In vivo identification of the retinal layer containing photopigments in OCT images through correlation with two-photon psychophysics.

Two-photon vision enables near-infrared light perception in humans. We investigate the possibility to utilize this phenomenon as an indicator of the location of the outer segments of photoreceptor cells in the OCT images. Since two-photon vision is independent on OCT imaging, it could provide external to OCT reference relative to which positions of retinal layers visible in OCT imaging could be measured. We show coincidence between OCT imaging of outer retinal layers and two-photon light perception. The experiment utilizes an intrinsic nonlinear process in the retina, two-photon absorption of light by visual photopigments, which triggers perception of near-infrared light. By shifting the focus of the imaging/stimulus beam, we link the peak efficiency of two-photon vision with the visibility of outer segments of photoreceptor cells, which can be seen as in vivo identification of a retinal layer containing visual photopigments in OCT images. Determination of the in-focus retinal layer is achieved by analysis of en face OCT image contrast. We discuss experimental methods and experimental factors that may influence two-photon light perception and the accuracy of the results. The limits of resolution are discussed in analysis of the one-photon and two-photon point spread functions.

Accurate calibration of beam trajectories in scanning optical imaging systems.

We present a calibration method for finding the coordinates of points in the trajectory of the scanning beam in flying-spot imaging devices. Our method is based on laterally translating the field of view on the imaging object plane by introducing additional beam deflections. We show that laterally translating the field of view provides a series of images whose relative translations are equal to the distances between the points in the scanning pattern to be calibrated. We show how these distances are mapped to the coordinates of the trajectory points. As an example, we demonstrate the calibration of the scanning patterns in an optical system with two independent microelectromechanical system based scanners. Our method profits from a large collection of distance measurements to find the trajectory coordinates, thereby minimizing the effect of random sources of uncertainty in the positions of points in the scanning pattern. We have found that we are capable of finding the coordinates of points in the scanning patterns with accuracy greater than the optical resolution of the imaging system.

High-resolution, ultrafast, wide-field retinal eye-tracking for enhanced quantification of fixational and saccadic motion.

We introduce a novel, noninvasive retinal eye-tracking system capable of detecting eye displacements with an angular resolution of 0.039 arcmin and a maximum velocity of 300°/s across an 8° span. Our system is designed based on a confocal retinal imaging module similar to a scanning laser ophthalmoscope. It utilizes a 2D MEMS scanner ensuring high image frame acquisition frequencies up to 1.24 kHz. In contrast with leading eye-tracking technology, we measure the eye displacements via the collection of the observed spatial excursions for all the times corresponding a full acquisition cycle, thus obviating the need for both a baseline reference frame and absolute spatial calibration. Using this approach, we demonstrate the precise measurement of eye movements with magnitudes exceeding the spatial extent of a single frame, which is not possible using existing image-based retinal trackers. We describe our retinal tracker, tracking algorithms and assess the performance of our system by using programmed artificial eye movements. We also demonstrate the clinical capabilities of our system with in vivo subjects by detecting microsaccades with angular extents as small as 0.028°. The rich kinematic ocular data provided by our system with its exquisite degree of accuracy and extended dynamic range opens new and exciting avenues in retinal imaging and clinical neuroscience. Several subtle features of ocular motion such as saccadic dysfunction, fixation instability and abnormal smooth pursuit can be readily extracted and inferred from the measured retinal trajectories thus offering a promising tool for identifying biomarkers of neurodegenerative diseases associated with these ocular symptoms.

Defocus vibrations in optical systems-considerations in reference to the human eye.

Experimental visual acuity (VA) of eight subjects was measured using the Freiburg vision test in a custom-made adaptive optics system. Measurements were conducted under one control and five defocus-induced conditions. In the defocus-induced conditions, 1 diopter of myopic defocus was added to the system using the Badal stage, and defocus vibrations with five different levels of amplitude were generated by a deformable mirror at 50 Hz. Computational simulations of the visual Strehl ratio (VSOTF) were performed using average aberrations of each subject recorded in the control condition. For the first time, to the best of our knowledge, it has been shown experimentally that both the simulated VSOTF and experimentally measured VA improve when defocus vibrations are added to a defocused eye.

Rotation asymmetry of the human sclera.

PURPOSE: To characterize the mean topographical shape of the human sclera of a normal eye. METHODS: Forty-five participants aged from 19 to 45 years and with no previous ocular surgeries were included in this study. Four three-dimensional (3D) corneo-scleral maps from both eyes were acquired using a corneo-scleral topographer (Eye Surface Profiler). For each 3D map, the sclera (maximum diameter of 16 mm) and cornea were automatically separated at the level of the limbus. The remaining 3D scleral ring was further fit to a quadratic function. The elevation difference between the original and fit data was calculated. For statistical analysis, sclera was divided into eight sectors: nasal, temporal, superior, inferior, supero-nasal, supero-temporal, inferior-nasal and inferior-temporal. In addition, sclera was separated as inner sclera (inner ring of 6-7-mm radius) and outer sclera (external ring of 7-8-mm radius). RESULTS: Horizontally, the nasal area of the sclera showed less elevation [mean (SD) -30 (52) µm (OD)] than the temporal area [mean (SD) 4 (47) µm (OD)], p < 0.001. Vertically, the inferior area of the sclera [mean (SD) 32 (72) µm (OD)] was slightly less elevated than the superior area (mean (SD) 36 (84) µm, but this difference was not statistically significant (p = 0.40). Besides, the asymmetry of the sclera was found to increase with radial distance from the corneal apex. No statistically significant difference was found between right and left eye. CONCLUSION: Human sclera is rotationally asymmetric, and its shape varies considerably between subjects.

Evaluating tear clearance rate with optical coherence tomography.

PURPOSE: To assess the early-phase of tear clearance rate (TCR) with anterior segment optical coherence tomography (OCT) and to determine the association between TCR and other clinical measures of the tear film in a group of young subjects with different levels of tear film quality. METHODS: TCR was classified as the percentage decrease of subject's inferior tear meniscus height 30s after instillation of 5µl 0.9% saline solution. Fifty subjects (32F and 18M) aged (mean±standard deviation) 25.5±4.3 years volunteered for the study. It consisted of a review of medical history, Ocular Surface Disease Index (OSDI) questionnaire, tear film osmolarity measurements, slit lamp examination and TCR estimation based on dynamic measurements of the lower tear meniscus with OCT. Estimates of TCR were contrasted against subject age and tear film measures commonly used for dry eye diagnosis, which includes OSDI score, fluorescein tear film break-up time (FBUT), tear meniscus height (TMH), blinking frequency, tear film osmolarity and corneal staining. RESULTS: The group mean TCR was 29±13% and 36±19% respectively after 30 and 60s margin after saline solution instillation. Statistically significant correlations were found between TCR and FBUT (r2=0.319, p<0.001), blinking frequency (r2=0.138, p<0.01), tear film osmolarity (r2=0.133, p<0.01) and subject's age (r2=0.095, p<0.05). CONCLUSIONS: Anterior segment optical coherence tomography allows following changes of tear meniscus morphology post saline solution instillation and evaluating the TCR. OCT based TCR might be used as additional measure of the lacrimal functional unit.

Corneo-scleral limbal changes following short-term soft contact lens wear.

PURPOSE: To assess whether short-term soft contact lens wear alters the anterior eye surface. METHODS: Twenty-two neophyte subjects wore soft contact lenses for a period of five hours. Topography based corneo-scleral limbal radius estimates were derived from height measurements acquired with a corneo-scleral profilometer. Additionally, central corneal thickness (CCT), anterior chamber depth (ACD), corneal curvature radius (R) and white-to-white (WTW) diameter were acquired with an OCT-assisted biometer. Measurements were obtained without lens wear (baseline), immediately after lens removal following five hours of wear and three hours after lens removal. RESULTS: Short-term soft contact lens wear significantly modifies corneo-scleral limbal radius (mean±SD: 130±74µm, p << 0.001) and the changes are repeatable. In contrast, the WTW diameter and R were not modified. ACD and CCT were significantly affected but no significant correlations were found between the increment of the limbal radius and the decrease in ACD and CCT. Limbal radius increment was reversed three hours after lens removal for 68% of the subjects but the time course of this reversal was not uniform. CONCLUSIONS: It is possible to accurately quantify limbal radius changes as a consequence of soft contact lens wear. The increment in the limbal diameter could reach over 0.5mm but that alteration does not correspond to changes in WTW diameter and it was not observable to the examiner using a slit lamp. Assessing topographical limbus after contact lens wear could be a tool to optimize the selection of the contact lens, from the perspective of anterior eye surface changes.

Automatic dynamic tear meniscus measurement in optical coherence tomography.

An image processing algorithm is developed for quantitative assessment of tear meniscus dynamics from continuous optical coherence tomography (OCT) measurements. Clinical utility of dynamic OCT tear meniscus measurement is assessed in studies of tear meniscus parameters. The results indicate that any apparent changes in the early post-blink phase meniscus parameters are essentially related to the longitudinal movements of the eye and not to the formation of tear meniscus corresponding to tear film build-up. Dynamic acquisition of tear film meniscus is essential for providing reliable estimates of its parameters such as height, depth, and area.

Optimization of 3D Poisson-Nernst-Planck model for fast evaluation of diverse protein channels.

We show the accuracy and applicability of our fast algorithmic implementation of a three-dimensional Poisson-Nernst-Planck (3D-PNP) flow model for characterizing different protein channels. Due to its high computational efficiency, our model can predict the full current-voltage characteristics of a channel within minutes, based on the experimental 3D structure of the channel or its computational model structure. Compared with other methods, such as Brownian dynamics, which currently needs a few weeks of the computational time, or even much more demanding molecular dynamics modeling, 3D-PNP is the only available method for a function-based evaluation of very numerous tentative structural channel models. Flow model tests of our algorithm and its optimal parametrization are provided for five native channels whose experimental structures are available in the protein data bank (PDB) in an open conductive state, and whose experimental current-voltage characteristics have been published. The channels represent very different geometric and structural properties, which makes it the widest test to date of the accuracy of 3D-PNP on real channels. We test whether the channel conductance, rectification, and charge selectivity obtained from the flow model, could be sufficiently sensitive to single-point mutations, related to unsignificant changes in the channel structure. Our results show that the classical 3D-PNP model, under proper parametrization, is able to achieve a qualitative agreement with experimental data for a majority of the tested characteristics and channels, including channels with narrow and irregular conductivity pores. We propose that although the standard PNP model cannot provide insight into complex physical phenomena due to its intrinsic limitations, its semiquantitative agreement is achievable for rectification and selectivity at a level sufficient for the bioinformatical purpose of selecting the best structural models with a great advantage of a very short computational time.