 ms to 2.0 ms at a rate of 0.53 volumes/s. Third, we demonstrate video-rate 4D microscopic imaging of a beating Xenopus embryo heart at a rate of 30 volumes/s. This high-speed and high-performance OCT system centered at 1300 nm suggests that it can be one of the most promising high-speed OCT platforms enabling a wide range of new scientific research, industrial, and clinical applications at speeds of 10 MHz.