Motion artifact and background noise suppression on optical microangiography frames using a naïve Bayes mask.

Optical coherence tomography (OCT) is a technique that allows for the three-dimensional (3D) imaging of small volumes of tissue (a few millimeters) with high resolution (∼10 µm). Optical microangiography (OMAG) is a method of processing OCT data, which allows for the extraction of the tissue vasculature with capillary resolution from the OCT images. Cross-sectional B-frame OMAG images present the location of the patent blood vessels; however, the signal-to-noise-ratio of these images can be affected by several factors such as the quality of the OCT system and the tissue motion artifact. This background noise can appear in the en face projection view image. In this work we propose to develop a binary mask that can be applied on the cross-sectional B-frame OMAG images, which will reduce the background noise while leaving the signal from the blood vessels intact. The mask is created by using a naïve Bayes (NB) classification algorithm trained with a gold standard image which is manually segmented by an expert. The masked OMAG images present better contrast for binarizing the image and quantifying the result without the influence of noise. The results are compared with a previously developed frequency rejection filter (FRF) method which is applied on the en face projection view image. It is demonstrated that both the NB and FRF methods provide similar vessel length fractions. The advantage of the NB method is that the results are applicable in 3D and that its use is not limited to periodic motion artifacts.

Analysis of cross-sectional image filters for evaluating nonaveraged optical microangiography images.

Optical microangiography (OMAG) is a method that enables the noninvasive extraction of blood vessels within biological tissues. OMAG B-frames are prone to noise; therefore, techniques such as B-frame averaging have been applied to reduce these effects. A drawback of this method is that the total acquisition time and amount of data collected are increased; hence, the data are susceptible to motion artifacts and decorrelation. In this paper we propose using an image filter on a nonaveraged OMAG B-frame to reduce its noise. Consequently, B-frames comparable to the averaged OMAG B-frame are obtained, while reducing the total acquisition and processing time. The method is tested with two different systems, a high-resolution spectral domain and a relatively low-resolution swept-source optical coherence tomography system. It is demonstrated that the weighted average filter produces the lowest B-frame error; however, all filters produce comparable results when quantifying the en face projection view image.

Fast synchronized dual-wavelength laser speckle imaging system for monitoring hemodynamic changes in a stroke mouse model.

In this Letter, we describe a newly developed synchronized dual-wavelength laser speckle contrast imaging system, which contains two cameras that are synchronously triggered to acquire data. The system can acquire data at a high spatiotemporal resolution (up to 500 Hz for ~1000×1000 pixels). A mouse model of stroke is used to demonstrate the capability for imaging the fast changes (within tens of milliseconds) in oxygenated and deoxygenated hemoglobin concentration, and the relative changes in blood flow in the mouse brain, through an intact cranium. This novel imaging technology will enable the study of fast hemodynamics and metabolic changes in vascular diseases.

Three-dimensional high-resolution imaging of gold nanorods uptake in sentinel lymph nodes.

In this paper, we demonstrate an application of a noninvasive imaging modality, photothermal optical coherence tomography (PT-OCT), for imaging gold nanorods (GNRs) uptake in sentinel lymph node (SLN) of mice in situ. This application enables us to obtain higher quality images of SLN structures due to the photothermal contrast properties of the GNRs. It is also demonstrated that GNRs accumulate differently within several SLN structures, and this uptake is time dependent. Finally, we determine the average concentration of GNRs within the whole SLN which is used to demonstrate uptake kinetics of the nanoparticles.

Inconsistent blood brain barrier disruption by intraarterial mannitol in rabbits: implications for chemotherapy.

The novel ability to quantify drug and tracer concentrations in vivo by optical means leads to the possibility of detecting and quantifying blood brain barrier (BBB) disruption in real-time by monitoring concentrations of chromophores such as Evan's Blue. In this study, experiments were conducted to assess the disruption of the BBB, by intraarterial injection of mannitol, in New Zealand white rabbits. Surgical preparation included: tracheotomy for mechanical ventilation, femoral and selective internal carotid artery (ICA) catheterizations, skull screws for monitoring electrocerebral activity, bilateral placement of laser Doppler probes and a small craniotomy for the placement of a fiber optic probe to determine tissue Evan's Blue dye concentrations. Evans Blue (6.5 mg/kg) was injected intravenously (IV) just before BBB disruption with intracarotid mannitol (25%, 8 ml/40 s). Brain tissue concentrations of the dye in mannitol-treated and control animals were monitored using the method of optical pharmacokinetics (OP) during the subsequent 60 min. Hemodynamic parameters, heart rate, blood pressure, and EKG remained stable throughout the experiments in both the control and the mannitol-treated group. Brain tissue concentrations of Evan's Blue and the brain:plasma Evan's Blue partition coefficient progressively increased during the period of observation. A wide variation in brain tissue Evan's Blue concentrations was observed in the mannitol group. The experiments demonstrate the feasibility of measuring tissue concentrations of Evan's Blue without invading the brain parenchyma, and in real-time. The data suggest that there are significant variations in the degree and duration of BBB disruption induced with intraarterial mannitol. The ability to optically monitor the BBB disruption in real-time could provide a feedback control for hypertonic disruption and/or facilitate dosage control for chemotherapeutic drugs that require such disruption.

Analysis of changes in reflectance measurements on biological tissues subjected to different probe pressures.

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.

Optical method for real-time monitoring of drug concentrations facilitates the development of novel methods for drug delivery to brain tissue.

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.

Real-Time Smart Textile-Based System to Monitor Pressure Offloading of Diabetic Foot Ulcers.

BACKGROUND: The lifetime risk of developing a diabetic foot ulcer (DFU) is at least 25%. A DFU carries a 50% risk for infection and at least 20% of those receive some form of amputation. The most significant parameter that prevents or delays ulcer healing is high plantar pressure. To improve the patient's healing process, the DFU's plantar pressure should remain cumulatively low. Therefore, a tool that continuously measures the DFU loading, and provides real-time feedback can improve the healing outcome. METHODS: We report the development of a system capable of continuously measuring the pressure, which could have applications to monitor DFU. The system contains a textile pressure sensor attached to a stretchable band, hardware that collects data and transmits them via Bluetooth to a phone, an app that gathers the data and stores them in the cloud, and a web dashboard that displays the data to the clinician. The sensor was characterized in vitro using the system, and the web-dashboard was developed and tested on simulated patient data. RESULTS: We demonstrate the feasibility of developing the system and characterize the pressure response of the device. As a result, we demonstrate a viable method for monitoring DFU off-loading in real time. CONCLUSIONS: The presented study demonstrates the feasibility to develop a simple, modular wearable system that opens up new possibilities for diabetic foot ulcer care by providing a way of monitoring the pressure under the ulcer in real time.

Cerebral hypoperfusion-assisted intra-arterial deposition of liposomes in normal and glioma-bearing rats.

BACKGROUND: Optimizing liposomal vehicles for targeted delivery to the brain has important implications for the treatment of brain tumors. The promise of efficient, brain-specific delivery of chemotherapeutic compounds via liposomal vehicles has yet to be achieved in clinical practice. Intra-arterial injection of specially designed liposomes may facilitate efficient delivery to the brain and to gliomas. OBJECTIVE: To test the hypothesis that cationic liposomes may be effectively delivered to both normal and glioma-bearing brain tissue utilizing a strategy of intra-arterial injection during transient cerebral hypoperfusion. METHODS: Cationic, anionic, and neutral liposomes were separately injected via the internal carotid artery of healthy rats during transient cerebral hypoperfusion. Rats bearing C6 gliomas were similarly injected with cationic liposomes. Liposomes were loaded with DilC18(5) dye whose concentrations can be measured by light absorbance and fluorescence methods. RESULTS: After intra-arterial injection, a robust uptake of cationic in comparison with anionic and neutral liposomes into brain parenchyma was observed by diffuse reflectance spectroscopy. Postmortem multispectral fluorescence imaging revealed that liposomal cationic charge was associated with more efficient delivery to the brain. Cationic liposomes were also readily observed within glioma tissue after intra-arterial injection. However, over time, cationic liposomes were retained longer and at higher concentrations in the surrounding, peritumoral brain than in the tumor core. CONCLUSION: This study demonstrates the feasibility of cationic liposome delivery to brain and glioma tissue after intra-arterial injection. Highly cationic liposomes directly delivered to the brain via an intracarotid route may represent an effective method for delivering antiglioma agents.

Functional evaluation of hemodynamic response during neural activation using optical microangiography integrated with dual-wavelength laser speckle imaging.

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.

Improved microcirculation imaging of human skin in vivo using optical microangiography with a correlation mapping mask.

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.

Platform to investigate aqueous outflow system structure and pressure-dependent motion using high-resolution spectral domain optical coherence tomography.

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.

Wide velocity range Doppler optical microangiography using optimized step-scanning protocol with phase variance mask.

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.

Changes in cochlear blood flow in mice due to loud sound exposure measured with Doppler optical microangiography and laser Doppler flowmetry.

In this work we determined the contributions of loud sound exposure (LSE) on cochlear blood flow (CoBF) in an in vivo anesthetized mouse model. A broadband noise system (20 kHz bandwidth) with an intensity of 119 dB SPL, was used for a period of one hour to produce a loud sound stimulus. Two techniques were used to study the changes in blood flow, a Doppler optical microangiography (DOMAG) system; which can measure the blood flow within individual cochlear vessels, and a laser Doppler flowmetry (LDF) system; which averages the blood flow within a volume (a hemisphere of ~1.5 mm radius) of tissue. Both systems determined that the blood flow within the cochlea is reduced due to the LSE stimulation.

Tracking dynamic microvascular changes during healing after complete biopsy punch on the mouse pinna using optical microangiography.

Optical microangiography (OMAG) and Doppler optical microangiography (DOMAG) are two non-invasive techniques capable of determining the tissue microstructural content, microvasculature angiography, and blood flow velocity and direction. These techniques were used to visualize the acute and chronic microvascular and tissue responses upon an injury in vivo. A tissue wound was induced using a 0.5 mm biopsy punch on a mouse pinna. The changes in the microangiography, blood flow velocity and direction were quantified for the acute (<30 min) wound response and the changes in the tissue structure and microangiography were determined for the chronic wound response (30 min-60 days). The initial wound triggered recruitment of peripheral capillaries, as well as redirection of main arterial and venous blood flow within 3 min. The complex vascular networks and new vessel formation were quantified during the chronic response using fractal dimension. The highest rate of wound closure occurred between days 8 and 22. The vessel tortuosity increased during this time suggesting angiogenesis. Taken together, these data signify that OMAG has the capability to track acute and chronic changes in blood flow, microangiography and structure during wound healing. The use of OMAG has great potential to improve our understanding of vascular and tissue responses to injury in order to develop more effective therapeutics.

Label-free imaging of blood vessel morphology with capillary resolution using optical microangiography.

Several tissue pathologies are correlated with changes in the blood vessel morphology and microcirculation that supplies the tissue. Optical coherence tomography (OCT) is an imaging technique that enables acquiring non-invasive three-dimensional images of biological structures with micrometer resolution. Optical microangiography (OMAG) is a method of processing OCT data which enables visualizing the three-dimensional blood vessel morphology within biological tissues. OMAG has high spatial resolution which allows visualizing single capillary vessels, and does not require the use of contrast agents. The intrinsic optical signals backscattered by the moving blood cells inside blood vessels are used as the contrast for which OMAG images are based on. In this paper, we discuss a brief review of the OMAG theory, and present some examples of applications for this technique.

Hemodynamic and morphological vasculature response to a burn monitored using a combined dual-wavelength laser speckle and optical microangiography imaging system.

A multi-functional imaging system capable of determining relative changes in blood flow, hemoglobin concentration, and morphological features of the blood vasculature is demonstrated. The system combines two non-invasive imaging techniques, a dual-wavelength laser speckle contrast imaging (2-LSI) and an optical microangiography (OMAG) system. 2-LSI is used to monitor the changes in the dynamic blood flow and the changes in the concentration of oxygenated (HbO), deoxygenated (Hb) and total hemoglobin (HbT). The OMAG system is used to acquire high resolution images of the functional blood vessel network. The vessel area density (VAD) is used to quantify the blood vessel network morphology, specifically the capillary recruitment. The proposed multi-functional system is employed to assess the blood perfusion status from a mouse pinna before and immediately after a burn injury. To our knowledge, this is the first non-invasive, non-contact and multifunctional imaging modality that can simultaneously measure variations of several blood perfusion parameters.

Effects of hypoxia on cochlear blood flow in mice evaluated using Doppler optical microangiography.

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.

Determining elastic properties of skin by measuring surface waves from an impulse mechanical stimulus using phase-sensitive optical coherence tomography.

The mechanical properties of skin are important tissue parameters that are useful for understanding skin patho-physiology, which can aid disease diagnosis and treatment. This paper presents an innovative method that employs phase-sensitive spectral-domain optical coherence tomography (PhS-OCT) to characterize the biomechanical properties of skin by measuring surface waves induced by short impulses from a home-made shaker. Experiments are carried out on single and double-layer agar-agar phantoms, of different concentrations and thickness, and on in vivo human skin, at the forearm and the palm. For each experiment, the surface wave phase-velocity dispersion curves were calculated, from which the elasticity of each layer of the sample was determined. It is demonstrated that the experimental results agree well with previous work. This study provides a novel combination of PhS-OCT technology with a simple and an inexpensive mechanical impulse surface wave stimulation that can be used to non-invasively evaluate the mechanical properties of skin in vivo, and may offer potential use in clinical situations.

Quantifying optical microangiography images obtained from a spectral domain optical coherence tomography system.

The blood vessel morphology is known to correlate with several diseases, such as cancer, and is important for describing several tissue physiological processes, like angiogenesis. Therefore, a quantitative method for characterizing the angiography obtained from medical images would have several clinical applications. Optical microangiography (OMAG) is a method for obtaining three-dimensional images of blood vessels within a volume of tissue. In this study we propose to quantify OMAG images obtained with a spectral domain optical coherence tomography system. A technique for determining three measureable parameters (the fractal dimension, the vessel length fraction, and the vessel area density) is proposed and validated. Finally, the repeatability for acquiring OMAG images is determined, and a new method for analyzing small areas from these images is proposed.