Multi-Scale Volumetric Dynamic Optoacoustic and Laser Ultrasound (OPLUS) Imaging Enabled by Semi-Transparent Optical Guidance.

Major biological discoveries are made by interrogating living organisms with light. However, the limited penetration of un-scattered photons within biological tissues limits the depth range covered by optical methods. Deep-tissue imaging is achieved by combining light and ultrasound. Optoacoustic imaging exploits the optical generation of ultrasound to render high-resolution images at depths unattainable with optical microscopy. Recently, laser ultrasound has been suggested as a means of generating broadband acoustic waves for high-resolution pulse-echo ultrasound imaging. Herein, an approach is proposed to simultaneously interrogate biological tissues with light and ultrasound based on layer-by-layer coating of silica optical fibers with a controlled degree of transparency. The time separation between optoacoustic and ultrasound signals collected with a custom-made spherical array transducer is exploited for simultaneous 3D optoacoustic and laser ultrasound (OPLUS) imaging with a single laser pulse. OPLUS is shown to enable large-scale anatomical characterization of tissues along with functional multi-spectral imaging of chromophores and assessment of cardiac dynamics at ultrafast rates only limited by the pulse repetition frequency of the laser. The suggested approach provides a flexible and scalable means for developing a new generation of systems synergistically combining the powerful capabilities of optoacoustics and ultrasound imaging in biology and medicine.

Real-time assessment of high-intensity focused ultrasound heating and cavitation with hybrid optoacoustic ultrasound imaging.

High-intensity focused ultrasound (HIFU) enables localized ablation of biological tissues by capitalizing on the synergistic effects of heating and cavitation. Monitoring of those effects is essential for improving the efficacy and safety of HIFU interventions. Herein, we suggest a hybrid optoacoustic-ultrasound (OPUS) approach for real-time assessment of heating and cavitation processes while providing an essential anatomical reference for accurate localization of the HIFU-induced lesion. Both effects could clearly be observed by exploiting the temperature dependence of optoacoustic (OA) signals and the strong contrast of gas bubbles in pulse-echo ultrasound (US) images. The differences in temperature increase and its rate, as recorded with a thermal camera for different HIFU pressures, evinced the onset of cavitation at the expected pressure threshold. The estimated temperatures based on OA signal variations were also within 10-20 % agreement with the camera readings for temperatures below the coagulation threshold (∼50 °C). Experiments performed in excised tissues as well as in a post-mortem mouse demonstrate that both heating and cavitation effects can be effectively visualized and tracked using the OPUS approach. The good sensitivity of the suggested method for HIFU monitoring purposes was manifested by a significant increase in contrast-to-noise ratio within the ablated region by > 10 dB and > 5 dB for the OA and US images, respectively. The hybrid OPUS-based monitoring approach offers the ease of handheld operation thus can readily be implemented in a bedside setting to benefit several types of HIFU treatments used in the clinics.

The AP-1 transcription factor Fosl-2 drives cardiac fibrosis and arrhythmias under immunofibrotic conditions.

Fibrotic changes in the myocardium and cardiac arrhythmias represent fatal complications in systemic sclerosis (SSc), however the underlying mechanisms remain elusive. Mice overexpressing transcription factor Fosl-2 (Fosl-2tg) represent animal model of SSc. Fosl-2tg mice showed interstitial cardiac fibrosis, disorganized connexin-43/40 in intercalated discs and deregulated expression of genes controlling conduction system, and developed higher heart rate (HR), prolonged QT intervals, arrhythmias with prevalence of premature ventricular contractions, ventricular tachycardias, II-degree atrio-ventricular blocks and reduced HR variability. Following stimulation with isoproterenol Fosl-2tg mice showed impaired HR response. In contrast to Fosl-2tg, immunodeficient Rag2-/-Fosl-2tg mice were protected from enhanced myocardial fibrosis and ECG abnormalities. Transcriptomics analysis demonstrated that Fosl-2-overexpression was responsible for profibrotic signature of cardiac fibroblasts, whereas inflammatory component in Fosl-2tg mice activated their fibrotic and arrhythmogenic phenotype. In human cardiac fibroblasts FOSL-2-overexpression enhanced myofibroblast signature under proinflammatory or profibrotic stimuli. These results demonstrate that under immunofibrotic conditions transcription factor Fosl-2 exaggerates myocardial fibrosis, arrhythmias and aberrant response to stress.

Volumetric optoacoustic neurobehavioral tracking of epileptic seizures in freely-swimming zebrafish larvae.

Fast three-dimensional imaging of freely-swimming zebrafish is essential to understand the link between neuronal activity and behavioral changes during epileptic seizures. Studying the complex spatiotemporal patterns of neuronal activity at the whole-brain or -body level typically requires physical restraint, thus hindering the observation of unperturbed behavior. Here we report on real-time volumetric optoacoustic imaging of aberrant circular swimming activity and calcium transients in freely behaving zebrafish larvae, continuously covering their motion across an entire three-dimensional region. The high spatiotemporal resolution of the technique enables capturing ictal-like epileptic seizure events and quantifying their propagation speed, independently validated with simultaneous widefield fluorescence recordings. The work sets the stage for discerning functional interconnections between zebrafish behavior and neuronal activity for studying fundamental mechanisms of epilepsy and in vivo validation of treatment strategies.

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.

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.

In vivo optoacoustic monitoring of percutaneous laser ablation of tumors in a murine breast cancer model.

Laser ablation (LA) is a promising approach for minimally invasive cancer treatments. Its in vivo applicability is often impeded by the lack of efficient monitoring tools that can help to minimize collateral tissue damage and aid in determining the optimal treatment end-points. We have devised a new, to the best of our knowledge, hybrid LA approach combining simultaneous volumetric optoacoustic (OA) imaging to monitor the lesion progression accurately in real time and 3D. Time-lapse imaging of laser ablation of solid tumors was performed in a murine breast cancer model in vivo by irradiation of subcutaneous tumors with a 100 mJ short-pulsed (${\sim}{5}\;{\rm ns}$∼5ns) laser operating at 1064 nm and 100 Hz pulse repetition frequency. Local changes in the OA signal intensity ascribed to structural alterations in the tumor vasculature were clearly observed, while the OA volumetric projections recorded in vivo appeared to correlate with cross sections of the excised tumors.

Endocardial irrigated catheter for volumetric optoacoustic mapping of radio-frequency ablation lesion progression.

Radiofrequency (RF) catheter ablation is widely employed for various minimally invasive procedures, including treatment of tumors, cardiac arrhythmias and varicose veins. Accurate real-time monitoring of the ablation treatments remains challenging with the existing clinical imaging modalities due to the lack of spatial or temporal resolution or insufficient tissue contrast for differentiating thermal lesions. Optoacoustic (OA) imaging has been recently suggested for monitoring temperature field and lesion progression during RF interventions. However, strong light absorption by standard metallic catheters hindered practical implementations of this approach. Herein, we introduce a new RF ablation catheter concept for combined RF ablation and OA lesion monitoring. The catheter tip encapsulates a multimode fiber bundle for OA excitation with near-infrared (NIR) light, whereas the electric current is conducted through the irrigation solution, thus avoiding direct exposure of the metallic parts to the excitation light. We optimized the catheter diameter and the saline flow rate in order to attain uniform and deep lesions. The newly introduced hybrid catheter design was successfully tested by real-time monitoring of the ablation process in smooth ventricle and rough atrium walls of a blood-filled ex vivo porcine heart, mimicking in vivo conditions in the clinical setting.

Acousto-Optic Catheter Tracking Sensor for Interventional MRI Procedures.

OBJECTIVE: The objective of this paper is to introduce an acousto-optic optical fiber sensor for tracking catheter position during interventional magnetic resonance imaging (MRI) to overcome RF induced heating of active markers. METHODS: The sensor uses a miniature coil coupled to a piezoelectric transducer, which is in turn mechanically connected to an optical fiber. The piezoelectric transducer converts the RF signal to acoustic waves in the optical fiber over a region including a fiber Bragg grating (FBG). The elastic waves in the fiber modulates the FBG geometry and hence the reflected light in the optical fiber. Since the coil is much smaller than the RF wavelength and the signal is transmitted on the dielectric optical fiber, the sensor effectively reduces RF induced heating risk. Proof of concept prototypes of the sensor are implemented using commercially available piezoelectric transducers and optical fibers with FBGs. The prototypes are characterized in a 1.5 T MRI system in comparison with an active tracking marker. RESULTS: Acousto-optical sensor shows linear response with flip angle and it can be used to detect signals from multiple coils for potential orientation detection. It has been successfully used to detect the position of a tacking coil in phantom in an imaging experiment. CONCLUSION: Acousto-optical sensing is demonstrated for tracking catheters during interventional MRI. Real-time operation of the sensor requires sensitivity improvements like using a narrow band FBG. SIGNIFICANCE: Acousto-optics provides a compact solution to sense RF signals in MRI with dielectric transmission lines.

Thin film based semi-active resonant marker design for low profile interventional cardiovascular MRI devices.

OBJECTIVES: A new microfabrication method to produce low profile radio frequency (RF) resonant markers on catheter shafts was developed. A semi-active RF resonant marker incorporating a solenoid and a plate capacitor was constructed on the distal shaft of a 5 Fr guiding catheter. The resulting device can be used for interventional cardiovascular MRI procedures. MATERIALS AND METHODS: Unlike current semi-active device visualization techniques that require rigid and bulky analog circuit components (capacitor and solenoid), we fabricated a low profile RF resonant marker directly on guiding the catheter surface by thin film metal deposition and electroplating processes using a modified physical vapor deposition system. RESULTS: The increase of the overall device profile thickness caused by the semi-active RF resonant marker (130 µm thick) was lowered by a factor of 4.6 compared with using the thinnest commercial non-magnetic and rigid circuit components (600 µm thick). Moreover, adequate visibility performance of the RF resonant marker in different orientations and overall RF safety were confirmed through in vitro experiments under MRI successfully. CONCLUSION: The developed RF resonant marker on a clinical grade 5 Fr guiding catheter will enable several interventional congenital heart disease treatment procedures under MRI.