The aqueous outflow system (AOS) is responsible for maintaining normal intraocular pressure (IOP) in the eye. Structures of the AOS have an active role in regulating IOP in healthy eyes and these structures become abnormal in the eyes with glaucoma. We describe a newly developed system platform to obtain high-resolution images of the AOS structures. By incorporating spectral domain optical coherence tomography (SD-OCT), the platform allows us to systematically control, image, and quantitate the responses of AOS tissue to pressure with a millisecond resolution of pulsed flow. We use SD-OCT to image radial limbal segments from the surface of the trabecular meshwork (TM) with a spatial resolution of ∼5 μm in ex vivo nonhuman primate eyes. We carefully insert a cannula into Schlemm’s canal (SC) to control both pressures and flow rates. The experimental results demonstrate the capability of the platform to visualize the unprecedented details of AOS tissue components comparable to that delivered by scanning electron microscopy, as well as to delineate the complex pressure-dependent relationships among the TM, structures within the SC, and collector channel ostia. The described technique provides a new means to characterize the anatomic and pressure-dependent relationships of SC structures, particularly the active motion of collagenous elements at collector channel ostia; such relationships have not previously been amenable to study. Experimental findings suggest that continuing improvements in the OCT imaging of the AOS may provide both insights into the glaucoma enigma and improvements in its management.
Optical microangiography based on optical coherence tomography (OCT) is prone to noise that arises from a static tissue region. Here, we propose a method that can significantly reduce this noise. The method is developed based on an approach that uses the magnitude information of OCT signals to produce tissue microangiograms, especially suitable for the case where a swept-source OCT system is deployed. By combined use of two existing OCT microangiography methods—ultrahigh-sensitive optical microangiography (UHS-OMAG) and correlation mapping OCT (cmOCT)—the final tissue microangiogram is generated by masking UHS-OMAG image using the binary representation of cmOCT image. We find that this process masks the residual static artifacts while preserving the vessel structures. The noise rejection capability of the masked approach (termed as mOMAG) is tested on a tissue-like flow phantom as well as an in vivo human skin tissue. Compared to UHS-OMAG and cmOCT, we demonstrate that the proposed method is capable of achieving improved signal-to-noise ratio in providing microcirculation images. Finally, we show its clinical potential by quantitatively assessing the vascular difference between a burn scar and a normal skin of human subject in vivo.
Evaluation of spatiotemporal hemodynamic and metabolic responses during neural activation is crucial in studying brain function. We explore the use of a noninvasive multifunctional optical imaging system to measure these responses in a mouse brain upon electrically stimulated neural activation, with the cranium left intact. The system is developed by integrating an optical microangiography (OMAG) imaging system with a dual-wavelength laser speckle imaging (DW-LSI) system. The DW-LSI, running at an image acquisition speed of ∼100 Hz, is used to extract the large-scale two-dimensional map, revealing the localized response of blood flow, hemoglobin concentration, and metabolic rate of oxygen change. Guided by DW-LSI, the OMAG is, however, used to image the response of individual blood vessels with its unique depth-resolved capability. We show that the integrated system is capable of investigating neural activation, thus is potentially valuable in the preclinical study of the mechanism of neurovascular coupling.
We propose a simple and optimized method for acquiring a wide velocity range of blood flow using Doppler optical microangiography. After characterizing the behavior of the scanner in the fast scan axis, a step-scanning protocol is developed by utilizing repeated A-scans at each step. Multiple velocity range images are obtained by the high-pass filtering and Doppler processing of complex signals between A-scans within each step with different time intervals. A phase variance mask is then employed to segment meaningful Doppler flow signals from noisy phase background. The technique is demonstrated by imaging in vivo mouse brain with skull left intact to provide bidirectional images of cerebral blood flow with high quality and wide velocity range.
Purpose. It is suspected that the abnormalities of aqueous outflow pump composed of trabecular meshwork (TM) and Schlemm’s canal (SC) results in the increased outflow resistance and then elevated intraocular pressure (IOP) in initial glaucoma. In order to explore the casual mechanism and the early diagnosis of glaucoma, the dynamic characterizations of aqueous outflow pump were explored.
Methods. As a functional extension of optical coherence tomography (OCT), tissue Doppler OCT (tissue-DOCT) method capable of measuring the slow tissue movement was developed. The tissue-DOCT imaging was conducted on the corneo-scleral limbus of 4 monkey eyes. The eye was mounted in an anterior segment holder, together with a perfusion system to control the mean IOP and to induce the cyclic IOP transients with amplitude 3 mm Hg at frequency 1 pulse/second. IOP was monitored on-line by a pressure transducer. Tissue-DOCT data and pressure data were recorded simultaneously. The IOP-transient induced Doppler velocity, displacement and strain rate of TM and the normalized area of SC were quantified at 7 different mean IOPs (5, 8, 10, 20, 30, 40, 50 mm Hg).
Results. The outflow system, including TM, SC and CCs, was visualized in the micro-structural imaging. The IOP-transient induced pulsatile TM movement and SC deformation were detected and quantified by tissue-DOCT. The TM movement was depth-dependent and the largest movement was located in the area closest to SC endothelium (SCE). Both the pulsations of TM and SC were found to be synchronous with the IOP pulse wave. At 8 mm Hg IOP, the global TM movement was around 0.65μm during one IOP transient. As IOP elevated, a gradual attenuation of TM movement and SC deformation was observed.
Conclusions. The observed pulsation of TM and SC induced by the pulsatile IOP transients was in good agreement with the predicated role of TM and SC acting as a biomechanical pump (pumping aqueous from anterior chamber into SC and from SC into CCs) in the aqueous outflow system. As the IOP elevated, the attenuated pulsation amplitude of the aqueous outflow pump indicated the failure of the mechanical pump and the increase of aqueous outflow resistance. The promising results revealed the potential of using the proposed tissue-DOCT for diagnosis and associated therapeutic guidance of the initial and progressive glaucoma process by monitoring the pulsation of the outflow pump.
Reduced cochlear blood flow (CoBF) is a main contributor to hearing loss. Studying CoBF has remained a challenge due to the lack of available tools. Doppler optical microangiography (DOMAG), a method to quantify single-vessel absolute blood flow, and laser Doppler flowmetry (LDF), a method for measuring the relative blood flow within a large volume of tissue, were used for determining the changes in CoBF due to systemic hypoxia in mice. DOMAG determined the change in blood flow in the apical turn (AT) with single-vessel resolution, while LDF averaged the change in the blood flow within a large volume of the cochlea (hemisphere with ∼1 to 1.5 mm radius). Hypoxia was induced by decreasing the concentration of oxygen-inspired gas, so that the oxygen saturation was reduced from >95% to ∼80% . DOMAG determined that during hypoxia the blood flow in two areas of the AT near and far from the helicotrema were increased and decreased, respectively. The LDF detected a decrease in blood flow within a larger volume of the cochlea (several turns averaged together). Therefore, the use of DOMAG as a tool for studying cochlear blood flow due to its ability to determine absolute flow values with single-vessel resolution was proposed.
Glaucoma is a blinding disease for which intraocular pressure (IOP) is the only treatable risk factor. The mean IOP is regulated through the aqueous outflow system, which contains the trabecular meshwork (TM). Considerable evidence indicates that trabecular tissue movement regulates the aqueous outflow and becomes abnormal during glaucoma; however, such motion has thus far escaped detection. The purpose of this study is to describe anovel use of a phase-sensitive optical coherence tomography (PhS-OCT) method to assess pulse-dependent TM movement. For this study, we used enucleated monkey eyes, each mounted in an anterior segment holder. A perfusion system was used to control the mean IOP as well as to provide IOP sinusoidal transients (amplitude 3 mmHg, frequency 1 pulse/second) in all experiments. Measurements were carried out at seven graded mean IOPs (5, 8, 10, 20, 30, 40, and 50 mm Hg). We demonstrate that PhS-OCT is sensitive enough to image/visualize TM movement synchronous with the pulse-induced IOP transients, providing quantitative measurements of dynamic parameters such as velocity, displacement, and strain rate that are important for assessing the biomechanical compliance of the TM. We find that the largest TM displacement is in the area closest to Schlemm's canal (SC) endothelium. While maintaining constant ocular pulse amplitude, an increase of mean IOP results in a decrease of TM displacement and mean size of the SC. These results demonstrate that the PhS-OCT is a useful imaging technique capable of assessing functional properties necessary to maintain IOP in a healthy range, offering a new diagnostic alternative for glaucoma.
Several imaging modalities such as optical coherence tomography, photothermal, photoacoustic and magnetic resonance imaging, are sensitive to different physical properties (i.e. scattering, absorption and magnetic) that can provide contrast within biological tissues. Usually exogenous agents are designed with specific properties to provide contrast for these imaging methods. In nano-biotechnology there is a need to combine several of these properties into a single contrast agent. This multifunctional contrast agent can then be used by various imaging techniques simultaneously or can be used to develop new imaging modalities. We reported and characterized a multifunctional nanoparticle, made from gold nanoshells, which exhibits scattering, photothermal, photoacoustic, and magnetic properties.
The concentration of photothermal (light-to-heat converters) compounds as a function of depth is determined in solid
agar gel phantoms. The system contains an 808nm pump laser, which excites the photothermal compound, and a phase
sensitive spectral domain optical coherence tomography system, which detects the changes in the optical pathlength of
the sample induced by the temperature increase. The derivation of the model is described, and its parameters are
empirically determined. The concentration of photothermal compounds are observed from double layer agar gel
phantoms.
A model that describes the concentration of photothermal (light-to-heat converters) compounds as a function of depth in a turbid medium is developed. The system consists of a pump laser (808 nm modulated at 400 Hz), which heats a photothermal compound, and a phase sensitive spectral domain optical coherence tomography system, which detects the changes in the optical path length of the sample induced by the temperature increase. The model is theoretically derived and the coefficients are empirically determined using solid homogeneous gel phantoms. The model is validated by reconstructing the concentration of a photothermal compound in thick single and double layer solid phantoms.
Spectral reflectance measurements of biological tissues have been studied for early diagnoses of several pathologies such as cancer. These measurements are often performed with a fiber optic probe in contact with the tissue surface. We report a study in which reflectance measurements are obtained in vivo from mouse thigh muscle while varying the contact pressure of the fiber optic probe. It is determined that the probe pressure is a variable that affects the local optical properties of the tissue. The reflectance spectra are analyzed with an analytical model that extracts the tissue optical properties and facilitates the understanding of underlying physiological changes induced by the probe pressure.
The understanding of drug delivery to organs, such as the brain, has been hampered by the inability to measure tissue drug concentrations in real time. We report an application of an optical spectroscopy technique that monitors in vivo the real-time drug concentrations in small volumes of brain tissue. This method will facilitate development of new protocols for delivery of drugs to treat brain cancers. The delivery of many anticancer drugs to the brain is limited by the presence of the blood-brain barrier (BBB). Mitoxantrone (MTX) is a water-soluble anticancer drug that poorly penetrates the BBB. It is preliminarily determined in an animal model that the brain tissue uptake of chemotherapy agents—in this demonstration, MTX—delivered intra-arterially is enhanced when the BBB is disrupted.
In the study of epithelial tissues and superficial cancers, it is often important to determine the optical properties of small
volumes of tissue, and at varying depths. A model that relates the reflectance spectrum to the optical properties of a
turbid medium at small source-detector separations is developed based on Monte Carlo simulations and experiments in
tissue phantoms. Four fiber probes are analyzed, for which each fiber probe presents a different tilt angle. Preliminary
results show good correlation between known optical properties in tissue phantoms, and the measured optical properties.
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