We investigate the influence of the OCT system resolution on high-quality en face corneal endothelial cell images in vivo, to allow for quantitative analysis of cell density. We vary the lateral resolution of the ultrahigh-resolution (UHR) OCT system (centered at 850 nm) by using different objectives, and the axial resolution by windowing the source spectrum. We are able to obtain a high-quality en face corneal endothelial cell map in vivo using UHR OCT for the first time. Quantitative analysis result of cell density from in vivo en face corneal endothelial cell map agrees with previously reported data.
A multi-functional OCT approach is used to identify different tissue types during the early development of spontaneous neovascularizations in the mouse retina based on their intrinsic optical properties.
OCT imaging in the super-resolution regime was investigated using simulations and experiments. Samples of known thickness in the range 46-163 nm were fabricated and imaged. Measurements of the tear film lipid layer were performed.
Corneal B-scan images and signal-to-noise ratio measurements using ultra-high resolution Spectral Domain Optical Coherence Tomography (SD-OCT) are reported. A comparison of results is obtained using a Ti:Sa laser and a supercontinuum optical source, is performed. Beside some differences in the SNR, the images are strikingly similar.
Dry eye syndrome is a highly prevalent disease of the ocular surface characterized by an instability of the tear film. Traditional methods used for the evaluation of tear film stability are invasive or show limited repeatability. Here we propose a new noninvasive approach to measure tear film thickness using an efficient delay estimator and ultrahigh resolution spectral domain OCT. Silicon wafer phantoms with layers of known thickness and group index were used to validate the estimator-based thickness measurement. A theoretical analysis of the fundamental limit of the precision of the estimator is presented and the analytical expression of the Cramér-Rao lower bound (CRLB), which is the minimum variance that may be achieved by any unbiased estimator, is derived. The performance of the estimator against noise was investigated using simulations. We found that the proposed estimator reaches the CRLB associated with the OCT amplitude signal. The technique was applied in vivo in healthy subjects and dry eye patients. Series of tear film thickness maps were generated, allowing for the visualization of tear film dynamics. Our results show that the central tear film thickness precisely measured in vivo with a coefficient of variation of about 0.65% and that repeatable tear film dynamics can be observed. The presented method has the potential of being an alternative to breakup time measurements (BUT) and could be used in clinical setting to study patients with dry eye disease and monitor their treatments.
René Werkmeister, Martin Vietauer, Corinna Knopf, Clemens Fürnsinn, Rainer Leitgeb, Herbert Reitsamer, Martin Gröschl, Gerhard Garhöfer, Walthard Vilser, Leopold Schmetterer
A wide variety of ocular diseases are associated with abnormalities in ocular circulation. As such, there is considerable interest in techniques for quantifying retinal blood flow, among which Doppler optical coherence tomography (OCT) may be the most promising. We present an approach to measure retinal blood flow in the rat using a new optical system that combines the measurement of blood flow velocities via Doppler Fourier-domain optical coherence tomography and the measurement of vessel diameters using a fundus camera-based technique. Relying on fundus images for extraction of retinal vessel diameters instead of OCT images improves the reliability of the technique. The system was operated with an 841-nm superluminescent diode and a charge-coupled device camera that could be operated at a line rate of 20 kHz. We show that the system is capable of quantifying the response of 100% oxygen breathing on the retinal blood flow. In six rats, we observed a decrease in retinal vessel diameters of 13.2% and a decrease in retinal blood velocity of 42.6%, leading to a decrease in retinal blood flow of 56.7%. Furthermore, in four rats, the response of retinal blood flow during stimulation with diffuse flicker light was assessed. Retinal vessel diameter and blood velocity increased by 3.4% and 28.1%, respectively, leading to a relative increase in blood flow of 36.2%. The presented technique shows much promise to quantify early changes in retinal blood flow during provocation with various stimuli in rodent models of ocular diseases in rats.
We present a method capable of measuring the total retinal blood flow in arteries and veins based on dual beam Fourierdomain Doppler optical coherence tomography (OCT) in combination with a fundus camera based Dynamic Vessel Analyzer. Incorporating a Dynamic vessel analyzer into the system not only gives a live image of the fundus – it also allows determining the vessels’ diameter precisely during the OCT measurement, which is necessary for the determination of the blood flow. While dual beam systems with fixed detection plane allow only vessels with certain orientations to be measured, the detection plane of our system can be rotated by 90°. This ensures that the blood’s velocity can be measured in all vessels around the optic nerve head. The results of the total blood flow measurements are in the same range as previously published data. Additionally, the high degree of conformity between the measured venous and arterial flow corroborated the system’s validity. For larger vessels, the logarithmic values of vessel diameter and blood flow were found to be related linearly with a regression coefficient of around 3, which is in accordance with Murray’s law. For smaller vessels (diameter below 60 μm), the values diverge from the linear dependence. The high sensitivity and the good agreement with published data suggest a high potential for examining the retinal blood flow in patients with ocular diseases.
We developed a three-beam Doppler optical coherence tomography (OCT) system that enables measurement of the velocity vector of moving particles in three-dimensions (3-D). The spatial orientation as well as the magnitude of motion can be determined without prior knowledge of the geometry of motion. The system combines three spectral-domain OCT interferometers whose sample beams are focused at the sample by a common focusing lens at three different angles. This provides three spatially independent velocity components simultaneously from which the velocity vector can be reconstructed. We demonstrate the system in a simple test object (rotating disc), a flow phantom, and for blood flow measurements in the retina of a healthy human subject. Measurements of blood flow at a venous bifurcation achieve a good agreement between in- and outflow and demonstrate the reliability of the method.
Low-coherence tissue interferometry is a technique for the depth-resolved measurement of ocular fundus pulsations. Whereas fundus pulsation amplitudes at preselected axial positions can readily be assessed by this method, coupling of the interferometer with a pulse oximeter additionally allows for the reconstruction of the time course of ocular fundus pulsation with respect to the cardiac cycle of the subject. For this purpose, the interferogram resulting from the superposition of waves reflected at the cornea and the ocular fundus is recorded synchronously with the plethysmogram. A new method for evaluating the time course of synthetic interferograms in combination with plethysmograms based on averaging several pulse periods has been developed. This technique allows for the analysis of amplitudes, time courses, and phase differences of fundus pulsations at preselected axial and transversal positions and for creating fundus pulsation movies. Measurements are performed in three healthy emmetropic subjects at angles from 0 deg to 18 deg to the axis of vision. Considerably different time courses, amplitudes, and phases with respect to the cardiac cycle are found at different angles. Data on ocular fundus pulsation obtained with this technique can—among other applications—be used to verify and to improve biomechanical models of the eye.
Traditional Doppler OCT is highly sensitive to motion artifacts due to the dependence on the Doppler angle. This limits its reproducibility in clinical practice. To overcome this limitation, we use a bidirectional technique with a novel rotating scanning scheme. The volume is probed simultaneously from two distinct illumination directions with variable controlled orientations, allowing reconstruction of the true flow velocity, independently of the vessel orientation. A Dove prism in the sample arm permits a rotation of the illumination directions that can be synchronized with the standard beam steering device. The principle is implemented with Swept Source OCT at 1060nm with 100,000 A-Scans/s. We apply the system to human retinal absolute blood velocity measurement by performing segment and circumpapillary time series scans around the optic nerve head. We also demonstrate microvasculature imaging by calculation of squared intensity differences between successive tomograms.
A device that allows for the measurement of ocular fundus pulsations at preselected axial positions of a subject's eye is presented. Unlike previously presented systems, which only allow for observation of the strongest reflecting retinal layer, our system enables the measurement of fundus pulsations at a preselected ocular layer. For this purpose the sample is illuminated by light of low temporal coherence. The layer is then selected by positioning one mirror of a Michelson interferometer according to the depth of the layer. The device contains a length measurement system based on partial coherence interferometry and a line scan charge-coupled device camera for recording and online inspection of the fringe system. In-vivo measurements in healthy humans are performed as proof of principle. The algorithms used for enhancing the recorded images are briefly introduced. The contrast of the observed interference pattern is evaluated for different positions of the measurement mirror and at various distances from the front surface of the cornea. The applications of such a system may be wide, including assessment of eye elongation during myopia development and blood-flow-related changes in intraocular volume.
A bidirectional FDOCT system capable of measuring absolute velocities of moving scatterers is described. In this setup the sample is illuminated with two differently polarized beams. These two probe beams impinging onto the sample at a known angle. The velocity estimation is independent of the exact direction of the velocity vector in the detection plane. Evaluation measurements were performed on a rotating disc driven at well defined velocities and tilted by various small angles around to π/2. Our results indicate a high correlation between pre-set and estimated velocities and the independency of these velocities from the tilting angle of the disc. Additional preliminary in vivo measurements proof the ability of this new method to measure absolute blood flow velocities in human retinal vessels.
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