Optimization of 3D Passive Acoustic Mapping Image Metrics: Impact of Sensor Geometry and Beamforming Approach.

Three-dimensional passive acoustic mapping (PAM) with matrix arrays typically suffers from high demands on the receiving electronics and high computational load. In our study, we investigated, both numerically and experimentally, the influence of matrix array aperture size, element count, and beamforming approaches on defined image metrics. With a numerical Vokurka model, matrix array acquisitions of cavitation signals were simulated. In the experimental part, two 32 × 32 matrix arrays with different pitches and aperture sizes were used. After being reconstructed into 3D cavitation maps, defined metrics were calculated for a quantitative comparison of experimental and numerical data. The numerical results showed that the enlargement of the aperture from 5 to 40 mm resulted in an improvement of the full width at half maximum (FWHM) by factors of 6 and 13 (in lateral and axial dimension, respectively). A larger array sparsity influenced the point spread function (PSF) only slightly, while the grating lobe level (GLL) remained more than 12 dB below the main lobe. These results were successfully experimentally confirmed. To further reduce the GLL caused by array sparsity, we adapted a non-linear filter from optoacoustics for use in PAM. In combination with the delay, multiply, sum, and integrate (DMSAI) algorithm, the GLL was decreased by 20 dB for 64-element reconstructions, resulting in levels that were identical to the fully populated matrix reconstruction levels.

Portable Ultrasound Research System for Use in Automated Bladder Monitoring with Machine-Learning-Based Segmentation.

We developed a new mobile ultrasound device for long-term and automated bladder monitoring without user interaction consisting of 32 transmit and receive electronics as well as a 32-element phased array 3 MHz transducer. The device architecture is based on data digitization and rapid transfer to a consumer electronics device (e.g., a tablet) for signal reconstruction (e.g., by means of plane wave compounding algorithms) and further image processing. All reconstruction algorithms are implemented in the GPU, allowing real-time reconstruction and imaging. The system and the beamforming algorithms were evaluated with respect to the imaging performance on standard sonographical phantoms (CIRS multipurpose ultrasound phantom) by analyzing the resolution, the SNR and the CNR. Furthermore, ML-based segmentation algorithms were developed and assessed with respect to their ability to reliably segment human bladders with different filling levels. A corresponding CNN was trained with 253 B-mode data sets and 20 B-mode images were evaluated. The quantitative and qualitative results of the bladder segmentation are presented and compared to the ground truth obtained by manual segmentation.

Application of Photoacoustic Methods and Confocal Microscopy for Monitoring of Therapeutic Response in Plaque Psoriasis.

Psoriasis is prone to relapses and requires long-term therapy that may induce a range of adverse effects; therefore, an efficient and early detection of relapses is desirable. In this study, photoacoustic imaging and confocal laser scanning microscopic (CLSM) methods were investigated for their suitability in psoriasis follow-up examinations. Using a high-resolution photoacoustic system, the vascular structures of 11 psoriatic patients and 6 healthy volunteers were investigated. No differences were detected with respect to the average vessel diameter and vasculature per unit volume in the tissue of healthy volunteers and non-lesional and lesional skin areas of psoriatic patients. By means of CLSM, the diameters of the dermal papillae of 6 volunteers and 6 psoriatic patients were determined. The diameters of the dermal papillae of the healthy volunteers (0.074 ± 0.006 mm) revealed no significant difference when compared to non-lesional skin areas of psoriatic patients (0.079 ± 0.005 mm). The results obtained for the lesions in psoriatic patients showed a significant difference (Wilcoxon test, p = 0.028) between the diameters of the dermal papillae of the lesional skin areas (0.114 ± 0.012 mm) and the non-lesional skin areas (0.079 ± 0.005 mm). Thus, CLSM can be applied for monitoring psoriasis follow-up examinations.

Contraction assessment of abdominal muscles using automated segmentation designed for wearable ultrasound applications.

PURPOSE: Wearable ultrasound devices can be used to continuously monitor muscle activity. One possible application is to provide real-time feedback during physiotherapy, to show a patient whether an exercise is performed correctly. Algorithms which automatically analyze the data can be of importance to overcome the need for manual assessment and annotations and speed up evaluations especially when considering real-time video sequences. They even could be used to present feedback in an understandable manner to patients in a home-use scenario. The following work investigates three deep learning based segmentation approaches for abdominal muscles in ultrasound videos during a segmental stabilizing exercise. The segmentations are used to automatically classify the contraction state of the muscles. METHODS: The first approach employs a simple 2D network, while the remaining two integrate the time information from the videos either via additional tracking or directly into the network architecture. The contraction state is determined by comparing measures such as muscle thickness and center of mass between rest and exercise. A retrospective analysis is conducted but also a real-time scenario is simulated, where classification is performed during exercise. RESULTS: Using the proposed segmentation algorithms, 71% of the muscle states are classified correctly in the retrospective analysis in comparison to 90% accuracy with manual reference segmentation. For the real-time approach the majority of given feedback during exercise is correct when the retrospective analysis had come to the correct result, too. CONCLUSION: Both retrospective and real-time analysis prove to be feasible. While no substantial differences between the algorithms were observed regarding classification, the networks incorporating the time information showed temporally more consistent segmentations. Limitations of the approaches as well as reasons for failing cases in segmentation, classification and real-time assessment are discussed and requirements regarding image quality and hardware design are derived.

Assessment of focused ultrasound stimulation to induce peripheral nerve activity and potential damage in vivo.

Introduction: Peripheral neuroprostheses are aimed to restore loss of sensory and motor functions by interfacing axons in the peripheral nerves. Most common interfaces in neuroprostheses are electrodes that establish electrical connection with peripheral axons. However, some challenges arise related to long-term functionality, durability, and body response. Recently, focused ultrasound stimulation (FUS) has emerged as a non-invasive approach to modulate the nervous system. However, it is controversial whether FUS can induce axon depolarization. Methods: We have assessed FUS applied in vivo to the rat peripheral nerve, with two objectives: first, to test whether FUS activates peripheral nerves under different stimulation conditions, and second, to evaluate if FUS inflicts damage to the nerve. FUS was delivered with three ultrasound transducers (Sonic Concept H115, H107, and H102) covering the largest set of parameters examined for FUS of peripheral nerves so far. Results: We did not obtain reliable evoked action potentials in either nerves or muscles, under any FUS condition applied, neither over the skin nor directly to the nerve exposed. Additional experiments ex vivo and in vivo on mice, confirmed this conclusion. When FUS stimulation was applied directly to the exposed sciatic nerve, neuromuscular function decreased significantly, and recovered one week later, except for FUS at 0.25 MHz. Histologically, degenerating nerve fibers were observed, with a tendency to be higher with the lower FUS frequency. Discussion: Past reports on the ability of ultrasound to stimulate the peripheral nerve are controversial. After testing a wide range of FUS conditions, we conclude that it is not a reliable and safe method for stimulating the peripheral nerve. Special consideration should be taken, especially when low-frequency FUS is applied, as it may lead to nerve damage.

High Performance Wearable Ultrasound as a Human-Machine Interface for Wrist and Hand Kinematic Tracking.

OBJECTIVE: Non-invasive human machine interfaces (HMIs) have high potential in medical, entertainment, and industrial applications. Traditionally, surface electromyography (sEMG) has been used to track muscular activity and infer motor intention. Ultrasound (US) has received increasing attention as an alternative to sEMG-based HMIs. Here, we developed a portable US armband system with 24 channels and a multiple receiver approach, and compared it with existing sEMG- and US-based HMIs on movement intention decoding. METHODS: US and motion capture data was recorded while participants performed wrist and hand movements of four degrees of freedom (DoFs) and their combinations. A linear regression model was used to offline predict hand kinematics from the US (or sEMG, for comparison) features. The method was further validated in real-time for a 3-DoF target reaching task. RESULTS: In the offline analysis, the wearable US system achieved an average [Formula: see text] of 0.94 in the prediction of four DoFs of the wrist and hand while sEMG reached a performance of [Formula: see text]= 0.60. In online control, the participants achieved an average 93% completion rate of the targets. CONCLUSION: When tailored for HMIs, the proposed US A-mode system and processing pipeline can successfully regress hand kinematics both in offline and online settings with performances comparable or superior to previously published interfaces. SIGNIFICANCE: Wearable US technology may provide a new generation of HMIs that use muscular deformation to estimate limb movements. The wearable US system allowed for robust proportional and simultaneous control over multiple DoFs in both offline and online settings.

Needs and Attitudes of Older Chronic Back Pain Patients towards a Wearable for Ultrasound Biofeedback during Stabilization Exercises: A Qualitative Analysis.

Chronic back pain has a high prevalence, especially in older adults, and seriously affects sufferers' quality of life. Segmental stabilization exercise (SSE) is often used during physiotherapy to enhance core stability. The execution of SSE requires the selective contraction of deep abdominal and back muscles. Motor learning can be supported using ultrasound imaging as visual biofeedback. ULTRAWEAR is a mobile ultrasound system that provides deep learning-based biofeedback on SSE execution, which is currently under development. We interviewed 15 older chronic back pain patients (CBPPs) to investigate their pain management behavior, experience with SSE, as well as their needs and requirements for ULTRAWEAR. We also gathered information about future-usage scenarios. CBPPs reported a high willingness to use the system as a feedback tool both in physiotherapeutic practices and at home. The automated detection and evaluation of muscle contraction states was highlighted as a major benefit of the system compared to the more subjective feedback provided by traditional methods such as palpation. The system to be developed was perceived as a helpful solution to support learning about SSE.

Evaluation of a Developed MRI-Guided Focused Ultrasound System in 7 T Small Animal MRI and Proof-of-Concept in a Prostate Cancer Xenograft Model to Improve Radiation Therapy.

Focused ultrasound (FUS) can be used to physiologically change or destroy tissue in a non-invasive way. A few commercial systems have clinical approval for the thermal ablation of solid tumors for the treatment of neurological diseases and palliative pain management of bone metastases. However, the thermal effects of FUS are known to lead to various biological effects, such as inhibition of repair of DNA damage, reduction in tumor hypoxia, and induction of apoptosis. Here, we studied radiosensitization as a combination therapy of FUS and RT in a xenograft mouse model using newly developed MRI-compatible FUS equipment. Xenograft tumor-bearing mice were produced by subcutaneous injection of the human prostate cancer cell line PC-3. Animals were treated with FUS in 7 T MRI at 4.8 W/cm2 to reach ~45 °C and held for 30 min. The temperature was controlled via fiber optics and proton resonance frequency shift (PRF) MR thermometry in parallel. In the combination group, animals were treated with FUS followed by X-ray at a single dose of 10 Gy. The effects of FUS and RT were assessed via hematoxylin-eosin (H&E) staining. Tumor proliferation was detected by the immunohistochemistry of Ki67 and apoptosis was measured by a TUNEL assay. At 40 days follow-up, the impact of RT on cancer cells was significantly improved by FUS as demonstrated by a reduction in cell nucleoli from 189 to 237 compared to RT alone. Inhibition of tumor growth by 4.6 times was observed in vivo in the FUS + RT group (85.3%) in contrast to the tumor volume of 393% in the untreated control. Our results demonstrated the feasibility of combined MRI-guided FUS and RT for the treatment of prostate cancer in a xenograft mouse model and may provide a chance for less invasive cancer therapy through radiosensitization.

Comparison of the optoacoustic signal generation efficiency of different nanoparticular contrast agents.

Optoacoustic imaging represents a new modality that allows noninvasive in vivo molecular imaging with optical contrast and acoustical resolution. Whereas structural or functional imaging applications such as imaging of vasculature do not require contrast enhancing agents, nanoprobes with defined biochemical binding behavior are needed for molecular imaging tasks. Since the contrast of this modality is based on the local optical absorption coefficient, all particle or molecule types that show significant absorption cross sections in the spectral range of the laser wavelength used for signal generation are suitable contrast agents. Currently, several particle types such as gold nanospheres, nanoshells, nanorods, or polymer particles are used as optoacoustic contrast agents. These particles have specific advantages with respect to their absorption properties, or in terms of biologically relevant features (biodegradability, binding to molecular markers). In the present study, a comparative analysis of the signal generation efficiency of gold nanorods, polymeric particles, and magnetite particles using a 1064 nm Nd:YAG laser for signal generation is described.

Antitumor necrosis factor-α antibody-coupled gold nanorods as nanoprobes for molecular optoacoustic imaging in arthritis.

Optoacoustic molecular imaging can provide spatially resolved information about the presence of molecular markers in vivo. We synthesized elongated gold nanorods having an absorption maximum in the range of 1064 nm modified with the antibodies infliximab and certolizumab for targeting TNF-α to detect inflammation in arthritic mouse knees. We showed an differential enhancement of optoacoustic signal amplitudes after the injection of infliximab-, but not certolizumab-modified and PEGylated control particles on arthritic and healthy control mice by using a fast-scanning optoacoustic imaging platform based on a pulsed Nd:YAG laser and a single focused ultrasound transducer. The excellent photoacoustic properties of the gold nanorods confirmed the overexpression of TNF-α in arthritic knees. Due to the uncomplicated coupling chemistry and the scalability of ultrasound-based imaging approaches, these results potentially allow a transfer to various preclinical and clinical applications. From the Clinical Editor: Gold nanorods were modified with TNF-α targeting antibodies and used to detect inflammation in arthritic mouse knees via optoaoustic imaging. A fast-scanning optoacoustic imaging platform based on a pulsed Nd:YAG laser and a single focused ultrasound transducer was utilized for imaging. The excellent photoacoustic properties of these gold nanorods confirmed the overexpression of TNF-α, paving the way towards further preclinical and future clinical applications.

Focused Ultrasound Treatment of a Spheroid In Vitro Tumour Model.

Focused ultrasound (FUS) is a non-invasive technique producing a variety of biological effects by either thermal or mechanical mechanisms of ultrasound interaction with the targeted tissue. FUS could bring benefits, e.g., tumour sensitisation, immune stimulation, and targeted drug delivery, but investigation of FUS effects at the cellular level is still missing. New techniques are commonly tested in vitro on two-dimensional (2D) monolayer cancer cell culture models. The 3D tumour model-spheroid-is mainly utilised to mimic solid tumours from an architectural standpoint. It is a promising method to simulate the characteristics of tumours in vitro and their various responses to therapeutic alternatives. This study aimed to evaluate the effects of FUS on human prostate and glioblastoma cancer tumour spheroids in vitro. The experimental follow-up enclosed the measurements of spheroid integrity and growth kinetics, DNA damage, and cellular metabolic activity by measuring intracellular ATP content in the spheroids. Our results showed that pulsed FUS treatment induced molecular effects in 3D tumour models. With the disruption of the spheroid integrity, we observed an increase in DNA double-strand breaks, leading to damage in the cancer cells depending on the cancer cell type.

A Novel Matrix-Array-Based MR-Conditional Ultrasound System for Local Hyperthermia of Small Animals.

OBJECTIVE: The goal of this work was to develop a novel modular focused ultrasound hyperthermia (FUS-HT) system for preclinical applications with the following characteristics: MR-compatible, compact probe for integration into a PET/MR small animal scanner, 3D-beam steering capabilities, high resolution focusing for generation of spatially confined FUS-HT effects. METHODS: For 3D-beam steering capabilities, a matrix array approach with 11 × 11 elements was chosen. For reaching the required level of integration, the array was mounted with a conductive backing directly on the interconnection PCB. The array is driven by a modified version of our 128 channel ultrasound research platform DiPhAS. The system was characterized using sound field measurements and validated using tissue-mimicking phantoms. Preliminary MR-compatibility tests were performed using a 7T Bruker MRI scanner. RESULTS: Four 11 × 11 arrays between 0.5 and 2 MHz were developed and characterized with respect to sound field properties and HT generation. Focus sizes between 1 and 4 mm were reached depending on depth and frequency. We showed heating by 4 °C within 60 s in phantoms. The integration concept allows a probe thickness of less than 12 mm. CONCLUSION: We demonstrated FUS-HT capabilities of our modular system based on matrix arrays and a 128 channel electronics system within a 3D-steering range of up to ±30°. The suitability for integration into a small animal MR could be demonstrated in basic MR-compatibility tests. SIGNIFICANCE: The developed system presents a new generation of FUS-HT for preclinical and translational work providing safe, reversible, localized, and controlled HT.

Preparation and biological evaluation of multifunctional PLGA-nanoparticles designed for photoacoustic imaging.

Nanoparticulate contrast agents for molecular imaging have attracted widespread interest for diagnostic applications with high resolution in medicine. Here we introduce polymer-based multifunctional nanoparticles exhibiting a near-infrared absorption in the range of the Nd:YAG laser wavelength of 1064 nm as a novel resorbable photoacoustic (PA) contrast system and report about their biological evaluation. Submicron-sized spherical nanoparticles with a high encapsulation efficiency (>87%) were created by incorporation of near-infrared dyes (IR5/IR26) in poly[(rac-lactide)-co-glycolide] (PLGA) with 50 mol% glycolide content via a specific spray-drying process in good yield (>75%). Subsequent application of a centrifugation protocol produced two different size fractions with diameters in the ranges 445-540 nm and 253-305 nm; these were further used for investigation of PA properties and cytotoxic effects. The prepared PLGA nanoparticles exhibited PA properties using a Nd:YAG laser-based system. After exposure of particle concentrations up to 10 µg·ml(-1) for 2 days no effects on viability, mitochondrial activity and proliferation, and cell death of human hepatocarcinoma cells and monkey kidney cells were observed. The excellent PA properties in combination with the positive biological results qualify the dye-loaded PLGA particles as promising candidates for a resorbable PA contrast system. FROM THE CLINICAL EDITOR: Photoacoustics (PA), a new modality, in which laser light is shined into tissue and absorbed by inherent proteins or synthetic particles is reflected back and received as ultrasound. This technique was shown to be effective with an erodible polymer particle containing near infrared dyes. In vitro, the PA properties of the PLGA particles persisted for 2 days in cell culture.

Enhancement of Acoustic Microscopy Lateral Resolution: A Comparison Between Deep Learning and Two Deconvolution Methods.

Scanning acoustic microscopy (SAM) provides high-resolution images of biological tissues. Since higher transducer frequencies limit penetration depth, image resolution enhancement techniques could help in maintaining sufficient lateral resolution without sacrificing penetration depth. Compared with existing SAM research, this work introduces two novelties. First, deep learning (DL) is used to improve lateral resolution of 180-MHz SAM images, comparing it with two deconvolution-based approaches. Second, 316-MHz images are used as ground truth in order to quantitatively evaluate image resolution enhancement. The samples used were mouse and rat brain sections. The results demonstrate that DL can closely approximate ground truth (NRMSE = 0.056 and PSNR = 28.4 dB) even with a relatively limited training set (four images, each smaller than 1 mm ×1 mm). This study suggests the high potential of using DL as a single image superresolution method in SAM.

Wave front analysis for enhanced time-domain beamforming of point-like targets in optoacoustic imaging using a linear array.

Using linear array transducers in combination with state-of-the-art multichannel electronics allows to perform optoacoustic imaging with frame rates only limited by the laser pulse repetition frequency and the acoustic time of flight. However, characteristic image artefacts resulting from the limited view and a lower SNR when compared to systems based on single-element focused transducers represent a burden for the clinical acceptance of the technology. In this paper, we present a new method for the improvement of image quality based on the analysis of the signal amplitudes along summation curves during the delay-and-sum beamforming process (DAS). The algorithm compares amplitude distributions along wave fronts with theoretical patterns from optoacoustic point sources. The method was validated on simulated and experimental phantom as well as in-vivo data. An improvement of the lateral resolution by more than a factor of two when comparing conventional DAS and our approach could be shown (numeric and experimental phantom data). For instance, on experimental data from a wire phantom, a PSF in the range of 0.18-0.22 mm was obtained with our approach against 0.48 mm for standard DAS. Furthermore, the SNR of a subcutaneous vessel 2.5 mm below the skin surface was improved by about 30 dB when compared to standard DAS.

Hybrid Photoacoustic/Ultrasound Tomograph for Real-Time Finger Imaging.

We report a target-enclosing, hybrid tomograph with a total of 768 elements based on capacitive micromachined ultrasound transducer technology and providing fast, high-resolution 2-D/3-D photoacoustic and ultrasound tomography tailored to finger imaging. A freely programmable ultrasound beamforming platform sampling data at 80 MHz was developed to realize plane wave transmission under multiple angles. A multiplexing unit enables the connection and control of a large number of elements. Fast image reconstruction is provided by GPU processing. The tomograph is composed of four independent and fully automated movable arc-shaped transducers, allowing imaging of all three finger joints. The system benefits from photoacoustics, yielding high optical contrast and enabling visualization of finger vascularization, and ultrasound provides morphologic information on joints and surrounding tissue. A diode-pumped, Q-switched Nd:YAG laser and an optical parametric oscillator are used to broaden the spectrum of emitted wavelengths to provide multispectral imaging. Custom-made optical fiber bundles enable illumination of the region of interest in the plane of acoustic detection. Precision in positioning of the probe in motion is ensured by use of a motor-driven guide slide. The current position of the probe is encoded by the stage and used to relate ultrasound and photoacoustic signals to the corresponding region of interest of the suspicious finger joint. The system is characterized in phantoms and a healthy human finger in vivo. The results obtained promise to provide new opportunities in finger diagnostics and establish photoacoustic/ultrasound-tomography in medical routine.

Calibrated Linear Array-Driven Photoacoustic/Ultrasound Tomography.

The anisotropic resolution of linear arrays, tools that are widely used in diagnostics, can be overcome by compounding approaches. We investigated the ability of a recently developed calibration and a novel algorithm to determine the actual radial transducer array distance and its misalignment (tilt) with respect to the center of rotation in a 2-D and 3-D tomographic setup. By increasing the time-of-flight accuracy, we force in-phase summation during the reconstruction. Our setup is composed of a linear transducer and a rotation and translation axis enabling multidimensional imaging in ultrasound and photoacoustic mode. Our approach is validated on phantoms and young mice ex vivo. The results indicate that application of the proposed analytical calibration algorithms prevents image artifacts. The spatial resolution achieved was 160 and 250 µm in photoacoustic mode of 2-D and 3-D tomography, respectively.

Optoacoustic imaging of subcutaneous microvasculature with a class one laser.

We developed a combined imaging platform allowing optoacoustic and ultrasound imaging based on a low energy laser and a handheld probe. The device is based on a sensitive single element 35-MHz focused transducer, a 2-D piezoscanner and a dual-wavelength switchable Nd:YAG laser. Acoustical detection and optical illumination are confocal for optimization of optoacoustic signal-to-noise ratio. The system allows to scan over a range up to 12 mm ×12 mm in xy-direction with an isotropic lateral resolution of about 90 µm. Although the device is a class 1 laser product having pulse energies in the range, in vivo images of subcutaneous microvasculature could be obtained from human skin with signal-to-noise levels as good as 20 dB.