Ultra-high isotropic resolution imaging of retinal structures was made possible with an adaptive optics system using dual deformable mirrors and a Fourier-domain optical coherence tomography (Fd-OCT) system with correction for longitudinal chromatic aberration. This system was used to image microscopic retinal structures of healthy as well as diseased retinas in vivo. The improved resolution and contrast enhanced visualization of morphological structures in the retina can be clearly seen. The benefits of this instrument are apparent from comparison of new images with those acquired using a previous generation AO-OCT instrument. Big change in the appearance of speckle field (reduction in speckle size) can be observed as well. Additionally, further improvements in volumetric data acquisition and image representation will be discussed. This includes creation of large Field of View (FOV) AO-OCT volume from multiple sub-volumes and its visualization. Also techniques and results of reducing speckle contrast by averaging multiple B-scans will be presented.
Two deformable mirrors (2DM) were used in an adaptive optics - optical coherence tomography (AO-OCT) system to
image in vivo microscopic retinal structures of healthy and diseased retinas. As a result, multiple morphological
structures not previously seen in vivo have been visualized. Among those presented are three-dimensional
representations of the fovea and optic nerve head (ONH), revealing cellular structures and micro-vasculature. Drusen in
macular degeneration and photoreceptor dystrophies are also presented. Different methods for displaying volumetric
AO-OCT data to facilitate visualization of certain morphological details are compared.
Adaptive optics (AO) coupled with ultra-fast spectral-domain optical coherence tomography (SD-OCT) has achieved the
necessary 3D resolution, sensitivity, and speed for imaging the microscopic retina at the cellular level. While this
technology has been rigorously applied to evaluating the 3D morphology of cone photoreceptors, similar detailed studies
of cell-sized structures in the inner retina have yet to be undertaken. In this paper, we improve the technical performance
of our AO ultrafast SD-OCT and investigate its use for imaging the microscopic inner retina, in particular the nerve fiber
layer (NFL) and retinal capillary network. To maximize lateral resolution within the inner retina, focus was controlled
with a high stroke, 37-actuator bimorph mirror (AOptix) that also served as the wavefront corrector of the AO. The AO
system operated at a closed-loop rate of 25 Hz. The SD-OCT sub-system consisted of a superluminescent diode (&lgr;= 842
nm, &Dgr;&lgr;= 50 nm) and a 512 pixel line scan charge-coupled device (CCD) that acquired 72,000 A-scans/sec. Three
different B-scan lengths (36, 60, and 120 A-scans/B-scan), which correspond to B-scan exposure durations of 0.5, 0.83,
and 1.67 ms, were evaluated to determine the maximum B-scan length that could be tolerated without noticeable loss in
image quality due to eye motion in the well fixated eye. Additional technical improvements included sub-pixel
registration to remove instrument error and axial registration of the volume images. Small volume images were acquired
at 2 and 7 degrees retinal eccentricity with focus systematically shifted through the retina. Small capillaries, some
approaching the smallest in the human eye, were readily detected with AO SD-OCT. Appearance of the nerve fiber layer
varied noticeably with depth. The most inner portion (presumably the inner limiting membrane) appeared as a thin
irregular surface with little characteristic speckle noise. Within the NFL, complex striation patterns (presumably NFL
bundles) were observed throughout the entire thickness with pattern density highest in the inner portion (~15 &mgr;m) and
large corrugations (~35 &mgr;m) at the interface with the ganglion cell layer below. Speckle noise was significant throughout
the NFL.
We evaluate a novel non-invasive technique for observing fast physiological processes, such as phototransduction,
in single photoreceptor cells in the living human eye. The method takes advantage of the interference of multiple
reflections within the outer segments of cones. This self-interference phenomenon is highly sensitive to phase
changes such as those caused by variations in refractive index and scatter within the photoreceptor cell. A high-speed
flood-illumination retina camera equipped with adaptive optics (AO) is used to observe this interference
pattern, and to monitor the changes in those patterns in response to visible stimuli. AO and high frame rates
are necessary for resolving individual cones and their fast temporal dynamics, respectively. Preliminary results
suggest that a frame rate of 192 fps, corresponding to a full field 1024x512 pixel rate of 100 MHz, may be sufficient
for observing these early stages of phototransduction. This pixel rate is at least 80 and 10 times faster than
current flood-illumination and SLO pixel rates, respectively. To our knowledge this is the first demonstration
of in vivo single photoreceptor functional imaging, and the first demonstration of in vivo optical detection of
phototransduction.
Ultrahigh axial resolution in adaptive optics - optical coherence tomography (AO-OCT) is fundamentally limited by the
intrinsic chromatic aberrations of the human eye. Variation in refractive index of the ocular media with wavelength
causes the spectral content of broadband light sources to focus at different depths in the retina for light entering the eye
and at the imaging detector for light exiting. This effect has not been previously reported for ultrahigh-resolution OCT
(without AO) likely because the effect is masked by the relatively long depth of focus dictated by the small pupils used
in these systems. With AO, the pupil size is much larger and depth of focus substantially narrower. As such the
chromatic aberrations of the eye can counteract the lateral resolution benefit of AO when used with broadband light
sources. To more fully tap the potential of AO-OCT, compensation of the eye's chromatic and monochromatic
aberrations must occur concurrently. One solution is to insert an achromatizing lens in front of the eye whose chromatic
aberrations are equal but opposite in sign to that of the eye. In this paper we evaluate the efficacy of a novel design that
uses a custom achromatizing lens placed near the fiber collimating optic. AO-OCT images are acquired on several
subjects with and without the achromatizing lens and in combination with two light sources of different spectral width.
The combination of the achromatizing lens and broadband light source yielded the sharpest images of the retina and the
smallest speckle.
Media considerations for a multiple-layered system using conventional thin films are presented in detail. The media are designed and fabricated for various geometries of thin film coatings to satisfy the optimum media condition minimizing inter-layer crosstalk for conventional reflective volumetric optical data storage systems.
We investigate a novel camera that incorporates adaptive optics (AO) and optical coherence tomography (OCT) to determine if it can achieve the necessary 3-D resolution, sensitivity, and speed for imaging individual cells in the living human retina. An AO spectral-domain OCT system was constructed that is based on a free-space Michelson interferometer design. The OCT sub-system consists of a broadband superluminescent diode whose beam passes through an astigmatic lens to form a line illumination pattern on the retina, which is then imaged onto the slit of an imaging spectrometer. The detector of the spectrometer is a scientific-grade areal CCD. Conventional flood illumination, also with AO, was integrated into the camera and provided confirmation of the focus position in the retina. Short bursts of narrow B-scans (100x560 microns) of the living retina were subsequently acquired at 500 Hz during dynamic compensation that corrected the most significant ocular aberrations across a dilated 6 mm pupil. Camera sensitivity (up to 94 dB) was sufficient for observing reflections from essentially all neural layers of the retina. The 3-D resolution of the B-scans (3.0x3.0x5.7 microns) is the highest reported to date in the living human eye. It was sufficient to observe the interface between the inner and outer segments of individual photoreceptor cells, resolved in both lateral and axial dimensions. The waveguiding nature of the photoreceptors is suggestive at multiple reflective sites. Micro-movements of the retina during short burst imaging allow averaging to reduce speckle contrast, but they appear insufficient for significant speckle reduction.
We report 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 consists of a Shack-Hartmann wavefront sensor and bimorph deformable mirror (AOptix) that
measure and correct the ocular and system aberrations at a closed-loop rate of 12 Hz. Unlike previous AO-OCT and AOSLO
instruments, the bimorph mirror was strategically positioned between the XY mechanical scanners and the subject's
eye so as to avoid beam distortion at the pupil plane, which is created when the mirror compensates for the refractive
error of the eye. This new configuration is evaluated empirically and with commercial ray tracing software. The SDOCT
system consists of a superluminescent diode and a 512 pixel line scan charge-coupled device (CCD) that acquires
75,000 A-scans/sec. This rate is more than two times faster than that previously reported. Retina motion artifiacts 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 distribution of bright spots observed
within C-scans at the inner segment / outer segment (IS/OS) junction 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 IS/OS
junction and either the plexiform layers or the layers immediately behind the OS posterior tips.
We demonstrate for the first time an adaptive optics (AO) spectral OCT retina camera that acquires with unprecedented 3D resolution (2.9 μm lateral; 5.5 μm axial) single shot B-scans of the living human retina. The camera centers on a Michelson interferometer that consists of a superluminescent diode for line illuminating the subject's retinal; voice coil translator for controlling the optical path length of the reference channel; and an imaging spectrometer that is cascaded with a 12-bit area CCD array. The imaging spectrometer was designed with negligible off-axis aberrations and was constructed from stock optical components. AO was integrated into the detector channel of the interferometer and dynamically compensated for most of the ocular aberration across a 6 mm pupil. Short bursts of B-scans, with 100 Ascans each, were successfully acquired at 1 msec intervals. Camera sensitivity was found sufficient to detect reflections from all major retinal layers. Individual outer segments of photoreceptors at different retinal eccentricities were observed in vivo. Periodicity of the outer segments matched cone spacing as measured from AO flood illuminated images of the same patches of retina.
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