The flow of erythrocytes in parafoveal capillaries was imaged in the living human eye with an adaptive optics near-confocal ophthalmoscope at a frame rate of 800 Hz with a low coherence near-infrared (NIR) light source. Spatiotemporal traces of the erythrocyte movement were extracted from consecutive images. Erythrocyte velocity was measured using custom software based on the Radon transform. The impact of imaging speed on velocity measurement was estimated using images of frame rates of 200, 400, and 800 Hz. The NIR light allowed for long imaging periods without visually stimulating the retina and disturbing the natural rheological state. High speed near-confocal imaging enabled direct and accurate measurement of erythrocyte velocity, and revealed a distinctively cardiac-dependent pulsatile velocity waveform of the erythrocyte flow in retinal capillaries, disclosed the impact of the leukocytes on erythrocyte motion, and provided new metrics for precise assessment of erythrocyte movement. The approach may facilitate new investigations on the pathophysiology of retinal microcirculation with applications for ocular and systemic diseases.

Normal physiological movements (e.g., respiration and heartbeat) induce eye motions during clinical measurements of human corneal biomechanical properties using optical coherence elastography (OCE). We quantified the effects of respiratory and cardiac-induced eye motions on clinical corneal OCE measurement precision and repeatability. Methods: Corneal OCE was performed using low-force, micro-air-pulse tissue stimulation and high-resolution phase-sensitive optical coherence tomography (OCT) imaging. Axial surface displacements of the corneal apex were measured (M-mode) at a 70-kHz sampling rate and three different stimulation pressures (20-60 Pa). Simultaneously, the axial corneal position was tracked with structural OCT imaging, while the heartrate and respiration were monitored over a 90 second period. Results: Respiratory-and cardiac-induced eye motions have distinctly lower frequency (0.1-1 Hz) and much greater amplitude (up to ± 50 μm movements) than air-pulseinduced corneal tissue deformations (∼250 Hz, <1 μm). The corneal displacements induced during OCE measurements in vivo were-0.41 ± 0.06 μm (n = 22 measurements, coefficient of variation [CV]: 14.6%) and-0.44 ± 0.07 μm (n = 50 measurements, CV: 15.9%), respectively, from two human subjects at 40 Pa stimulation pressure. Observed variation in corneal tissue displacements were not associated with tissue stimulation magnitude, or the amplitude of physiologically induced axial eye motion. Conclusions: The microsecond timescale and submicron tissue displacements observed during corneal OCE measurements are separable from normal involuntary physiological movements, such as the oculocardiac pulse and respiratory movements. Translational Relevance: This work advances innovations in biomedical imaging and engineering for clinical diagnostic applications for soft-tissue biomechanical testing.

PurposeContinuous and rapid eye movement causes significant intraframe distortion in adaptive optics high resolution retinal imaging. To minimize this artifact, we developed a high speed adaptive optics line scan confocal retinal imaging system.MethodsA high speed line camera was employed to acquire retinal image and custom adaptive optics was developed to compensate the wave aberration of the human eye’s optics. The spatial resolution and signal to noise ratio were assessed in model eye and in living human eye. The improvement of imaging fidelity was estimated by reduction of intra-frame distortion of retinal images acquired in the living human eyes with frame rates at 30 frames/second (FPS), 100 FPS, and 200 FPS.ResultsThe device produced retinal image with cellular level resolution at 200 FPS with a digitization of 512×512 pixels/frame in the living human eye. Cone photoreceptors in the central fovea and rod photoreceptors near the fovea were resolved in three human subjects in normal chorioretinal health. Compared with retinal images acquired at 30 FPS, the intra-frame distortion in images taken at 200 FPS was reduced by 50.9% to 79.7%.ConclusionsWe demonstrated the feasibility of acquiring high resolution retinal images in the living human eye at a speed that minimizes retinal motion artifact. This device may facilitate research involving subjects with nystagmus or unsteady fixation due to central vision loss.