Insights into Sickle Cell Disease through the Retinal Microvasculature: Adaptive Optics Scanning Light Ophthalmoscopy Correlates of Clinical OCT Angiography.

Purpose: Clinical OCT angiography (OCTA) of the retinal microvasculature offers a quantitative correlate to systemic disease burden and treatment efficacy in sickle cell disease (SCD). The purpose of this study was to use the higher resolution of adaptive optics scanning light ophthalmoscopy (AOSLO) to elucidate OCTA features of parafoveal microvascular compromise identified in SCD patients. Design: Case series of 11 SCD patients and 1 unaffected control. Participants: A total of 11 eyes of 11 SCD patients (mean age, 33 years; range, 23-44; 8 female, 3 male) and 1 eye of a 34-year-old unaffected control. Methods: Ten sequential 3 × 3 mm parafoveal OCTA full vascular slab scans were obtained per eye using a commercial spectral domain OCT system (Avanti RTVue-XR; Optovue). These were used to identify areas of compromised perfusion near the foveal avascular zone (FAZ), designated as regions of interest (ROIs). Immediately thereafter, AOSLO imaging was performed on these ROIs to examine the cellular details of abnormal perfusion. Each participant was imaged at a single cross-sectional time point. Additionally, 2 of the SCD patients were imaged prospectively 2 months after initial imaging to study compromised capillary segments across time and with treatment. Main Outcome Measures: Detection and characterization of parafoveal perfusion abnormalities identified using OCTA and resolved using AOSLO imaging. Results: We found evidence of abnormal blood flow on OCTA and AOSLO imaging among all 11 SCD patients with diverse systemic and ocular histories. Adaptive optics scanning light ophthalmoscopy imaging revealed a spectrum of phenomena, including capillaries with intermittent blood flow, blood cell stasis, and sites of thrombus formation. Adaptive optics scanning light ophthalmoscopy imaging was able to resolve single sickled red blood cells, rouleaux formations, and blood cell-vessel wall interactions. OCT angiography and AOSLO imaging were sensitive enough to document improved retinal perfusion in an SCD patient 2 months after initiation of oral hydroxyurea therapy. Conclusions: Adaptive optics scanning light ophthalmoscopy imaging was able to reveal the cellular details of perfusion abnormalities detected using clinical OCTA. The synergy between these clinical and laboratory imaging modalities presents a promising avenue in the management of SCD through the development of noninvasive ocular biomarkers to prognosticate progression and measure the response to systemic treatment.

Efficacy of CRISPR-Based Gene Editing in a Sickle Cell Disease Patient as Measured through the Eye.

Sickle cell disease (SCD) exists on a phenotypic spectrum with variable genetic expressivity, making it difficult to assess an individual patient's risk of complications at any particular point in time. Current and emerging SCD treatments, including CRISPR-based gene editing, result in a variable proportion of affected red blood cells (RBCs) still vulnerable to sickling. Clinical serological indicators of disease such as hemoglobin, indirect bilirubin, and reticulocyte count and clinical metrics including number of emergency department visits and hospitalizations over time often fall short in their ability to objectively quantify ischemic disease activity and efficacy of treatments. Clearly, better clinical biomarkers are needed. The rapidly developing field of oculomics leverages the transparent nature of the ocular tissue to directly study the retinal microvasculature in order to characterize the status of systemic diseases. In this case report, we demonstrate the ability of optical coherence tomography angiography (OCT-A) to detect and measure micro-occlusive events within the retinal capillary bed before and after RBC exchange transfusion and following CRISPR-based gene editing, as an indicator of systemic ischemic disease activity and measure of treatment efficacy. The implications of these findings are discussed.

Imaging of vitreous cortex hyalocyte dynamics using non-confocal quadrant-detection adaptive optics scanning light ophthalmoscopy in human subjects.

Vitreous cortex hyalocytes are resident macrophage cells that help maintain the transparency of the media, provide immunosurveillance, and respond to tissue injury and inflammation. In this study, we demonstrate the use of non-confocal quadrant-detection adaptive optics scanning light ophthalmoscopy (AOSLO) to non-invasively visualize the movement and morphological changes of the hyalocyte cell bodies and processes over 1-2 hour periods in the living human eye. The average velocity of the cells 0.52 ± 0.76 µm/min when sampled every 5 minutes and 0.23 ± 0.29 µm/min when sampled every 30 minutes, suggesting that the hyalocytes move in quick bursts. Understanding the behavior of these cells under normal physiological conditions may lead to their use as biomarkers or suitable targets for therapy in eye diseases such as diabetic retinopathy, preretinal fibrosis and glaucoma.

3-D OCT angiographic evidence of Anti-VEGF therapeutic effects on retinal capillary hemangioma.

PURPOSE: To report the impact of intravitreal anti-vascular endothelial growth factor (VEGF) therapy on a retinal capillary hemangioma (RCH) using clinical OCT angiography (OCT-A) in addition to standard imaging modalities. OBSERVATIONS: A 25-year-old male patient with Von Hippel-Lindau (VHL) disease presented with a history of bilateral RCH. No view was present in the right eye. Examination of the left eye revealed six peripheral RCH, the smallest of which was temporal to the macula with active exudation. This RCH was thought to be the source of cystoid macular edema (CME) involving the fovea, and therefore, the source of vision decline. 11 injections of 1.25mg of Bevacizumab EA across 14-month was given. Comparison of the pre- and post-treatment OCT-A at the temporal RCH showed a reduction of CME and regression of RCH. CONCLUSION: Anti-VEGF therapy appeared to stabilize the visual acuity and produce partial regression of RCH. It offers a safe option when visual acuity is threatened. OCT and OCT-A have the ability to document the impact of antiangiogenic therapy on RCH. 3D renderings of OCT-A offer enhanced sensitivity to recognition of structural and functional changes of RCH which may prove useful for monitoring treatment response.

Quantification of intermittent retinal capillary perfusion in sickle cell disease.

Pathophysiology of sickle cell disease (SCD) features intermittent vaso-occlusion of microcirculatory networks that facilitate ischemic damage. Past research has, however, relied on static images to characterize this active disease state. This study develops imaging metrics to more fully capture dynamic vascular changes, quantifying intermittent retinal capillary perfusion in unaffected controls and SCD patients using sequential optical coherence tomography angiography (OCT-A) scans. The results reveal significant dynamic variation of capillary perfusion in SCD patients compared to controls. This measurement of vaso-occlusive burden in patients would provide utility in monitoring of the disease state and in evaluating treatment efficacy.

Fundus albipunctatus photoreceptor microstructure revealed using adaptive optics scanning light ophthalmoscopy.

PURPOSE: Fundus albipunctatus is an inherited cause of congenital stationary night blindness. The objective of this report is to describe structural changes occurring in a macular phenotype of a novel RDH5 mutation producing fundus albipunctatus using high-resolution in vivo imaging. A 62-year-old male with longstanding night blindness underwent imaging and genetic evaluation. High-resolution images of the photoreceptor mosaic were compared to those of a healthy subject. Results of a comprehensive ophthalmic evaluation and genetic testing with imaging including fundus photography, spectral-domain optical coherence tomography (OCT), fluorescein angiography (FA), OCT angiography (OCT-A), and adaptive optics scanning light ophthalmoscopy (AOSLO) are described. OBSERVATIONS: The patient presented with visual acuity of 20/25 in both eyes and longstanding poor dark adaptation. Anterior segment examination was unremarkable. Fundoscopy revealed well circumscribed bilateral perifoveal mottling and atrophy in both eyes. Discrete white-yellow flecks were present beyond the vascular arcades extending to the far periphery. Genetic testing revealed a novel compound heterozygous RDH5 mutation (c.388C > T, p.Gln130*; c.665T > C, p.Leu222Pro). OCT demonstrated perifoveal photoreceptor and outer retinal irregularities, which corresponded to a window defect with late staining on FA. OCT-A demonstrated normal retinal vasculature with patchy areas of non-perfusion in the choriocapillaris. Macular abnormalities in both eyes were imaged using AOSLO to assess cone and rod photoreceptor architecture. While clinical features are consistent with a primary rod disorder, confocal AOSLO showed a paucity of normal cones with a small spared central island in both eyes. Rods appeared larger and more irregular throughout the macula. Non-confocal split detection AOSLO imaging revealed the presence of cone inner segments in dark regions of confocal imaging, indicating some degree of photoreceptor preservation. CONCLUSIONS AND IMPORTANCE: The AOSLO imaging of this particular macular phenotype of fundus albipunctatus demonstrates some of the structural photoreceptor abnormalities that occur in this condition, adding insight to the variable presentation of RDH5 retinopathy. The presence of preserved inner segment architecture suggests the possibility that gene therapy could play a future role in treating this condition.

Imaging of Macrophage-Like Cells in Living Human Retina Using Clinical OCT.

Purpose: To image retinal macrophages at the vitreoretinal interface in the living human retina using a clinical optical coherence tomography (OCT) device. Methods: Eighteen healthy controls and three patients with retinopathies were imaged using a clinical spectral-domain OCT. In controls, 10 sequential scans were collected at three different locations: (1) ∼9 degrees temporal to the fovea, (2) the macula, and (3) the optic nerve head (ONH). Intervisit repeatability was evaluated by imaging the temporal retina twice on the same day and 3 days later. Only 10 scans at the temporal retina were obtained from each patient. A 3-µm OCT reflectance (OCT-R) slab located above the inner limiting membrane (ILM) surface was averaged. Results: In controls, ramified macrophage-like cells with regular spatial separation were visualized in the temporal and ONH OCT-R images; however, cell structures were not resolvable at the macula. Interim changes in cell position suggestive of cell translocation were observed between images collected on the same day and those collected 3 days later. There was considerable variation in cell density and nearest-neighbor distance (NND) across controls. Mean ± SD cell densities measured at the temporal and ONH were 78 ± 23 cells/mm2 and 57 ± 16 cells/mm2, respectively. Similarly, mean ± SD NNDs measured at the temporal and ONH were 74.3 ± 13.3 µm and 93.3 ± 20.0 µm, respectively. Nonuniform spatial distribution and altered morphology of the cells were identified in patients with retinopathies. Conclusions: Our findings showed regular spatial separation and ramified morphology of macrophage-like cells on the ILM surface with cell translocation over time in controls. Their distribution and morphology suggest an origin of macrophage-like cells such as microglia or hyalocytes.

Automated quantification of choriocapillaris anatomical features in ultrahigh-speed optical coherence tomography angiograms.

In vivo visualization and quantification of choriocapillaris vascular anatomy is a fundamental step in understanding the relation between choriocapillaris degradation and atrophic retinopathies, including geographic atrophy. We describe a process utilizing ultrahigh-speed swept-source optical coherence tomography and a custom-designed "local min-max normalized masking" algorithm to extract in vivo anatomical metrics of the choriocapillaris. We used a swept-source optical coherence tomography system with a 1.6 MHz A-scan rate to image healthy retinas. With the postprocessing algorithm, we reduced noise, optimized visibility of vasculature, and skeletonized the vasculature within the images. These skeletonizations were in 89 % agreement with those made by skilled technicians and were, on average, completed in 18.6 s as compared to the 5.6 h technicians required. Anatomy within the processed images and skeletonizations was analyzed to identify average values ( mean ± SD ) of flow void radius ( 9.8 ± 0.7 µm ), flow void area ( 749 ± 110 µm 2 ), vessel radius ( 5.0 ± 0.3 µm ), branch-point to branch-point vessel length ( 26.8 ± 1.1 µm ), and branches per branch-point ( 3.1 ± 0.1 ) . To exemplify the uses of this tool a retina with geographic atrophy was imaged and processed to reveal statistically significant ( p < 0.05 ) increases in flow void radii and decreases in vessel radii under atrophic lesions as compared to atrophy-free regions on the same retina. Our results demonstrate a new avenue for quantifying choriocapillaris anatomy and studying vasculature changes in atrophic retinopathies.

Vascular Response to Sildenafil Citrate in Aging and Age-Related Macular Degeneration.

Age-related macular degeneration (AMD) - the leading cause of vision loss in the elderly - share many risks factors as atherosclerosis, which exhibits loss of vascular compliance resulting from aging and oxidative stress. Here, we attempt to explore choroidal and retinal vascular compliance in patients with AMD by evaluating dynamic vascular changes using live ocular imaging following treatment with oral sildenafil citrate, a phosphodiesterase type 5 (PDE5) inhibitor and potent vasodilator. Enhanced-depth imaging optical coherence tomography (EDI-OCT) and OCT angiography (OCT-A) were performed on 46 eyes of 23 subjects, including 15 patients with non-exudative AMD in one eye and exudative AMD in the fellow eye, and 8 age-matched control subjects. Choroidal thickness, choroidal vascularity, and retinal vessel density were measured across the central macula at 1 and 3 hours after a 100 mg oral dose of sildenafil citrate. Baseline choroidal thickness was 172.1 ± 60.0 µm in non-exudative AMD eyes, 196.4 ± 89.8 µm in exudative AMD eyes, and 207.4 ± 77.7 µm in control eyes, with no difference between the 3 groups (P = 0.116). After sildenafil, choroidal thickness increased by 6.0% to 9.0% at 1 and 3 hours in all groups (P = 0.001-0.014). Eyes from older subjects were associated with choroidal thinning at baseline (P = 0.005) and showed less choroidal expansion at 1 hour and 3 hours after sildenafil (P = 0.001) regardless of AMD status (P = 0.666). The choroidal thickening appeared to be primarily attributed to expansion of the stroma rather than luminal component. Retinal vascular density remained unchanged after sildenafil in all 3 groups (P = 0.281-0.587). Together, our studies suggest that vascular response of the choroid to sildenafil decreases with age, but is not affected by the presence of non-exudative or exudative AMD, providing insight into changes in vessel compliance in aging and AMD.

Megahertz-rate optical coherence tomography angiography improves the contrast of the choriocapillaris and choroid in human retinal imaging.

Angiographic imaging of the human eye with optical coherence tomography (OCT) is becoming an increasingly important tool in the scientific investigation and clinical management of several blinding diseases, including age-related macular degeneration and diabetic retinopathy. We have observed that OCT angiography (OCTA) of the human choriocapillaris and choroid with a 1.64 MHz A-scan rate swept-source laser yields higher contrast images as compared to a slower rate system operating at 100 kHz. This result is unexpected because signal sensitivity is reduced when acquisition rates are increased, and the incident illumination power is kept constant. The contrast of angiography images generated by acquiring multiple sequential frames and calculating the variation caused by blood flow, however, appears to be improved significantly when lower-contrast images are taken more rapidly. To demonstrate that the acquisition rate plays a role in the quality improvement, we have imaged five healthy subjects with a narrow field of view (1.2 mm) OCTA imaging system using two separate swept-source lasers of different A-line rates and compared the results quantitatively using the radially-averaged power spectrum. The average improvement in the contrast is 23.0% (+/-7.6%). Although the underlying cause of this enhancement is not explicitly determined here, we speculate that the higher-speed system suppresses the noise contribution from eye motion in subjects and operates with an inter-scan time that better discriminates the flow velocities present in the choroid and choriocapillaris. Our result informs OCT system developers on the merits of ultrahigh-speed acquisition in functional imaging applications.

Functional retinal imaging using adaptive optics swept-source OCT at 1.6 MHz.

Objective optical assessment of photoreceptor function may permit earlier diagnosis of retinal disease than current methods such as perimetry, electrophysiology, and clinical imaging. In this work, we describe an adaptive optics (AO) optical coherence tomography (OCT) system designed to measure functional responses of single cones to visible stimuli. The OCT subsystem consisted of a raster-scanning Fourier-domain mode-locked laser that acquires A scans at 1.64 MHz with a center wavelength of 1063 nm and an AO system operating in closed-loop. Analysis of serial volumetric images revealed phase changes of cone photoreceptors consistent with outer segment elongation and proportional to stimulus intensity, as well as other morphological changes in the outer segment and retinal pigment epithelium.

Intraframe motion correction for raster-scanned adaptive optics images using strip-based cross-correlation lag biases.

In retinal raster imaging modalities, fixational eye movements manifest as image warp, where the relative positions of the beam and retina change during the acquisition of single frames. To remove warp artifacts, strip-based registration methods-in which fast-axis strips from target images are registered to a reference frame-have been applied in adaptive optics (AO) scanning light ophthalmoscopy (SLO) and optical coherence tomography (OCT). This approach has enabled object tracking and frame averaging, and methods have been described to automatically select reference frames with minimal motion. However, inconspicuous motion artifacts may persist in reference frames and propagate themselves throughout the processes of registration, tracking, and averaging. Here we test a previously proposed method for removing movement artifacts in reference frames, using biases in stripwise cross-correlation statistics. We applied the method to synthetic retinal images with simulated eye motion artifacts as well as real AO-SLO images of the cone mosaic and volumetric AO-OCT images, both affected by eye motion. In the case of synthetic images, the method was validated by direct comparison with motion-free versions of the images. In the case of real AO images, performance was validated by comparing the correlation of uncorrected images with that of corrected images, by quantifying the effect of motion artifacts on the image power spectra, and by qualitative examination of AO-OCT B-scans and en face projections. In all cases, the proposed method reduced motion artifacts and produced more faithful images of the retina.

Challenges and advantages in wide-field optical coherence tomography angiography imaging of the human retinal and choroidal vasculature at 1.7-MHz A-scan rate.

We present noninvasive, three-dimensional, depth-resolved imaging of human retinal and choroidal blood circulation with a swept-source optical coherence tomography (OCT) system at 1065-nm center wavelength. Motion contrast OCT imaging was performed with the phase-variance OCT angiography method. A Fourier-domain mode-locked light source was used to enable an imaging rate of 1.7 MHz. We experimentally demonstrate the challenges and advantages of wide-field OCT angiography (OCTA). In the discussion, we consider acquisition time, scanning area, scanning density, and their influence on visualization of selected features of the retinal and choroidal vascular networks. The OCTA imaging was performed with a field of view of 16 deg (5 mm×5 mm) and 30 deg (9 mm×9 mm). Data were presented in en face projections generated from single volumes and in en face projection mosaics generated from up to 4 datasets. OCTA imaging at 1.7 MHz A-scan rate was compared with results obtained from a commercial OCTA instrument and with conventional ophthalmic diagnostic methods: fundus photography, fluorescein, and indocyanine green angiography. Comparison of images obtained from all methods is demonstrated using the same eye of a healthy volunteer. For example, imaging of retinal pathology is presented in three cases of advanced age-related macular degeneration.

The Properties of Outer Retinal Band Three Investigated With Adaptive-Optics Optical Coherence Tomography.

Purpose: Optical coherence tomography's (OCT) third outer retinal band has been attributed to the zone of interdigitation between RPE cells and cone outer segments. The purpose of this paper is to investigate the structure of this band with adaptive optics (AO)-OCT. Methods: Using AO-OCT, images were obtained from two subjects. Axial structure was characterized by measuring band 3 thickness and separation between bands 2 and 3 in segmented cones. Lateral structure was characterized by correlation of band 3 with band 2 and comparison of their power spectra. Band thickness and separation were also measured in a clinical OCT image of one subject. Results: Band 3 thickness ranged from 4.3 to 6.4 µm. Band 2 correlations ranged between 0.35 and 0.41 and power spectra of both bands confirmed peak frequencies that agree with histologic density measurements. In clinical images, band 3 thickness was between 14 and 19 µm. Measurements of AO-OCT of interband distance were lower than our corresponding clinical OCT measurements. Conclusions: Band 3 originates from a structure with axial extent similar to a single surface. Correlation with band 2 suggests an origin within the cone photoreceptor. These two observations indicate that band 3 corresponds predominantly to cone outer segment tips (COST). Conventional OCT may overestimate both the thickness of band 3 and outer segment length.

Long working distance OCT with a compact 2f retinal scanning configuration for pediatric imaging.

Young and/or autistic children cannot be imaged with tabletop or handheld optical coherence tomography (OCT) because of their lack of attention and fear of large objects close to their face. We demonstrate a prototype retinal swept-source OCT system with a long working distance (from the last optical element to the subject's eye) to facilitate pediatric imaging. To reduce the number of optical elements and axial length compared to the traditional 4f telescope, we employ a compact 2f retinal scanning configuration and achieve a working distance of 350 mm with a 16° OCT field of view. We test our prototype system on pediatric and adult subjects.

Comparison of amplitude-decorrelation, speckle-variance and phase-variance OCT angiography methods for imaging the human retina and choroid.

We compared the performance of three OCT angiography (OCTA) methods: speckle variance, amplitude decorrelation and phase variance for imaging of the human retina and choroid. Two averaging methods, split spectrum and volume averaging, were compared to assess the quality of the OCTA vascular images. All data were acquired using a swept-source OCT system at 1040 nm central wavelength, operating at 100,000 A-scans/s. We performed a quantitative comparison using a contrast-to-noise (CNR) metric to assess the capability of the three methods to visualize the choriocapillaris layer. For evaluation of the static tissue noise suppression in OCTA images we proposed to calculate CNR between the photoreceptor/RPE complex and the choriocapillaris layer. Finally, we demonstrated that implementation of intensity-based OCT imaging and OCT angiography methods allows for visualization of retinal and choroidal vascular layers known from anatomic studies in retinal preparations. OCT projection imaging of data flattened to selected retinal layers was implemented to visualize retinal and choroidal vasculature. User guided vessel tracing was applied to segment the retinal vasculature. The results were visualized in a form of a skeletonized 3D model.

Intrasurgical Human Retinal Imaging With Manual Instrument Tracking Using a Microscope-Integrated Spectral-Domain Optical Coherence Tomography Device.

PURPOSE: To characterize the first in-human intraoperative imaging using a custom prototype spectral-domain microscope-integrated optical coherence tomography (MIOCT) device during vitreoretinal surgery with instruments in the eye. METHODS: Under institutional review board approval for a prospective intraoperative study, MIOCT images were obtained at surgical pauses with instruments held static in the vitreous cavity and then concurrently with surgical maneuvers. Postoperatively, MIOCT images obtained at surgical pauses were compared with images obtained with a high-resolution handheld spectral-domain OCT (HHOCT) system with objective endpoints, including acquisition of images acceptable for analysis and identification of predefined macular morphologic or pathologic features. RESULTS: Human MIOCT images were successfully obtained before incision and during pauses in surgical maneuvers. MIOCT imaging confirmed preoperative diagnoses, such as epiretinal membrane, full-thickness macular hole, and vitreomacular traction and demonstrated successful achievement of surgical goals. MIOCT and HHOCT images obtained at surgical pauses in two cohorts of five patients were comparable with greater than or equal to 80% correlation in 80% of patients. Real-time video-imaging concurrent with surgical manipulations enabled, for the first time using this device, visualization of dynamic instrument-retina interaction with targeted OCT tracking. CONCLUSION: MIOCT is successful for imaging at surgical pauses and for real-time image guidance with implementation of targeted OCT tracking. Even faster acquisition speeds are currently being developed with incorporation of a swept-source MIOCT engine. Further refinements and investigations will be directed toward continued integration for real-time volumetric imaging of surgical maneuvers. TRANSLATIONAL RELEVANCE: Ongoing development of seamless MIOCT systems will likely transform surgical visualization, approaches, and decision-making.

Preclinical evaluation and intraoperative human retinal imaging with a high-resolution microscope-integrated spectral domain optical coherence tomography device.

PURPOSE: The authors have recently developed a high-resolution microscope-integrated spectral domain optical coherence tomography (MIOCT) device designed to enable OCT acquisition simultaneous with surgical maneuvers. The purpose of this report is to describe translation of this device from preclinical testing into human intraoperative imaging. METHODS: Before human imaging, surgical conditions were fully simulated for extensive preclinical MIOCT evaluation in a custom model eye system. Microscope-integrated spectral domain OCT images were then acquired in normal human volunteers and during vitreoretinal surgery in patients who consented to participate in a prospective institutional review board-approved study. Microscope-integrated spectral domain OCT images were obtained before and at pauses in surgical maneuvers and were compared based on predetermined diagnostic criteria to images obtained with a high-resolution spectral domain research handheld OCT system (HHOCT; Bioptigen, Inc) at the same time point. Cohorts of five consecutive patients were imaged. Successful end points were predefined, including ≥80% correlation in identification of pathology between MIOCT and HHOCT in ≥80% of the patients. RESULTS: Microscope-integrated spectral domain OCT was favorably evaluated by study surgeons and scrub nurses, all of whom responded that they would consider participating in human intraoperative imaging trials. The preclinical evaluation identified significant improvements that were made before MIOCT use during human surgery. The MIOCT transition into clinical human research was smooth. Microscope-integrated spectral domain OCT imaging in normal human volunteers demonstrated high resolution comparable to tabletop scanners. In the operating room, after an initial learning curve, surgeons successfully acquired human macular MIOCT images before and after surgical maneuvers. Microscope-integrated spectral domain OCT imaging confirmed preoperative diagnoses, such as full-thickness macular hole and vitreomacular traction, and demonstrated postsurgical changes in retinal morphology. Two cohorts of five patients were imaged. In the second cohort, the predefined end points were exceeded with ≥80% correlation between microscope-mounted OCT and HHOCT imaging in 100% of the patients. CONCLUSION: This report describes high-resolution MIOCT imaging using the prototype device in human eyes during vitreoretinal surgery, with successful achievement of predefined end points for imaging. Further refinements and investigations will be directed toward fully integrating MIOCT with vitreoretinal and other ocular surgery to image surgical maneuvers in real time.

Unprocessed real-time imaging of vitreoretinal surgical maneuvers using a microscope-integrated spectral-domain optical coherence tomography system.

BACKGROUND: We have recently developed a microscope-integrated spectral-domain optical coherence tomography (MIOCT) device towards intrasurgical cross-sectional imaging of surgical maneuvers. In this report, we explore the capability of MIOCT to acquire real-time video imaging of vitreoretinal surgical maneuvers without post-processing modifications. METHODS: Standard 3-port vitrectomy was performed in human during scheduled surgery as well as in cadaveric porcine eyes. MIOCT imaging of human subjects was performed in healthy normal volunteers and intraoperatively at a normal pause immediately following surgical manipulations, under an Institutional Review Board-approved protocol, with informed consent from all subjects. Video MIOCT imaging of live surgical manipulations was performed in cadaveric porcine eyes by carefully aligning B-scans with instrument orientation and movement. Inverted imaging was performed by lengthening of the reference arm to a position beyond the choroid. RESULTS: Unprocessed MIOCT imaging was successfully obtained in healthy human volunteers and in human patients undergoing surgery, with visualization of post-surgical changes in unprocessed single B-scans. Real-time, unprocessed MIOCT video imaging was successfully obtained in cadaveric porcine eyes during brushing of the retina with the Tano scraper, peeling of superficial retinal tissue with intraocular forceps, and separation of the posterior hyaloid face. Real-time inverted imaging enabled imaging without complex conjugate artifacts. CONCLUSIONS: MIOCT is capable of unprocessed imaging of the macula in human patients undergoing surgery and of unprocessed, real-time, video imaging of surgical maneuvers in model eyes. These capabilities represent an important step towards development of MIOCT for efficient, real-time imaging of manipulations during human surgery.

The use of optical coherence tomography in intraoperative ophthalmic imaging.

Optical coherence tomography (OCT) has transformed diagnostic ophthalmic imaging but until recently has been limited to the clinic setting. The development of spectral-domain OCT (SD-OCT), with its improved speed and resolution, along with the development of a handheld OCT scanner, enabled portable imaging of patients unable to sit in a conventional tabletop scanner. This handheld SD-OCT unit has proven useful in examinations under anesthesia and, more recently, in intraoperative imaging of preoperative and postoperative manipulations. Recently, several groups have pioneered the development of novel OCT modalities, such as microscope-mounted OCT systems. Although still immature, the development of these systems is directed toward real-time imaging of surgical maneuvers in the intraoperative setting. This article reviews intraoperative imaging of the posterior and anterior segment using the handheld SD-OCT and recent advances toward real-time microscope-mounted intrasurgical imaging.