Zhangyi Zhong

Variation of cone photoreceptor packing density with retinal eccentricity and age.

PURPOSE To study the variation of cone photoreceptor packing density across the retina in healthy subjects of different ages. METHODS High-resolution adaptive optics scanning laser ophthalmoscope (AOSLO) systems were used to systematically image the retinas of two groups of subjects of different ages. Ten younger subjects (age range, 22-35 years) and 10 older subjects (age range, 50-65 years) were tested. Strips of cone photoreceptors, approximately 12° × 1.8° long were imaged for each of the four primary retinal meridians: superior, inferior, nasal, and temporal. Cone photoreceptors within the strips were counted, and cone photoreceptor packing density was calculated. Statistical analysis (three-way ANOVA) was used to calculate the interaction for cone photoreceptor packing density between age, meridian, and eccentricity. RESULTS As expected, cone photoreceptor packing density was higher close to the fovea and decreased with increasing retinal eccentricity from 0.18 to 3.5 mm (∼0.6-12°). Older subjects had approximately 75% of the cone density at 0.18 mm (∼0.6°), and this difference decreased rapidly with eccentricity, with the two groups having similar cone photoreceptor packing densities beyond 0.5 mm retinal eccentricity on average. CONCLUSIONS Cone packing density in the living human retina decreases as a function of age within the foveal center with the largest difference being found at our most central measurement site. At all ages, the retina showed meridional difference in cone densities, with cone photoreceptor packing density decreasing faster with increasing eccentricity in the vertical dimensions than in the horizontal dimensions.

Adaptive optics scanning laser ophthalmoscope with integrated wide-field retinal imaging and tracking

We have developed a new, unified implementation of the adaptive optics scanning laser ophthalmoscope (AOSLO) incorporating a wide-field line-scanning ophthalmoscope (LSO) and a closed-loop optical retinal tracker. AOSLO raster scans are deflected by the integrated tracking mirrors so that direct AOSLO stabilization is automatic during tracking. The wide-field imager and large-spherical-mirror optical interface design, as well as a large-stroke deformable mirror (DM), enable the AOSLO image field to be corrected at any retinal coordinates of interest in a field of >25 deg. AO performance was assessed by imaging individuals with a range of refractive errors. In most subjects, image contrast was measurable at spatial frequencies close to the diffraction limit. Closed-loop optical (hardware) tracking performance was assessed by comparing sequential image series with and without stabilization. Though usually better than 10 μm rms, or 0.03 deg, tracking does not yet stabilize to single cone precision but significantly improves average image quality and increases the number of frames that can be successfully aligned by software-based post-processing methods. The new optical interface allows the high-resolution imaging field to be placed anywhere within the wide field without requiring the subject to re-fixate, enabling easier retinal navigation and faster, more efficient AOSLO montage capture and stitching.

In vivo measurement of erythrocyte velocity and retinal blood flow using adaptive optics scanning laser ophthalmoscopy

In vivo measurement of retinal blood flow is obtained by measuring the blood velocity of erythrocytes and lumen diameters of the blood vessels using an adaptive optics scanning laser ophthalmoscope. Erythrocyte velocity is measured by tracking erythrocytes moving across a horizontal scanning line. This approach provides high temporal bandwidth measurements, allowing the fluctuation of blood flow during cardiac cycles to be measured. The technique is most applicable to medium-sized blood vessels.

Foveal Avascular Zone and Its Relationship to Foveal Pit Shape

Purpose. To investigate the retinal microvasculature at the fovea and peripheral retina in humans using an adaptive optics scanning laser ophthalmoscope (AOSLO) and to examine the association of foveal avascular zone (FAZ) and foveal pit morphology. Methods. Retinal imaging of the foveal capillary network was performed on 11 subjects (15 eyes; age range 20 to 54 years) with an AOSLO. Standard deviation maps of the AOSLO images were generated from ∼10 to 30 frames, producing high-resolution maps delineating the complete capillary distribution of the retina. Foveal pit morphology was investigated in the same subjects by using a spectral domain optical coherence tomography. In an additional subject, only a relatively large retinal vasculature map was obtained using AOSLO. Results. A well-demarcated FAZ was seen in 11 subjects tested with foveal capillary imaging. There was considerable individual variation in the size and shape of the FAZ. The mean FAZ area and mean FAZ effective diameter were 0.33 mm2 and 622 &mgr;m, respectively. Foveal thickness was found to be negatively correlated with the FAZ effective diameter. Conclusions. The structure of the capillary network could be evaluated in the fovea and parafovea using our approach. We find that a smaller FAZ is associated with a narrower foveal pit opening and a thicker fovea.

Noninvasive measurements and analysis of blood velocity profiles in human retinal vessels.

PURPOSE To quantitatively model the changes in blood velocity profiles for different cardiac phases in human retinal vessels. METHODS An adaptive optics scanning laser ophthalmoscope (AOSLO) was used to measure blood velocity profiles in three healthy subjects. Blood velocity was measured by tracking erythrocytes moving across a scanning line. From the radial position of the cells within the lumen, the blood velocity profile was computed. The cardiac pulsatility was recorded with a cardiac signal monitor. RESULTS The shape of the blood velocity profile in retinal arteries changed systematically during the cardiac cycle, with the flattest profile occurring during the diastolic phase. The measured blood velocity profiles were typically flatter than the commonly assumed parabolic shape. The flatness increased with decreasing vessel size. For the large veins (>80 μm), the ratio of the centerline velocity to the cross-sectional average velocity was between 1.50 and 1.65. This ratio decreased to 1.36 in the smallest vein studied (32 μm). Velocity profiles downstream from a venous confluence showed two peaks at 120 μm from the confluence, but a single velocity peak 500 μm downstream from the confluence. CONCLUSIONS The cardiac cycle influences the blood flow velocity profiles systematically in retinal arteries but not in veins. Parabolic flow was not found in even the largest vessels studied, and deviations from parabolic flow increased in smaller vessels. The measurements are sensitive enough to measure the dual-humped blood velocity profile at a vein confluence.

A Clinical Planning Module for Adaptive Optics Slo Imaging

Purpose. To develop a clinical planning module (CPM) to improve the efficiency of imaging subjects with a steerable wide-field adaptive optics scanning laser ophthalmoscope (AOSLO) and to evaluate the performance of this module by imaging the retina in healthy and diseased eyes. Methods. We developed a software-based CPM with two submodules: a navigation module and a montage acquisition module. The navigation module guides the AOSLO to image identified retinal regions from a clinical imaging platform using a matrix-based mapping between the two. The montage acquisition module systematically moves the AOSLO steering mirrors across the retina in predefined patterns. The CPM was calibrated using a model eye and tested on five normal subjects and one patient with a retinal nerve fiber layer defect. Results. Within the central ±7° from the fixation target, the CPM can direct the AOSLO beam to the desired regions with localization errors of <0.3°. The navigation error increases with eccentricity, and larger errors (up to 0.8°) were evident for regions beyond 7°. The repeatability of CPM navigation was tested on the same locations from two subjects. The localization errors between trials on different days did not differ significantly (p > 0.05). The region with a size of approximately 13° × 10° can be imaged in about 30 min. An approximately 12° × 4.5° montage of the diseased region from a patient was imaged in 18 min. Conclusions. We have implemented a clinical planning module to accurately guide the AOSLO imaging beam to desired locations and to quickly acquire high-resolution AOSLO montages. The approach is not only friendly for patients and clinicians but also convenient to relate the imaging data between different imaging platforms.

The relationship between peripapillary crescent and axial length: implications for differential eye growth

We evaluated the relationship between the size of the peripapillary crescent and the axial length (AL) of the eye as well as the fine structure of the peripapillary crescent in selected eyes. Infrared fundus imaging and spectral domain optical coherence tomography (SDOCT) (Spectralis HRA+OCT, Heidelberg Engineering, Germany) centered at the fovea were performed on 72 healthy adults. On the infrared fundus images, we measured (a) the distance between the foveola and the temporal edge of the optic disc (FOD) and (b) the distance between the foveola and the temporal edge of the peripapillary crescent (FOC) (if present). A peripapillary crescent presented at the nasal margin of the disc in 64% of the subjects. The FOD and FOC were 4.22mm±0.46 and 3.97mm±0.25, respectively. Only the FOD was significantly correlated with axial length. As AL increased by 10%, the FOD increased by 13%, the outer neural retina only expanded by 4% (as indicated by the FOC). This result emphasizes that retinal stretching may not mirror scleral growth, and the existence in some eyes of a difference between the photoreceptor margin and retinal pigment epithelium (RPE) margin suggests that within the retina there could be slippage during eye growth.

Local flicker stimulation evokes local retinal blood velocity changes

We investigated the effect of localized visual stimulation on human retinal blood velocity using an adaptive optics scanning laser ophthalmoscope (AOSLO). To measure the blood velocity response, the AOSLO scanning raster was moved over the target arteries and red blood cell velocity was measured. Localized visual stimuli were delivered by projecting flicker patterns inside or outside the target arterys downstream region. The blood velocity increased in the presence of a flicker stimulus in the downstream region but not when outside the downstream region. The blood velocity increased more with larger area of stimulation. This increase was significant even when the stimulus was smaller than 600 μm × 600 μm. These findings suggest that when the retina regulates its blood flow to metabolic demands, it regulates blood velocity in the vascular system selectively, according to activity of neurons within its field of influence.

Dual electro-optical modulator polarimeter based on adaptive optics scanning laser ophthalmoscope

We constructed a high speed and high-resolution Stokes vector retinal imaging polarimeter with dual electro-optical modulators based on adaptive optics scanning laser ophthalmoscope. By varying the voltages on the EO crystals line by line, we were able to measure over 500,000 Stokes vectors per second. We used this system in three human subjects demonstrating the capability of the system to be employed in vivo. The high speed effectively decreases the adverse impact of eye motion induced errors in polarization calculations, improving the contrast of retinal structures based on their polarization properties.

Adapting AO systems for clinical Imaging

Modern adaptive optics systems for retinal imaging represent a blend between optical design and software control. The problems faced for real-world clinical imaging include the need to obtain high quality data rapidly in less than ideal conditions, including variable size and shape moving pupils, the need to control low order aberrations of 10’s of microns while also correcting high order aberrations to RMS values on the order of 20-40 ums.