Tail artifact removal in OCT angiography images of rodent cortex.

Optical coherence tomography angiography (OCTA) is a surging non-invasive, label-free, in vivo volumetric imaging method, currently being translated to clinical ophthalmology and becoming popular in neuroscience. Despite its attractiveness, there is an inherent issue of using OCT angiograms for quantitative cerebrovascular studies: The dynamic scattering of moving erythrocytes within pial vasculature creates tail-like artifacts that shadow the capillary vessels in the deeper layers of cortex. This false flow effect is relatively benign for qualitative visualization purposes, but it might have a significant impact on quantitative interpretation of angiographic results. In this work, we propose a simple image processing method to remove these tail artifacts in depth-resolved OCTA images using an adaptive enface mask generated with OCT structural images. We demonstrate the effectiveness of our method by comparing vessel densities and vessel similarities of depth-resolved OCT angiograms in a stroke study in a rodent model, in vivo. Thanks to the ability of seeing through the tails of pial vessels, capillary vessels beneath these vessels could be recovered to some extend in the deeper layers of mouse cerebral cortex, leading to a more accurate quantification. Tail artifact removed enface OCT angiogram of deeper layer in vivo mouse cortex.

Optical coherence tomography based microangiography provides an ability to longitudinally image arteriogenesis in vivo.

BACKGROUND: Arteriogenesis describes the active growth of the pre-existing collateral arterioles, which is a crucial tissue-saving process in occlusive vascular diseases. NEW METHOD: We propose to use optical coherence tomography (OCT)-based microangiography (OMAG) to monitor arteriogenesis following artery transection in mouse ear and focal stroke in mouse brain. RESULTS: Our longitudinal mouse ear study shows that the growth phase of arteriogenesis, indicated by changes in collateral vessel diameter and velocity, occurs between 12 and 24h after vessel transection. Additionally, the magnitude of local inflammation is consistent with the time course of arteriogenesis, judging by the tissue thickness measurement and lymphatic vessel signals in OCT. In the mouse brain study, collateral vessel morphology, blood flow velocity and directionality are identified, and an activation of the collateral flow at the arteriolo-arteriolar anastomoses (AAA) is observed during stroke. COMPARISON WITH EXISTING METHODS: In comparison with histology and fluorescence imaging, OCT/OMAG is completely non-invasive and capable of producing consistent results of longitudinal changes in collateral vessel morphology and vasodynamics. CONCLUSION: OCT/OMAG is a promising imaging tool for longitudinal study of collateral vessel remodeling in small animals. This technique can be applied in guiding the in vivo experiments of arteriogenesis stimulation to treat occlusive vascular diseases, including stroke.

Impaired Collateral Flow Compensation During Chronic Cerebral Hypoperfusion in the Type 2 Diabetic Mice.

BACKGROUND AND PURPOSE: The presence of collaterals is associated with a reduced risk of stroke and transient ischemic attack in patients with steno-occlusive carotid artery disease. Although metabolic syndrome negatively impacts collateral status, it is unclear whether and to what extent type 2 diabetes mellitus affects cerebral collateral flow regulation during hypoperfusion. METHODS: We examined the spatial and temporal changes of the leptomeningeal collateral flow and the flow dynamics of the penetrating arterioles in the distal middle cerebral artery and anterior cerebral artery branches over 2 weeks after unilateral common carotid artery occlusion (CCAO) using optical coherent tomography in db/+ and db/db mice. We also assessed the temporal adaptation of the circle of Willis after CCAO by measuring circle of Willis vessel diameters. RESULTS: After unilateral CCAO, db/db mice exhibited diminished leptomeningeal collateral flow compensation compared with db/+ mice, which coincided with a reduced dilation of distal anterior cerebral artery branches, leading to reduced flow not only in pial vessels but also in penetrating arterioles bordering the distal middle cerebral artery and anterior cerebral artery. However, no apparent cell death was detected in either strain of mice during the first week after CCAO. db/db mice also experienced a more severe early reduction in the vessel diameters of several ipsilateral main feeding arteries in the circle of Willis, in addition to a delayed post-CCAO adaptive response by 1 to 2 weeks, compared with db/+ mice. CONCLUSIONS: Type 2 diabetes mellitus is an additional risk factor for hemodynamic compromise during cerebral hypoperfusion, which may increase the severity and the risk of stroke or transient ischemic attack.

Automated segmentation and enhancement of optical coherence tomography-acquired images of rodent brain.

BACKGROUND: Optical coherence tomography (OCT) is a non-invasive optical imaging method that has proven useful in various fields such as ophthalmology, dermatology and neuroscience. In ophthalmology, significant progress has been made in retinal layer segmentation and enhancement of OCT images. There are also segmentation algorithms to separate epidermal and dermal layers in OCT-acquired images of human skin. NEW METHOD: We describe simple image processing methods that allow automatic segmentation and enhancement of OCT images of rodent brain. RESULTS: We demonstrate the effectiveness of the proposed methods for OCT-based microangiography (OMAG) and tissue injury mapping (TIM) of mouse cerebral cortex. The results show significant improvement in image contrast, delineation of tissue injury, allowing visualization of different layers of capillary beds. COMPARISON WITH EXISTING METHODS: Previously reported methods for other applications are yet to be used in neuroscience due to the complexity of tissue anatomy, unique physiology and technical challenges. CONCLUSIONS: OCT is a promising tool that provides high resolution in vivo microvascular and structural images of rodent brain. By automatically segmenting and enhancing OCT images, structural and microvascular changes in mouse cerebral cortex after stroke can be monitored in vivo with high contrast.

Intervolume analysis to achieve four-dimensional optical microangiography for observation of dynamic blood flow.

We demonstrate in vivo volumetric optical microangiography at ∼ 200 volumes/s by the use of 1.6 MHz Fourier domain mode-locking swept source optical coherence tomography and an effective 36 kHz microelectromechanical system (MEMS) scanner. We propose an intervolume analysis strategy to contrast the dynamic blood flow signal from the static tissue background. The proposed system is demonstrated by imaging cerebral blood flow in mice in vivo. For the first time, imaging speed, sensitivity, and temporal resolution become possible for a direct four-dimensional observation of microcirculations within live body parts.

OCT-based label-free in vivo lymphangiography within human skin and areola.

Due to the limitations of current imaging techniques, visualization of lymphatic capillaries within tissue in vivo has been challenging. Here, we present a label-free high resolution optical coherence tomography (OCT) based lymphangiography (OLAG) within human skin in vivo. OLAG enables rapid (~seconds) mapping of lymphatic networks, along with blood vessel networks, over 8 mm x 8 mm of human skin and 5 mm x 5 mm of human areola. Moreover, lymphatic system's response to inflammation within human skin is monitored throughout an acne lesion development over 7 days. The demonstrated results promise OLAG as a revolutionary tool in the clinical research and treatment of patients with pathologic conditions such as cancer, diabetes, and autoimmune diseases.

Review of optical coherence tomography based angiography in neuroscience.

The brain is a complex ecosystem, consisting of multiple layers and tissue compartments. To facilitate the understanding of its function and its response to neurological insults, a fast in vivo imaging tool with a micron-level resolution, which can provide a field of view at a few millimeters, is desirable. Optical coherence tomography (OCT) is a noninvasive method for imaging three-dimensional biological tissues with high resolution ([Formula: see text]) and without a need for contrast agents. Recent development of OCT-based angiography has started to shed some new light on cerebral hemodynamics in neuroscience. We give an overview of the recent developments of OCT-based imaging techniques for neuroscience applications in rodents. We summarize today's technological alternatives for OCT-based angiography for neuroscience and provide a discussion of challenges and opportunities. Moreover, a summary of OCT angiography studies for stroke, traumatic brain injury, and subarachnoid hemorrhage cases on rodents is provided.

Potential use of OCT-based microangiography in clinical dermatology.

BACKGROUND: Optical coherence tomography (OCT) is a revolutionary imaging technique used commonly in ophthalmology, and on the way to become clinically viable alternative in dermatology due to its capability of acquiring histopathology level details of in vivo tissue, non-invasively. In this study, we demonstrate the capabilities of OCT-based microangiography in detecting high resolution, three-dimensional structural, and microvascular features of in vivo human skin with various conditions. METHODS: A swept-source OCT system that operates on a central wavelength of 1310 nm with an A-line rate of 100 kHz is used in this study. We apply optical microangiography (OMAG) technique to visualize the structural and microvascular changes in tissue. RESULTS: OMAG images provide detailed visualization of functional microvasculature of healthy human skin from cheek and forehead areas, abnormal skin conditions from face, chest and belly. Moreover, OMAG is capable of monitoring the progress of wound healing on human skin from arm, delivering unprecedented detail of microstructural and microvascular information during longitudinal wound healing process. CONCLUSION: The presented results promise the clinical use of OCT angiography, aiming to treat prevalent cutaneous diseases, by detecting blood perfusion, and structural changes within human skin, in vivo.

In vivo tissue injury mapping using optical coherence tomography based methods.

An injury causes changes in the optical attenuation coefficient (OAC) of a light beam traveling inside a tissue. We report a method called tissue injury mapping (TIM), which utilizes a noninvasive in vivo optical coherence tomography approach to generate an OAC and microvascular map of the injured tissue. Using TIM, the infarct region development in a mouse cerebral cortex during a stroke is visualized. Moreover, we demonstrate the changes in human facial skin structure and microvasculature during an acne lesion development from initiation to scarring. The results indicate that TIM may be used to aid in the characterization and the treatment of various diseases by enabling a high-resolution detection of tissue structural and microvascular changes.

Lymphatic response to depilation-induced inflammation in mouse ear assessed with label-free optical lymphangiography.

BACKGROUND AND OBJECTIVES: Optical microangiography (OMAG) is a noninvasive technique capable of imaging 3D microvasculature. OMAG-based optical lymphangiography has been developed for 3D visualization of lymphatic vessels without the need for exogenous contrast agents. In this study, we utilize the optical lymphangiography to investigate dynamic changes in lymphatic response within skin tissue to depilation-induced inflammation by using mouse ear as a simple tissue model. MATERIALS AND METHODS: A spectral-domain optical coherence tomography (OCT) system is used in this study to acquire volumetric images of mouse ear. The system operates under the ultrahigh-sensitive OMAG scanning protocol with five repetitions for each B frame. An improved adaptive-threshold-based method is proposed to segment lymphatic vessels from OCT microstructure images. Depilation is achieved by placing hair removal lotion on mouse ear pinna for 5 minutes. Three acquisitions are made before depilation, 3-minute and 30-minute post-depilation, respectively. RESULTS: Right after the application of depilation lotion on the skin, we observe that the blind-ended sacs of initial lymphatics are mainly visible in a specific area of the normal tissue. At 5 minutes, more collecting lymphatic vessels start to form, evidenced by their valve structure that only exists in collecting lymphatic vessels. The lymphangiogenesis is almost completed within 8 minutes in the inflammatory tissue. CONCLUSIONS: Our experimental results demonstrate that the OMAG-based optical lymphangiography has great potential to improve the understanding of lymphatic system in response to various physiological conditions, thus would benefit the development of effective therapeutics.

Vasodynamics of pial and penetrating arterioles in relation to arteriolo-arteriolar anastomosis after focal stroke.

Changes in blood perfusion in highly interconnected pial arterioles provide important insights about the vascular response to ischemia within brain. The functional role of arteriolo-arteriolar anastomosis (AAA) in regulating blood perfusion through penetrating arterioles is yet to be discovered. We apply a label-free optical microangiography (OMAG) technique to evaluate the changes in vessel lumen diameter and red blood cell velocity among a large number of pial and penetrating arterioles within AAA abundant region overlaying the penumbra in the parietal cortex after a middle cerebral artery occlusion (MCAO). In comparison with two-photon microscopy, the OMAG technique makes it possible to image a large number of vessels in a short period of time without administering exogenous contrast agents during a time-constrained MCAO experiment. We compare vasodynamics in penetrating arterioles at various locations. The results show that the MCA connected penetrating arterioles close to a strong AAA dilate, while those belonging to a region away from AAAs constrict in various degrees. These results suggest AAAs play a major role in supporting the active dilation of the penetrating arterioles, thus compensating a significant amount of blood to the ischemic region, whereas the poor blood perfusion occurs at the regions away from AAA connections, leading to ischemia.

High resolution imaging of acne lesion development and scarring in human facial skin using OCT-based microangiography.

BACKGROUND AND OBJECTIVE: Acne is a common skin disease that often leads to scarring. Collagen and other tissue damage from the inflammation of acne give rise to permanent skin texture and microvascular changes. In this study, we demonstrate the capabilities of optical coherence tomography-based microangiography in detecting high-resolution, three-dimensional structural, and microvascular features of in vivo human facial skin during acne lesion initiation and scar development. MATERIALS AND METHODS: A real time swept source optical coherence tomography system is used in this study to acquire volumetric images of human skin. The system operates on a central wavelength of 1,310 nm with an A-line rate of 100 kHz, and with an extended imaging range (∼12 mm in air). The system uses a handheld imaging probe to image acne lesion on a facial skin of a volunteer. We utilize optical microangiography (OMAG) technique to evaluate the changes in microvasculature and tissue structure. RESULTS: Thanks to the high sensitivity of OMAG, we are able to image microvasculature up to capillary level and visualize the remodeled vessels around the acne lesion. Moreover, vascular density change derived from OMAG measurement is provided as an alternative biomarker for the assessment of human skin diseases. In contrast to other techniques like histology or microscopy, our technique made it possible to image 3D tissue structure and microvasculature up to 1.5 mm depth in vivo without the need of exogenous contrast agents. CONCLUSIONS: The presented results are promising to facilitate clinical trials aiming to treat acne lesion scarring, as well as other prevalent skin diseases, by detecting cutaneous blood flow and structural changes within human skin in vivo.

MEMS scanning micromirror for optical coherence tomography.

This paper describes an endoscopic-inspired imaging system employing a micro-electromechanical system (MEMS) micromirror scanner to achieve beam scanning for optical coherence tomography (OCT) imaging. Miniaturization of a scanning mirror using MEMS technology can allow a fully functional imaging probe to be contained in a package sufficiently small for utilization in a working channel of a standard gastroesophageal endoscope. This work employs advanced image processing techniques to enhance the images acquired using the MEMS scanner to correct non-idealities in mirror performance. The experimental results demonstrate the effectiveness of the proposed technique.

Capillary blood flow imaging within human finger cuticle using optical microangiography.

We report non-invasive 3D imaging of capillary blood flow within human finger cuticle by the use of Doppler optical microangiography (DOMAG) and ultra-high sensitive optical microangiography (UHS-OMAG) techniques. Wide velocity range DOMAG method is applied to provide red blood cell (RBC) axial velocity mapping in capillary loops with ranges of ±0.9 mm/s and ±0.3 mm/s. Additionally, UHS-OMAG technique is engineered to acquire high resolution image of capillary morphology. The presented results are promising to facilitate clinical trials of treatment and diagnosis of various diseases such as diabetes, Raynaud's phenomenon, and connective tissue diseases by quantifying cutaneous blood flow changes within human finger cuticle.

In vivo blood flow imaging of inflammatory human skin induced by tape stripping using optical microangiography.

Vasculature response is a hallmark for most inflammatory skin disorders. Tape stripping on human skin causes a minor inflammation which leads to changes in microvasculature. In this study, optical microangiography (OMAG), noninvasive volumetric microvasculature in vivo imaging method, has been used to track the vascular responses after tape stripping. Vessel density has been quantified and used to correlate with the degree of skin irritation. The proved capability of OMAG technique in visualizing the microvasculature network under inflamed skin condition can play an important role in clinical trials of treatment and diagnosis of inflammatory skin disorders.

Application of thinned-skull cranial window to mouse cerebral blood flow imaging using optical microangiography.

In vivo imaging of mouse brain vasculature typically requires applying skull window opening techniques: open-skull cranial window or thinned-skull cranial window. We report non-invasive 3D in vivo cerebral blood flow imaging of C57/BL mouse by the use of ultra-high sensitive optical microangiography (UHS-OMAG) and Doppler optical microangiography (DOMAG) techniques to evaluate two cranial window types based on their procedures and ability to visualize surface pial vessel dynamics. Application of the thinned-skull technique is found to be effective in achieving high quality images for pial vessels for short-term imaging, and has advantages over the open-skull technique in available imaging area, surgical efficiency, and cerebral environment preservation. In summary, thinned-skull cranial window serves as a promising tool in studying hemodynamics in pial microvasculature using OMAG or other OCT blood flow imaging modalities.

Does optical microangiography provide accurate imaging of capillary vessels?: validation using multiphoton microscopy.

Optical microangiography (OMAG) has been extensively utilized to study three-dimensional tissue vasculature in vivo. However, with the limited image resolution (∼10 µm ) of the commonly used systems, some concerns were raised: (1) whether OMAG is capable of providing the imaging of capillary vessels that are of an average diameter of ∼6 µm ; (2) if yes, whether OMAG can provide meaningful quantification of vascular density within the scanned tissue volume. Multiphoton microscopy (MPM) is capable of depth-resolved high-resolution (∼1 µm ) imaging of biological tissue structures. With externally labeled plasma, the vascular network including single capillaries can be well visualized. We compare the vascular images of in vivo mouse brain acquired by both OMAG and MPM systems. We found that within the penetration depth range of the MPM system, OMAG is able to accurately visualize blood vessels including capillaries. Although the resolution of OMAG may not be able to 100% resolve two closely packed tiny capillaries in tissue, it is still capable of visualizing most of the capillaries because there are interstitial tissue spaces between them. We believe our validation results reinforce the application of OMAG in microvasculature-related studies.

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.