We recently reported on a new swept source polarization sensitive optical coherence tomography system and its application to skin imaging [Biomed. Opt. Express 3, 2987 (2012)]. In some of the tomographic images, two skin layers were labeled wrongly (interchanged). We present figures with corrected labeling.
(170.4500) Optical coherence tomography; (230.5440) Polarization-selective devices; (170.4580) Optical diagnostics for medicine
We correct an error in the methods reported in our previous paper [Biomed. Opt. Express 3, 2752
(2012)]; the liquid used for the final step of specimen preparation was 100% mineral oil, not a
mixture of oil and glycerol.
(170.6900) Three-dimensional microscopy; (170.6930) Tissue; (110.0113) Imaging through turbid media
We present a novel application of optical microangiography (OMAG) imaging technique for
visualization of depth-resolved vascular network within retina and choroid as well as measurement of
total retinal blood flow in mice. A fast speed spectral domain OCT imaging system at 820nm with a
line scan rate of 140 kHz was developed to image the posterior segment of eyes in mice. By applying
an OMAG algorithm to extract the moving blood flow signals out of the background tissue, we are able
to provide true capillary level imaging of the retinal and choroidal vasculature. The microvascular
patterns within different retinal layers are presented. An en face Doppler OCT
approach [Srinivasan et al., Opt Express 18, 2477 (2010)] was adopted for retinal blood
flow measurement. The flow is calculated by integrating the axial blood flow velocity over the
vessel area measured in an en face plane without knowing the blood vessel angle.
Total retinal blood flow can be measured from both retinal arteries and veins. The results indicate
that OMAG has the potential for qualitative and quantitative evaluation of the microcirculation in
posterior eye compartments in mouse models of retinopathy and neovascularization.
(170.4500) Optical coherence tomography; (170.3880) Medical and biological imaging
An interesting method to measure and correct chromatic magnification offsets in a
multi-wavelength retinal imaging microscope was recently reported [Harmening et al., Biomed. Opt.
Express 3, 2066 (2012)]. These values were in part related to the ocular transverse
chromatic aberration (TCA). This method could be potentially used in the future to overcome the
fundamental limitation to estimate the eye’s TCA with ophthalmoscopic (double-pass)
(170.4460) Ophthalmic optics and devices; (330.5370) Physiological optics
Abstract: The authors provide corrections to tabular data and a figure, which do not alter the conclusions of the original paper [Biomed. Opt. Express 2, 2068 (2011)].
(170.1470) Blood or tissue constituent monitoring; (170.3660) Light propagation in tissues; (170.3890) Medical optics instrumentation
Correction of the eye’s monochromatic aberrations using adaptive optics (AO) can improve the resolution of in vivo mouse retinal images [Biss et al., Opt. Lett. 32(6), 659 (2007) and Alt et al., Proc. SPIE 7550, 755019 (2010)], but previous attempts have been limited by poor spot quality in the Shack-Hartmann wavefront sensor (SHWS). Recent advances in mouse eye wavefront sensing using an adjustable focus beacon with an annular beam profile have improved the wavefront sensor spot quality [Geng et al., Biomed. Opt. Express 2(4), 717 (2011)], and we have incorporated them into a fluorescence adaptive optics scanning laser ophthalmoscope (AOSLO). The performance of the instrument was tested on the living mouse eye, and images of multiple retinal structures, including the photoreceptor mosaic, nerve fiber bundles, fine capillaries and fluorescently labeled ganglion cells were obtained. The in vivo transverse and axial resolutions of the fluorescence channel of the AOSLO were estimated from the full width half maximum (FWHM) of the line and point spread functions (LSF and PSF), and were found to be better than 0.79 μm ± 0.03 μm (STD)(45% wider than the diffraction limit) and 10.8 μm ± 0.7 μm (STD)(two times the diffraction limit), respectively. The axial positional accuracy was estimated to be 0.36 μm. This resolution and positional accuracy has allowed us to classify many ganglion cell types, such as bistratified ganglion cells, in vivo.
(170.4460) Ophthalmic optics and devices; (110.1080) Active or adaptive optics; (330.7324) Visual optics, comparative animal models
We correct an error in our previous paper [Biomed. Opt. Express 2, 1218 (2011)] which led to an erroneous conclusion that a dispersive optical delay line (DODL) used in a swept source optical coherence tomography (SSOCT) system generated a pure phase modulation allowing for complex conjugate artifact removal in Fourier domain OCT via optical heterodyning. We now understand that an alternate phenomenon known as coherence revival was responsible for the observed phase modulation, while the DODL provided a compact means of generating a large group delay with readily adjustable group velocity dispersion compensation.
(170.4500) Optical coherence tomography; (230.4110) Modulators
In July 2011 a new concept of a closed microfluidic system equipped with a fixed micropipette, optical tweezers and a UV-Vis spectrometer was presented [Biomed. Opt. Express 2, 2299 (2011)]. Figure 1 showed falsely oriented mirrors. To clarify the design of the setup, this erratum presents a correct schematic.
(350.4855) Optical tweezers or optical manipulation; (170.3880) Medical and biological imaging; (300.1030) Absorption; (280.2490) Flow diagnostics; (220.4000) Microstructure fabrication; (110.0180) Microscopy
The Monte Carlo (MC) method is a popular approach to modeling photon propagation inside general turbid media, such as human tissue. Progress had been made in the past year with the independent proposals of two mesh-based Monte Carlo methods employing ray-tracing techniques. Both methods have shown improvements in accuracy and efficiency in modeling complex domains. A recent paper by Shen and Wang [Biomed. Opt. Express 2, 44 (2011)] reported preliminary results towards the cross-validation of the two mesh-based MC algorithms and software implementations, showing a 3–6 fold speed difference between the two software packages. In this comment, we share our views on unbiased software comparisons and discuss additional issues such as the use of pre-computed data, interpolation strategies, impact of compiler settings, use of Russian roulette, memory cost and potential pitfalls in measuring algorithm performance. Despite key differences between the two algorithms in handling of non-tetrahedral meshes, we found that they share similar structure and performance for tetrahedral meshes. A significant fraction of the observed speed differences in the mentioned article was the result of inconsistent use of compilers and libraries.
(170.3660) Light propagation through tissue; (170.5280) Photon migration; (170.7050) Turbid media
We compare the accuracy of TIM-OS and MMCM in response to the recent analysis made by Fang [Biomed. Opt. Express 2, 1258 (2011)]. Our results show that the tetrahedron-based energy deposition algorithm used in TIM-OS is more accurate than the node-based energy deposition algorithm used in MMCM.
(170.3660) Light propagation in tissues
We demonstrated a novel approach of imaging the anterior segment including the ciliary muscle
using combined and synchronized two spectral domain optical coherence tomography devices (SD-OCT).
In one SD-OCT, a Complementary Metal-Oxide-Semiconductor Transistor (CMOS) camera and an alternating
reference arm was used to image the anterior segment from the cornea to the lens. Another SD-OCT for
imaging the ciliary muscle was equipped with a light source with a center wavelength of 1,310 nm and
a bandwidth of 75 nm. Repeated measurements were performed under relaxed and 4.00 D accommodative
stimulus states in six eyes from 6 subjects. We also imaged dynamic changes in the anterior segment
in one eye during accommodation. The biometry of the anterior segment and the ciliary muscle was
obtained. The combined system appeared to be capable to simultaneously real-time image the biometry
of the anterior segment, including the ciliary muscle, in vivo during
(170.4500) Optical coherence tomography; (170.3880) Medical and biological imaging; (170.4580) Optical diagnostics for medicine; (330.4460) Ophthalmic optics and devices; (330.7322) Visual optics, accommodation
Knowledge of myocardial fiber architecture is essential towards understanding heart functions. We demonstrated in this study a method to map cardiac muscle structure using the local optical axis obtained from polarization-sensitive optical coherence tomography (PSOCT). An algorithm was developed to extract the true local depth-resolved optical axis, retardance, and diattenuation from conventional round-trip results obtained in a Jones matrix-based PSOCT system. This method was applied to image the myocardial fiber orientation in a bovine heart muscle sample.
(110.4500) Optical coherence tomography; (230.5440) Polarization-selective devices
We describe a novel dual-modality imaging approach that integrates diffuse optical tomography (DOT) and photoacoustic imaging (PAI) through a miniaturized handheld probe based on microelectromechanical systems (MEMS) scanning mirror. We validate this dual-modal DOT/PAI approach using extensive phantom experiments, and demonstrate its application for tumor imaging using tumor-bearing mice systematically injected with targeted contrast agents.
(110.6960) Tomography; (170.0110) Imaging systems; (170.5120) Photoacoustic imaging
In this paper, we present and validate a new method for optical properties recovery of turbid media with slab geometry. This method is an iterative method that compares diffuse reflectance and transmittance, measured using integrating spheres, with those obtained using the known algorithm MCML. The search procedure is based in the evolution of a population due to selection of the best individual, i.e., using a genetic algorithm. This new method includes several corrections such as non-linear effects in integrating spheres measurements and loss of light due to the finite size of the sample. As a potential application and proof-of-principle experiment of this new method, we use this new algorithm in the recovery of optical properties of blood samples at different degrees of coagulation.
(170.3660) Light propagation in tissues; (170.5280) Photon migration; (290.7050) Turbid media
Time-resolved (TR) techniques provide a means of discriminating photons based on their time-of-flight. Since early arriving photons have a lower probability of probing deeper tissue than photons with long time-of-flight, time-windowing has been suggested as a method for improving depth sensitivity. However, TR measurements also contain instrument contributions (instrument-response-function, IRF), which cause temporal broadening of the measured temporal point-spread function (TPSF) compared to the true distribution of times-of-flight (DTOF). The purpose of this study was to investigate the influence of the IRF on the depth sensitivity of TR measurements. TPSFs were acquired on homogeneous and two-layer tissue-mimicking phantoms with varying optical properties. The measured IRF and TPSFs were deconvolved using a stable algorithm to recover the DTOFs. The microscopic Beer-Lambert law was applied to the TPSFs and DTOFs to obtain depth-resolved absorption changes. In contrast to the DTOF, the latest part of the TPSF was not the most sensitive to absorption changes in the lower layer, which was confirmed by computer simulations. The improved depth sensitivity of the DTOF was illustrated in a pig model of the adult human head. Specifically, it was shown that dynamic absorption changes obtained from the late part of the DTOFs recovered from TPSFs acquired by probes positioned on the scalp were similar to absorption changes measured directly on the brain. These results collectively demonstrate that this method improves the depth sensitivity of TR measurements by removing the effects of the IRF.
(170.3660) Light propagation in tissues; (170.3890) Medical optics instrumentation
The effect of task-related extracerebral circulatory changes on diffuse optical tomography (DOT) of brain activation was evaluated using experimental data from 14 healthy human subjects and computer simulations. Total hemoglobin responses to weekday-recitation, verbal-fluency, and hand-motor tasks were measured with a high-density optode grid placed on the forehead. The tasks caused varying levels of mental and physical stress, eliciting extracerebral circulatory changes that the reconstruction algorithm was unable to fully distinguish from cerebral hemodynamic changes, resulting in artifacts in the brain activation images. Crosstalk between intra- and extracranial layers was confirmed by the simulations. The extracerebral effects were attenuated by superficial signal regression and depended to some extent on the heart rate, thus allowing identification of hemodynamic changes related to brain activation during the verbal-fluency task. During the hand-motor task, the extracerebral component was stronger, making the separation less clear. DOT provides a tool for distinguishing extracerebral components from signals of cerebral origin. Especially in the case of strong task-related extracerebral circulatory changes, however, sophisticated reconstruction methods are needed to eliminate crosstalk artifacts.
(170.0110) Imaging systems; (170.1470) Blood or tissue constituent monitoring; (170.3880) Medical and biological imaging; (170.6960) Tomography
Enhanced Depth Imaging (EDI) optical coherence tomography (OCT) provides high-definition cross-sectional images of the choroid in vivo, and hence is used in many clinical studies. However, the quantification of the choroid depends on the manual labelings of two boundaries, Bruch’s membrane and the choroidal-scleral interface. This labeling process is tedious and subjective of inter-observer differences, hence, automatic segmentation of the choroid layer is highly desirable. In this paper, we present a fast and accurate algorithm that could segment the choroid automatically. Bruch’s membrane is detected by searching the pixel with the biggest gradient value above the retinal pigment epithelium (RPE) and the choroidal-scleral interface is delineated by finding the shortest path of the graph formed by valley pixels using Dijkstra’s algorithm. The experiments comparing automatic segmentation results with the manual labelings are conducted on 45 EDI-OCT images and the average of Dice’s Coefficient is 90.5%, which shows good consistency of the algorithm with the manual labelings. The processing time for each image is about 1.25 seconds.
(100.0100) Image processing; (110.4500) Optical coherence tomography; (100.2960) Image analysis; (170.4470) Ophthalmology
In vivo three-dimensional (3-D) anterior segment biometry before and after cataract surgery was analyzed by using custom high-resolution high-speed anterior segment spectral domain Optical Coherence Tomography (OCT). The system was provided with custom algorithms for denoising, segmentation, full distortion correction (fan and optical) and merging of the anterior segment volumes (cornea, iris, and crystalline lens or IOL), to provide fully quantitative data of the anterior segment of the eye. The method was tested on an in vitro artificial eye with known surfaces geometry at different orientations and demonstrated on an aging cataract patient in vivo. Biometric parameters CCT, ACD/ILP, CLT/ILT Tilt and decentration are retrieved with a very high degree of accuracy. IOL was placed 400 μm behind the natural crystalline lens, The IOL was aligned with a similar orientation of the natural lens (2.47 deg superiorly), but slightly lower amounts (0.77 deg superiorly). The IOL was decentered superiorly (0.39 mm) and nasally (0.26 mm).
(110.4500) Optical coherence tomography; (120.6650) Surface measurements, figure; (120.4640) Optical instruments; (120.4800) Optical standards and testing; (110.6880) Three-dimensional image acquisition; (330.7327) Visual optics, ophthalmic instrumentation
A technique was developed for assaying axonal transport in retinal ganglion cells using 2
µl injections of 1% cholera toxin b-subunit conjugated to AlexaFluor488 (CTB). In
vivo retinal and post-mortem brain imaging by confocal scanning laser ophthalmoscopy and
post-mortem microscopy were performed. The transport of CTB was sensitive to colchicine, which
disrupts axonal microtubules. The bulk rates of transport were determined to be approximately
80–90 mm/day (anterograde) and 160 mm/day (retrograde). Results demonstrate that axonal
transport of CTB can be monitored in vivo in the rodent anterior visual pathway, is
dependent on intact microtubules, and occurs by active transport mechanisms.
(170.2655) Functional monitoring and imaging; (170.3880) Medical and biological imaging
The OSA Board of Editors informs the community of its policy and procedures with regards to
plagiarism. This is an update of the original editorial published in April 2005.
(000.1200) Announcements, awards, news, and organizational activities; (000.5360) Physics literature and publications
Swept source/Fourier domain OCT is demonstrated for in vivo imaging of the rodent eye. Using
commercial swept laser technology, we developed a prototype OCT imaging system for small animal
ocular imaging operating in the 1050 nm wavelength range at an axial scan rate of 100 kHz with ~6
µm axial resolution. The high imaging speed enables volumetric imaging with high axial scan
densities, measuring high flow velocities in vessels, and repeated volumetric imaging over time. The
1050 nm wavelength light provides increased penetration into tissue compared to standard commercial
OCT systems at 850 nm. The long imaging range enables multiple operating modes for imaging the
retina, posterior eye, as well as anterior eye and full eye length. A registration algorithm using
orthogonally scanned OCT volumetric data sets which can correct motion on a per A-scan basis is
applied to compensate motion and merge motion corrected volumetric data for enhanced OCT image
quality. Ultrahigh speed swept source OCT is a promising technique for imaging the rodent eye,
proving comprehensive information on the cornea, anterior segment, lens, vitreous, posterior
segment, retina and choroid.
(170.3880) Medical and biological imaging; (170.4500) Optical coherence tomography; (170.4470) Ophthalmology
A combined time-domain fluorescence and hemoglobin diffuse optical tomography (DOT) system and the image reconstruction methods are proposed for enhancing the reliability of breast-dedicated optical measurement. The system equipped with two pulsed laser diodes at wavelengths of 780 nm and 830 nm that are specific to the peak excitation and emission of the FDA-approved ICG agent, and works with a 4-channel time-correlated single photon counting device to acquire the time-resolved distributions of the light re-emissions at 32 boundary sites of tissues in a tandem serial-to-parallel mode. The simultaneous reconstruction of the two optical (absorption and scattering) and two fluorescent (yield and lifetime) properties are achieved with the respective featured-data algorithms based on the generalized pulse spectrum technique. The performances of the methodology are experimentally assessed on breast-mimicking phantoms for hemoglobin- and fluorescence-DOT alone, as well as for fluorescence-guided hemoglobin-DOT. The results demonstrate the efficacy of improving the accuracy of hemoglobin-DOT based on a priori fluorescence localization.
(170.3880) Medical and biological imaging; (170.6960) Tomography; (170.6920) Time-resolved imaging; (170.3010) Image reconstruction techniques
We demonstrate vertical cross-sectional (XZ-plane) images of near-infrared (NIR) fluorescence with a handheld dual axes confocal endomicroscope that reveals specific binding of a Cy5.5-labeled peptide to pre-malignant colonic mucosa. This view is perpendicular to the tissue surface, and is similar to that used by pathologists. The scan head is 10 mm in outer diameter (OD), and integrates a one dimensional (1-D) microelectromechanical systems (MEMS) X-axis scanner and a bulky lead zirconate titanate (PZT) based Z-axis actuator. The microscope images in a raster-scanning pattern with a ±6 degrees (mechanical) scan angle at ~3 kHz in the X-axis (fast) and up to 10 Hz (0–400 μm) in the Z-axis (slow). Vertical cross-sectional fluorescence images are collected with a transverse and axial resolution of 4 and 5 μm, respectively, over a field-of-view of 800 μm (width) × 400 μm (depth). NIR vertical cross-sectional fluorescence images of fresh mouse colonic mucosa demonstrate histology-like imaging performance with this miniature instrument.
(170.1790) Confocal microscopy; (170.3880) Medical and biological imaging; (170.4580) Optical diagnostics for medicine; (170.5810) Scanning microscopy
Minimally invasive, specific measurement of cellular energy metabolism is crucial for
understanding cerebral pathophysiology. Here, we present high-resolution, in
vivo observations of autofluorescence lifetime as a biomarker of cerebral energy
metabolism in exposed rat cortices. We describe a customized two-photon imaging system with
time correlated single photon counting detection and specialized software for modeling
multiple-component fits of fluorescence decay and monitoring their transient behaviors.
In vivo cerebral NADH fluorescence suggests the presence of four distinct
components, which respond differently to brief periods of anoxia and likely indicate different
enzymatic formulations. Individual components show potential as indicators of specific
molecular pathways involved in oxidative metabolism.
(170.0170) Medical optics and biotechnology; (180.4315) Nonlinear microscopy; (170.3650) Lifetime-based sensing
Currently, researchers and clinicians lack achromatized endomicroscope objectives that are as narrow as biopsy needles. We present a proof-of-concept prototype that validates the optical design of an NA0.4 objective. The objective, built with plastic lenses, has a 0.9 mm clear aperture and is achromatized from 452 nm to 623 nm. The objective’s measured Strehl ratio is 0.74 ± 0.05 across a 250 μm FOV. We perform optical sectioning via structured illumination through the objective while capturing fluorescence images of breast carcinoma cells stained with proflavine and cresyl violet. This technology has the potential to improve optical biopsies and provide the next step forward in cancer diagnostics.
(080.3620) Lens system design; (220.0220) Optical design and fabrication; (170.2520) Fluorescence microscopy; (170.3880) Medical and biological imaging; (220.1920) Diamond machining