Reviews

Optical Coherence Tomography: History, Current

Status, and Laboratory Work
Michelle L. Gabriele,1,2,3 Gadi Wollstein,1 Hiroshi Ishikawa,1,2 Larry Kagemann,1,2
Juan Xu,1 Lindsey S. Folio,1 and Joel S. Schuman1,2,3,4

Optical coherence tomography (OCT) imaging has become allows backscattered tissue intensity levels to be detected from
widespread in ophthalmology over the past 15 years, because different depths in the tissue sample. This approach is referred
of its ability to visualize ocular structures at high resolution. to as time-domain (TD)-OCT because time-encoded signals are
This article reviews the history of OCT imaging of the eye, its obtained directly. Several improvements in OCT hardware
current status, and the laboratory work that is driving the have been introduced since the first commercial TD-OCT sys-
future of the technology. (Invest Ophthalmol Vis Sci. 2011;52: tem became available. Better axial resolution11–13 and in-
2425–2436) DOI:10.1167/iovs.10-6312 creased scanning speed14 –23 are the two main advancements
that have recently become incorporated into commercial sys-

O ptical coherence tomography (OCT) has advanced con-

siderably since it was first applied to the eye.1–7 It is an
extension of a technique called low-coherence interferometry,
tems.
The implementation of broadband light sources into OCT
systems11 improved the axial resolution from ⬃10 ␮m to as
which was initially applied to ophthalmology for in vivo mea- high as 2 ␮m in tissue.24 Acquisition speed has improved
surements of eye axial length.1 At the time of introduction, it considerably by detecting backscattering signals in the fre-
was used to obtain in vivo optical cross sections of the anterior quency domain,14 –23 which means backscattered depth infor-
segment,6 as well as retinal diseases, such as macular detach- mation at a given location can be collected without the move-
ment, macular hole, epiretinal membrane, macular edema, and ment of a reference mirror. Frequency information is acquired
idiopathic central serous chorioretinopathy.4 OCT cross sec- with a broad-bandwidth light source, charge-coupled device
tions were also used to evaluate the optic disc and retinal (CCD) camera, and a spectrometer14,17,18,20 or by sweeping a
layers5,8 such as the retinal nerve fiber layer (RNFL).9 Scan narrow-bandwidth source through a broad range of frequen-
patterns that enabled reproducible measurements were devel- cies with a photodetector.16,21–23 The approach that incorpo-
oped,10 and these eventually became incorporated into a com- rates a broadband light source is often referred to as spectral-
mercial system, which had an axial resolution of ⬃10 ␮m. domain (SD)-OCT, whereas the latter is termed swept-source
The first clinical system was limited to a scanning speed of (SS)-OCT. In both approaches, intensity profiles (A-scans) are
400 axial scans (A-scans)/s because of a physical constraint: a obtained using a Fourier-transform of the detected frequencies,
moving reference mirror. OCT uses low coherence interferom- and this facilitates rapid A-scan collection. In addition to im-
etry to obtain A-scan intensity profiles, and the process re- proved scanning speed, frequency-domain-OCT also offers the
quires light to be split and sent to both a reference arm with a advantage of higher detection sensitivity—that is, it exhibits
mirror and to the sample. Provided the path length to the higher signal-to-noise, given a perfect reflector.23,25 A summary
reference mirror and tissue match to within the coherence of OCT detection techniques can be seen in Table 1.
length of the light source, when the reflected beams recom- With these speed and sensitivity improvements, it is now
bine, interference occurs. Intensity information, in the form of feasible to collect volumetric (three-dimensional; 3D) scans of
a reflectivity profile in depth, can be extracted from the inter- tissue, whereas in the past, the amount of time required to do
ference profile. Changing the location of the reference mirror this would have been prohibitive. Broadband volumetric reti-
nal imaging with SD-OCT at speeds of up to 312,500 A-scans/s26
and SS-OCT at 249,000 A-scans/s27 have been demonstrated.
From the 1Department of Ophthalmology, UPMC Eye Center, Eye To date, most clinical systems operate at an acquisition rate of
and Ear Institute, Ophthalmology and Visual Science Research Center, ⬃27,000 A-scans/s and an axial resolution of 5 to 6 ␮m. A
University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania; summary of the current commercially available retinal imaging
the 2Department of Bioengineering, Swanson School of Engineering systems is presented in Table 2.
and the 4McGowan Institute for Regenerative Medicine, University of
Pittsburgh, Pittsburgh, Pennsylvania; and the 3Center for the Neural
Basis of Cognition, University of Pittsburgh and Carnegie Mellon Uni- CURRENT STATUS OF OCT IN THE CLINIC
versity, Pittsburgh, Pennsylvania.
Supported in part by National Institutes of Health Grants R01- Glaucoma
EY13178 and P30-EY08098; The Eye and Ear Foundation, Pittsburgh,
PA; and an unrestricted grant from Research to Prevent Blindness, New Conventionally, the most common scan patterns in TD-OCT
York, NY. glaucoma imaging were a 3.4 mm scan around the optic nerve
Submitted for publication July 30, 2010; revised October 20, 2010; head (ONH) and six equally spaced radial scans through the
accepted November 13, 2010. macula (6 mm) and optic nerve (4 mm). RNFL thickness is
Disclosure: M.L. Gabriele, None; G. Wollstein, Carl Zeiss Med- obtained via automated RNFL segmentation in the circumpap-
itec (F), Optovue (F), P; H. Ishikawa, P; P.L. Kagemann, None; J. Xu,
P; L.S. Folio, None; J.S. Schuman, Carl Zeiss Meditec (F), P
illary scan protocol, whereas macular thickness (internal limit-
Corresponding author: Joel S. Schuman, UPMC Eye Center, De- ing membrane [ILM] to the photoreceptor inner segment-outer
partment of Ophthalmology, University of Pittsburgh School of Medi- [IS–OS] segment junction) is automatically segmented in the
cine, 203 Lothrop Street, Eye and Ear Institute, Suite 816, Pittsburgh, macular scan pattern. The optic nerve scan is used to obtain
PA 15213; [email protected]. cup area, disc area, cup diameter, disc diameter, and rim area.

Investigative Ophthalmology & Visual Science, April 2011, Vol. 52, No. 5
Copyright 2011 The Association for Research in Vision and Ophthalmology, Inc. 2425

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TABLE 1. Comparison of TD-, SD-, and SS-OCT Devices

Ophthalmic System
Light Source Commercially Available? Primary Advantages Primary Disadvantages

TD-OCT Broadband width Yes Intensity information acquired in time Moving reference mirror required
domain; no complex conjugate image limiting acquisition rate
SD-OCT Broadband width Yes No moving reference mirror required; Noticeable signal drop-off with depth
higher sensitivity than TD-OCT; high
scanning speed and axial resolution
have been attained
SS-OCT Narrow band, swept No No moving reference mirror required; Most ophthalmic systems operating
through broad range Higher sensitivity than TD-OCT; very at longer wavel engths (␭ ⫽ 1⫺1.3
high scanning speeds can be attained; ␮m), with lower axial resolution
minimal signal drop-off with depth

These ONH parameters are obtained automatically: the soft- (e.g., 6.0 ⫻ 6.0 ⫻ 2.0 mm, centered on the ONH) after acqui-
ware detects the ONH margin/RPE tips, but the user can sition, at a diameter of 3.4 mm centered on the ONH (Fig. 1).
modify the location if the ONH margin detection algorithm is This method has been shown to have higher reproducibility
inaccurate. than the conventional TD-OCT 3.4 mm scan circle, where
The ONH and RNFL scan protocols have been used since the image is acquired along the circle only.36 One explana-
TD-OCT became commercially available, and RNFL and ONH tion for the improved performance is that, with TD-OCT,
parameters have been shown to differ between glaucomatous scan placement is dependent on the user and can be vari-
and healthy eyes.9,28 –33 The glaucoma-discriminating ability, able, but with SD-OCT, the circle can be consistently placed
measured by the area under receiver operating characteristic in the same location by using landmarks within the 3D
curves (AROC) of RNFL (AROC ⫽ 0.94) and disc parameters volume.
(e.g., rim area AROC ⫽ 0.97), has been reported to be higher Although sampling 3D volumes after acquisition may be an
than macular volume and thickness (AROC, both 0.80).34 A effective way of summarizing RNFL measurements, it is doing
similar glaucoma-discriminating ability is seen in comparing so at a cost: data outside the 3.4-mm band are not being used.
TD-OCT imaging and SD-OCT imaging when similar parameters Subjectively, wedge defects and global thinning may be easy to
are examined.35 However, it may be possible to further im- spot, but subtle changes or deviations from normal outside the
prove glaucoma discrimination using parameters obtained 3.4-mm sampling band may be missed. One way of addressing
from 3D scanning. With the commercialization of rapidly scan- this is to create an RNFL thickness map, which consists of all
ning SD-OCT systems, 3D volumes of tissue are now easily thickness measurements outside of the ONH. From this, thick-
acquired. A 3D dataset not only allows a quantitative analysis ness measurements from one subject can be compared to
from more locations but, once a volume has been collected, population thickness measurements. To date, however, com-
OCT fundus (en face) images can be obtained by integrating mercial software is available for looking at deviation from
A-scans.20 These can be used for a subjective assessment of normal, but there is no quantitative assessment using all avail-
signal quality and to assist with evaluating and/or correcting able RNFL information.
eye motion that may have occurred throughout the scan. The Different approaches have been proposed for quantifying
OCT fundus image also allows registration of OCT cross sec- 3D data. 3D RNFL thickness has been analyzed in terms of a
tions to precise retinal locations. thickness profile as distance from the ONH increased.37 In
Acquisition of 3D datasets has led to the advancement of healthy eyes, the slope of RNFL thickness increases near the
software methods for efficiently analyzing and summarizing margin of the ONH, peaks, and then decreases with increasing
these vast amounts of data. One method of obtaining RNFL distance from the ONH center in all but the nasal quadrant,
thickness measurements has been sampling the 3D volume which linearly decreases starting from the disc margin. An-

TABLE 2. Description of Commercially Available SD-OCT Systems

Device (Manufacturer) Description

3D-OCT 2000 (Topcon, Tokyo, Japan) SD-OCT and high-resolution fundus camera; axial resolution, 5 ␮m; A-scan
acquisition rate, 27 kHz.
Bioptigen SD-OCT (Bioptigen, Research Triangle Park, NC) Designed for both clinical and research use and includes a hand-held
probe and microscope setup; axial resolution, 4 ␮m; A-scan acquisition
rate, 20 kHz
Cirrus HD-OCT (Carl Zeiss Meditec, Dublin, CA) Software includes guided progression analysis for glaucoma progression
detection; axial resolution, 5 ␮m; A-scan acquisition rate, 27 kHz.
RTVue-100 (Optovue, Fremont, CA) Offers multiple scanning protocols for glaucoma detection, including
ganglion cell complex analysis; axial resolution, 5 ␮m; A-scan
acquisition rate, 26 kHz.
SOCT Copernicus (Optopol, Zawiercie, Poland) Software includes progression analysis software that incorporates disk
damage likelihood scale, asymmetry between the discs, and RNFL
thickness; axial resolution, 6 ␮m; A-scan acquisition rate, 27 kHz.
Spectral OCT SLO (Opko, Miami, FL) Combines SD-OCT, scanning laser ophthalmoscopy, and microperimetry.
Axial resolution, 6 ␮m, A-scan acquisition rate, 27 kHz.
Spectralis OCT (Heidelberg Engineering, Heidelberg, Germany) High-speed SD-OCT device with eye-tracking, fluorescein angiography,
ICG angiography, and autofluorescence. Axial resolution, 4 ␮m; A-scan
acquisition rate, 40 kHz.

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 IOVS, April 2011, Vol. 52, No. 5 Optical Coherence Tomography 2427

FIGURE 1. Top: OCT fundus images;

bottom: RNFL thickness maps ob-
tained by segmenting RNFL at all loca-
tions outside of the optic nerve, for a
healthy and glaucoma subject. Outer
red circle: the location of a resampled
3.4-mm peripapillary cross-section. Im-
ages acquired with Cirrus HD-OCT;
200 ⫻ 200 A-scans, 6 ⫻ 6 mm.

other approach (Ishikawa H et al. IOVS 2009;50:ARVO E-Ab- data to superpixels (4 ⫻ 4 adjacent sampling points) and
stract 3328) exploits 3D macular data, which have been sum- compares these superpixels to a normative thickness super-
marized using segmentation of the inner retinal complex (IRC: pixel dataset. By condensing measurements into superpixels, it
retinal ganglion cell layer [RGC], inner plexiform layer [IPL], is less likely that small imaging artifacts or algorithm failure will
inner nuclear layer [INL]; Fig. 2).38 This approach reduces IRC have an effect.

FIGURE 2. Left: SD-OCT macular

thickness map (inner retinal com-
plex); right: cross-sectional image
showing automated segmentation
results for one frame of the 3D vol-
ume. Image acquired with Cirrus HD-
OCT; 200 ⫻ 200 A-scans, 6 ⫻ 6 mm);
Blue lines: from inner to outer retina,
indicate the outer border of the retinal
nerve fiber layer, outer border of the
inner plexiform layer, outer border of
the outer plexiform layer and RPE.

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One commercially available system has developed an ap- using polarization-sensitive (PS)-OCT47– 49 are two techniques
proach for summarizing macular data called the Ganglion Cell under development that may improve the diagnosis and mon-
Complex (GCC; RTVue, Optovue, Inc) which consists of es- itoring of glaucoma. These are further described in the Preclin-
sentially the same layers as the IRC: the RGC (retinal ganglion ical and Laboratory Studies section of this review.
cell bodies), RNFL (RGC axons) and IPL (RGC dendrites). The
GCC measurements are then directly compared to a normative Retina
database and thickness difference and significance maps are The macular scan pattern discussed above—six radial macular
available (Fig. 3). scans, 6 mm long, spaced 30o apart— has traditionally been
While a comparison to a normal population may reveal used in TD-OCT imaging to assess retinal parameters such as
differences, structural changes may be occurring while a pa- total retinal thickness and the IRC. Three-dimensional imaging;
tient remains within normal limits and therefore go unde- however, has revolutionized the examination of retinal dis-
tected. Ideally, a longitudinal comparison could be made for a ease.50 –55 An examination of the 3D structure of the retina, as
given individual to look for subtle structural changes attributed opposed to just six radial scans, may make subtle structural
to disease progression. One approach, proposed by Kim changes apparent. For example, using high-resolution 3D im-
et al.,39 is to allow compatibility between TD-OCT and 3D OCT aging to observe the photoreceptor IS–OS junctions may be an
device iterations. Since TD-OCT devices have been commer- indicator of visual outcomes after macular hole surgery.56 –58
cially available longer than 3D imaging systems, years of patient At present, one application of 3D OCT of imaging retinal
information may be available. The method presented by Kim et diseases that has considerable clinical potential is surgical plan-
al. resamples a 3D-OCT dataset for every possible 3.4-mm ning and the evaluation of surgical outcomes. The use of OCT
circular scan location within the boundaries of the 3D-OCT for planning an access point to release the hyaloid for vitrec-
volume. It then uses cross correlation between these virtual tomy using the six radial scan pattern in TD-OCT has been
circular scans and the TD-OCT 3.4-mm scan to automatically described.59 Although this was effective for minimizing trac-
match the TD-OCT scan circle location within the volume. tion forces on the macula during surgery, a detailed 3D map of
To longitudinally compare 3D volumes, however, image the hyaloid membrane and subhyaloid space could further
registration techniques must be developed to spatially align 3D inform the clinician. Falker-Radler et al.60 used 3D imaging to
scans before they can be compared. This may be accomplished visualize the vitreomacular interface in subjects who were
using cross-correlation,40,41 or by using landmarks within the undergoing surgery for epiretinal membrane. Others have used
OCT fundus image, such as blood vessels.42,43 Eye motion OCT for evaluation of structure after surgery for macular
during acquisition has been shown to alter scan location,44 – 46 hole57,58,61 and vitreomacular traction.52,62– 64 The use of 3D
and the effect of eye motion is visible on OCT en face images imaging for surgical preparation and evaluation of surgical
as discontinuous blood vessels. Detecting and correcting blood outcomes has the potential to improve with the use of longer
vessel location to align the OCT fundus can help correct eye wavelength imaging, which is described later in this review.
motion,42 which may be useful for cross-sectional analysis. Automated segmentation of structures of interest, when possi-
Longer wavelength imaging (⬃1-␮m center wavelength) of ble, may provide objective measurements to clinicians for pre-
the lamina cribrosa27 and birefringence imaging of the RNFL and postsurgical evaluation.

FIGURE 3. Ganglion cell complex

thickness and significance maps for
a healthy (left) and glaucoma
(right) subject. The ganglion cell
complex includes cell bodies, ax-
ons and dendrites of retinal gan-
glion cells. Images acquired with
RTVue-100: 1 horizontal B-scan,
and 15 vertical B-scans (separated
by 0.5 mm). All B-scans consisted of
933 A-scans; 7 ⫻ 6 mm scan area.

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 IOVS, April 2011, Vol. 52, No. 5 Optical Coherence Tomography 2429

Quantification of thickness is possible in certain dis- shows an example C-mode section taken after aligning a mac-
eases,65,66 especially with early stage changes.67,68 The repro- ular SD-OCT 3D volume to the RPE. Moving axially, past the
ducibility of SD-OCT retinal thickness measurements is higher retina and RPE, enables visualization of the choroidal blood
in than that of TD-OCT.69 Thickness has been shown to cor- vessels. The choroidal vessels are not apparent in the corre-
relate with best corrected visual acuity in diabetic macular sponding SD-OCT fundus image because of highly reflective
edema70 and ERM.71 Although thickness may be a clinically layers superficial to the choroid. The C-mode provides alterna-
useful correlate of visual function, there are cases and diseases tive viewing perspective for many retinal diseases, such as
in which no correlation to thickness is seen, and thus clinicians cystoid macular edema, central serous retinopathy, vitreoreti-
should exercise caution in interpretation of thickness measure- nal traction, and age-related macular degeneration,77 and it can
ments.72 improve the visualization of their pathologic features.
Drusen volume may be a predictor of progression of age- It is also possible to image the choroid by focusing the
related macular degeneration,65 and efforts are under way for illuminating OCT beam deeper and moving the choroid closer
automated assessment.66 Although accurate quantification of to 0 delay.80 In addition, longer wavelength imaging at ⬃1
volumetric tissue changes will assist with longitudinal monitor- ␮m81 allows for deeper penetration of light into the retina and
ing of disease, fully automated segmentation may not be reli- choroid. A combination of these approaches may improve the
able because of shadowing from fluid in the retina73,74 or current understanding of choroidal diseases.
because of pathologic events that disrupt normal retinal struc- Correcting ocular aberrations with adaptive-optics (AO)-
tures, such as macular hole, subretinal fluid, pigment epithe- OCT82 may also provide a unique viewing perspective for
lium detachment, and others.75,76 retinal diseases. This technique has been applied to view
In cases in which fully automated segmentation fails, C- photoreceptors83 and RNFL.84 The utility of longer wave-
mode visualization of structures may augment subjective anal- length imaging and AO-OCT is under investigation and is
yses.77 A 3D volume of data can be sectioned in any plane after described in the Preclinical and Laboratory Studies section
acquisition, and for C-mode visualization, data are sectioned below.
perpendicular to the retina. The section can be of any thick-
ness, so structures embedded within a volume can be exposed. Anterior Segment
Often, since the true structure of the retina is curved, exact
perpendicular sections slice through several layers simultane- Anterior segment OCT (AS-OCT) provides structural informa-
ously78,79; thus, aligning the volume to structures such as the tion of the cornea and anterior chamber without contacting
ILM or RPE assists with isolating structures of interest.77 Figure 4 the eye, offering an ease of image acquisition and a consider-

FIGURE 4. Top left: Macular SD-OCT

fundus image; top right: cross-sec-
tion through the fovea. Bottom left:
C-mode of choroidal vessels; bottom
right: based on a slab of thickness
indicated by the three horizontal
white lines after aligning to the RPE.
Image acquired with Cirrus HD-OCT;
200 ⫻ 200 A-scans, 6 ⫻ 6 mm).

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able advantage over ultrasound biomicroscopy (UBM). While it tures anterior and posterior to the iris, Schlemm’s canal, tra-
cannot be used to image deep structures such as the ciliary becular meshwork, and the scleral spur,107 as well as the
body, as UBM can, AS-OCT has higher axial resolution (5–10 anterior chamber angle.108
␮m for AS-OCT compared with 25 ␮m for UBM).85 While SS-OCT systems at any wavelength are not yet com-
It is possible to acquire high-resolution images of the sclera, mercially available for clinicians, in part due to the cost of the
angle, and iris with AS-OCT imaging at longer wavelengths (1.3 light source, there is clinical potential for such devices. No
␮m).86 High-resolution images of the anterior chamber angle signal drop-off with depth in SS-OCT, in combination with
can also be obtained with 850-nm systems, and this has led to deeper penetration from longer wavelengths, may improve
the visualization of the trabecular meshwork and Schlemm’s delineation of the outer retina, RPE, and choroid thereby en-
canal.87– 89 hancing the performance of segmentation algorithms. In addi-
Raster scanning and radial scanning of the cornea have been tion, high-speed 1.3-␮m imaging may expand the use of ante-
used to measure thickness,90 resulting in reliable pachymetric rior segment OCT imaging.
mapping.91,92 Pachymetric measurements obtained with AS-
OCT may assist with planning or follow-up of LASIK patients93
Adaptive Optics OCT
or may be used to diagnose keratoconus.94
In addition to its potential benefits in the evaluation of the Ophthalmic systems that employ adaptive optics (AO) dynam-
anterior chamber angle and cornea, AS-OCT has also been ically adjust their optical characteristics to compensate for
shown to be applicable in the assessment of lens thickness in monochromatic aberrations that occur naturally in the eye. AO
phakic eyes95 or intracorneal ring placement.96 This can pro- was initially proposed109 and later used by astronomers to
vide an alternate, noncontact, method of pre- and postsurgical correct distortions of light passing through the atmosphere.110
assessment. In 1997, AO was demonstrated in the eye by Liang et al.,111
who used a Hartmann-Shack wavefront sensor and a deform-
able mirror to correct contrast sensitivity and improve quality
PRECLINICAL AND LABORATORY STUDIES of vision for human subjects and to obtain higher resolution
Swept-Source OCT and Longer images with an AO fundus camera. Shortly thereafter, individ-
ual cone mosaics were imaged.112
Wavelength Imaging AO-OCT was first reported by Miller et al.82 in 2003 to
As previously discussed, SS-OCT obtains time-encoded spectral improve transverse resolution. Uncorrected, conventional
information by sweeping a narrow-bandwidth laser through a OCT beams 1 mm in diameter have a transverse resolution
broad optical spectrum. Backscattered intensity is detected limited to ⬃15 to 20 ␮m.113 This makes it difficult to visualize
with a photodetector. This process is in contrast to SD-OCT, individual cellular structures. One way to improve transverse
which uses a broad bandwidth light source and detects the resolution is to increase the numerical aperture, which in
interference spectra with a CCD camera and spectrometer. practice means increasing the diameter of the OCT beam
The use of spectrometer-based SD-OCT has become wide- entering the eye, since this would decrease the spot size on the
spread in the clinic, but there are some benefits to photode- retina. However, the theoretical diffraction-limited resolution
tector-based SS-OCT systems. Similar to SD-OCT, SS-OCT offers cannot be attained due to ocular aberrations114 that occur
speed and sensitivity advantages over TD-OCT.23,27 To date, when the pupil is dilated.115,116 AO-OCT measures and cor-
speeds of up to 249,000A-scans/s have been attained in the rects these aberrations using wavefront sensing and deform-
eye.27 Therefore, eye motion artifacts are greatly reduced com- able mirrors, thereby minimizing spot size and improving trans-
pared with TD-OCT.22 verse resolution. It should also be noted that aberrations can be
One advantage of SS-OCT over SD-OCT is that it does not dependent on the bandwidth of the light source used for OCT
require a CCD camera and spectrometer and instead uses a imaging,116 and these may be improved using an achromatizing
simpler photodetector.27 A drawback to camera-based SD-OCT lens.117
detection is a drop-off in signal with depth of scanning because Ultrahigh (axial)-resolution AO-OCT was introduced in
of the finite pixel size of the CCD camera.25,97 Although this 2004, improving transverse resolution to 5 to 10 ␮m in the
can be improved by reducing the camera pixel size,97 it in- retina.113 Zhang et al.118 developed an AO SD-OCT system and
creases the complexity and therefore the cost of the CCD saw an enhancement of the photoreceptor IS–OS junction in
array. A noticeable drop-off in signal with depth typically does vivo with AO. C-mode sectioning of 3D datasets have also
not occur with SS-OCT imaging due to the narrow bandwidth facilitated the visualization of axon bundles in the RNFL84 and
of the light source.23,97 cone photoreceptor mosaics from healthy subjects,83,119 and
At this time, one disadvantage of SS-OCT is that most sys- subjects with structural abnormalities120 and optic neuropa-
tems are now operating at longer wavelengths (␭ ⫽ 1–1.3 ␮m), thies.121
with very few studies demonstrating SS-OCT in the 800 nm One disadvantage of AO imaging is that the depth of focus
range.98,99 Water absorption limits the usable bandwidth at 1 is narrow, which means focusing simultaneously at different
and 1.3 ␮m99 and this limits the axial resolution; the water depths is difficult. For example, photoreceptors, located deep
absorption window at 850 nm is larger, so higher axial resolu- in the retina, and superficial retinal ganglion cells cannot be
tion can be achieved. brought into focus at the same time. It may be possible to
While axial resolution at longer wavelengths may not be as address this limitation by scanning in depth and varying the
fine as at 850 nm, there are advantages to using ⬃1- and 1.3-␮m focal plane122 or by stitching together volumes.123 Another
sources. Posterior segment imaging using ⬃1-␮m (1040 –1060- limitation to AO imaging is that the field of view is restricted to
nm)81,100,101 center wavelengths has allowed deeper penetra- approximately 1° to 3°; the use of an eye-tracking system to
tion into the retina, optic nerve head, and choroid,81,102 which acquire a series of neighboring scans and gradually build up an
may be beneficial for imaging choroidal vessels, lamina cri- image covering a larger volume may provide one solution to
brosa, and diseases such as choroidal neovascularization.103 this limitation.124
The water absorption window at 1.3 ␮m offers even deeper A potential advantage of improved lateral resolution with
penetration of light and may be useful for cornea and anterior AO-OCT is improved understanding of normal and pathologic
segment imaging.16,22,23,104 –106 Anterior chamber imaging at retinal function in vivo. AO may also help to improve the
1310 nm has been applied to visualize anterior segment struc- overall quality of images obtained from eyes that have more

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aberrations. Enhanced lateral resolution and improved image device has also been demonstrated for use in patients under-
quality may then lead to better performance of automated going vitrectomy, after removing either the ILM or epiretinal
segmentation algorithms and assist with disease diagnosis and membrane, to better visualize the macular disease.142 It is
follow-up. possible that the development of an intraoperative approach
may be further improved using a projection of a virtual OCT
Polarization-Sensitive OCT image over the surgical site and within the line of sight of the
Polarization-sensitive (PS) OCT detects polarization changes in surgeon,143 but the means of implementing this technique for
circularly polarized light.125 PS-OCT was initially applied to surgery still has to be investigated.
characterize the birefringence of tooth enamel,126 skin,127 and
cartilage.128 In 2001, PS-OCT was first used in the eye129 to Animal OCT Imaging
measure birefringence of the RNFL in rhesus monkeys. RNFL
birefringence was measured in humans by Cense et al.47,48 and The noninvasive nature of image acquisition, together with the
Yamanari et al.,49 who found that, unlike RNFL thickness, commercialization of systems optimized for laboratory use has
birefringence does not change as a function of increasing resulted in a recent increase in the number of animal studies
radius from the ONH.48 It does, however, vary by sector using OCT. Two- and three-dimensional scanning with OCT is
around the ONH, with higher birefringence in thicker areas.48 appealing, because the same animals can be observed over
Because birefringence may change with disease, RNFL birefrin- time in vivo, making longitudinal studies of ocular structures
gence obtained with OCT may eventually provide an additional possible without the need to kill animals at various time points
indicator of glaucomatous change. The utility of PS-OCT in and obtain histologic sections. Not only does this method
glaucoma detection and monitoring is currently under investi- reduce the number of animals needed for experiments, it is
gation. In addition to measuring RNFL birefringence, a longer also superior to cross-sectional experiments that require differ-
wavelength (␭ ⫽ 1.3 ␮m) PS-OCT system has been used to ent animals for different time points. The following briefly
observe the anterior chamber in subjects after glaucoma sur- summarizes recent studies using OCT in animals, in small to
gery.108 A swept-source PS-OCT system at a 1-␮m center wave- large animal models.
length was used to image sclera and lamina cribrosa,130 which The eyes of small animal models commonly used in devel-
may provide insight into structural changes occurring in the opmental biology, such as xenopus laevis larvae144 and ze-
ONH in glaucoma. brafish embryos145 have successfully been imaged with OCT.
Polarization of the RPE may be important in the detection of Rodent imaging with OCT is becoming increasingly popular,
macular disease.131,132 Gotzinger et al.132 developed a segmen- given their relatively low cost and short lifespan and therefore
tation algorithm based on what they refer to as the “polariza- shorter time for disease progression. In addition, many trans-
tion scrambling effect ” of the RPE, which provides an alterna- genic models are easy for researchers to access. OCT has been
tive to conventional intensity-based quantification. A combined AO used to study ocular dimensions146 and characterize normal
PS-OCT system was later used to measure RPE polarization eye growth147 as well as growth of eyes in mouse models of
scrambling.133 Conventional PS-OCT was used to observe sub- myopia.148 Mouse models of retinal degeneration have been
jects with AMD,134,135 where abnormal birefringence was co- imaged using TD-OCT,149,150 and healthy and degenerative
localized with exudative lesions. mice with SD-OCT.151–158
PS-OCT has been used for anterior segment imaging to Recently, methods for automatically obtaining measure-
measure corneal birefringence,136,137 and these measurements ments from mouse OCT images have been presented.159,160
were used to compensate for corneal birefringence in retinal Images taken in an anesthetized mouse, held in place using a
imaging.138 A difference in polarization in healthy corneas stage with a glass coverslip to neutralize the strong refractive
versus those with keratoconus was demonstrated in vitro, power of the mouse cornea were shown to be reproducible.159
suggesting that PS-OCT may eventually provide insight into This indicates that 3D SD-OCT imaging of the mouse retina may
corneal diseases in vivo.139 be useful for longitudinal studies of retinal structure in mice.
The aforementioned studies indicate that PS-OCT offers an Rats also provide an interesting platform for studying struc-
alternative approach for detecting changes of optical proper- tural changes in the retina and optic nerve in response to injury
ties in tissue. If it can be established that a change in birefrin- or disease. Given their larger eyes, it is less complicated to
gence occurs before tissue thinning or thickening, it may allow focus on the retina than in the mouse. Retina and optic nerve
earlier detection and the opportunity for earlier intervention. imaging has been demonstrated in rat models of retinal degen-
eration,151 retinal vein occlusion,161 retinal ganglion cell de-
Eye-Tracking OCT Systems generation post nerve-crush injury162,163 and elevated intraoc-
ular pressure,164 suggesting that there is also potential for rats
Subject eye motion can alter the intended location of an OCT to be used for longitudinal studies with OCT.
scan. Attempts to correct eye motion by using postprocessing The eyes of larger animal models, such as chickens with
methods are under development, but real-time eye-tracking retinal degeneration,165 have been imaged. Researchers have
systems may provide an alternate method of avoiding eye also used OCT to examine birds of prey,166 pigs,167,168 cats,169
motion artifacts.46 Menke et al.140 showed that an SD-OCT and rabbits.170 –178 These animals have eyes that are compara-
system with built-in eye-tracking can provide reproducible ble in size to the human eye, which means large modifications
measurements, but it is yet to be shown whether this yields of the OCT system optics are not necessary.
higher reproducibility or better sensitivity and specificity than Nonhuman primate models are especially appealing for
devices without eye-tracking systems. studies with OCT, since their ocular size and structure closely
match those of the human eye. Nonhuman primate imaging
OCT Systems for Surgical Guidance may provide novel insight into the mechanical damage to the
As described earlier in this review, OCT is already being used RNFL and ONH associated with increased IOP, as is often seen
for surgical planning and follow-up. In addition, there has been in glaucoma. PS-OCT has been used to look at the birefringence
progress in the development of intraoperative OCT systems. of the RNFL,129,179,180 and RNFL thickness was measured in
Intraoperative OCT was first demonstrated in anterior segment eyes with unilateral, laser-induced ocular hypertension.181,182
surgery, where a 1310-nm system was coupled to an operating Strouthidis et al.183 examined 3D SD-OCT images of the optic
microscope.141 The use of a handheld OCT retinal imaging nerve in nonhuman primate eyes. They visualized the termina-

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 2432 Gabriele et al. IOVS, April 2011, Vol. 52, No. 5

tion of Bruch’s membrane, border tissue, and the anterior 10. Schuman JS, Pedut-Kloizman T, Hertzmark E, et al. Reproducibil-
scleral canal opening and showed that these structures corre- ity of nerve fiber layer thickness measurements using optical
lated to disc photos184 and histology.185 This set the stage for coherence tomography. Ophthalmology. 1996;103:1889 –1898.
a study of alterations in the ONH that are due to increased 11. Drexler W, Morgner U, Kartner FX, et al. In vivo ultrahigh-
intraocular pressure.186 resolution optical coherence tomography. Opt Lett. 1999;24:
Ultimately, longitudinal OCT studies of small and large ani- 1221–1223.
mals may help evaluate the efficacy of pharmacologic agents, 12. Lim H, Jiang Y, Wang Y, Huang YC, Chen Z, Wise FW. Ultrahigh-
resolution optical coherence tomography with a fiber laser
stem cell therapies, surgical intervention, and retinal prosthet-
source at 1 microm. Opt Lett. 2005;30:1171–1173.
ics while reducing the required number of animals. OCT has
13. Unterhuber A, Povazay B, Bizheva K, et al. Advances in broad
the potential to provide a better understanding of disease bandwidth light sources for ultrahigh resolution optical coher-
development and progression in transgenic and other models ence tomography. Phys Med Biol. 2004;49:1235–1246.
of disease, which may eventually translate to improved clinical 14. Wojtkowski M, Leitgeb R, Kowalczyk A, Bajraszewski T, Fercher
assessment and understanding of disease. A. In vivo human retinal imaging by Fourier-domain optical co-
herence tomography. J Biomed Opt. 2002;7:457– 463.
15. Fercher A, Hitzenberger C, Kamp G, Elzaiat S. Measurement of
intraocular distances by backscattering spectral interferometry.
CONCLUSION Opt Commun. 1995;117:43– 48.
16. Choma MA, Hsu K, Izatt JA. Swept source optical coherence
The use of OCT imaging in ophthalmology has increased tomography using an all-fiber 1300-nm ring laser source.
steadily in recent years, in part due to technological improve- J Biomed Opt. 2005;10:44009.
ments such as scanning speed, sensitivity and resolution. The 17. de Boer JF, Cense B, Park BH, Pierce MC, Tearney GJ, Bouma BE.
field continues to grow and transform the way glaucoma and Improved signal-to-noise ratio in spectral-domain compared with
retinal diseases are monitored. With 3D imaging, there are new time-domain optical coherence tomography. Opt Lett. 2003;28:
ways to visualize pathologic features and corresponding chal- 2067–2069.
lenges to overcome. Current approaches, such as RNFL thick- 18. Leitgeb R, Wojtkowski M, Kowalczyk A, Hitzenberger CK, Sticker
ness maps and C-mode visualization, attempt to summarize M, Fercher AF. Spectral measurement of absorption by spectro-
structural information. These methods are not necessarily op- scopic frequency-domain optical coherence tomography. Opt
timized for efficient cross-sectional and longitudinal analysis, Lett. 2000;25:820 – 822.
but with technological improvements such as longer wave- 19. Nassif N, Cense B, Park BH, et al. In vivo human retinal imaging
length imaging, SS-OCT, AO-OCT, and PS-OCT on the horizon, by ultrahigh-speed spectral domain optical coherence tomogra-
even more structural detail will be available. Translating these phy. Opt Lett. 2004;29:480 – 482.
techniques to the clinic has already begun and many could 20. Wojtkowski M, Srinivasan V, Fujimoto JG, et al. Three-dimen-
eventually be made available to the clinician. A combined sional retinal imaging with high-speed ultrahigh-resolution opti-
cal coherence tomography. Ophthalmology. 2005;112:1734 –
slit-lamp/OCT system that allows the clinician to access struc-
1746.
tural information during a routine examination may eventually
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