Wide-Field Three-Dimensional Depth-Invariant Cellular-Resolution Imaging of the Human Retina.

Three-dimensional (3D) cellular-resolution imaging of the living human retina over a large field of view will bring a great impact in clinical ophthalmology, potentially finding new biomarkers for early diagnosis and improving the pathophysiological understanding of ocular diseases. While hardware-based and computational adaptive optics (AO) optical coherence tomography (OCT) have been developed to achieve cellular-resolution retinal imaging, these approaches support limited 3D imaging fields, and their high cost and intrinsic hardware complexity limit their practical utility. Here, this work demonstrates 3D depth-invariant cellular-resolution imaging of the living human retina over a 3 × 3 mm field of view using the first intrinsically phase-stable multi-MHz retinal swept-source OCT and novel computational defocus and aberration correction methods. Single-acquisition imaging of photoreceptor cells, retinal nerve fiber layer, and retinal capillaries is presented across unprecedented imaging fields. By providing wide-field 3D cellular-resolution imaging in the human retina using a standard point-scan architecture routinely used in the clinic, this platform proposes a strategy for expanded utilization of high-resolution retinal imaging in both research and clinical settings.

Using the dynamic forward scattering signal for optical coherence tomography based blood flow quantification.

To our knowledge, all existing optical coherence tomography approaches for quantifying blood flow, whether Doppler-based or decorrelation-based, analyze light that is back-scattered by moving red blood cells (RBCs). This work investigates the potential advantages of basing these measurements on light that is forward-scattered by RBCs, i.e., by looking at the signals back-scattered from below the vessel. We show experimentally that flowmetry based on forward-scattering is insensitive to vessel orientation for vessels that are approximately orthogonal to the imaging beam. We further provide proof-of-principle demonstrations of dynamic forward-scattering (DFS) flowmetry in human retinal and choroidal vessels.

Treatment Effects in Retinal Angiomatous Proliferation Imaged with OCT Angiography.

PURPOSE: This prospective case series is aimed at exploring optical coherence tomographic angiography (OCT-A) as a treatment monitoring tool in patients treated for retinal angiomatous proliferation (RAP). METHODS: Twelve treatment-naïve RAP patients were included, with a median age of 79 years (range 65-90). Patients were imaged with an experimental 1,040-nm swept-source phase-resolved OCT-A instrument before and after treatment. Treatment consisted of either intravitreal bevacizumab or triamcinolone injections with or without photodynamic therapy (PDT). Abnormal blood flow after treatment was graded as increased, unchanged, decreased, or resolved. RESULTS: OCT-A images before and after treatment could be obtained in 9 patients. The median follow-up period was 10 weeks (range 5-19). After various treatments, the RAP lesion resolved in 7 patients, in 1 patient the OCT-A depicted decreased flow in the lesion, and 1 patient showed unchanged abnormal blood flow. Monotherapy with intravitreal bevacizumab injections resolved RAP in 1 out of 2 patients. Combined therapy of bevacizumab with PDT resolved RAP in 6 out of 7 patients. CONCLUSIONS: OCT-A visualized resolution of abnormal blood flow in 7 out of 9 RAP patients after various short-term treatment sequences. OCT-A may become an important noninvasive monitoring tool for optimizing treatment strategies in RAP patients.

Self-interference fluorescence microscopy with three-phase detection for depth-resolved confocal epi-fluorescence imaging.

Three-dimensional confocal fluorescence imaging of in vivo tissues is challenging due to sample motion and limited imaging speeds. In this paper a novel method is therefore presented for scanning confocal epi-fluorescence microscopy with instantaneous depth-sensing based on self-interference fluorescence microscopy (SIFM). A tabletop epi-fluorescence SIFM setup was constructed with an annular phase plate in the emission path to create a spectral self-interference signal that is phase-dependent on the axial position of a fluorescent sample. A Mach-Zehnder interferometer based on a 3 × 3 fiber-coupler was developed for a sensitive phase analysis of the SIFM signal with three photon-counter detectors instead of a spectrometer. The Mach-Zehnder interferometer created three intensity signals that alternately oscillated as a function of the SIFM spectral phase and therefore encoded directly for the axial sample position. Controlled axial translation of fluorescent microsphere layers showed a linear dependence of the SIFM spectral phase with sample depth over axial image ranges of 500 µm and 80 µm (3.9 × Rayleigh range) for 4 × and 10 × microscope objectives respectively. In addition, SIFM was in good agreement with optical coherence tomography depth measurements on a sample with indocyanine green dye filled capillaries placed at multiple depths. High-resolution SIFM imaging applications are demonstrated for fluorescence angiography on a dye-filled capillary blood vessel phantom and for autofluorescence imaging on an ex vivo fly eye.

Parallel line scanning ophthalmoscope for retinal imaging.

A parallel line scanning ophthalmoscope (PLSO) is presented using a digital micromirror device (DMD) for parallel confocal line imaging of the retina. The posterior part of the eye is illuminated using up to seven parallel lines, which were projected at 100 Hz. The DMD offers a high degree of parallelism in illuminating the retina compared to traditional scanning laser ophthalmoscope systems utilizing scanning mirrors. The system operated at the shot-noise limit with a signal-to-noise ratio of 28 for an optical power measured at the cornea of 100 µW. To demonstrate the imaging capabilities of the system, the macula and the optic nerve head of a healthy volunteer were imaged. Confocal images show good contrast and lateral resolution with a 10°×10° field of view.

Angiography of the retina and the choroid with phase-resolved OCT using interval-optimized backstitched B-scans.

In conventional phase-resolved OCT blood flow is detected from phase changes between successive A-scans. Especially in high-speed OCT systems this results in a short evaluation time interval. This method is therefore often unable to visualize complete vascular networks since low flow velocities cause insufficient phase changes. This problem was solved by comparing B-scans instead of successive A-scans to enlarge the time interval. In this paper a detailed phase-noise analysis of our OCT system is presented in order to calculate the optimal time intervals for visualization of the vasculature of the human retina and choroid. High-resolution images of the vasculature of a healthy volunteer taken with various time intervals are presented to confirm this analysis. The imaging was performed with a backstitched B-scan in which pairs of small repeated B-scans are stitched together to independently control the time interval and the imaged lateral field size. A time interval of ≥ 2.5 ms was found effective to image the retinal vasculature down to the capillary level. The higher flow velocities of the choroid allowed a time interval of 0.64 ms to reveal its dense vasculature. Finally we analyzed depth-resolved histograms of volumetric phase-difference data to assess changes in amount of blood flow with depth. This analysis indicated different flow regimes in the retina and the choroid.

Phase-stabilized optical frequency domain imaging at 1-µm for the measurement of blood flow in the human choroid.

In optical frequency domain imaging (OFDI) the measurement of interference fringes is not exactly reproducible due to small instabilities in the swept-source laser, the interferometer and the data-acquisition hardware. The resulting variation in wavenumber sampling makes phase-resolved detection and the removal of fixed-pattern noise challenging in OFDI. In this paper this problem is solved by a new post-processing method in which interference fringes are resampled to the exact same wavenumber space using a simultaneously recorded calibration signal. This method is implemented in a high-speed (100 kHz) high-resolution (6.5 µm) OFDI system at 1-µm and is used for the removal of fixed-pattern noise artifacts and for phase-resolved blood flow measurements in the human choroid. The system performed close to the shot-noise limit (<1dB) with a sensitivity of 99.1 dB for a 1.7 mW sample arm power. Suppression of fixed-pattern noise artifacts is shown up to 39.0 dB which effectively removes all artifacts from the OFDI-images. The clinical potential of the system is shown by the detection of choroidal blood flow in a healthy volunteer and the detection of tissue reperfusion in a patient after a retinal pigment epithelium and choroid transplantation.

Clinical validation of point-source corneal topography in keratoplasty.

PURPOSE: To validate the clinical performance of point-source corneal topography (PCT) in postpenetrating keratoplasty (PKP) eyes and to compare it with conventional Placido-based topography. METHODS: Corneal elevation maps of the anterior corneal surface were obtained from 20 post-PKP corneas using PCT (VU topographer, prototype; VU University Medical Center, Amsterdam, The Netherlands) and Placido-based topography (Keratron, Optikon 2000, Rome, Italy). Corneal surface parameters are calculated in terms of radius and asphericity. Corneal aberrations were characterized using standard Zernike convention. An artificial surface with quadrafoil feature (SUMIPRO, Almelo, The Netherlands) was measured and used as a reference to assess instrument performance compared with the gold standard. RESULTS: The differences (mean ± std of PCT - Placido) found between the two types of topographers in measurements of post-PKP eyes are 0.02 ± 0.21 mm (p=0.64) for radius of curvature, 0.14 ± 0.49 (p=0.23) for asphericity, -0.19 ± 1.67 µm (p=0.61) for corneal astigmatism, -0.25 ± 1.34 µm (p=0.41) for corneal coma, 0.23 ± 0.82 µm (p=0.23) for corneal trefoil, and 0.15 ± 0.28 µm (p=0.02) for corneal quadrafoil. The PCT measured the artificial surface more accurate (rms error 0.16 µm; 0.12 eq. Dpt.) than the Placido-based topographer (rms error 1.50 µm; 1.15 eq. Dpt.). CONCLUSIONS: PCT is more accurate than Placido-based topography in measuring quadrafoil aberration.

Forward ray tracing for image projection prediction and surface reconstruction in the evaluation of corneal topography systems.

A forward ray tracing (FRT) model is presented to determine the exact image projection in a general corneal topography system. Consequently, the skew ray error in Placido-based topography is demonstrated. A quantitative analysis comparing FRT-based algorithms and Placido-based algorithms in reconstructing the front surface of the cornea shows that arc step algorithms are more sensitive to noise (imprecise). Furthermore, they are less accurate in determining corneal aberrations particularly the quadrafoil aberration. On the other hand, FRT-based algorithms are more accurate and more precise showing that point to point corneal topography is superior compared to its Placido-based counterpart.

A Neural Network Approach to Quantify Blood Flow from Retinal OCT Intensity Time-Series Measurements.

Many diseases of the eye are associated with alterations in the retinal vasculature that are possibly preceded by undetected changes in blood flow. In this work, a robust blood flow quantification framework is presented based on optical coherence tomography (OCT) angiography imaging and deep learning. The analysis used a forward signal model to simulate OCT blood flow data for training of a neural network (NN). The NN was combined with pre- and post-processing steps to create an analysis framework for measuring flow rates from individual blood vessels. The framework's accuracy was validated using both blood flow phantoms and human subject imaging, and across flow speed, vessel angle, hematocrit levels, and signal-to-noise ratio. The reported flow rate of the calibrated NN framework was measured to be largely independent of vessel angle, hematocrit levels, and measurement signal-to-noise ratio. In vivo retinal flow rate measurements were self-consistent across vascular branch points, and approximately followed a predicted power-law dependence on the vessel diameter. The presented OCT-based NN flow rate estimation framework addresses the need for a robust, deployable, and label-free quantitative retinal blood flow mapping technique.

Artifact Rates for 2D Retinal Nerve Fiber Layer Thickness Versus 3D Neuroretinal Rim Thickness Using Spectral-Domain Optical Coherence Tomography.

Purpose: To compare the rates of clinically significant artifacts for two-dimensional peripapillary retinal nerve fiber layer (RNFL) thickness versus three-dimensional (3D) neuroretinal rim thickness using spectral-domain optical coherence tomography (SD-OCT). Methods: Only one eye per patient was used for analysis of 120 glaucoma patients and 114 normal patients. For RNFL scans and optic nerve scans, 15 artifact types were calculated per B-scan and per eye. Neuroretinal rim tissue was quantified by the minimum distance band (MDB). Global MDB neuroretinal rim thicknesses were calculated before and after manual deletion of B-scans with artifacts and subsequent automated interpolation. A clinically significant artifact was defined as one requiring manual correction or repeat scanning. Results: Among glaucomatous eyes, artifact rates per B-scan were significantly more common in RNFL scans (61.7%, 74 of 120) compared to B-scans in neuroretinal rim volume scans (20.9%, 1423 of 6820) (95% confidence interval [CI], 31.6-50.0; P < 0.0001). For clinically significant artifact rates per eye, optic nerve scans had significantly fewer artifacts (15.8% of glaucomatous eyes, 13.2% of normal eyes) compared to RNFL scans (61.7% of glaucomatous eyes, 25.4% of normal eyes) (glaucoma group: 95% CI, 34.1-57.5, P < 0.0001; normal group: 95% CI, 1.3-23.3, P = 0.03). Conclusions: Compared to the most commonly used RNFL thickness scans, optic nerve volume scans less frequently require manual correction or repeat scanning to obtain accurate measurements. Translational Relevance: This paper illustrates the potential for 3D OCT algorithms to improve in vivo imaging in glaucoma.

Subretinal Fibrosis Detection Using Polarization Sensitive Optical Coherence Tomography.

Purpose: Subretinal fibrosis (SRFib) is an important cause of permanent loss-of-vision diseases with submacular neovascularization, but a reliable diagnostic method is currently missing. This study uses polarization-sensitive optical coherence tomography (PS-OCT) to detect SRFib within retinal lesions by measurement of its birefringent collagen fibers. Methods: Twenty-five patients were enrolled with retinal pathology in one or both eyes containing (1) suspected SRFib, (2) lesions suspected not to be fibrotic, or (3) lesions with doubtful presence of SRFib. All eyes were evaluated for SRFIb using conventional diagnostics by three retinal specialists. PS-OCT images were visually evaluated for SRFib based on cumulative phase retardation, local birefringence, and optic axis uniformity. Results: Twenty-nine eyes from 22 patients were scanned successfully. In 13 eyes, SRFib was diagnosed by all retinal specialists; of these, 12 were confirmed by PS-OCT and one was inconclusive. In nine eyes, the retinal specialists expected no SRFib, which was confirmed by PS-OCT in all cases. In seven eyes, the retinal specialists' evaluations were inconsistent with regard to the presence of SRFib. PS-OCT confirmed the presence of SRFib in four of these eyes and the absence of SRFib in two eyes and was inconclusive in one eye. Conclusions: In 21 out of 22 eyes, PS-OCT confirmed the evaluation of retinal specialists regarding the presence of SRFib. PS-OCT provided additional information to distinguish SRFib from other tissues within subretinal neovascular lesions in 6 out of 7 eyes. Translational Relevance: PS-OCT can identify and quantify SRFib in doubtful cases for which a reliable diagnosis is currently lacking.

Artifact Rates for 2D Retinal Nerve Fiber Layer Thickness Versus 3D Retinal Nerve Fiber Layer Volume.

Purpose: To compare artifact rates in two-dimensional (2D) versus three-dimensional (3D) retinal nerve fiber layer (RNFL) scans using Spectralis optical coherence tomography (OCT). Methods: Thirteen artifact types in 2D and 3D RNFL scans were identified in 106 glaucomatous eyes and 95 normal eyes. Artifact rates were calculated per B-scan and per eye. In 3D volume scans, artifacts were counted only for the 97 B-scans used to calculate RNFL parameters for the 2.5-3.5-mm annulus. 3D RNFL measurements were calculated twice, once before and again after deletion of B-scans with artifacts and subsequent automated interpolation. Results: For 2D scans, artifacts were present in 58.5% of B-scans (62 of 106) in glaucomatous eyes. For 3D scans, a mean of 35.4% of B-scans (34.3 of 97 B-scans per volume scan) contained an artifact in 106 glaucomatous eyes. For 3D data of glaucoma patients, mean global RNFL thickness values were similar before and after interpolation (77.0 ± 11.6 µm vs. 75.1 ± 11.2 µm, respectively; P = 0.23). Fewer clinically significant artifacts were noted in 3D RNFL scans, where only 7.5% of glaucomatous eyes (8 of 106) and 0% of normal eyes (0 of 95) had artifacts, compared to 2D RNFL scans, where 58.5% of glaucomatous eyes (62 of 106) and 14.7% of normal eyes (14 of 95) had artifacts. Conclusions: Compared to 2D RNFL scans, 3D RNFL volume scans less often require manual correction to obtain accurate measurements. Translational Relevance: 3D RNFL volume scans have fewer clinically significant artifacts compared to 2D RNFL thickness scans.

Performance in specular reflection and slit-imaging corneal topography.

PURPOSE: Assessment of the relative performance in measuring corneal shape and corneal aberrations for two specular reflection topographers: Keratron Placido Ring Topographer, VU Topographer, and two slit-lamp imaging instruments: Orbscan II and Topcon SL-45 Scheimpflug. METHODS: Corneal height maps of the anterior corneal surface were obtained from a group of 34 subjects with all four instruments; posterior corneal surface height maps were only obtained with the two slit-lamp imaging instruments. Corneal surface shapes are calculated in terms of radius of curvature and asphericity fitting an aspheric model. Wave aberrations for the anterior corneal surface and the total cornea are determined up to and including sixth order Zernike convention by means of ray tracing. RESULTS: Clinical relevant differences were observed for radius of curvature of the anterior corneal surface, where the slit-imaging instruments measure higher values (mean difference = 0.05 mm, p < 0.05) and anterior corneal astigmatism for which the Orbscan II measures higher values than the VU Topographer [mean difference = 0.174 microm (0.134 Equivalent Diopters), p < 0.01]. Small significant differences were observed for asphericity and spherical aberration of the anterior corneal surface; however, these are not clinically relevant. Clinically relevant differences were also observed for posterior radius (difference = 0.135 mm p < 0.001), total corneal astigmatism (difference = 0.207 microm (0.159 Equivalent Diopters), p = 0.001), and central corneal thickness (CCT) (difference = -18.6 microm, p < 0.001). The differences found for total corneal coma and trefoil were not clinical relevant. Furthermore, the precision of the specular reflection topographers is superior to that of the slit-lamp instruments by at least a factor of two. CONCLUSIONS: For traditional spectacle and contact lens applications, the corneal topographers are interchangeable except for measuring anterior radius of curvature. However, for more modern techniques as customized corneal refractive surgery, the subtle differences (e.g., total corneal astigmatism and CCT) between the instruments are clinically relevant.

Quantitative depolarization measurements for fiber-based polarization-sensitive optical frequency domain imaging of the retinal pigment epithelium.

A full quantitative evaluation of the depolarization of light may serve to assess concentrations of depolarizing particles in the retinal pigment epithelium and to investigate their role in retinal diseases in the human eye. Optical coherence tomography and optical frequency domain imaging use spatial incoherent averaging to compute depolarization. Depolarization depends on accurate measurements of the polarization states at the receiver but also on the polarization state incident upon and within the tissue. Neglecting this dependence can result in artifacts and renders depolarization measurements vulnerable to birefringence in the system and in the sample. In this work, we discuss the challenges associated with using a single input polarization state and traditional depolarization metrics such as the degree-of-polarization and depolarization power. We demonstrate quantitative depolarization measurements based on Jones vector synthesis and polar decomposition using fiber-based polarization-sensitive optical frequency domain imaging of the retinal pigment epithelium in a human eye.

Three-Dimensional Optical Coherence Tomography Imaging For Glaucoma Associated With Boston Keratoprosthesis Type I and II.

PRECIS: Three-dimensional (3D) spectral domain optical coherence tomography (OCT) volume scans of the optic nerve head (ONH) and the peripapillary area are useful in the management of glaucoma in patients with a type I or II Boston Keratoprosthesis (KPro). PURPOSE: The purpose of this study was to report the use of spectral domain OCT in the management of glaucoma in patients with a type I or II Boston KPro. MATERIALS AND METHODS: This study is an observational case series. Four consecutive patients with KPro implants were referred for glaucoma evaluation. A comprehensive eye examination was performed which included disc photography, visual field testing, and high-density spectral domain OCT volume scans of the ONH and the peripapillary area. 2D and 3D parameters were calculated using custom-designed segmentation algorithms developed for glaucoma management. RESULTS: Spectral domain OCT parameters provided useful information in the diagnosis and management of 4 KPro patients. OCT parameters which can be used in KPro patients included 2D retinal nerve fiber layer (RNFL) thickness, 3D peripapillary RNFL volume, 3D peripapillary retinal thickness and volume, 3D cup volume, and 3D neuroretinal rim thickness and volume. In 3 of 4 cases where the traditional 2D RNFL thickness scan was limited by artifacts, 3D spectral domain OCT volume scans provided useful quantitative objective measurements of the ONH and peripapillary region. Therefore, 3D parameters derived from high-density volume scans as well as radial scans of the ONH can be used to overcome the limitations and artifacts associated with 2D RNFL thickness scans. CONCLUSIONS: Spectral domain OCT volume scans offer the possibility to enhance the evaluation of KPro patients with glaucoma by using both 2D and 3D diagnostic parameters that are easily obtained in a clinic setting.

Diagnostic Capability of 3D Peripapillary Retinal Volume for Glaucoma Using Optical Coherence Tomography Customized Software.

PRéCIS:: The diagnostic capability of peripapillary retinal volume is similar to peripapillary retinal nerve fiber layer thickness for diagnosing glaucoma, but with fewer artifacts. PURPOSE: To compare the diagnostic capability of 3-dimensional peripapillary retinal volume (RV) versus 2-dimensional peripapillary retinal nerve fiber layer (RNFL) thickness for open-angle glaucoma. PATIENTS AND METHODS: A retrospective cross-sectional analysis was conducted. A total of 180 subjects (113 open-angle glaucoma, 67 normal participants) had spectral domain optical coherence tomography volume scans and RNFL thickness measurements. Peripapillary RV values were calculated using a custom-designed program with 4 circumpapillary annuli (CA): CA1 had circle diameters of 2.5 and 3.5 mm; CA2, 3 and 4 mm; CA3, 3.5 and 4.5 mm; and CA4, 4 and 5 mm. Area under the receiver operating characteristic curves were calculated for global, quadrant, and octant regions for RV (CA1 to CA4) and RNFL thickness. Pair-wise comparisons were conducted. Artifacts rates were determined. RESULTS: Mean age was 62.7±15.4 years, and 47.8% (86/180) were male. Among RV measurements, best diagnostic performances were for the smallest 2 annuli for inferior RV (CA1: 0.964, CA2: 0.955). Of the 4 annuli, CA1 had the highest diagnostic performance. Of specific regions, the inferior RV quadrant had the highest performance across CA1 to CA4. Peripapillary RV had similar diagnostic capability compared with RNFL thickness (P>0.05). The artifact rate per B-scan for RV was 6.0%, which was significantly lower compared with 2-dimensional RNFL thickness in the same patient population (32.2%, P<0.0001). CONCLUSIONS: The diagnostic capability of RV is similar to RNFL thickness for perimetric open-angle glaucoma, but RV had fewer artifacts compared with RNFL thickness.

Complex differential variance angiography with noise-bias correction for optical coherence tomography of the retina.

Complex differential variance (CDV) provides phase-sensitive angiographic imaging for optical coherence tomography (OCT) with immunity to phase-instabilities of the imaging system and small-scale axial bulk motion. However, like all angiographic methods, measurement noise can result in erroneous indications of blood flow that confuse the interpretation of angiographic images. In this paper, a modified CDV algorithm that corrects for this noise-bias is presented. This is achieved by normalizing the CDV signal by analytically derived upper and lower limits. The noise-bias corrected CDV algorithm was implemented into an experimental 1 µm wavelength OCT system for retinal imaging that used an eye tracking scanner laser ophthalmoscope at 815 nm for compensation of lateral eye motions. The noise-bias correction improved the CDV imaging of the blood flow in tissue layers with a low signal-to-noise ratio and suppressed false indications of blood flow outside the tissue. In addition, the CDV signal normalization suppressed noise induced by galvanometer scanning errors and small-scale lateral motion. High quality cross-section and motion-corrected en face angiograms of the retina and choroid are presented.

Optic axis mapping with catheter-based polarization-sensitive optical coherence tomography.

Birefringence offers an intrinsic contrast mechanism related to the microstructure and arrangement of fibrillary tissue components. Here we present a reconstruction strategy to recover not only the scalar amount of birefringence but also its optic axis orientation as a function of depth in tissue from measurements with catheter-based polarization sensitive optical coherence tomography. A polarization symmetry constraint, intrinsic to imaging in the backscatter direction, facilitates the required compensation for wavelength-dependent transmission through system elements, the rotating catheter, and overlying tissue layers. Applied to intravascular imaging of coronary atherosclerosis in human patients, the optic axis affords refined interpretation of plaque architecture.

In vivo retinal imaging for fixational eye motion detection using a high-speed digital micromirror device (DMD)-based ophthalmoscope.

Retinal motion detection with an accuracy of 0.77 arcmin corresponding to 3.7 µm on the retina is demonstrated with a novel digital micromirror device based ophthalmoscope. By generating a confocal image as a reference, eye motion could be measured from consecutively measured subsampled frames. The subsampled frames provide 7.7 millisecond snapshots of the retina without motion artifacts between the image points of the subsampled frame, distributed over the full field of view. An ophthalmoscope pattern projection speed of 130 Hz enabled a motion detection bandwidth of 65 Hz. A model eye with a scanning mirror was built to test the performance of the motion detection algorithm. Furthermore, an in vivo motion trace was obtained from a healthy volunteer. The obtained eye motion trace clearly shows the three main types of fixational eye movements. Lastly, the obtained eye motion trace was used to correct for the eye motion in consecutively obtained subsampled frames to produce an averaged confocal image correct for motion artefacts.