Stefan Zotter

Visualization of microvasculature by dual-beam phase-resolved Doppler optical coherence tomography

We present a dual-beam Doppler optical coherence tomography system for visualizing the microvasculature within the retina. The sample arm beams from two identical spectral domain optical coherence tomography (SD-OCT) systems are combined such that there is a small horizontal offset between them at the retina. Thereby we record two tomograms which are slightly separated in time. Phase-resolved Doppler analysis is performed between these two data sets. This system allows blood capillary imaging with high flow sensitivity and variable velocity range. To demonstrate the performance of our system we present images of the microvascular network around the fovea and around the optic nerve head of the human eye.

Adaptive optics SLO/OCT for 3D imaging of human photoreceptors in vivo

We present a new instrument that is capable of imaging human photoreceptors in three dimensions. To achieve high lateral resolution, the system incorporates an adaptive optics system. The high axial resolution is achieved through the implementation of optical coherence tomography (OCT). The instrument records simultaneously both, scanning laser ophthalmoscope (SLO) and OCT en-face images, with a pixel to pixel correspondence. The information provided by the SLO is used to correct for transverse eye motion in post-processing. In order to correct for axial eye motion, the instrument is equipped with a high speed axial eye tracker. In vivo images of foveal cones as well as images recorded at an eccentricity from the fovea showing cones and rods are presented.

In vitro and in vivo three-dimensional velocity vector measurement by three-beam spectral-domain Doppler optical coherence tomography.

Abstract. 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.

High-speed polarization sensitive optical coherence tomography scan engine based on Fourier domain mode locked laser

We report on a new swept source polarization sensitive optical coherence tomography scan engine that is based on polarization maintaining (PM) fiber technology. The light source is a Fourier domain mode locked laser with a PM cavity that operates in the 1300 nm wavelength regime. It is equipped with a PM buffer stage that doubles the fundamental sweep frequency of 54.5 kHz. The fiberization allows coupling of the scan engine to different delivery probes. In a first demonstration, we use the system for imaging human skin at an A-scan rate of 109 kHz. The system illuminates the sample with circularly polarized light and measures reflectivity, retardation, optic axis orientation, and Stokes vectors simultaneously. Furthermore, depolarization can be quantified by calculating the degree of polarization uniformity (DOPU). The high scanning speed of the system enables dense sampling in both, the x- and y-direction, which provides the opportunity to use 3D evaluation windows for DOPU calculation. This improves the spatial resolution of DOPU images considerably.

Large-field high-speed polarization sensitive spectral domain OCT and its applications in ophthalmology

We present a novel spectral domain polarization sensitive OCT system (PS-OCT) that operates at an A-scan rate of 70 kHz and supports scan angles of up to 40° × 40°. The high-speed imaging allows the acquisition of up to 1024 × 250 A-scans per 3D scan, which, together with the large field of view, considerably increases the informative value of the images. To demonstrate the excellent performance of the new PS-OCT system, we imaged several healthy volunteers and patients with various diseases such as glaucoma, AMD, Stargardt’s disease, and albinism. The results are compared with clinically established methods such as scanning laser polarimetry and autofluorescence.

Measuring retinal nerve fiber layer birefringence, retardation, and thickness using wide-field, high-speed polarization sensitive spectral domain OCT.

PURPOSE We presented a novel polarization sensitive optical coherence tomography (PS-OCT) system for measuring retinal nerve fiber layer (RNFL) birefringence, retardation, and thickness, and report on the repeatability of acquiring these quantities. METHODS A new PS-OCT system, measuring at 840 nm, was developed that supports scan angles of up to 40° × 40° with an A-scan rate of 70 kHz. To test the performance and reproducibility, we measured 10 eyes of 5 healthy human volunteers five times each. All volunteers were imaged further with scanning laser polarimetry (SLP). The obtained RNFL birefringence, retardation, and thickness maps were averaged, and standard deviation maps were calculated. For quantitative comparison between the new PS-OCT and SLP, a circumpapillary evaluation within 2 annular segments (superior and inferior to the optic disc) was performed. RESULTS High quality RNFL birefringence, retardation, and thickness maps were obtained. Within the superior and inferior segments, the mean retardation for individual eyes ranged from 20° to 28.9° and 17.2° to 28.2°, respectively. The quadrant precision over the 5 consecutive measurements for each subject, calculated for the average retardation obtained within the superior and inferior quadrants ranged from 0.16° to 0.69°. The mean birefringence ranged from 0.106°/μm to 0.141°/μm superior and 0.101°/μm to 0.135°/μm inferior, with a quadrant precision of 0.001°/μm to 0.007°/μm. The mean RNFL thickness varied from 114 to 150 μm superior, and 111 to 140.9 μm inferior (quadrant precision ranged from 3.6 to 11.9 μm). CONCLUSIONS The new PS-OCT system showed high image quality and reproducibility, and, therefore, might be a valuable tool for glaucoma diagnosis.

Automated measurement of choroidal thickness in the human eye by polarization sensitive optical coherence tomography

We present a new method to automatically segment the thickness of the choroid in the human eye by polarization sensitive optical coherence tomography (PS-OCT). A swept source PS-OCT instrument operating at a center wavelength of 1040 nm is used. The segmentation method is based entirely on intrinsic, tissue specific polarization contrast mechanisms. In a first step, the anterior boundary of the choroid, the retinal pigment epithelium, is segmented based on depolarization. In a second step, the choroid-sclera interface is found by using the birefringence of the sclera. The method is demonstrated in five healthy eyes. The mean repeatability (standard deviation) of thickness measurement was found to be 18.3 µm.

Retinal polarization-sensitive optical coherence tomography at 1060 nm with 350 kHz A-scan rate using an Fourier domain mode locked laser

Abstract. We present a novel, high-speed, polarization-sensitive, optical coherence tomography set-up for retinal imaging operating at a central wavelength of 1060 nm which was tested for in vivo imaging in healthy human volunteers. We use the system in combination with a Fourier domain mode locked laser with active spectral shaping which enables the use of forward and backward sweep in order to double the imaging speed without a buffering stage. With this approach and with a custom designed data acquisition system, we show polarization-sensitive imaging with an A-scan rate of 350 kHz. The acquired three-dimensional data sets of healthy human volunteers show different polarization characteristics in the eye, such as depolarization in the retinal pigment epithelium and birefringence in retinal nerve fiber layer and sclera. The increased speed allows imaging of large volumes with reduced motion artifacts. Moreover, averaging several two-dimensional frames allows the generation of high-definition B-scans without the use of an eye-tracking system. The increased penetration depth of the system, which is caused by the longer probing beam wavelength, is beneficial for imaging choroidal and scleral structures and allows automated segmentation of these layers based on their polarization characteristics.

Characterization of Stargardt Disease Using Polarization- Sensitive Optical Coherence Tomography and Fundus Autofluorescence Imaging

PURPOSE To identify disease-specific changes in Stargardt disease (STGD) based on imaging with polarization-sensitive spectral-domain optical coherence tomography (PS-OCT) and to compare structural changes with those visible on blue light fundus autofluorescence (FAF) imaging. METHODS Twenty-eight eyes of 14 patients diagnosed with STGD were imaged using a novel high-speed, large-field PS-OCT system and FAF (excitation 488 nm, emission > 500 nm). The ophthalmoscopic phenotype was classified into three groups. ABCA4 mutation testing detected 15 STGD alleles, six of which harbor novel mutations. RESULTS STGD phenotype 1 (12 eyes) showed sharply delineated areas of absent RPE signal on RPE segmentation B-scans of PS-OCT correlating with areas of hypofluorescence on FAF. Adjacent areas of irregular fluorescence correlated with an irregular RPE segmentation line with absence of overlaying photoreceptor layers. Eyes characterized on OCT by a gap in the subfoveal outer segment layer (foveal cavitation) showed a normal RPE segmentation line on PS-OCT. Hyperfluorescent flecks on FAF in phenotype 2 STGD (8 eyes) were identified as clusters of depolarizing material at the level of the RPE. Distribution of flecks could be depicted on RPE elevation maps. An increased amount of depolarizing material in the choroid was characteristic for STGD Phenotype 3 (8 eyes). CONCLUSIONS PS-OCT together with FAF identified characteristic patterns of changes in different stages of the disease. PS-OCT is a promising new tool for diagnosis and evaluation of future treatment modalities in STGD.

Motion artifact and speckle noise reduction in polarization sensitive optical coherence tomography by retinal tracking

We present a novel polarization sensitive optical coherence tomography (PS-OCT) system with an integrated retinal tracker. The tracking operates at up to 60 Hz, correcting PS-OCT scanning positions during the acquisition to avoid artifacts caused by eye motion. To demonstrate the practical performance of the system, we imaged several healthy volunteers and patients with AMD both with B-scan repetitions for frame averaging and with 3D raster scans. Under large retinal motions with up to 1 mm amplitude at 0.5 ~a few Hz frequency range, motion artifact suppression in the PS-OCT images as well as standard deviation noise reduction in the frame averaged retardation images are presented.