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Purpose: To visualize in vivo and quantify the thickness of the posterior corneal layers: the acellular pre-Descemet's layer (PDL), Descemet's membrane (DM), and endothelium (END) in healthy subjects, using ultrahigh-resolution optical coherence tomography (UHR-OCT)., Methods: A research-grade, 800-nm UHR-OCT system with 0.95-μm axial resolution in corneal tissue was used to image in vivo the posterior cornea in healthy subjects. The system offers approximately 98 dB sensitivity for 680 μW optical power incident on the cornea and 34,000 A-scans/s image acquisition rate. This study comprised 20 healthy subjects, aged 20 to 60 years. The thickness of the PDL, DM, and END layers was measured both with a custom, automatic segmentation algorithm and manually., Results: The boundaries and structure of the posterior corneal layers were clearly visible in the UHR-OCT images. The average thickness was measured to be 6.6 ± 1.4 μm (PDL), 10.4 ± 2.9 μm (DM), and 4.8 ± 0.4 μm (END), which agrees well with published data from ex vivo studies. Both the END and DM thickness showed minor spatial variations, whereas the PDL showed up to 2× thickness change for different locations on the same cross-sectional corneal image or over the entire imaged region of the cornea., Conclusions: Our data indicate that all three layers of the posterior cornea can be clearly visualized in vivo and their thicknesses measured precisely with UHR-OCT. Although the PDL thickness showed large spatial variations, the thickness of the DM and END layers was consistent over the entire imaged region of the cornea.

Purpose: Moderately elevated intraocular pressure (IOP) is a risk factor for open-angle glaucoma. Some patients suffer glaucoma despite clinically measured normal IOPs. Fluctuations in IOP may have a significant role since IOPs are higher during sleep and inversion activities. Controlled transient elevations of IOPs in rats over time lead to optic nerve structural changes that are similar to the early changes observed in constant chronic models of glaucoma. Because early intervention decreases glaucoma progression, this study was done to determine if early physiological changes to the retina could be detected with noninvasive electrophysiological and optical imaging tests during moderately elevated IOP., Methods: Intraocular pressures were raised to moderately high levels (35 mm Hg) in one eye of Sprague-Dawley rats while the other (control) eye was untreated. One group of rats underwent scotopic threshold response (STR) and electroretinogram (ERG) testing, while another 3 groups underwent optical coherence tomography (OCT) imaging, Western blot, or histologic evaluation., Results: The amplitudes of the STR and ERG responses in eyes with moderately elevated IOPs were enhanced compared to the values before IOP elevation, and compared to untreated contralateral eyes. Structural changes to the optic nerve also occurred during IOP elevation., Conclusions: Although ischemic IOP elevations are well-known to globally reduce components of the scotopic ERG, acute elevation in rats to levels often observed in untreated glaucoma patients caused an increase in these parameters. Further exploration of these phenomena may be helpful in better understanding the mechanisms mediating early retinal changes during fluctuating or chronically elevated IOP.

Purpose: To demonstrate the ability of high speed, ultrahigh-resolution optical coherence tomography (UHR-OCT) to measure and characterize in vivo visual stimulus-specific pupil dynamics in birds., Methods: Ten two-week old White Leghorn (Gallus gallus domesticus) chickens were imaged in this study. The chickens were dark-adapted for 1 hour and anesthetized with 2% isoflurane prior to the imaging procedure. Blue, green, and red single flash visual stimuli of 7 ms duration were used to evoke pupillary responses. UHR-OCT cross-sectional images of the pupil were acquired prior, during, and for several seconds after the visual stimuli onset. Images were processed with a novel custom automatic algorithm, designed to determine the pupil diameter changes over time., Results: Results from this study show that the pupillary constriction begins with the onset of the visual stimuli; however, maximum pupil constriction occurs ∼150 ms later. No statistically significant variation in the timing of the maximum pupillary constriction was observed for stimuli of different colors. However, significant variation was observed in the maximum pupil constriction amplitudes, between red-green and red-blue stimuli, but not between blue-green stimuli. Furthermore, the magnitude of the maximum pupil constriction decreased monotonically with time under isoflurane anesthesia., Conclusions: We demonstrated, for the first time, measurements of visually evoked pupillary dynamics in animals using high speed UHR-OCT. The results suggest dependence of the pupillary dynamics on the color of the visual stimulus, and adverse effects of isoflurane anesthesia on the visually evoked pupillary responses in chickens.

Purpose: To provide in vivo quantitative assessment of sodium iodate-induced retinal damage in a rat model of outer retinal degeneration using ultrahigh resolution optical coherence tomography (UHR-OCT)., Methods: Outer retinal degeneration was induced in four female Long Evans rats via tail vein injection of sodium iodate (40 mg/kg). Changes in the thickness and optical reflectivity of individual retinal layers were extracted using a semi-automatic segmentation algorithm and were assessed in vivo at 6 hours, days 1, 3, and 7, and up to 3 months post injection with UHR-OCT. Hematoxylin and eosin (H&E) histology was used to confirm the morphologic changes observed in the UHR-OCT images., Results: UHR-OCT tomograms showed progressive structural damage in the rat retina over time, such as swelling, thinning, complete disintegration of individual retinal layers, and clustering of highly reflective cellular debris. Photoreceptor swelling was observed 6 hours after injection of sodium iodate, followed by progressive structural decomposition of the outer retina. At 3 months post injection, the outer retina was completely disintegrated, and the inner nuclear layer (INL) was in direct contact with the choroid. Changes in the reflectivity of individual retinal layers were observed over time and correlated well with the morphologic changes., Conclusions: UHR-OCT permits in vivo, noninvasive, longitudinal, quantitative assessment of the progressive changes in retinal morphology and optical reflectivity in a sodium iodate rodent model of outer retinal degeneration.

Purpose: To evaluate the change in thickness of the anterior, stromal, and posterior corneal laminae in response to hypoxia-induced corneal swelling, by means of ultrahigh-resolution optical coherence tomography (UHR-OCT)., Methods: A UHR-OCT system, operating in the 1060-nm range, was used to acquire in vivo cross-sectional images of human cornea with a 3.2x10-microm (axial x lateral) resolution in corneal tissue. Corneal edema was induced by inserting a thick, positive-powered, soft contact lens, over which the eye was closed and patched for 3 hours. Tomograms were acquired from eight non-contact-lens wearers. Baseline images were obtained before inducing corneal edema, immediately after removal of the patch and the lens, and then every 15 minutes for approximately 2 hours. All images were postprocessed with a segmentation algorithm to identify the laminae visible in the image. The apical thickness of the laminae (epithelium [EPI], epithelial-Bowman's membrane [Ep-BM] complex, stroma, and endothelial-Descemet's membrane [En-DM] complex) were determined at each time interval., Results: There was an interaction between time after removal of the hypoxic stimulus and deswelling of the layers (RM-ANOVA; P<0.001). The epithelial and stromal thickness reduced significantly with time (P=0.001; P<0.001, respectively), whereas the Ep-BM and En-DM complexes did not (P>0.50). All layers except the En-DM complex exhibited a biphasic pattern of recovery., Conclusions: UHR-OCT showed regional differences in swelling due to hypoxic provocation. On removal of the hypoxic stimulus, the rate of recovery varied between layers, and all layers except the En-DM complex exhibited a biphasic recovery.