This PDF file contains the front matter associated with SPIE Proceedings Volume 9693, including the Title Page, Copyright information, Table of Contents, and Conference Committee listing.

We present a three beam optical Doppler tomography (ODT) technique suitable for 3-D velocity and flow measurements to evaluate total retinal blood circulation from and to the optic nerve head (ONH). The system consists of three independent ODT channels. Superluminescent diodes with a central wavelength of 840 nm and a spectral bandwidth of 50 nm were used. The sources are coupled to collimators resting in a specially designed mount to ensure a well-defined beam geometry, necessary for the full reconstruction of the three dimensional velocity vector. The reconstruction works without prior knowledge on the vessel geometry, which is normally required for ODT systems with less than three beams. The beams share a common bulk optics Michelson interferometer, while the detection comprises three identical spectrometers with a line scan rate of 50 kHz. 20 eyes of healthy volunteers were imaged with the 3 beam ODT, employing a circular scan pattern around the ONH. The mean total blood flow was calculated for arteries (47.1 ± 2.4 μl/min (mean ± SD)) and veins (47.1 ± 2.7 μl/min μl/min) independently. The two results showed no significant difference (paired t-test, p < 0.96), rendering both equally reliable for total flow measurements. Furthermore the reproducibility of the method was evaluated for the total flow and flow, velocities within each individual vessel of 6 eyes. The average variation for total flow measurements is sufficiently low to detect deviations of ~ 6% indicating high precision of the proposed method.

To investigate the application of optical microangiography (OMAG) in living human eye. Patients with different macular diseases were recruited, including diabetic retinopathy (DR), geographic atrophy (GA), retinitis pigmentosa (RP), and venous occlusion, et al. Wide field OCT angiography images can be generated by montage scanning protocol based on the tracking system. OMAG algorithm based on complex differentiation was used to extract the blood flow and removed the bulk motion by 2D cross-correlation method. The 3D angiography was segmented into 3 layers in the retina and 2 layers in the choroid. The en-face maximum projection was used to obtain 2-dimensional angiograms of different layers coded with different colors. Flow and structure images were combined for cross-sectional view. En face OMAG images of different macular diseases showed a great agreement with FA. Meanwhile, OMAG gave more distinct vascular network visions that were less affected by hemorrhage and leakage. The MAs were observed in both superficial and middle retinal layers based on OMAG angiograms in different layers of DR patients. The contour line of FAZ was extracted as well, which can be quantitative the retinal diseases. For GA patient, the damage of RPE layer enhanced the penetration of light and enabled the acquisition of choriocapillaries and choroidal vessels. The wide field OMAG angiogram enabled the capability of capturing the entire geographic atrophy. OMAG provides depth-resolved information and detailed vascular images of DR and GA patients, providing a better visualization of vascular network compared to FA.

Ophthalmic surgeons manipulate micron-scale tissues using stereopsis through an operating microscope and instrument shadowing for depth perception. While ophthalmic microsurgery has benefitted from rapid advances in instrumentation and techniques, the basic principles of the stereo operating microscope have not changed since the 1930’s. Optical Coherence Tomography (OCT) has revolutionized ophthalmic imaging and is now the gold standard for preoperative and postoperative evaluation of most retinal and many corneal procedures. We and others have developed initial microscope-integrated OCT (MIOCT) systems for concurrent OCT and operating microscope imaging, but these are limited to 2D real-time imaging and require offline post-processing for 3D rendering and visualization. Our previously presented 4D MIOCT system can record and display the 3D surgical field stereoscopically through the microscope oculars using a dual-channel heads-up display (HUD) at up to 10 micron-scale volumes per second. In this work, we show that 4D MIOCT guidance improves the accuracy of depth-based microsurgical maneuvers (with statistical significance) in mock surgery trials in a wet lab environment. Additionally, 4D MIOCT was successfully performed in 38/45 (84%) posterior and 14/14 (100%) anterior eye human surgeries, and revealed previously unrecognized lesions that were invisible through the operating microscope. These lesions, such as residual and potentially damaging retinal deformation during pathologic membrane peeling, were visualized in real-time by the surgeon. Our integrated system provides an enhanced 4D surgical visualization platform that can improve current ophthalmic surgical practice and may help develop and refine future microsurgical techniques.

Conventional optical coherence tomography (OCT) systems have working distances of about 25 mm, and require cooperative subjects to immobilize and fixate on a target. Handheld OCT probes have also been demonstrated for successful imaging of pre-term infants and neonates up to ~1 year old. However, no technology yet exists for OCT in young children due to their lack of attention and inherent fear of large objects close to their face. In this work, 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 footprint and weight of the system compared to the conventional 4f scheme, a novel 2f scanning configuration was implemented to achieve a working distance of 348mm with a +/- 8° scanning angle prior to cornea. Employing two custom-designed lenses, the system design resolution was nearly diffraction limited throughout a -8D to +5D refractive corrections. A fixation target displayed on a LCD monitor and an iris camera were used to facilitate alignment and imaging. Our prototype was tested in consented adult subjects and has the potential to facilitate imaging of young children. With this apparatus, young children could potentially sit comfortably in caretaker’s lap while viewing entertainment on the fixation screen designed to direct their gaze into the imaging apparatus.

Hand-held wide-field contact color fundus photography is currently the standard method to acquire diagnostic images of children during examination under anesthesia and in the neonatal intensive care unit. The recent development of portable non-contact hand-held OCT retinal imaging systems has proved that OCT is of tremendous help to complement fundus photography in the management of pediatric patients. Currently, there is no commercial or research system that combines color wide-field digital fundus and OCT imaging in a contact-fashion. The contact of the probe with the cornea has the advantages of reducing motion experienced by the photographer during the imaging and providing fundus and OCT images with wider field of view that includes the periphery of the retina. In this study we produce proof of concept for a contact-type hand-held unit for simultaneous color fundus and OCT live view of the retina of pediatric patients. The front piece of the hand-held unit consists of a contact ophthalmoscopy lens integrating a circular light guide that was recovered from a digital fundus camera for pediatric imaging. The custom-made rear piece consists of the optics to: 1) fold the visible aerial image of the fundus generated by the ophthalmoscopy lens on a miniaturized level board digital color camera; 2) conjugate the eye pupil to the galvanometric scanning mirrors of an OCT delivery system. Wide-field color fundus and OCT images were simultaneously obtained in an eye model and sequentially obtained on the eye of a conscious 25 year-old human subject with healthy retina.

We demonstrate a novel optical coherence tomography system specifically developed and validated for clinical imaging of retinoblastoma tumors in pediatric patients. The existing treatment options for this malignant tumor of the retina aim at reduction of tumor (re)growth risks, and vision preservation. The choice of optimal treatment strongly depends on skilled and detailed clinical assessment. Currently, the patients at risk are periodically monitored with retinal imaging for possible morphological changes over time, and new tumor seedings, as the existing real-time diagnostic tools are limited. Three-dimensional visualization of tissue layer and microvasculature at improved axial and lateral resolution of interference-based OCT imaging provides sensitivity for detection of vital tumor tissue concurrent with local treatment. Our METC-approved system accommodates for the range of optical parameters of infants’ eyes, and uses the 1050nm wavelength to access the deeper choroid layers of retina. The prototype is designed for patients in supine position under general anesthesia, where ergonomic handheld module is connected to fiber-based optical setup via umbilical cord. The system conforms to clinical safety requirements, including fully isolated low-voltage electric circuit. Focusing is performed with a mechanically tunable lens, where resolution is 6 µm axially, and varies with focusing at 10-18µm laterally. We will present optical design, performance limitations, and results of the ongoing clinical study, including the increased OCT diagnostic sensitivity in three dimensions in comparison with the established clinical imaging modalities. We will discuss images of early, active, and treated tumors, as well as follow-up on patients after local and systemic treatments.

The purpose of this study was to test the suitability of three available camera technologies (desktop, portable, and iphone based) for imaging comatose children who presented with clinical symptoms of malaria. Ultimately, the results of the project would form the basis for a design of a future camera to screen for malaria retinopathy (MR) in a resource challenged environment. The desktop, portable, and i-phone based cameras were represented by the Topcon, Pictor Plus, and Peek cameras, respectively. These cameras were tested on N=23 children presenting with symptoms of cerebral malaria (CM) at a malaria clinic, Queen Elizabeth Teaching Hospital in Malawi, Africa. Each patient was dilated for binocular indirect ophthalmoscopy (BIO) exam by an ophthalmologist followed by imaging with all three cameras. Each of the cases was graded according to an internationally established protocol and compared to the BIO as the clinical ground truth. The reader used three principal retinal lesions as markers for MR: hemorrhages, retinal whitening, and vessel discoloration. The study found that the mid-priced Pictor Plus hand-held camera performed considerably better than the lower price mobile phone-based camera, and slightly the higher priced table top camera. When comparing the readings of digital images against the clinical reference standard (BIO), the Pictor Plus camera had sensitivity and specificity for MR of 100% and 87%, respectively. This compares to a sensitivity and specificity of 87% and 75% for the i-phone based camera and 100% and 75% for the desktop camera. The drawback of all the cameras were their limited field of view which did not allow complete view of the periphery where vessel discoloration occurs most frequently. The consequence was that vessel discoloration was not addressed in this study. None of the cameras offered real-time image quality assessment to ensure high quality images to afford the best possible opportunity for reading by a remotely located specialist.

Scanning Laser Ophthalmoscopy (SLO) utilizes the eye’s cornea and lens as the imaging objective. Thus, in a perfect eye the Numerical Aperture (NA) defined by the eye’s fully dilated pupil would limit the resolution that can be achieved. However, the eye’s ocular aberrations “blur the image”, thereby limiting the pupil size and NA for aberrationless SLO imaging. By combining SLO with Adaptive Optics (AO), AO-SLO systems can correct for ocular aberrations even over large pupil sizes and allow diffraction limited resolution in both axial and lateral directions. Here, we evaluate the effects of ocular aberrations on the performance of mouse retinal imaging with SLO and AO-SLO systems. To achieve this we first measured the RMS error of the wavefront aberrations in two populations of mice of different ages. Then, we simulated the imaging PSF and resulting lateral resolution that could be achieved with varied input beam diameter (NA) at the mouse pupil, assuming the presence of ocular aberrations (simulation of SLO imaging) and no aberrations (simulation of perfect AO-SLO imaging). The SLO system performance along with different imaging beam sizes was further assessed by computing the PSF Strehl Ratio. Finally, the advantages and limitations of SLO and AO-SLO retinal imaging in mice are discussed based on presented results.

Chorioretinal blood vessel morphology in mice is of great interest to researchers studying eye disease mechanisms in animal models. Two leading retinal imaging modalities -- Optical Coherence Tomography (OCT) and Scanning Laser Ophthalmoscopy (SLO) -- have offered much insight into vascular morphology and blood flow. OCT “flow-contrast” methods have provided detailed mapping of vascular morphology with micrometer depth resolution, while OCT Doppler methods have enabled the measurement of local flow velocities. SLO remains indispensable in studying blood leakage, microaneurysms, and the clearance time of contrast agents of different sizes. In this manuscript we present results obtained with a custom OCT/SLO system applied to visualize the chorioretinal vascular morphology of pigmented C57Bl/6J and albino nude (Nu/Nu) mice. Blood perfusion maps of choroidal vessels and choricapillaris created by OCT and SLO are presented, along with detailed evaluation of different OCT imaging parameters, including the use of the scattering contrast agent Intralipid. Future applications are discussed.

Intrinsic optical signal (IOS) imaging is a promising noninvasive method for advanced study and diagnosis of eye diseases. Before pursuing clinical applications, more IOS studies employing animal models are necessary to establish the relationship between IOS distortions and eye diseases. Ample mouse models are available for investigating the relationship between IOS distortions and eye diseases. However, in vivo IOS imaging of mouse retinas is challenging due to the small ocular lens (compared to frog eyes) and inevitable eye movements. We report here in vivo IOS imaging of mouse retinas using a custom-designed functional OCT. The OCT system provided high resolution (3 μm) and high speed (up to 500 frames/s) imaging of mouse retinas. An animal holder equipped with a custom designed ear bar and bite bar was used to minimize eye movement due to breathing and heartbeats. Residual eye movement in OCT images was further compensated by accurate image registration. Dynamic OCT imaging revealed rapid IOSs from photoreceptor outer segments immediately (<10 ms) after the stimulation delivery, and unambiguous IOS changes were also observed from inner retinal layers with delayed time courses compared to that of photoreceptor IOSs.

Patient motion artifacts are an important source of data irregularities in OCT imaging. With longer duration OCT scans – as is needed for large wide field of view scans or increased scan density – motion artifacts become increasingly problematic. Strategies to mitigate these motion artifacts are then necessary to ensure OCT data integrity. A popular strategy for reducing motion artifacts in OCT images is to capture two orthogonally oriented volumetric scans containing uncorrelated motion and subsequently reconstructing a motion-free volume by combining information from both datasets. While many different variations of this registration approach have been proposed, even the most recent methods might not be suitable for wide FOV OCT scans which can be lacking in features away from the optic nerve head or arcades. To address this problem, we propose a two-stage motion correction algorithm for wide FOV OCT volumes. In the first step, X and Y axes motion is corrected by registering OCT summed voxel projections (SVPs). To achieve this, we introduce a method based on a custom variation of the dense optical flow technique which is aware of the motion free orientation of the scan. Secondly, a depth (Z axis) correction approach based on the segmentation of the retinal layer boundaries in each B-scan using graph-theory and dynamic programming is applied. This motion correction method was applied to wide field retinal OCT volumes (approximately 80° FOV) of 3 subjects with substantial reduction in motion artifacts.

Quantitative evaluation of optical properties of choroid and sclera are performed by multifunctional optical coherence tomography. Five normal eyes, five glaucoma eyes and one choroidal atrophy eye are examined. The refractive error was found to be correlated with choroidal birefringence, polarization uniformity, and flow in addition to scleral birefringence among normal eyes. The significant differences were observed between the normal and the glaucoma eyes, as for choroidal polarization uniformity, flow and scleral birefringence. An automatic segmentation algorithm of retinal pigment epithelium and chorioscleral interface based on multifunctional signals is also presented.

The aim of this project was to investigate the possibility of using OCT optic nerve head 3D information captured with a Topcon OCT 2000 device for detection of the shortest distance between the inner limit of the retina and the central limit of the pigment epithelium around the circumference of the optic nerve head. The shortest distance between these boundaries reflects the nerve fiber layer thickness and measurement of this distance is interesting for follow-up of glaucoma.

The present study aimed to analyze the clinical usefulness of the thinnest cross section of the nerve fibers in the optic nerve head averaged over the circumference of the optic nerve head. 3D volumes of the optic nerve head of the same eye was captured at two different visits spaced in time by 1-4 weeks, in 13 subjects diagnosed with early to moderate glaucoma. At each visit 3 volumes containing the optic nerve head were captured independently with a Topcon OCT- 2000 system. In each volume, the average shortest distance between the inner surface of the retina and the central limit of the pigment epithelium around the optic nerve head circumference, PIMD-Average [0;2π], was determined semiautomatically. The measurements were analyzed with an analysis of variance for estimation of the variance components for subjects, visits, volumes and semi-automatic measurements of PIMD-Average [0;2π]. It was found that the variance for subjects was on the order of five times the variance for visits, and the variance for visits was on the order of 5 times higher than the variance for volumes. The variance for semi-automatic measurements of PIMD-Average [0;2π] was 3 orders of magnitude lower than the variance for volumes. A 95 % confidence interval for mean PIMD-Average [0;2π] was estimated to 1.00 ±0.13 mm (D.f. = 12). The variance estimates indicate that PIMD-Average [0;2π] is not suitable for comparison between a onetime estimate in a subject and a population reference interval. Cross-sectional independent group comparisons of PIMD-Average [0;2π] averaged over subjects will require inconveniently large sample sizes. However, cross-sectional independent group comparison of averages of within subject difference between baseline and follow-up can be made with reasonable sample sizes. Assuming a loss rate of 0.1 PIMD-Average [0;2π] per year and 4 visits per year it was found that approximately 18 months follow up is required before a significant change of PIMDAverage [0;2π] can be observed with a power of 0.8. This is shorter than what has been observed both for HRT measurements and automated perimetry measurements with a similar observation rate. It is concluded that PIMDAverage [0;2π] has the potential to detect deterioration of glaucoma quicker than currently available primary diagnostic instruments. To increase the efficiency of PIMD-Average [0;2π] further, the variation among visits within subject has to be reduced.

Ophthalmic optical coherence tomography (OCT) is a powerful tool which provides high resolution three dimensional (3D) volumetric image of human retina. However, the measurement data of OCT suffer motion artifact due to the involuntary eye motion during data acquisition. To eliminate this motion artifact and provide the true shape of retinal image, an eye motion corrected OCT imaging method based on Lissajous scan pattern is proposed in this paper. In this approach, we adopted Lissajous scan pattern for 3D-OCT imaging and developed motion correction algorithm. To verify the effectiveness of this method, we compare our method with single raster scan method by the experimental results. The experimental results show that the eye motion can be corrected by our method effectively.

Scanning laser ophthalmoscopy (SLO) and optical coherence tomography (OCT) benefit clinical diagnostic imaging in ophthalmology by enabling in vivo noninvasive en face and volumetric visualization of retinal structures, respectively. Spectrally encoding methods enable confocal imaging through fiber optics and reduces system complexity. Previous applications in ophthalmic imaging include spectrally encoded confocal scanning laser ophthalmoscopy (SECSLO) and a combined SECSLO-OCT system for image guidance, tracking, and registration. However, spectrally encoded imaging suffers from speckle noise because each spectrally encoded channel is effectively monochromatic. Here, we demonstrate in vivo human retinal imaging using a swept source spectrally encoded scanning laser ophthalmoscope and OCT (SSSESLO- OCT) at 1060 nm. SS-SESLO-OCT uses a shared 100 kHz Axsun swept source, shared scanner and imaging optics, and are detected simultaneously on a shared, dual channel high-speed digitizer. SESLO illumination and detection was performed using the single mode core and multimode inner cladding of a double clad fiber coupler, respectively, to preserve lateral resolution while improving collection efficiency and reducing speckle contrast at the expense of confocality. Concurrent en face SESLO and cross-sectional OCT images were acquired with 1376 x 500 pixels at 200 frames-per-second. Our system design is compact and uses a shared light source, imaging optics, and digitizer, which reduces overall system complexity and ensures inherent co-registration between SESLO and OCT FOVs. En face SESLO images acquired concurrent with OCT cross-sections enables lateral motion tracking and three-dimensional volume registration with broad applications in multivolume OCT averaging, image mosaicking, and intraoperative instrument tracking.

Scanning laser ophthalmoscopy is a confocal imaging technique that allows high-contrast imaging of retinal structures. Rapid, involuntary eye movements during image acquisition are known to cause artefacts and high-speed imaging of the retina is crucial to avoid them. To reach higher imaging speeds we propose to illuminate the retina with multiple parallel lines simultaneously within the whole field of view (FOV) instead of a single focused line that is raster-scanned. These multiple line patterns were generated with a digital micro-mirror device (DMD) and by shifting the line pattern, the whole FOV is scanned. The back-scattered light from the retinal layers is collected via a beam-splitter and imaged onto an area camera. After every pattern from the sequence is projected, the final image is generated by combining these back-reflected illumination patterns. Image processing is used to remove the background and out-of-focus light. Acquired pattern images are stacked, pixels sorted according to intensity and finally bottom layer of the stack is subtracted from the top layer to produce confocal image. The obtained confocal images are rich in structure, showing the small blood vessels around the macular avascular zone and the bow tie of Henle's fiber layer in the fovea. In the optic nerve head images the large arteries/veins, optic cup rim and cup itself are visualized. Images have good contrast and lateral resolution with a 10°×10° FOV. The initial results are promising for the development of high-speed retinal imaging using spatial light modulators such as the DMD.

In our previous reports we demonstrated a novel Fourier domain optical coherence tomography method, Master Slave optical coherence tomography (MS-OCT), that does not require resampling of data and can deliver en-face images from several depths simultaneously. While ideally suited for delivering information from a selected depth, the MS-OCT has been so far inferior to the conventional FFT based OCT in terms of time of producing cross section images. Here, we demonstrate that by taking advantage of the parallel processing capabilities offered by the MS-OCT method, cross-sectional OCT images of the human retina can be produced in real-time by assembling several T-scans from different depths. We analyze the conditions that ensure a real-time B-scan imaging operation, and demonstrate in-vivo real-time images from human fovea and the optic nerve, of comparable resolution and sensitivity to those produced using the traditional Fourier domain based method.

High-resolution optical coherence tomography (OCT) is of critical importance to disease diagnosis because it is capable of providing detailed microstructural information of the biological tissues. However, a compromise usually has to be made between its spatial resolutions and sensitivity due to the suboptimal spectral response of the system components, such as the linear camera, the dispersion grating, and the focusing lenses, etc. In this study, we demonstrate an OCT system that achieves both high spatial resolutions and enhanced sensitivity through utilizing a spectrally encoded source. The system achieves a lateral resolution of 3.1 μm and an axial resolution of 2.3 μm in air; when with a simple dispersive prism placed in the infinity space of the sample arm optics, the illumination beam on the sample is transformed into a line source with a visual angle of 10.3 mrad. Such an extended source technique allows a ~4 times larger maximum permissible exposure (MPE) than its point source counterpart, which thus improves the system sensitivity by ~6dB. In addition, the dispersive prism can be conveniently switched to a reflector. Such flexibility helps increase the penetration depth of the system without increasing the complexity of the current point source devices. We conducted experiments to characterize the system’s imaging capability using the human fingertip in vivo and the swine eye optic never disc ex vivo. The higher penetration depth of such a system over the conventional point source OCT system is also demonstrated in these two tissues.

Corneal biomechanical properties are influenced by several factors, including intraocular pressure, corneal thickness, and viscoelastic responses. Corneal thickness is directly proportional to tissue hydration and can influence corneal stiffness, but there is no consensus on the magnitude or direction of this effect. We evaluated the influence of corneal hydration on dynamic surface deformation responses using optical coherence elastography (OCE). Fresh rabbit eyes (n=10) were prepared by removing the corneal epithelium and dropping with 0.9% saline every 5 minutes for 1 hour, followed by 20% dextran solution every 5 minutes for one hour. Corneal thickness was determined from structural OCT imaging and OCE measurements were performed at baseline and every 20 minutes thereafter. Micron-scale deformations were induced at the apex of the corneal tissue using a spatially-focused (150μm) short-duration (<1ms) air-pulse delivery system. These dynamic tissue responses were measured non-invasively with a phase-stabilized swept source OCT system. The tissue surface deformation response (Relaxation Rate: RR) was quantified as the time constant, over which stimulated tissue recovered from the maximum deformation amplitude. Elastic wave group velocity (GV) was also quantified and correlated with change in corneal thickness due to hydration process. Corneal thickness rapidly increased and remained constant following epithelium removal and changed little thereafter. Likewise, corneal stiffness changed little over the first hour and then decreased sharply after Dextran application (thickness: -46% [-315/682 μm]; RR: - 24% [-0.7/2.88 ms^-1]; GV: -19% [-0.6/3.2 m/s]). Corneal thickness and corneal stiffness (RR) were well correlated (R² = .66). Corneal biomechanical properties are highly correlated with tissue hydration over a wide range of corneal thickness and these changes in corneal stiffness are quantifiable using OCE.

Keratoconus is a degenerative disorder of the eye characterized by human cornea thinning and morphological change to a more conical shape. Current diagnosis of this disease relies on topographic imaging of the cornea. Early and differential diagnosis is difficult. In keratoconus, mechanical properties are found to be compromised. A clinically available invasive technique capable of measuring the mechanical properties of the cornea is of significant importance for understanding the mechanism of keratoconus development and improve detection and intervention in keratoconus. The capability of Brillouin imaging to detect local longitudinal modulus in human cornea has been demonstrated previously. We report our non-contact, non-invasive, clinically viable Brillouin imaging system engineered to evaluate mechanical properties human cornea in vivo. The system takes advantage of a highly dispersive 2-stage virtually imaged phased array (VIPA) to detect weak Brillouin scattering signal from biological samples. With a 1.5-mW light beam from a 780-nm single-wavelength laser source, the system is able to detect Brillouin frequency shift of a single point in human cornea less than 0.3 second, at a 5μm/30μm lateral/axial resolution. Sensitivity of the system was quantified to be ~ 10 MHz. A-scans at different sample locations on a human cornea with a motorized human interface. We imaged both normal and keratoconic human corneas with this system. Whereas no significantly difference were observed outside keratocnic cones compared with normal cornea, a highly statistically significantly decrease was found in the cone regions.

The biomechanical properties of the cornea have a profound influence on its health and function. Rose bengal/green light corneal collagen cross-linking (RGX) has been proposed as an alternative to UV-A Riboflavin collagen cross-linking (UV-CXL) for treatment of keratoconus. However, the effects of RGX on the biomechanical properties of the cornea are not as well understood as UV-CXL. In this work, we demonstrate the feasibility of quantifying the viscoelasticity of the rabbit cornea before and after RGX using a noncontact method of phase-stabilized swept source optical coherence elastography (PhS-SSOCE) and finite element modeling (FEM). Viscoelastic FE models of the corneas were constructed to simulate the elastic wave propagation based on the OCE measurements. In addition, the effect of the fluid-structure interface (FSI) between the corneal posterior surface and aqueous humor on the elastic wave group velocity was also investigated. The effect of the FSI was first validated by OCE measurements and FEM simulations on contact lenses, and the OCE and FEM results were in good agreement. The Young’s modulus of the rabbit cornea before RGX was assessed as E=80 kPa, and the shear viscosity was η=0.40 Pa•s at an intraocular pressure (IOP) of 15 mmHg. After RGX, the Young’s modulus increased to E=112 kPa and shear viscosity decreased to η=0.37 Pa•s. Both the corneal OCE experiments and the FE simulations also demonstrated that the FSI significantly reduced the group velocity of the elastic wave, and thus, the FSI should be considered when determining the biomechanical properties of the cornea.

Keratoconus is an ophthalmic disease in which the cornea acquires an abnormal conical shape that prevents the correct focusing on the retina, causing visual impairment. The late diagnosis of keratoconus is among the principal causes of corneal transplantation surgery. In this study, we characterize the morphology of keratoconic corneas by means of the correlation of SHG images, finding that keratoconus can be diagnosed by analyzing the inclination of lamellae below Bowman’s membrane. In addition, imaging performed with both sagittal and “en face” geometry demonstrated that this morphological features can be highlighted both ex vivo and in vivo.

The use of a Prosthetic Replacement of the Ocular Surface Environment (PROSE) device is a revolutionary treatment for military patients that have lost their eyelids due to 3rd degree facial burns and for civilians who suffer from a host of corneal diseases. However, custom manual fitting is often a protracted painful, inexact process that requires multiple fitting sessions. Training for new practitioners is a long process. Automated methods to measure the complete corneal and scleral topology would provide a valuable tool for both clinicians and PROSE device manufacturers and would help streamline the fitting process. PSI has developed an ocular anterior-segment profiler based on Optical Coherence Tomography (OCT), which provides a 3D measure of the surface of the sclera and cornea. This device will provide topography data that will be used to expedite and improve the fabrication process for PROSE devices. OCT has been used to image portions of the cornea and sclera and to measure surface topology for smaller contact lenses [1-3]. However, current state-of-the-art anterior eye OCT systems can only scan about 16 mm of the eye’s anterior surface, which is not sufficient for covering the sclera around the cornea. In addition, there is no systematic method for scanning and aligning/stitching the full scleral/corneal surface and commercial segmentation software is not optimized for the PROSE application. Although preliminary, our results demonstrate the capability of PSI’s approach to generate accurate surface plots over relatively large areas of the eye, which is not currently possible with any other existing platform. Testing the technology on human volunteers is currently underway at Boston Foundation for Sight.

As there is no clinically available instrument to systematically and reliably determine the photosensitivity thresholds of patients with dry eyes, blepharospasms, migraines, traumatic brain injuries, and genetic disorders such as Achromatopsia, retinitis pigmentosa and other retinal dysfunctions, a computer-controlled optoelectronics system was designed. The BPEI Photosensitivity System provides a light stimuli emitted from a bi-cupola concave, 210 white LED array with varying intensity ranging from 1 to 32,000 lux. The system can either utilize a normal or an enhanced testing mode for subjects with low light tolerance. The automated instrument adjusts the intensity of each light stimulus. The subject is instructed to indicate discomfort by pressing a hand-held button. Reliability of the responses is tracked during the test. The photosensitivity threshold is then calculated after 10 response reversals. In a preliminary study, we demonstrated that subjects suffering from Achromatopsia experienced lower photosensitivity thresholds than normal subjects. Hence, the system can safely and reliably determine the photosensitivity thresholds of healthy and light sensitive subjects by detecting and quantifying the individual differences. Future studies will be performed with this system to determine the photosensitivity threshold differences between normal subjects and subjects suffering from other conditions that affect light sensitivity.

Light sensation relies on photoisomerization of chromophores in rod and cone photoreceptor cells. Spectral sensitivity of these photoreceptor cells in the retina is determined by the absorption spectra of their pigments which covers a range from 400 nm to above 700 nm. Regardless the mechanism leading to visual pigment isomerization, light sensation is triggered every time visual pigment molecules change their conformation. Thus, two-photon absorption (TPA) should produce the same result (visual sensation) as single photon absorption of light. This observation was positively verified and published by our group. During human psychophysics experiments, we found that humans can perceive light in the infrared (IR) range as colors that match half of the wavelength of the applied laser beam. Other experiments and theoretical research, such as mouse electrophysiology, biochemical studies of TPA in rhodopsin or molecular modeling studies, confirmed that visual sensation can be triggered by TPA. There are few publications describing human near infrared (NIR) perception and no formal proposals to use this phenomenon to improve ophthalmic diagnosis and monitor treatment. Here we report that the use of novel instrumentation revealed that the sensitivity threshold for NIR vision depends on age.

Purpose: To objectively assess visual field (VF) defects and retinal cell function in healthy subjects and patients with macular degeneration using a chromatic multifocal pupillometer. Methods: A multifocal chromatic pupillometer (MCP) was used to record pupillary responses (PR) of 17 healthy subjects and 5 Best Vitelliform macular dystrophy patients. Blue and red light stimuli (peak 485nm and 620nm, respectively) were presented at light intensities of 400 and 1000 cd/m2, respectively at 76 different points in a 16.2 degree VF. The PR of patients were compared with their findings on Humphrey's 24-2 perimetry, optical coherence tomography and the PR obtained from healthy subjects. Results: Patients demonstrated reduced percentage of pupillary contraction and slower maximal contraction velocity, more than two standard errors (SE) away from the mean of healthy subjects in response to red light in majority of VF locations. In response to blue light, the percentage of pupillary contraction was lower (by over two SE) compared with normal controls only in central locations. The latency of maximal contraction velocity was shorter in patients compared with healthy subjects in response to both colors. Conclusions: This study demonstrated the advantage of using MCP-based objective VF to assess central scotoma in macular degeneration. Our finding also suggests that chromatic perimetry may differentiate between PR mediated by cones and rods, and can specifically detect defects in macular cones. Different parameters of PR such as latency of maximal contraction velocity may shed light on the pathophysiology of different blinding diseases.

The purpose of this project is to design and evaluate a system that will enable objective assessment of the optical accommodative response in real-time while acquiring axial biometric information. The system combines three sub-systems which were integrated and mounted on a joystick x-y-z adjustable modified slit-lamp base to facilitate alignment and data acquisition: (1) a Shack-Hartmann wavefront sensor for dynamic refraction measurement, provided software calculates sphere, cylinder and axis values, (2) an extended-depth Optical Coherence Tomography (OCT) system using an optical switch records high-resolution cross-sectional images across the length of the eye, from which, dynamic axial biometry (corneal thickness, anterior chamber depth, crystalline lens thickness and vitreous depth) can be extracted, and (3) a modified dual-channel accommodation stimulus unit based on the Badal optometer for providing a step change in accommodative stimulus. The prototypal system is capable of taking simultaneous measurements of both the optical and the mechanical response of lens accommodation. These measurements can provide insight into correlating changes in lens shape with changes in lens power and ocular refraction and ultimately provide a more comprehensive understanding of accommodation, presbyopia and an objective assessment of presbyopia correction techniques.

In this paper we discuss the problem of children myopia control and how extended depth of focus ophthalmic solutions can assist in resolving this problem.

Laser radiation entering the eye has the potential of damaging the retina. As an inflammatory response, the proteins can rush to the lesion site created by laser exposure. We explore the hypothesis if these proteins can be detected non-invasively. In this preliminary study, we developed a new brief-case size dynamic light scattering instrument to detect these proteins in-vivo in the rabbit vitreous. The results were validated with bio-chemical analysis.

Developing a one-second automatic glaucoma treatment using trans-scleral laser trabeculoplasty (LTP) without a gonioscopy lens Purpose: Developing an LTP device for delivering multiple simultaneous trans-scleral applications of low energy laser irradiation to the trabecular meshwork (TM) for reducing Intraocular Pressure (IOP). Methods: Concept proof: A randomized, masked, controlled one was performed on open angle glaucoma patients. The control group underwent conventional SLT (100 laser spots through a gonioscope for 360 degrees directly on the TM). The trial group underwent irradiation by the same laser at the same irradiation parameters on the sclera overlying the TM. Topical glaucoma therapy was not changed during the 12 months trial. Feasibility trial: Using optimized laser parameters, 60 discrete applications were administered on similar locations of patients’ sclera. Results: Concept proof: Trans-scleral applications: (N=15), IOP decrease from 20.21 mmHg before treatment to 16.00 (27.1%) at one year. The corresponding numbers for the control group (n=15), were 21.14 mmHg and 14.30 (23.4%). There was no statistical difference between the two groups in IOP reduction. The complications rate was significantly higher in the control group. Trial 2: IOP was reduced from an of 25.3 mmHg to 19.3 (23.7%) in the 11 patients. Conclusions: Laser coherency, lost in tissue transmission, is not required for the therapeutic effect. The new method will possibly enable treatment of angle closure glaucoma as well as simultaneous applications of all laser spots to the sclera. When used conjointly with target acquisition, will make feasible an automatic glaucoma treatment in less than one second.

Unlike conventional photocoagulation, non-damaging retinal laser therapy (NRT) limits laser-induced heating to stay below the retinal damage threshold and therefore requires careful dosimetry. Without the adverse effects associated with photocoagulation, NRT can be applied to critical areas of the retina and repeatedly to manage chronic disorders. Although the clinical benefits of NRT have been demonstrated, the mechanism of therapeutic effect and width of the therapeutic window below damage threshold are not well understood. Here, we measure activation of heat shock response via laser-induced hyperthermia as one indication of cellular response. A 577 nm laser is used with the Endpoint Management (EpM) user interface, a titration algorithm, to set experimental pulse energies relative to a barely visible titration lesion. Live/dead staining and histology show that the retinal damage threshold in rabbits is at 40% of titration energy on EpM scale. Heat shock protein 70 (HSP70) expression in the retinal pigment epithelium (RPE) was detected by whole-mount immunohistochemistry after different levels of laser treatment. We show HSP70 expression in the RPE beginning at 25% of titration energy indicating that there is a window for NRT between 25% and 40% with activation of the heat shock protein expression in response to hyperthermia. HSP70 expression is also seen at the perimeter of damaging lesions, as expected based on a computational model of laser heating. Expression area for each pulse energy setting varied between laser spots due to pigmentation changes, indicating the relatively narrow window of non-damaging activation and highlighting the importance of proper titration.

Photocoagulation is a laser treatment widely used for the therapy of several retinal diseases. Intra- and inter-individual variations of the ocular transmission, light scattering and the retinal absorption makes it impossible to achieve a uniform effective exposure and hence a uniform damage throughout the therapy. A real-time monitoring and control of the induced damage is highly requested. Here, an approach to realize a real time optical feedback using dynamic speckle analysis is presented. A 532 nm continuous wave Nd:YAG laser is used for coagulation. During coagulation, speckle dynamics are monitored by a coherent object illumination using a 633nm HeNe laser and analyzed by a CMOS camera with a frame rate up to 1 kHz. It is obvious that a control system needs to determine whether the desired damage is achieved to shut down the system in a fraction of the exposure time. Here we use a fast and simple adaption of the generalized difference algorithm to analyze the speckle movements. This algorithm runs on a FPGA and is able to calculate a feedback value which is correlated to the thermal and coagulation induced tissue motion and thus the achieved damage. For different spot sizes (50-200 μm) and different exposure times (50-500 ms) the algorithm shows the ability to discriminate between different categories of retinal pigment epithelial damage ex-vivo in enucleated porcine eyes. Furthermore in-vivo experiments in rabbits show the ability of the system to determine tissue changes in living tissue during coagulation.

Some of the most commonly performed surgical operations in the world, including laser-assisted in-situ keratomileusis (LASIK), lens replacement (e.g. cataract surgery), and keratoplasty (cornea transplant), now employ therapeutic infrared femtosecond lasers (FSLs) for their extreme precision, low energy delivered into tissue and advanced ablation characteristics. Although the widely exploited applications of FSLs in medical therapeutics offer significant benefits, FSLs must generate very high intensities in order to achieve optical breakdown, the predominant tissue ablative mechanism, which can also stimulate nonlinear optical effects such as harmonic generation, an effect that generates coherent visible and UV light in the case of second- (SHG) and third-harmonic generation (THG), respectively. In order to improve the understanding of HG in corneal tissue, the effect of FSL polarization and pulse energy were investigated. FSL stimulated SHG intensity in corneal tissue was measured as the laser polarization was rotated 360 degrees. Further, the pulse energy at the SHG wavelength were measured for single FSL pulses as the pulse energy at the fundamental wavelength was varied through a range of clinically relevant values. The results of this study revealed SHG intensity oscillated with laser polarization, having a variation greater than 20%. This relationship seems to due to the intrinsic anisotropy of collagen fibril hyperpolarizability, not related to tissue birefringence. SHG pulse energy measurements showed an increase in SHG pulse energy with increasing FSL pulse energy, however conversion efficiency decreased. This may be related to the dynamic relationship between optical breakdown leading to tissue destruction and HG evolution.

Keratitis associated with Pseudomonas aeruginosa is difficult to manage. Treatment includes antibiotic eye drops, however, some strains of Pseudomonas aeruginosa are resistant. Current research efforts are focused on finding alternative and adjunct therapies to treat multi-drug resistant bacteria. One promising alternate technique is photodynamic therapy (PDT). The purpose of this study was to evaluate the effect of riboflavin- and rose bengal-mediated PDT on Pseudomonas aeruginosa keratitis isolates in vitro. Two isolates (S+U- and S-U+) of Pseudomonas aeruginosa were derived from keratitis patients and exposed to five experimental groups: (1) Control (dark, UV-A irradiation, 525nm irradiation); (2) 0.1% riboflavin (dark, UV-A irradiation); and (3) 0.1% rose bengal, (4) 0.05% rose bengal and (5) 0.01% rose bengal (dark, 525nm irradiation). Three days after treatment, in dark conditions of all concentration of riboflavin and rose bengal showed no inhibition in both S+U- and S-U+ strains of Pseudomonas aeruginosa. In 0.1% and 0.05% rose bengal irradiated groups, for both S+U- and S-U+ strains, there was complete inhibition of bacterial growth in the central 50mm zone corresponding to the diameter of the green light source. These in vitro results suggest that rose bengal photodynamic therapy may be an effective adjunct treatment for Pseudomonas aeruginosa keratitis.

Retinal pigment epithelium (RPE) cells are vital to health of the outer retina, but are often compromised in ageing and major ocular diseases that lead to blindness. Early manifestation of RPE disruption occurs at the cellular level, and while biomarkers at this scale hold considerable promise, RPE cells have proven extremely challenging to image in the living human eye. We present a novel method based on optical coherence tomography (OCT) equipped with adaptive optics (AO) that overcomes the associated technical obstacles. The method takes advantage of the 3D resolution of AO-OCT, but more critically sub-cellular segmentation and registration that permit organelle motility to be used as a novel contrast mechanism. With this method, we successfully visualized RPE cells and characterized their 3D reflectance profile in every subject and retinal location (3° and 7° temporal to the fovea) imaged to date. We have quantified RPE packing geometry in terms of cell density, cone-to-RPE ratio, and number of nearest neighbors using Voronoi and power spectra analyses. RPE cell density (cells/mm²) showed no significant difference between 3° (4,892±691) and 7° (4,780±354). In contrast, cone-to- RPE ratio was significantly higher at 3° (3.88±0.52:1) than 7° (2.31± 0.23:1). Voronoi analysis also showed most RPE cells have six nearest neighbors, which was significantly larger than the next two most prevalent associations: five and seven. Averaged across the five subjects, prevalence of cells with six neighbors was 51.4±3.58% at 3°, and 54.58±3.01% at 7°. These results are consistent with histology and in vivo studies using other imaging modalities.

It is well known that patient-specific ocular aberrations limit imaging resolution in the human retina. Previously, hardware adaptive optics (HAO) has been employed to measure and correct these aberrations to acquire high-resolution images of various retinal structures. While the resulting aberration-corrected images are of great clinical importance, clinical use of HAO has not been widespread due to the cost and complexity of these systems. We present a technique termed computational adaptive optics (CAO) for aberration correction in the living human retina without the use of hardware adaptive optics components. In CAO, complex interferometric data acquired using optical coherence tomography (OCT) is manipulated in post-processing to adjust the phase of the optical wavefront. In this way, the aberrated wavefront can be corrected. We summarize recent results in this technology for retinal imaging, including aberration-corrected imaging in multiple retinal layers and practical considerations such as phase stability and image optimization.

We imaged the retina using the Indiana Adaptive Optics Scanning Laser Ophthalmoscope (AOSLO). Our system uses two deformable mirrors to provide en face, high-resolution images of retinal structures at a 28 Hz frame rate. The wavelength of the sensor light was 850 nm and the imaging wavelength was 820 nm at 50 and 120 W respectively. The confocal pinhole was located in a position conjugate with the retina allowed us to segment one retina plane. Two different confocal apertures of 75 m and 100 m (1.5 and 2 times the Airy disk size) were used to provide different amounts of confocal or scattered light. The imaging area was 1.4 x 1.2 deg which corresponds roughly to 400 x 350 m. Using the large stroke deformable mirror, which provides the focusing capability of the confocal system, we imaged the same location at different planes. We moved from superficial layers to the retinal pigment epithelium in 0.3 D increments. The range of adjustments included the subjectively best overall image, and focal planes anterior and posterior to this. We imaged 10 subjects at approximately 7.5 deg temporal from the fovea. A video of individual frames was taken, and the individual frames were dewarped, aligned, and averaged. We measured 10 bright and 10 dim cones for each subject at the 10 depths, with brightness groupings based subjectively on the most superficial location. The function for amount of light reflected differed for the two groups of cones. Reflectivity varied as a function of depth.

In vivo photoreceptor imaging has enhanced the way vision scientists and ophthalmologists understand the retinal structure, function, and etiology of numerous retinal pathologies. However, the complexity and large footprint of current systems capable of resolving photoreceptors has limited imaging to patients who are able to sit in an upright position and fixate for several minutes. Unfortunately, this excludes an important fraction of patients including bedridden patients, small children, and infants. Here, we show that our dual-modality, high-resolution handheld probe with a weight of only 94 g is capable of visualizing photoreceptors in supine children. Our device utilizes a microelectromechanical systems (MEMS) scanner and a novel telescope design to achieve over an order of magnitude reduction in size compared to similar systems. The probe has a 7° field of view and a lateral resolution of 8 µm. The optical coherence tomography (OCT) system has an axial resolution of 7 µm and a sensitivity of 101 dB. High definition scanning laser ophthalmoscopy (SLO) and OCT images were acquired from children ranging from 14 months to 12 years of age with and without pathology during examination under anesthesia in the operating room. Parafoveal cone imaging was shown using the SLO arm of this device without adaptive optics using a 3° FOV for the first time in children under 4 years old. This work lays the foundation for pediatric research, which will improve understanding of retinal development, maldevelopment and early onset of diseases at the cellular level during the beginning stages of human growth.

We present retinal photoreceptor imaging with a line-field parallel spectral domain OCT modality, utilizing a commercially available 2D CMOS detector array operating at and imaging speed of 500 B-scans/s. Our results demonstrate for the first time in vivo structural and functional retinal assessment with a line-field OCT setup providing sufficient sensitivity, lateral and axial resolution and 3D acquisition rates in order to resolve individual photoreceptor cells. The setup comprises a Michelson interferometer illuminated by a broadband light source, where a line-focus is formed via a cylindrical lens and the back-propagated light from sample and reference arm is detected by a 2D array after passing a diffraction grating. The spot size of the line-focus on the retina is 5μm, which corresponds to a PSF of 50μm and an oversampling factor of 3.6 at the detector plane, respectively. A full 3D stack was recorded in only 0.8 s. We show representative enface images, tomograms and phase-difference maps of cone photoreceptors with a lateral FOV close to 2°. The high-speed capability and the phase stability due to parallel illumination and detection may potentially lead to novel structural and functional diagnostic tools on a cellular and microvascular imaging level. Furthermore, the presented system enables competitive imaging results as compared to respective point scanning modalities and facilitates utilizing software based digital aberration correction algorithms for achieving 3D isotropic resolution across the full FOV.

Forward scattered light from the anterior segment of the human eye can be measured by Shack-Hartmann (SH) wavefront aberrometers with limited visual angle. We propose a novel Point Spread Function (PSF) reconstruction algorithm based on SH measurements with a novel measurement devise to overcome these limitations. In our optical setup, we use a Digital Mirror Device as variable field stop, which is conventionally a pinhole suppressing scatter and reflections. Images with 21 different stop diameters were captured and from each image the average subaperture image intensity and the average intensity of the pupil were computed. The 21 intensities represent integral values of the PSF which is consequently reconstructed by derivation with respect to the visual angle. A generalized form of the Stiles-Holladay-approximation is fitted to the PSF resulting in a stray light parameter Log(IS). Additionaly the transmission loss of eye is computed. For the proof of principle, a study on 13 healthy young volunteers was carried out. Scatter filters were positioned in front of the volunteer’s eye during C-Quant and scatter measurements to generate straylight emulating scatter in the lens. The straylight parameter is compared to the C-Quant measurement parameter Log(ISC) and scatter density of the filters SDF with a partial correlation. Log(IS) shows significant correlation with the SDF and Log(ISC). The correlation is more prominent between Log(IS) combined with the transmission loss and the SDF and Log(ISC). Our novel measurement and reconstruction technique allow for objective stray light analysis of visual angles up to 4 degrees.

We introduce a system for rapidly measuring the intraocular distances of human eyes in vivo with high sensitivity by using Fourier domain low-coherence interferometry. The system mainly consisting of a rapid focus displacement unit and a reference arm which has a variable optical path length. This system is capable of providing a complete biometrical assessment of a human eye in a single measurement procedure, including cornea thickness, anterior chamber depth, lens thickness, and axial length. The system is experimentally verified by measuring the four parameters of a human eye in vivo.

An optical system that conjugates the patient’s pupil to the plane of a Hartmann-Shack (HS) wavefront sensor has been simulated using optical design software. And an optical bench prototype is mounted using mechanical eye device, beam splitter, illumination system, lenses, mirrors, mirrored prism, movable mirror, wavefront sensor and camera CCD. The mechanical eye device is used to simulate aberrations of the eye. From this device the rays are emitted and travelled by the beam splitter to the optical system. Some rays fall on the camera CCD and others pass in the optical system and finally reach the sensor. The eye models based on typical in vivo eye aberrations is constructed using the optical design software Zemax. The computer-aided outcomes of each HS images for each case are acquired, and these images are processed using customized techniques. The simulated and real images for low order aberrations are compared using centroid coordinates to assure that the optical system is constructed precisely in order to match the simulated system. Afterwards a simulated version of retinal images is constructed to show how these typical eyes would perceive an optotype positioned 20 ft away. Certain personalized corrections are allowed by eye doctors based on different Zernike polynomial values and the optical images are rendered to the new parameters. Optical images of how that eye would see with or without corrections of certain aberrations are generated in order to allow which aberrations can be corrected and in which degree. The patient can then “personalize” the correction to their own satisfaction. This new approach to wavefront sensing is a promising change in paradigm towards the betterment of the patient-physician relationship.

Cataract is one of the most common degenerative diseases that causes blindness. Careful quantification of lens biomechanical properties can greatly assist in early detection of the disease as well as personalization of treatment procedures. In this study, we utilize a phase-sensitive optical coherence elastography (OCE) system to assess the effects of the cold cataract on the biomechanical properties of porcine crystalline lens in vitro. Relaxation rates of air puff induced elastic waves were measured on the same crystalline lens with and without cold cataract. Results demonstrate that the relaxation rate and, thus, associated elasticity of the porcine lens, increased due to the presence of cold cataract.

Purpose: Fluorescence lifetime imaging ophthalmoscopy (FLIO) provides in vivo metabolic mapping of the ocular fundus. Changes in FLIO have been found in e.g. diabetes patients. The influence of short term metabolic changes caused by blood glucose level changes on is unknown. Aim of this work is the detection of short-term changes in fundus autofluorescence lifetime during an oral glucose tolerance test. Methods: FLIO was performed in 10 healthy volunteers (29±4 years, fasting for 12h) using a scanning laser ophthalmoscope (30° fundus, 34μm resolution, excitation with 473nm diode laser with 70 ps pulses at 80 MHz repetition rate, detection in two spectral channels 500-560nm (ch1) and 560-720nm (ch2) using the timecorrelated single photon counting method). The blood glucose level (BGL) was measured by an Accu-Chek® Aviva self-monitoring device. Before and after a glucose drink (300ml solution, containing 75g of glucose (Accu-Chek® Dextrose O.G.T.), BGL and FLIO were measured every 15min. The FLIMX software package was applied to compute the average fluorescence lifetime τ on the inner ring of the ETDRS grid using a modified 3-exponential approach. Results: The results are given as mean ± standard deviation over all volunteers in ch1. Baseline measurement: BGL: 5.3±0.4 mmol/l, τ₁: 49±6ps. A significant reduction (α=5%; Wilcoxon rank-sum test) in τ₁ is detected after 15min (BGL: 8.4±1.1 mmol/l, τ₁: 44±5ps) and after 90min (BGL: 6.3±1.4 mmol/l, τ₁: 41±5ps). Results of ch2 show smaller reductions in the fluorescence lifetimes over time.

The biomechanical properties of the cornea are critical factors which determine its health and subsequent visual acuity. Keratoconus is a structural degeneration of the cornea which can diminish vision quality. Riboflavin/UV-A corneal collagen cross-linking (UV-CXL) is an emerging treatment that increases the stiffness of the cornea and improves its ability to resist further degeneration. While UV-CXL has shown great promise for effective therapy of the keratoconus, there are concerns associated with the UV irradiation, such as keratocyte cytotoxicity. Rose-bengal/green light corneal collagen cross-linking (RGX) has been proposed as an alternative to UV-CXL. Because of the high absorbance of the rose-bengal dye at green wavelengths, the treatment time is significantly shorter than with UV-CXL. Moreover, because green light is used in lieu of UV irradiation, there are no cytotoxic side-effects. In this study, noncontact optical coherence elastography (OCE) was used to compare the outcomes of UV-CXL and RGX treatment in rabbit cornea. Low-amplitude (micrometer scale) elastic waves were induced by a focused air-pulse loading system. The elastic wave propagation was then imaged by a phase-stabilized swept source OCE (PhS-SSOCE) system. The changes in the viscoelasticity of the corneas were quantified by a previously developed modified Rayleigh Lamb frequency model. The depth-resolved micro-scale phase-velocity distribution in the cornea was used to reveal the depth-wise heterogeneity before and after both cross-linking techniques. Our results show that UV-CXL and RGX increased the stiffness of the corneas by ~54% and ~5% while reducing the viscosity by ~42% and ~17%, respectively. The depth-wise phase velocities showed that UV-CXL affected the anterior ~1/3 of the corneas, while RGX only affected the anterior ~1/7 of the corneas.

Optical Coherence Tomography (OCT) is a non-invasive 3 dimensional optical imaging modality that enables high resolution cross sectional imaging in biological tissues and materials. Its high axial and lateral resolution combined with high sensitivity, imaging depth and wide field of view makes it suitable for wide variety of high resolution medical imaging applications at clinically relevant speed. With the advent of swept source lasers, the imaging speed of OCT has increased considerably in recent years. OCT has been used in ophthalmology to study dynamic changes occurring in the cornea and iris, thereby providing physiological and pathological changes that occur within the anterior segment structures such as in glaucoma, during refractive surgery, lamellar keratoplasty and corneal diseases. In this study, we assess the changes in corneal thickness in the anterior segment of the eye during wound healing process in a rat corneal burn model following stem cell therapy using high speed swept source OCT.

As the world’s population ages, cataract-induced visual dysfunction and blindness is on the increase. This is a significant global problem. The most common symptoms of cataracts are glared and blurred vision. Usually, people with cataract have trouble seeing and reading at distance or in low light and also their color perception is altered. Furthermore, cataract is a sneaky disease as it is usually a very slow but progressive process, which creates adaptation so that patients find it difficult to recognize. All this can be very difficult to explain, so we built and tested an optical device to help doctors giving comprehensive answers to the patients’ symptoms. This device allows visualizing how cataract impairs vision mimicking the optical degradation of the crystalline related cataracts. This can be a valuable optical tool for medical education as well as to provide a method to illustrate the patients how cataract progression process will affect their vision.

Cataract, a clouding of the crystalline eye lens, is the leading cause of blindness. It can effectively be treated by cataract surgery, where the clouded lens is replaced by an artificial intraocular lens (IOL). Postoperative healing processes can cause a displacement of the IOL, which further leads to the fact that the quality of vision is deteriorated. Studies have shown that the imaging quality of high sophisticated IOL designs is more sensitive to lens displacements than simpler designs. The effects of IOL displacements are not well represented and tested within the current IOL test standard ISO 11979-2. This fact leads to the necessity to develope new test standards for novel and more sophisticated IOL designs. In this paper we present an improved model eye, which extends the current standard in three main aspects: First, the eye-model is very close to the physiology of the human eye. Second, electromechanic drives allow an automatic and precise simulation of postoperative lens tilts and decentrations, and finally in addition to standard conform MTF analysis, in the proposed setup also wavefront aberrations are measured. The latter reveals specific image aberrations caused by lens displacements. The model eye allows to objectively analyze the displacement tolerance of various IOL designs. The functionality of this novel setup is tested by measuring a spherical and an aspheric IOL design. Additionally, for comparison, IOLs that were already investigated with a previous version of the presented model eye are used for analysis. Measurements results reveal improvements compared to the previous version of the model eye and a functional prototype for wavefront measurement.

The aim of this study was to determine the potential of a novel diode-pumped Er:YAG laser for phacoemulsification in basic experimental investigations. An appropriate experimental setup was created, including a translation stage for sample movement, a sample holder, a water spray for sample humidification and a surgical microscope with a CCD camera for video documentation. The analysis of the laser cuts and histological sections was done by light microscopy. As samples porcine eye lenses hardened by formalin were used. In ablation experiments with different spot diameters and radiant powers and a constant repetition rate ν_r = 200 Hz the maximum ablation depths of (4.346 ± 0.044) mm have reached at (Ø = 480 μm, Φ = 24.15 W) with a maximum extend of thermal damage of (0.165 ± 0.030) mm. The average ablation efficiency is 0.241 mm³/J. With a spot diameter of 308 μm the maximum ablation depth is (4.238 ± 0.040) mm at 24.65 W with a mean ablation efficiency of 0.293 mm3/J. The extend of the thermally damaged region is (0.171 ± 0.024) mm at this laser power. Using a sapphire cylinder with a diameter of 412 μm (length 38.5 mm) in direct tissue contact with water spray for sample humidification the ablation depth reaches (1.017 ± 0.074) mm at 4.93 W and (1.840 ± 0.092) mm at 9.87 W with a mean efficiency of 0.261 mm3/J. A thermal damage zone of (0.064 ±0.024) mm at 9.87 W was measured. Additionally, at this high power, a progressive contamination and destruction of the cylinder end facet was observed. In conclusion, the investigations show that the diode-pumped Er:YAG laser has considerable potential for cataract surgery.

The accuracy of the estimation of optical aberrations by measuring the distorted wave front using a Hartmann-Shack wave front sensor (HSWS) is mainly dependent upon the measurement accuracy of the centroid of the focal spot. The most commonly used methods for centroid estimation such as the brightest spot centroid; first moment centroid; weighted center of gravity and intensity weighted center of gravity, are generally applied on the entire individual sub-apertures of the lens let array. However, these processes of centroid estimation are sensitive to the influence of reflections, scattered light, and noise; especially in the case where the signal spot area is smaller compared to the whole sub-aperture area. In this paper, we give a comparison of performance of the commonly used centroiding methods on estimation of optical aberrations, with and without the use of some pre-processing steps (thresholding, Gaussian smoothing and adaptive windowing). As an example we use the aberrations of the human eye model. This is done using the raw data collected from a custom made ophthalmic aberrometer and a model eye to emulate myopic and hyper-metropic defocus values up to 2 Diopters. We show that the use of any simple centroiding algorithm is sufficient in the case of ophthalmic applications for estimating aberrations within the typical clinically acceptable limits of a quarter Diopter margins, when certain pre-processing steps to reduce the impact of external factors are used.