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The Shack-Hartmann wavefront sensor (SHWS) spots upon which ocular aberration measurements depend have poor quality in mice due to light reflected from multiple retinal layers. We have designed and implemented a SHWS that can favor light from a specific retinal layer and measured monochromatic aberrations in 20 eyes from 10 anesthetized C57BL/6J mice. Using this instrument, we show that mice are myopic, not hyperopic as is frequently reported. We have also measured longitudinal chromatic aberration (LCA) of the mouse eye and found that it follows predictions of the water-filled schematic mouse eye. Results indicate that the optical quality of the mouse eye assessed by measurement of its aberrations is remarkably good, better for retinal imaging than the human eye. The dilated mouse eye has a much larger numerical aperture (NA) than that of the dilated human eye (0.5 NA vs. 0.2 NA), but it has a similar amount of root mean square (RMS) higher order aberrations compared to the dilated human eye. These measurements predict that adaptive optics based on this method of wavefront sensing will provide improvements in retinal image quality and potentially two times higher lateral resolution than that in the human eye.

Wavefront sensor noise and fidelity place a fundamental limit on achievable image quality in current adaptive optics ophthalmoscopes. Additionally, the wavefront sensor ‘beacon’ can interfere with visual experiments. We demonstrate real-time (25 Hz), wavefront sensorless adaptive optics imaging in the living human eye with image quality rivaling that of wavefront sensor based control in the same system. A stochastic parallel gradient descent algorithm directly optimized the mean intensity in retinal image frames acquired with a confocal adaptive optics scanning laser ophthalmoscope (AOSLO). When imaging through natural, undilated pupils, both control methods resulted in comparable mean image intensities. However, when imaging through dilated pupils, image intensity was generally higher following wavefront sensor-based control. Despite the typically reduced intensity, image contrast was higher, on average, with sensorless control. Wavefront sensorless control is a viable option for imaging the living human eye and future refinements of this technique may result in even greater optical gains.

Wavefront sensor noise and fidelity place a fundamental limit on achievable image quality in current adaptive optics ophthalmoscopes. Additionally, the wavefront sensor ‘beacon’ can interfere with visual experiments. We demonstrate real-time (25 Hz), wavefront sensorless adaptive optics imaging in the living human eye with image quality rivaling that of wavefront sensor based control in the same system. A stochastic parallel gradient descent algorithm directly optimized the mean intensity in retinal image frames acquired with a confocal adaptive optics scanning laser ophthalmoscope (AOSLO). When imaging through natural, undilated pupils, both control methods resulted in comparable mean image intensities. However, when imaging through dilated pupils, image intensity was generally higher following wavefront sensor-based control. Despite the typically reduced intensity, image contrast was higher, on average, with sensorless control. Wavefront sensorless control is a viable option for imaging the living human eye and future refinements of this technique may result in even greater optical gains.

We implemented a Lagrange-multiplier (LM)-based damped least-squares (DLS) control algorithm in a woofer-tweeter dual deformable-mirror (DM) adaptive optics scanning laser ophthalmoscope (AOSLO). The algorithm uses data from a single Shack-Hartmann wavefront sensor to simultaneously correct large-amplitude low-order aberrations by a woofer DM and small-amplitude higher-order aberrations by a tweeter DM. We measured the in vivo performance of high resolution retinal imaging with the dual DM AOSLO. We compared the simultaneous LM-based DLS dual DM controller with both single DM controller, and a successive dual DM controller. We evaluated performance using both wavefront (RMS) and image quality metrics including brightness and power spectrum. The simultaneous LM-based dual DM AO can consistently provide near diffraction-limited in vivo routine imaging of human retina.

The directional sensitivity of the retina, known as the Stiles-Crawford effect (SCE), originates from the waveguide property of photoreceptors. This effect has been extensively studied in normal and pathologic eyes using highly customized optical instrumentation. Here we investigate a new approach based on a Shack-Hartmann wavefront sensor (SHWS), a technology that has been traditionally employed for measuring wave aberrations (phase) of the eye and is available in clinics. Using a modified research-grade SHWS, we demonstrate in five healthy subjects and at four retinal eccentricities that intensity information can be readily extracted from the SHWS measurement and the spatial distribution of which is consistent with that produced by the optical SCE. The technique is found sufficiently sensitive even at near-infrared wavelengths where the optical SCE is faint. We demonstrate that the optical SCE signal is confined to the core of the SHWS spots with the tails being diffuse and non-directional, suggesting cones fail to recapture light that is multiply scattered in the retina. The high sensitivity of the SHWS to the optical SCE raises concern as to how this effect, intrinsic to the retina, may impact the SHWS measurement of ocular aberrations.

Current vision science adaptive optics systems use near infrared wavefront sensor ‘beacons’ that appear as red spots in the visual field. Colored fixation targets are known to influence the perceived color of macroscopic visual stimuli(Jameson, D. and Hurvich, L. M., 1967. Fixation-light bias: an unwanted by-product of fixation control. Vis. Res. 7, 805 – 809.), suggesting that the wavefront sensor beacon may also influence perceived color for stimuli displayed with adaptive optics. Despite its importance for proper interpretation of adaptive optics experiments on the fine scale interaction of the retinal mosaic and spatial and color vision, this potential bias has not yet been quantified or addressed. Here we measure the impact of the wavefront sensor beacon on color appearance for dim, monochromatic point sources in 5 subjects. The presence of the beacon altered color reports both when used as a fixation target as well as when displaced in the visual field with a chromatically neutral fixation target. This influence must be taken into account when interpreting previous experiments and new methods of adaptive correction should be used in future experiments using adaptive optics to study color.

In vivo imaging of the mouse retina using visible and near infrared wavelengths does not achieve diffraction-limited resolution due to wavefront aberrations induced by the eye. Considering the pupil size and axial dimension of the eye, it is expected that unaberrated imaging of the retina would have a transverse resolution of 2 μm. Higher-order aberrations in retinal imaging of human can be compensated for by using adaptive optics. We demonstrate an adaptive optics system for in vivo imaging of fluorescent structures in the retina of a mouse, using a microelectromechanical system membrane mirror and a Shack–Hartmann wavefront sensor that detects fluorescent wavefront.

The performance of a MEMS (micro-electro-mechanical-system) segmented deformable mirror was evaluated in an adaptive optics (AO) scanning laser ophthalmoscope. The tested AO mirror (Iris AO, Inc, Berkeley, CA) is composed of 37 hexagonal segments that allow piston/tip/tilt motion up to 5 μm stroke and ±5 mrad angle over a 3.5 mm optical aperture. The control system that implements the closed-loop operation employs a 1:1 matched 37-lenslet Shack-Hartmann wavefront sensor whose measurements are used to apply modal corrections to the deformable mirror. After a preliminary evaluation of the AO mirror optical performance, retinal images from 4 normal subjects over a 0.9°x0.9° field size were acquired through a 6.4 mm ocular pupil, showing resolved retinal features at the cellular level. Cone photoreceptors were observed as close as 0.25 degrees from the foveal center. In general, the quality of these images is comparable to that obtained using deformable mirrors based on different technologies.

We have combined Fourier-domain optical coherence tomography (FD-OCT) with a closed-loop adaptive optics (AO) system using a Hartmann-Shack wavefront sensor and a bimorph deformable mirror. The adaptive optics system measures and corrects the wavefront aberration of the human eye for improved lateral resolution (~4 μm) of retinal images, while maintaining the high axial resolution (~6 μm) of stand alone OCT. The AO-OCT instrument enables the three-dimensional (3D) visualization of different retinal structures in vivo with high 3D resolution (4×4×6 μm). Using this system, we have demonstrated the ability to image microscopic blood vessels and the cone photoreceptor mosaic.

We report the first observations of the three-dimensional morphology of cone photoreceptors in the living human retina. Images were acquired with a high-speed adaptive optics (AO) spectral-domain optical coherence tomography (SD-OCT) camera. The AO system consisted of a Shack-Hartmann wavefront sensor and bimorph mirror (AOptix) that measured and corrected the ocular and system aberrations at a closed-loop rate of 12 Hz. The bimorph mirror was positioned between the XY mechanical scanners and the subject’s eye. The SD-OCT system consisted of a superluminescent diode and a 512 pixel line scan charge-coupled device (CCD) that acquired 75,000 A-scans/s. This rate is more than two times faster than that previously reported. Retinal motion artifacts were minimized by quickly acquiring small volume images of the retina with and without AO compensation. Camera sensitivity was sufficient to detect reflections from all major retinal layers. The regular distribution of bright spots observed within C-scans at the inner segment / outer segment (IS/OS) junctions and at the posterior tips of the OS were found to be highly correlated with one another and with the expected cone spacing. No correlation was found between the posterior tips of the OS and the other retinal layers examined, including the retinal pigment epithelium.

Many next-generation adaptive optics (AO) systems for vision will have two deformable mirrors (DMs) instead of one: a high-stroke, low-resolution mirror (the woofer) and a low-stroke, high-resolution mirror (the tweeter). We developed a zonal wavefront-control algorithm and validated it using simulations. Rather than separating the woofer and tweeter corrections into two independent control processes or using a modal decomposition, the algorithm we proposed uses wavefront slope measurements from a single Shack–Hartmann wavefront sensor to generate control signals for both deformable mirrors within a single zonal control. A Lagrange multiplier is chosen to integrate two DMs into a single-DM wavefront control, and a damped least-squares control is employed to suppress the correlation between the two DMs.

The monochromatic optical aberrations of the eye degrade retinal image quality. Any significant aberrations during postnatal development could contribute to infants’ immature visual performance and provide signals for the control of eye growth. Aberrations of human infant eyes from 5 to 7 weeks old were compared with those of adult subjects using a model of an adultlike infant eye that accounted for differences in both eye and pupil size. Data were collected using the COAS Shack-Hartmann wavefront sensor. The results demonstrate that the higher order aberrations of the 5-to-7-week-old eye are less than a factor of 2 greater than predicted for an adultlike infant eye of this age. The data are discussed in the context of infants’ visual performance and the signals available for controlling growth of the eye.

Peripheral vision and off-axis aberrations not only play an important role in daily visual tasks but may also influence eye growth and refractive development. Thus it is important to measure off-axis wavefront aberrations of human eyes objectively. To achieve efficient measurement, we incorporated a double-pass scanning system with a Shack Hartmann wavefront sensor (SHWS) to develop a scanning Shack Hartmann aberrometer (SSHA). The prototype SSHA successfully measured the off-axis wavefront aberrations over +/− 15 degree visual field within 7 seconds. In two validation experiments with a wide angle model eye, it measured change in defocus aberration accurately (<0.02μm, 4mm pupil) and precisely (<0.03μm, 4mm pupil). A preliminary experiment with a human subject suggests its feasibility in clinical applications.

We have developed an adaptive optics photoacoustic microscope (AO-PAM) for high-resolution imaging of biological tissues, especially the retina. To demonstrate the feasibility of AO-PAM we first designed the AO system to correct the wavefront errors of the illuminating light of PAM. The aberrations of the optical system delivering the illuminating light to the sample in PAM was corrected with a close-loop AO system consisting of a 141-element MEMS-based deformable mirror (DM) and a Shack-Hartmann (SH) wavefront sensor operating at 15 Hz. The photoacoustic signal induced by the illuminating laser beam was detected by a custom-built needle ultrasonic transducer. When the wavefront errors were corrected by the AO system, the lateral resolution of PAM was measured to be better than 2.5 μm using a low NA objective lens. We tested the system on imaging ex vivo ocular samples, e.g., the ciliary body and retinal pigment epithelium (RPE) of a pig eye. The AO-PAM images showed significant quality improvement. For the first time we were able to resolve single RPE cells with PAM.

We have developed an adaptive optics photoacoustic microscope (AO-PAM) for high-resolution imaging of biological tissues, especially the retina. To demonstrate the feasibility of AO-PAM we first designed the AO system to correct the wavefront errors of the illuminating light of PAM. The aberrations of the optical system delivering the illuminating light to the sample in PAM was corrected with a close-loop AO system consisting of a 141-element MEMS-based deformable mirror (DM) and a Shack-Hartmann (SH) wavefront sensor operating at 15 Hz. The photoacoustic signal induced by the illuminating laser beam was detected by a custom-built needle ultrasonic transducer. When the wavefront errors were corrected by the AO system, the lateral resolution of PAM was measured to be better than 2.5 µm using a low NA objective lens. We tested the system on imaging ex vivo ocular samples, e.g., the ciliary body and retinal pigment epithelium (RPE) of a pig eye. The AO-PAM images showed significant quality improvement. For the first time we were able to resolve single RPE cells with PAM.

Inhomogeneity in thick biological specimens results in poor imaging by light microscopy, which deteriorates as the focal plane moves deeper into the specimen. Here, we have combined selective plane illumination microscopy (SPIM) with wavefront sensor adaptive optics (wao). Our waoSPIM is based on a direct wavefront measure using a Hartmann-Shack wavefront sensor and fluorescent beads as point source emitters. We demonstrate the use of this waoSPIM method to correct distortions in three-dimensional biological imaging and to improve the quality of images from deep within thick inhomogeneous samples.

PURPOSE:

To determine whether the spatial demodulation processing of Shack-Hartmann images is suitable for extracting wavefront gradients for ocular wavefront sensors.

METHODS:

We developed a custom software program to implement the spatial demodulation technique. To test the algorithm’s performance, we generated simulated spot images and obtained an eye examination image. We generated a collection of simulated aberrated spot images corresponding to: astigmatic wavefront (-5.00 -2.00 × 17), highly aberrated defocus (±20.00 diopters [D]), high-resolution defocus (-0.01 D), and third-order aberrations (trefoil and coma). The eye examination image and its measured Zernike coefficients were obtained from a Shack-Hartmann ocular aberrations system. We evaluated the output from the algorithm in terms of comparing the results to the known Zernike coefficients (for the simulated images) or the previously measured Zernike coefficients (for the eye examination image).

RESULTS:

The spatial demodulation algorithm was able to correctly recover the aberrations to better than 1/100 (0.01) D for the simulated spot images. The processing of the eye examination image yielded results within approximately 1/4 (0.25) D to the values provided by the Shack-Hartmann system.

CONCLUSIONS:

From the set of simulated images and the eye examination image used to test the spatial demodulation technique, it appears that the method is suitable for application in ocular wavefront aberrations Shack-Hartmann systems. The method appears capable of accurately processing high levels of aberrations (±20.00 D) as well as providing high resolution as evidenced by finding the -0.01 D defocus. The method may be especially well suited for processing highly aberrated wavefronts.

In order to determine light aberrations, Shack-Hartmann optical wavefront sensors make use of microlens arrays (MLA) to divide the incident light into small parts and focus them onto image planes. In this paper, we present the design and fabrication of long focal length MLA with various shapes and arrangements based on a double layer structure for optical wavefront sensing applications. A longer focal length MLA could provide high sensitivity in determining the average slope across each microlens under a given wavefront, and spatial resolution of a wavefront sensor is increased by numbers of microlenses across a detector. In order to extend focal length, we used polydimethysiloxane (PDMS) above MLA on a glass substrate. Because of small refractive index difference between PDMS and MLA interface (UV-resin), the incident light is less refracted and focused in further distance. Other specific focal lengths could also be realized by modifying the refractive index difference without changing the MLA size. Thus, the wavefront sensor could be improved with better sensitivity and higher spatial resolution.

An optical analysis is developed to separate forward light scatter of the human eye from the conventional wavefront aberrations in a double pass optical system. To quantify the separate contributions made by these micro- and macro-aberrations, respectively, to the spot image blur in the Shark-Hartmann aberrometer, we develop a metric called radial variance for spot blur. We prove an additivity property for radial variance that allows us to distinguish between spot blurs from macro-aberrations and micro-aberrations. When the method is applied to tear break-up in the human eye, we find that micro-aberrations in the second pass accounts for about 87% of the double pass image blur in the Shack-Hartmann wavefront aberrometer under our experimental conditions.

The smallest foveal cones can now be visualized using adaptive optics scanning laser ophthalmoscopy. Cone density is not affected by eye length inside the foveola.

Purpose.

Adaptive optics scanning laser ophthalmoscopy (AOSLO) under optimized wavefront correction allows for routine imaging of foveal cone photoreceptors. The intersubject variability of foveal cone density was measured and its relation to eye length evaluated.

Methods.

AOSLO was used to image 18 healthy eyes with axial lengths from 22.86 to 28.31 mm. Ocular biometry and an eye model were used to estimate the retinal magnification factor. Individual cones in the AOSLO images were labeled, and the locations were used to generate topographic maps representing the spatial distribution of density. Representative retinal (cones/mm2) and angular (cones/deg2) cone densities at specific eccentricities were calculated from these maps.

Results.

The entire foveal cone mosaic was resolved in four eyes, whereas the cones within 0.03 mm eccentricity remained unresolved in most eyes. The preferred retinal locus deviated significantly (P < 0.001) from the point of peak cone density for all except one individual. A significant decrease in retinal density (P < 0.05) with increasing axial length was observed at 0.30 mm eccentricity but not closer. Longer, more myopic eyes generally had higher angular density near the foveal center than the shorter eyes, but by 1°, this difference was nullified by retinal expansion, and so angular densities across all eyes were similar.

Conclusions.

The AOSLO can resolve the smallest foveal cones in certain eyes. Although myopia causes retinal stretching in the fovea, its effect within the foveola is confounded by factors other than cone density that have high levels of intersubject variability.

A retinal imaging instrument that integrates adaptive optics (AO), scanning laser ophthalmoscopy (SLO), and retinal tracking components was built and tested. The system uses a Hartmann-Shack wave-front sensor (HS-WS) and MEMS-based deformable mirror (DM) for AO-correction of high-resolution, confocal SLO images. The system includes a wide-field line-scanning laser ophthalmoscope for easy orientation of the high-magnification SLO raster. The AO system corrected ocular aberrations to <0.1 μm RMS wave-front error. An active retinal tracking with custom processing board sensed and corrected eye motion with a bandwidth exceeding 1 kHz. We demonstrate tracking accuracy down to 6 μm RMS for some subjects (typically performance: 10–15 μm RMS). The system has the potential to become an important tool to clinicians and researchers for vision studies and the early detection and treatment of retinal diseases.

Maximum-likelihood (ML) estimation in wavefront sensing requires careful attention to all noise sources and all factors that influence the sensor data. We present detailed probability density functions for the output of the image detector in a wavefront sensor, conditional not only on wavefront parameters but also on various nuisance parameters. Practical ways of dealing with nuisance parameters are described, and final expressions for likelihoods and Fisher information matrices are derived. The theory is illustrated by discussing Shack–Hartmann sensors, and computational requirements are discussed. Simulation results show that ML estimation can significantly increase the dynamic range of a Shack–Hartmann sensor with four detectors and that it can reduce the residual wavefront error when compared with traditional methods.

PURPOSE

To evaluate the influence of spherical aberration on contrast sensitivity using adaptive optics.

SETTING

Vision Science and Advanced Retinal Imaging Laboratory, Department of Ophthalmology & Vision Science, University of California, Davis Medical Center, Sacramento, California, USA.

METHODS

Contrast sensitivity at 8 cycles per degree was evaluated using an adaptive optics system that permitted aberrations to be measured with a Shack-Hartman wavefront sensor and controlled by a 109 actuator continuous-surface deformable mirror that was at a plane conjugate to the observer’s pupil. Vertical Gabor patches were viewed through a 6.3 mm diameter pupil conjugate aperture. Contrast sensitivity was measured with the deformable mirror set to produce 1 of 5 spherical aberration profiles (−0.2 to +0.2 μm). Contrast sensitivity over the range of spherical aberration was fitted with a polynomial function.

RESULTS

Three observers (age 21 to 24 years) participated. The measured total mean spherical aberration resulting from the spherical aberration profiles produced by the deformable mirror was between −0.15 μm and +0.25 μm. The peak contrast sensitivity of this function for the 3 observers combined occurred at +0.06 μm of spherical aberration. The peak contrast sensitivity was also achieved with positive spherical aberration for observer (mean 0.09).

CONCLUSION

There was intersubject variability in the measurements; however, the average visual performance was best with the introduction of a small positive spherical aberration.

We present an iterative technique for improving adaptive optics (AO) wavefront correction for retinal imaging, called the Adaptive-Influence-Matrix (AIM) method. This method is based on the fact that the deflection-to-voltage relation of common deformable mirrors used in AO are nonlinear, and the fact that in general the wavefront errors of the eye can be considered to be composed of a static, non-zero wavefront error (such as the defocus and astigmatism), and a time-varying wavefront error. The aberrated wavefront is first corrected with a generic influence matrix, providing a mirror compensation figure for the static wavefront error. Then a new influence matrix that is more accurate for the specific static wavefront error is calibrated based on the mirror compensation figure. Experimental results show that with the AIM method the AO wavefront correction accuracy can be improved significantly in comparison to the generic AO correction. The AIM method is most useful in AO modalities where there are large static contributions to the wavefront aberrations.

Purpose.

The purpose of this study is to determine the ability of single-value metrics of retinal image quality of the eye to predict visual performance as measured by high (HC) and low (LC) -contrast acuity at photopic (P) and mesopic (M) light levels in eyes with 20/17 and better visual acuity.

Methods.

Forty-nine normal subjects in good health ranging in age from 21.8 to 62.6 with 20/17 or better monocular high-contrast logarithm of the minimum angle of resolution (logMAR) acuity served as subjects. Wavefront error through the 10th Zernike radial order over a 7-mm pupil was measured on each test eye using a custom-built Shack/Hartmann wavefront sensor. For each eye, 31 different single-value retinal image quality metrics were calculated. Visual acuity was measured using HC (95%) and LC (11%) logMAR at photopic (270 cd/m2) and mesopic (0.75 cd/m2) light levels. To determine the ability of each metric of retinal image quality to predict each type of logMAR acuity (P HC, P LC, M HC, and M LC), each acuity measure was regressed against each optical quality metric.

Results.

The ability of the metrics of retinal image quality to predict logMAR acuity improved as luminance and/or contrast is lowered. The best retinal image quality metric (logPFSc) accounted for 2.6%, 15.1%, 27.6%, and 40.0% of the variance in P HC, P LC, M HC, and M LC logMAR acuity, respectively.

Conclusions.

In eyes with 20/17 and better P HC acuity, P HC logMAR acuity is insensitive to variations in retinal image quality compared with M LC logMAR acuity. Retinal image quality becomes increasingly predictive of logMAR acuity as contrast and/or luminance is decreased. Everyday life requires individuals to function over a large range of contrast and luminance levels. Clinically, the impact of retinal image quality as a function of luminance and contrast is readily measurable in a time-efficient manner with M LC logMAR acuity charts.