Multispectral Optoacoustic Tomography Enables In Vivo Anatomical and Functional Assessment of Human Tendons.

Tendon injuries resulting from accidents and aging are increasing globally. However, key tendon functional parameters such as microvascularity and oxygen perfusion remain inaccessible via the currently available clinical diagnostic tools, resulting in disagreements on optimal treatment options. Here, a new noninvasive method for anatomical and functional characterization of human tendons based on multispectral optoacoustic tomography (MSOT) is reported. Healthy subjects are investigated using a hand-held scanner delivering real-time volumetric images. Tendons in the wrist, ankle, and lower leg are imaged in the near-infrared optical spectrum to utilize endogenous contrast from Type I collagen. Morphology of the flexor carpi ulnaris, carpi radialis, palmaris longus, and Achilles tendons are reconstructed in full. The functional roles of the flexor digitorium longus, hallicus longus, and the tibialis posterior tendons have been visualized by dynamic tracking during toe extension-flexion motion. Furthermore, major vessels and microvasculature near the Achilles tendon are localized, and the global increase in oxygen saturation in response to targeted exercise is confirmed by perfusion studies. MSOT is shown to be a versatile tool capable of anatomical and functional tendon assessments. Future studies including abnormal subjects can validate the method as a viable noninvasive clinical tool for tendinopathy management and healing monitoring.

Noninvasive Tracking of Embryonic Cardiac Dynamics and Development with Volumetric Optoacoustic Spectroscopy.

Noninvasive monitoring of cardiac development can potentially prevent cardiac anomalies in adulthood. Mouse models provide unique opportunities to study cardiac development and disease in mammals. However, high-resolution noninvasive functional analyses of murine embryonic cardiac models are challenging because of the small size and fast volumetric motion of the embryonic heart, which is deeply embedded inside the uterus. In this study, a real time volumetric optoacoustic spectroscopy (VOS) platform for whole-heart visualization with high spatial (100 µm) and temporal (10 ms) resolutions is developed. Embryonic heart development on gestational days (GDs) 14.5-17.5 and quantify cardiac dynamics using time-lapse-4D image data of the heart is followed. Additionally, spectroscopic recordings enable the quantification of the blood oxygenation status in heart chambers in a label-free and noninvasive manner. This technology introduces new possibilities for high-resolution quantification of embryonic heart function at different gestational stages in mammalian models, offering an invaluable noninvasive method for developmental biology.

Hybrid spherical array for combined volumetric optoacoustic and B-mode ultrasound imaging.

Optoacoustic (OA) imaging has achieved tremendous progress with state-of-the-art systems providing excellent functional and molecular contrast, centimeter scale penetration into living tissues, and ultrafast imaging performance, making it highly suitable for handheld imaging in the clinics. OA can greatly benefit from efficient integration with ultrasound (US) imaging, which remains the routine method in bedside clinical diagnostics. However, such integration has not been straightforward since the two modalities typically involve different image acquisition strategies. Here, we present a new, to our knowledge, hybrid optoacoustic ultrasound (OPUS) imaging approach employing a spherical array with dedicated segments for each modality to enable volumetric OA imaging merged with conventional B-mode US. The system performance is subsequently showcased in healthy human subjects. The new OPUS approach hence represents an important step toward establishing OA in point-of-care diagnostic settings.

Model-based correction of rapid thermal confounds in fluorescence neuroimaging of targeted perturbation.

Significance: An array of techniques for targeted neuromodulation is emerging, with high potential in brain research and therapy. Calcium imaging or other forms of functional fluorescence imaging are central solutions for monitoring cortical neural responses to targeted neuromodulation, but often are confounded by thermal effects that are inter-mixed with neural responses. Aim: Here, we develop and demonstrate a method for effectively suppressing fluorescent thermal transients from calcium responses. Approach: We use high precision phased-array 3 MHz focused ultrasound delivery integrated with fiberscope-based widefield fluorescence to monitor cortex-wide calcium changes. Our approach for detecting the neural activation first takes advantage of the high inter-hemispheric correlation of resting state Ca2+ dynamics and then removes the ultrasound-induced thermal effect by subtracting its simulated spatio-temporal signature from the processed profile. Results: The focused 350 µm-sized ultrasound stimulus triggered rapid localized activation events dominated by transient thermal responses produced by ultrasound. By employing bioheat equation to model the ultrasound heat deposition, we can recover putative neural responses to ultrasound. Conclusions: The developed method for canceling transient thermal fluorescence quenching could also find applications with optical stimulation techniques to monitor thermal effects and disentangle them from neural responses. This approach may help deepen our understanding of the mechanisms and macroscopic effects of ultrasound neuromodulation, further paving the way for tailoring the stimulation regimes toward specific applications.

Universal Real-Time Adaptive Signal Compression for High-Frame-Rate Optoacoustic Tomography.

Optoacoustic tomography (OAT) has recently been advanced toward ultrafast volumetric imaging frame rates in the kilohertz range. As a result, excessive data processing and storage capacity requirements are increasingly being imposed on the imaging systems. OAT data commonly exhibit significant sparsity across the spatial, temporal or spectral domains, which facilitated the development of compressed sensing algorithms exploiting various sparse acquisition and under-sampling schemes to reduce data rates. However, performance of compressed sensing critically depends on a priori knowledge on the type of acquired data and/or imaged object, commonly resulting in lack of general applicability and unpredictable image quality. In this work, we report on a fast adaptive OAT data compression framework operating on fully sampled tomographic data. It is based on a wavelet packet transform that maximizes the data compression ratio according to the desired signal energy loss. A dedicated reconstruction method was further developed that efficiently renders images directly from the compressed data. Up to 1000x compression ratios were achieved while providing efficient control over the resulting image quality from arbitrary datasets exhibiting diverse spatial, temporal and spectral characteristics. Our approach enables faster and longer acquisitions and facilitates long-term storage of large OAT datasets.

Dual-Mode Volumetric Optoacoustic and Contrast Enhanced Ultrasound Imaging With Spherical Matrix Arrays.

Spherical matrix arrays represent an advantageous tomographic detection geometry for non-invasive deep tissue mapping of vascular networks and oxygenation with volumetric optoacoustic tomography (VOT). Hybridization of VOT with ultrasound (US) imaging remains difficult with this configuration due to the relatively large inter-element pitch of spherical arrays. We suggest a new approach for combining VOT and US contrast-enhanced 3D imaging employing injection of clinically-approved microbubbles. Power Doppler (PD) and US localization imaging were enabled with a sparse US acquisition sequence and model-based inversion based on infimal convolution of total variation (ICTV) regularization. In vitro experiments in tissue-mimicking phantoms and in living mouse brain demonstrate the powerful capabilities of the new dual-mode imaging approach attaining 80 µm spatial resolution and a more than 10 dB signal to noise improvement with respect to a classical delay and sum beamformer. Microbubble localization and tracking allowed for flow velocity mapping up to 40 mm/s.

High-resolution fluorescence-guided transcranial ultrasound mapping in the live mouse brain.

Understanding the physiological impact of transcranial ultrasound in rodent brains may offer an important preclinical model for human scale magnetic resonance­guided focused ultrasound methods. However, precision tools for high-resolution transcranial ultrasound targeting and real-time in vivo tracking of its effects at the mouse brain scale are currently lacking. We report a versatile bidirectional hybrid fluorescence-ultrasound (FLUS) system incorporating a 0.35-mm precision spherical-phased array ultrasound emission with a fiberscope-based wide-field fluorescence imaging. We show how the marriage between cortex-wide functional imaging and targeted ultrasound delivery can be used to transcranially map previously undocumented localized fluorescence events caused by reversible thermal processes and perform high-speed large-scale recording of neural activity induced by focused ultrasound. FLUS thus naturally harnesses the extensive toolbox of fluorescent tags and ultrasound's localized bioeffects toward visualizing and causally perturbing a plethora of normal and pathophysiological processes in the living murine brain.

Ultrafast four-dimensional imaging of cardiac mechanical wave propagation with sparse optoacoustic sensing.

Propagation of electromechanical waves in excitable heart muscles follows complex spatiotemporal patterns holding the key to understanding life-threatening arrhythmias and other cardiac conditions. Accurate volumetric mapping of cardiac wave propagation is currently hampered by fast heart motion, particularly in small model organisms. Here we demonstrate that ultrafast four-dimensional imaging of cardiac mechanical wave propagation in entire beating murine heart can be accomplished by sparse optoacoustic sensing with high contrast, ∼115-µm spatial and submillisecond temporal resolution. We extract accurate dispersion and phase velocity maps of the cardiac waves and reveal vortex-like patterns associated with mechanical phase singularities that occur during arrhythmic events induced via burst ventricular electric stimulation. The newly introduced cardiac mapping approach is a bold step toward deciphering the complex mechanisms underlying cardiac arrhythmias and enabling precise therapeutic interventions.

Deep learning of image- and time-domain data enhances the visibility of structures in optoacoustic tomography.

Images rendered with common optoacoustic system implementations are often afflicted with distortions and poor visibility of structures, hindering reliable image interpretation and quantification of bio-chrome distribution. Among the practical limitations contributing to artifactual reconstructions are insufficient tomographic detection coverage and suboptimal illumination geometry, as well as inability to accurately account for acoustic reflections and speed of sound heterogeneities in the imaged tissues. Here we developed a convolutional neural network (CNN) approach for enhancement of optoacoustic image quality which combines training on both time-resolved signals and tomographic reconstructions. Reference human finger data for training the CNN were recorded using a full-ring array system that provides optimal tomographic coverage around the imaged object. The reconstructions were further refined with a dedicated algorithm that minimizes acoustic reflection artifacts induced by acoustically mismatch structures, such as bones. The combined methodology is shown to outperform other learning-based methods solely operating on image-domain data.

LightSpeed: A Compact, High-Speed Optical-Link-Based 3D Optoacoustic Imager.

Wide-scale adoption of optoacoustic imaging in biology and medicine critically depends on availability of affordable scanners combining ease of operation with optimal imaging performance. Here we introduce LightSpeed: a low-cost real-time volumetric handheld optoacoustic imager based on a new compact software-defined ultrasound digital acquisition platform and a pulsed laser diode. It supports the simultaneous signal acquisition from up to 192 ultrasound channels and provides a hig-bandwidth direct optical link (2x 100G Ethernet) to the host-PC for ultra-high frame rate image acquisitions. We demonstrate use of the system for ultrafast (500Hz) 3D human angiography with a rapidly moving handheld probe. LightSpeed attained image quality comparable with a conventional optoacoustic imaging systems employing bulky acquisition electronics and a Q-switched pulsed laser. Our results thus pave the way towards a new generation of compact, affordable and high-performance optoacoustic scanners.

Spherical Array System for High-Precision Transcranial Ultrasound Stimulation and Optoacoustic Imaging in Rodents.

Ultrasound can be delivered transcranially to ablate brain tissue, open the blood-brain barrier, or affect neural activity. Transcranial focused ultrasound in small rodents is typically done with low-frequency single-element transducers, which results in unspecific targeting and impedes the concurrent use of fast neuroimaging methods. In this article, we devised a wide-angle spherical array bidirectional interface for high-resolution parallelized optoacoustic imaging and transcranial ultrasound (POTUS) delivery in the same target regions. The system operates between 3 and 9 MHz, allowing to generate and steer focal spots with widths down to [Formula: see text] across a field of view covering the entire mouse brain, while the same array is used to capture high-resolution 3-D optoacoustic data in real time. We showcase the system's versatile beam-forming capacities as well as volumetric optoacoustic imaging capabilities and discuss its potential to noninvasively monitor brain activity and various effects of ultrasound emission.

Multifocal structured illumination optoacoustic microscopy.

Optoacoustic (OA) imaging has the capacity to effectively bridge the gap between macroscopic and microscopic realms in biological imaging. High-resolution OA microscopy has so far been performed via point-by-point scanning with a focused laser beam, thus greatly restricting the achievable imaging speed and/or field of view. Herein we introduce multifocal structured illumination OA microscopy (MSIOAM) that attains real-time 3D imaging speeds. For this purpose, the excitation laser beam is shaped to a grid of focused spots at the tissue surface by means of a beamsplitting diffraction grating and a condenser and is then scanned with an acousto-optic deflector operating at kHz rates. In both phantom and in vivo mouse experiments, a 10 mm wide volumetric field of view was imaged with 15 Hz frame rate at 28 µm spatial resolution. The proposed method is expected to greatly aid in biological investigations of dynamic functional, kinetic, and metabolic processes across multiple scales.

Acoustic Scattering Mediated Single Detector Optoacoustic Tomography.

Optoacoustic image formation is conventionally based upon ultrasound time-of-flight readings from multiple detection positions. Herein, we exploit acoustic scattering to physically encode the position of optical absorbers in the acquired signals, thus reducing the amount of data required to reconstruct an image from a single waveform. This concept is experimentally tested by including a random distribution of scatterers between the sample and an ultrasound detector array. Ultrasound transmission through a randomized scattering medium was calibrated by raster scanning a light-absorbing microparticle across a Cartesian grid. Image reconstruction from a single time-resolved signal was then enabled with a regularized model-based iterative algorithm relying on the calibration signals. The signal compression efficiency is facilitated by the relatively short acquisition time window needed to capture the entire scattered wave field. The demonstrated feasibility to form an image using a single recorded optoacoustic waveform paves a way to the development of faster and affordable optoacoustic imaging systems.

Optoacoustic imaging at kilohertz volumetric frame rates.

State-of-the-art optoacoustic tomographic imaging systems have been shown to attain three-dimensional (3D) frame rates of the order of 100 Hz. While such a high volumetric imaging speed is beyond reach for other bio-imaging modalities, it may still be insufficient to accurately monitor some faster events occurring on a millisecond scale. Increasing the 3D imaging rate is usually hampered by the limited throughput capacity of the data acquisition electronics and memory used to capture vast amounts of the generated optoacoustic (OA) data in real time. Herein, we developed a sparse signal acquisition scheme and a total-variation-based reconstruction approach in a combined space-time domain in order to achieve 3D OA imaging at kilohertz rates. By continuous monitoring of freely swimming zebrafish larvae in a 3D region, we demonstrate that the new approach enables significantly increasing the volumetric imaging rate by using a fraction of the tomographic projections without compromising the reconstructed image quality. The suggested method may benefit studies looking at ultrafast biological phenomena in 3D, such as large-scale neuronal activity, cardiac motion, or freely behaving organisms.

Influence of the absorber dimensions on wavefront shaping based on volumetric optoacoustic feedback.

The recently demonstrated control over light distribution through turbid media based on real-time three-dimensional optoacoustic feedback has offered promising prospects to interferometrically focus light within scattering objects. Nevertheless, the focusing capacity of the feedback-based approach is strongly conditioned by the number of optical modes (speckle grains) enclosed in the volume that can be resolved with the optoacoustic imaging system. In this Letter, we experimentally tested the light intensity enhancement achieved with optoacoustic feedback measurements from different sizes of absorbing microparticles. The importance of the obtained results is discussed in the context of potential signal enhancement at deep locations within a scattering medium where the effective speckle grain sizes approach the minimum values dictated by optical diffraction.

Volumetric real-time tracking of peripheral human vasculature with GPU-accelerated three-dimensional optoacoustic tomography.

Optoacoustic tomography provides a unique possibility for ultra-high-speed 3-D imaging by acquiring complete volumetric datasets from interrogation of tissue by a single nanosecond-duration laser pulse. Yet, similarly to ultrasound, optoacoustics is a time-resolved imaging method, thus, fast 3-D imaging implies real-time acquisition and processing of high speed data from hundreds of detectors simultaneously, which presents significant technological challenges. Herein we present a highly efficient graphical processing unit (GPU) framework for real-time reconstruction and visualization of 3-D tomographic optoacoustic data. By utilizing a newly developed 3-D optoacoustic scanner, which simultaneously acquires signals with a handheld 256-element spherical ultrasonic array system, we further demonstrate tracking of deep tissue human vasculature rendered at a rate of 10 volumetric frames per second. The flexibility provided by the handheld hardware design, combined with the real-time operation, makes the developed platform highly usable for both clinical imaging practice and small animal research applications.

Surgical approach to the management of Brucella endocarditis.

OBJECTIVE: Brucella endocarditis is a rare complication of Brucella infection; however, it is the major cause of deaths in those infected with this disease. In this study, we aim to discuss the results of seven cases who underwent surgery for Brucella endocarditis in our clinic using the knowledge gathered through the literature. METHODS: We reviewed seven patients with Brucella endocarditis, who underwent surgery in our department between October 1990 and April 2007. Brucella endocarditis was diagnosed by physical examination, laboratory findings, serological tests, blood culture, transthoracic and trans-oesophageal echocardiography. All cases underwent surgery after 4-6 weeks of medical therapy. Antimicrobial treatment was maintained for an average of 6 months after surgery. The mean follow-up was 27.4 months. RESULTS: The mean age was 30 years (range, 5-47 years). Four of the patients were male. Of the cases, aortic valve replacement (AVR) was performed in three, mitral valve replacement (MVR) was performed in three and combined aortic and mitral valve replacement (AVR+MVR) was performed in one patient. Pericardial tube drainage was done in one patient because of pericardial effusion and cardiac tamponade that developed 13 days after surgery. One (14.3%) of our patients died 15 days after surgery. The others were discharged. CONCLUSIONS: We concluded that medical and surgical treatment had to be performed simultaneously for the successful management of Brucella endocarditis, a fatal complication of Brucella infection.