Microcirculatory Tissue Beds Research Articles

Speckle Variance Photoacoustic Microscopy for Microhemodynamic Imaging.

Relying on the strong optical absorption of hemoglobin to pulsed laser energy, photoacoustic microscopy provides morphological and functional information on microvasculature label-freely. Here, we propose speckle variance photoacoustic microscopy (SV-PAM), which harnesses intrinsic imaging contrast from temporal-varied photoacoustic signals of moving red blood cells in blood vessels, for recovering three-dimension hemodynamic images down to capillary-level resolution within the microcirculatory tissue beds in vivo. Calculating the speckle variance of consecutive photoacoustic B-scan frames acquired at the same lateral position enables accurate identification of blood perfusion and occlusion, which provides interpretations of dynamic blood flow in the microvasculature, in addition to the microvascular anatomic structures. We demonstrate high-resolution hemodynamic imaging of vascular occlusion and reperfusion in the microvasculature of mice ears in vivo. The results suggest that our SV-PAM is potentially invaluable for biomedical hemodynamic investigations, for example, imaging ischemic stroke and hemorrhagic stroke.

Complex-based OCT angiography algorithm recovers microvascular information better than amplitude- or phase-based algorithms in phase-stable systems

Optical coherence tomography angiography (OCTA) is increasingly becoming a popular inspection tool for biomedical imaging applications. By exploring the amplitude, phase and complex information available in OCT signals, numerous algorithms have been proposed that contrast functional vessel networks within microcirculatory tissue beds. However, it is not clear which algorithm delivers optimal imaging performance. Here, we investigate systematically how amplitude and phase information have an impact on the OCTA imaging performance, to establish the relationship of amplitude and phase stability with OCT signal-to-noise ratio (SNR), time interval and particle dynamics. With either repeated A-scan or repeated B-scan imaging protocols, the amplitude noise increases with the increase of OCT SNR; however, the phase noise does the opposite, i.e. it increases with the decrease of OCT SNR. Coupled with experimental measurements, we utilize a simple Monte Carlo (MC) model to simulate the performance of amplitude-, phase- and complex-based algorithms for OCTA imaging, the results of which suggest that complex-based algorithms deliver the best performance when the phase noise is < ~40 mrad. We also conduct a series of in vivo vascular imaging in animal models and human retina to verify the findings from the MC model through assessing the OCTA performance metrics of vessel connectivity, image SNR and contrast–to–noise ratio. We show that for all the metrics assessed, the complex-based algorithm delivers better performance than either the amplitude- or phase–based algorithms for both the repeated A-scan and the B-scan imaging protocols, which agrees well with the conclusion drawn from the MC simulations.

Microvascular imaging and monitoring of human oral cavity lesions in vivo by swept-source OCT-based angiography.

We report the development of optical coherence tomography- (OCT) based angiography (OCTA) to image blood flow within microcirculatory tissue beds in human oral cavity in vivo with a field of view at 10mm×10mm. Three-dimensional (3D) structural and vascular images of labial mucosa tissue are obtained at a single 3D acquisition. Pathologic mucosal sites with mouth ulcers are examined using the OCT tomograms and angiograms, upon which to monitor the lesion healing process over a period of 2weeks. Quantitative metrics of the capillary loop density within the lamina propria layer are evaluated, providing statistically significant difference between healthy and diseased conditions over time. Furthermore, tissue anatomy and vessel morphology of other susceptible sites to ulcer, such as tongue, alveolar mucosa, and labial frenulum, are also imaged to demonstrate the promise of the proposed method as a clinically useful tool for the diagnosis and monitoring of therapeutic treatment of oral tissue abnormalities.

Optical coherence tomography based angiography [Invited

Optical coherence tomography (OCT)-based angiography (OCTA) provides in vivo, three-dimensional vascular information by the use of flowing red blood cells as intrinsic contrast agents, enabling the visualization of functional vessel networks within microcirculatory tissue beds non-invasively, without a need of dye injection. Because of these attributes, OCTA has been rapidly translated to clinical ophthalmology within a short period of time in the development. Various OCTA algorithms have been developed to detect the functional micro-vasculatures in vivo by utilizing different components of OCT signals, including phase-signal-based OCTA, intensity-signal-based OCTA and complex-signal-based OCTA. All these algorithms have shown, in one way or another, their clinical values in revealing micro-vasculatures in biological tissues in vivo, identifying abnormal vascular networks or vessel impairment zones in retinal and skin pathologies, detecting vessel patterns and angiogenesis in eyes with age-related macular degeneration and in skin and brain with tumors, and monitoring responses to hypoxia in the brain tissue. The purpose of this paper is to provide a technical oriented overview of the OCTA developments and their potential pre-clinical and clinical applications, and to shed some lights on its future perspectives. Because of its clinical translation to ophthalmology, this review intentionally places a slightly more weight on ophthalmic OCT angiography.

Wide-field imaging of retinal vasculature using optical coherence tomography-based microangiography provided by motion tracking.

Optical coherence tomography (OCT)-based optical microangiography (OMAG) is a high-resolution, noninvasive imaging technique capable of providing three-dimensional in vivo blood flow visualization within microcirculatory tissue beds in the eye. Although the technique has demonstrated early clinical utility by imaging diseased eyes, its limited field of view (FOV) and the sensitivity to eye motion remain the two biggest challenges for the widespread clinical use of the technology. Here, we report the results of retinal OMAG imaging obtained from a Zeiss Cirrus 5000 spectral domain OCT system with motion tracking capability achieved by a line scan ophthalmoscope (LSO). The tracking LSO is able to guide the OCT scanning, which minimizes the effect of eye motion in the final results. We show that the tracking can effectively correct the motion artifacts and remove the discontinuities and distortions of vascular appearance due to microsaccade, leading to almost motion-free OMAG angiograms with good repeatability and reliability. Due to the robustness of the tracking LSO, we also show the montage scan protocol to provide unprecedented wide field retinal OMAG angiograms. We experimentally demonstrate a 12 x 16 mm² retinal OMAG angiogram acquired from a volunteer, which is the widest FOV retinal vasculature imaging up to now in the community.

4D optical coherence tomography-based micro-angiography achieved by 16-MHz FDML swept source

We demonstrate the use of an ultra-high-speed swept-source optical coherence tomography (OCT) to achieve optical micro-angiography (OMAG) of microcirculatory tissue beds in vivo. The system is based on a 1310-nm Fourier domain mode-locking (FDML) laser with 1.6-MHz A-line rate, providing a frame rate of 3.415KHz, an axial resolution of ∼10 μm and signal to noise ratio of 102dB. Motion from blood flow causes change in OCT signals between consecutive B-frames acquired at the same location. Intensity-based inter-frame subtraction algorithm is applied to extract blood flow from tissue background without any motion correction. We demonstrate the capability of this 1.6-MHz OCT system for 4D OMAG of in vivo tissue at a volume rate of 4.7 volumes/s (volume size: 512×200×720 voxels).

Segmentation and quantification of blood vessels for OCT-based micro-angiograms using hybrid shape/intensity compounding

Optical coherence tomography (OCT) based microangiography is capable of visualizing 3D functional blood vessel networks within microcirculatory tissue beds in vivo. To provide the quantitative information of vasculature from the microangiograms such as vessel diameter and morphology, it is necessary to develop efficient vessel segmentation algorithms. In this paper, we propose to develop a hybrid Hessian/intensity based method to segment and quantify shape and diameter of the blood vessels innervating capillary beds that are imaged by functional OCT in vivo. The proposed method utilizes multi-scale Hessian filters to segment tubular structures such as blood vessels, but compounded by the intensity-based segmentation method to mitigate the limitations of Hessian filters' sensitivity to the selection of scale parameters. Such compounding segmentation scheme takes advantage of the morphological nature of Hessian filters while correcting for the scale parameter selection by intensity-based segmentation. The proposed algorithm is tested on a wound healing model and its performance of segmentation vessels is quantified by a publicly available manual segmentation dataset. We believe that this method will play an important role in the quantification of micro-angiograms for microcirculation research in ophthalmology and diagnosing retinal eye diseases involved with microcirculation.

Super-resolution spectral estimation of optical micro-angiography for quantifying blood flow within microcirculatory tissue beds in vivo

In this paper, we propose a super-resolution spectral estimation technique to quantify microvascular hemodynamics using optical microangiography (OMAG) based on optical coherence tomography (OCT). The proposed OMAG technique uses both amplitude and phase information of the OCT signals which makes it sensitive to the axial and transverse flows. The scanning protocol for the proposed method is identical to three-dimensional ultrahigh sensitive OMAG, and is applicable for in vivo measurements. In contrast to the existing capillary flow quantification methods, the proposed method is less sensitive to tissue motion and does not have aliasing problems due fast flow within large blood vessels. This method is analogous to power Doppler in ultrasonography and estimates the number of red blood cells passing through the beam as opposed to the velocity of the particles. The technique is tested both qualitatively and quantitatively by using OMAG to image microcirculation within mouse ear flap in vivo.

Uniform enhancement of optical micro-angiography images using Rayleigh contrast-limited adaptive histogram equalization.

Optical microangiography is an imaging technology that is capable of providing detailed functional blood flow maps within microcirculatory tissue beds in vivo. Some practical issues however exist when displaying and quantifying the microcirculation that perfuses the scanned tissue volume. These issues include: (I) Probing light is subject to specular reflection when it shines onto sample. The unevenness of the tissue surface makes the light energy entering the tissue not uniform over the entire scanned tissue volume. (II) The biological tissue is heterogeneous in nature, meaning the scattering and absorption properties of tissue would attenuate the probe beam. These physical limitations can result in local contrast degradation and non-uniform micro-angiogram images. In this paper, we propose a post-processing method that uses Rayleigh contrast-limited adaptive histogram equalization to increase the contrast and improve the overall appearance and uniformity of optical micro-angiograms without saturating the vessel intensity and changing the physical meaning of the micro-angiograms. The qualitative and quantitative performance of the proposed method is compared with those of common histogram equalization and contrast enhancement methods. We demonstrate that the proposed method outperforms other existing approaches. The proposed method is not limited to optical microangiography and can be used in other image modalities such as photo-acoustic tomography and scanning laser confocal microscopy.

Label-free 3D imaging of microstructure, blood, and lymphatic vessels within tissue beds in vivo

This Letter reports the use of an ultrahigh resolution optical microangiography (OMAG) system for simultaneous 3D imaging of microstructure and lymphatic and blood vessels without the use of an exogenous contrast agent. An automatic algorithm is developed to segment the lymphatic vessels from the microstructural images based on the fact that the lymph fluid is optically transparent. An OMAG system is developed that utilizes a broadband supercontinuum light source, providing an axial resolution of 2.3 μm and lateral resolution of 5.8 μm, capable of resolving the capillary vasculature and lymphatic vessels innervating microcirculatory tissue beds. Experimental demonstration is performed by showing detailed 3D lymphatic and blood vessel maps, coupled with morphology, within mouse ears in vivo.

Full range complex ultrahigh sensitive optical microangiography

This Letter presents a useful method that combines the full range complex Fourier domain optical coherence tomography (OCT) with the ultrahigh sensitive optical microangiography (OMAG) to achieve full range complex imaging of blood flow within microcirculatory tissue beds in vivo. We propose to use the fast scanning axis to realize the full range complex imaging, while using the slow axis to achieve OMAG imaging of blood flow. We demonstrate the proposed method by using a high speed 1310 nm OCT/OMAG system running at 92 kHz line scan rate to image the flow phantoms in vitro, and the blood flows in tissue beds in vivo.

Ultrahigh sensitive optical microangiography reveals depth-resolved microcirculation and its longitudinal response to prolonged ischemic event within skeletal muscles in mice

The primary pathophysiology of peripheral arterial disease is associated with impaired perfusion to the muscle tissue in the lower extremities. The lack of effective pharmacologic treatments that stimulate vessel collateralization emphasizes the need for an imaging method that can be used to dynamically visualize depth-resolved microcirculation within muscle tissues. Optical microangiography (OMAG) is a recently developed label-free imaging method capable of producing three-dimensional images of dynamic blood perfusion within microcirculatory tissue beds at an imaging depth of up to ∼2 mm, with an unprecedented imaging sensitivity of blood flow at ∼4 μm∕s. In this paper, we demonstrate the utility of OMAG in imaging the detailed blood flow distributions, at a capillary-level resolution, within skeletal muscles of mice. By use of the mouse model of hind-limb ischemia, we show that OMAG can assess the time-dependent changes in muscle perfusion and perfusion restoration along tissue depth. These findings indicate that OMAG can represent a sensitive, consistent technique to effectively study pharmacologic therapies aimed at promoting the growth and development of collateral vessels.

Biophotonics Modalities for High-Resolution Imaging of Microcirculatory Tissue Beds Using Endogenous Contrast: A Review on Present Scenario and Prospects

The microcirculation is a complex system, and the visualization of microcirculation has great significance in improving our understanding of pathophysiological processes in various disease conditions, in both clinical and fundamental studies. A range of techniques are available or emerging for investigating different aspect of the microcirculation in animals and humans. This paper reviews the recent developments in the field of high-resolution and high-sensitive optical imaging of microcirculatory tissue beds, emphasizing technologies that utilize the endogenous contrast mechanism. Optical imaging techniques such as intravital microscopy, Capillaroscopy, laser Doppler perfusion imaging, laser speckle perfusion imaging, polarization spectroscopy, photo-acoustic tomography, and various implementations of optical coherence tomography based on Doppler and speckle contrast imaging are presented together with their prospectives and challenges.

Label-free in vivo optical imaging of functional microcirculations within meninges and cortex in mice

Abnormal microcirculation within meninges is common in many neurological diseases. There is a need for an imaging method that is capable of monitoring dynamic meningeal microcirculations, preferably decoupled from cortical blood flow. Optical microangiography (OMAG) is a recently developed label-free imaging method capable of producing 3D images of dynamic blood perfusion within micro-circulatory tissue beds at an imaging depth up to ∼2 mm, with an unprecedented imaging sensitivity to blood flow at ∼4 μm/s. In this paper, we demonstrate the utility of OMAG in imaging the detailed blood flow distributions, at a capillary level resolution, within the meninges and cortex in mice with the cranium left intact. Using a thrombotic mouse model, we show that the OMAG can yield longitudinal measurements of meningeal vascular responses to the insult and can decouple these responses from those in the cortex, giving valuable information regarding the localized hemodynamics along with the dynamic formation of thrombotic event. The results indicate that OMAG can be a useful tool to study therapeutic strategies in preclinical animal models in order to mitigate various pathologies that are mainly related to the meningeal circulations.

Optical Microangiography: A Label-Free 3-D Imaging Technology to Visualize and Quantify Blood Circulations Within Tissue Beds In Vivo

Optical microangiography (OMAG) is a recently developed volumetric imaging technique that is capable of producing 3D images of dynamic blood perfusion within microcirculatory tissue beds in vivo. The imaging contrast of OMAG image is based on the intrinsic optical scattering signals backscattered by the moving blood cells in patent blood vessels, thus it is a label free imaging technique. In this paper, I will first discuss its recent developments that use a constant modulation frequency introduced in the spectral interferograms to achieve the blood perfusion imaging. I will then introduce its latest development that utilizes the inherent blood flow to modulate the spectral interferograms to realize the blood perfusion imaging. Finally, examples of using OMAG to delineate the dynamic blood perfusion, down to capillary level resolution, within living tissues are given, including cortical blood perfusion in the brain of small animals and blood flow within human retina and choroids.

Doppler optical micro-angiography for volumetric imaging of vascular perfusion in vivo

We propose a Doppler optical micro-angiography (DOMAG) method to image flow velocities of the blood flowing in functional vessels within microcirculatory tissue beds in vivo. The method takes the advantages of recently developed optical micro-angiography (OMAG) technology, in which the endogenous optical signals backscattered from the moving blood cells are isolated from those originated from the tissue background, i.e., the tissue microstructures. The phase difference between adjacent A scans of OMAG flow signals is used to evaluate the flow velocity, similar to phase-resolved Doppler optical coherence tomography (PRDOCT). To meet the requirement of correlation between adjacent A scans in using the phase resolved technique to evaluate flow velocity, an ideal tissue-sample background (i.e., optically homogeneous tissue sample) is digitally reconstructed to replace the signals that represent the heterogeneous features of the static sample that are rejected in the OMAG flow images. Because of the ideal optical-homogeneous sample, DOMAG is free from the characteristic texture pattern noise due to the heterogeneous property of sample, leading to dramatic improvement of the imaging performance. A series of phantom flow experiments are performed to evaluate quantitatively the improved imaging performance. We then conduct in vivo experiments on a mouse brain to demonstrate that DOMAG is capable of quantifying the flow velocities within cerebrovascular network, down to capillary level resolution. Finally, we compare the in vivo imaging performance of DOMAG with that of PRDOCT, and show that DOMAG delivers at least 15-fold increase over the PRDOCT method in terms of the lower limit of flow velocity that can be detected.