Miniaturized MR-compatible ultrasound system for real-time monitoring of acoustic effects in mice using high-resolution MRI.

Visualization of focused ultrasound in high spatial and temporal resolution is crucial for accurately and precisely targeting brain regions noninvasively. Magnetic resonance imaging (MRI) is the most widely used noninvasive tool for whole-brain imaging. However, focused ultrasound studies employing high-resolution (> 9.4 T) MRI in small animals are limited by the small size of the radiofrequency (RF) volume coil and the noise sensitivity of the image to external systems such as bulky ultrasound transducers. This technical note reports a miniaturized ultrasound transducer system packaged directly above a mouse brain for monitoring ultrasound-induced effects using high-resolution 9.4 T MRI. Our miniaturized system integrates MR-compatible materials with electromagnetic (EM) noise reduction techniques to demonstrate echo-planar imaging (EPI) signal changes in the mouse brain at various ultrasound acoustic intensities. The proposed ultrasound-MRI system will enable extensive research in the expanding field of ultrasound therapeutics.

Multifocal skull-compensated transcranial focused ultrasound system for neuromodulation applications based on acoustic holography.

Transcranial focused ultrasound stimulation is a promising therapeutic modality for human brain disorders because of its noninvasiveness, long penetration depth, and versatile spatial control capability through beamforming and beam steering. However, the skull presents a major hurdle for successful applications of ultrasound stimulation. Specifically, skull-induced focal aberration limits the capability for accurate and versatile targeting of brain subregions. In addition, there lacks a fully functional preclinical neuromodulation system suitable to conduct behavioral studies. Here, we report a miniature ultrasound system for neuromodulation applications that is capable of highly accurate multiregion targeting based on acoustic holography. Our work includes the design and implementation of an acoustic lens for targeting brain regions with compensation for skull aberration through time-reversal recording and a phase conjugation mirror. Moreover, we utilize MEMS and 3D-printing technology to implement a 0.75-g lightweight neuromodulation system and present in vivo characterization of the packaged system in freely moving mice. This preclinical system is capable of accurately targeting the desired individual or multitude of brain regions, which will enable versatile and explorative behavior studies using ultrasound neuromodulation to facilitate widespread clinical adoption.

General-Purpose Ultrasound Neuromodulation System for Chronic, Closed-Loop Preclinical Studies in Freely Behaving Rodents.

Transcranial focused ultrasound stimulation (tFUS) is an effective noninvasive treatment modality for brain disorders with high clinical potential. However, the therapeutic effects of ultrasound neuromodulation are not widely explored due to limitations in preclinical systems. The current preclinical studies are head-fixed, anesthesia-dependent, and acute, limiting clinical translatability. Here, this work reports a general-purpose ultrasound neuromodulation system for chronic, closed-loop preclinical studies in freely behaving rodents. This work uses microelectromechanical systems (MEMS) technology to design and fabricate a small and lightweight transducer capable of artifact-free stimulation and simultaneous neural recording. Using the general-purpose system, it can be observed that state-dependent ultrasound neuromodulation of the prefrontal cortex increases rapid eye movement (REM) sleep and protects spatial working memory to REM sleep deprivation. The system will allow explorative studies in brain disease therapeutics and neuromodulation using ultrasound stimulation for widespread clinical adoption.

Transcranial focused ultrasound stimulation with high spatial resolution.

BACKGROUND: Low-intensity transcranial focused ultrasound stimulation is a promising candidate for noninvasive brain stimulation and accurate targeting of brain circuits because of its focusing capability and long penetration depth. However, achieving a sufficiently high spatial resolution to target small animal sub-regions is still challenging, especially in the axial direction. OBJECTIVE: To achieve high axial resolution, we designed a dual-crossed transducer system that achieved high spatial resolution in the axial direction without complex microfabrication, beamforming circuitry, and signal processing. METHODS: High axial resolution was achieved by crossing two ultrasound beams of commercially available piezoelectric curved transducers at the focal length of each transducer. After implementation of the fixture for the dual-crossed transducer system, three sets of in vivo animal experiments were conducted to demonstrate high target specificity of ultrasound neuromodulation using the dual-crossed transducer system (n = 38). RESULTS: The full-width at half maximum (FWHM) focal volume of our dual-crossed transducer system was under 0.52 µm3. We report a focal diameter in both lateral and axial directions of 1 mm. To demonstrate successful in vivo brain stimulation of wild-type mice, we observed the movement of the forepaws. In addition, we targeted the habenula and verified the high spatial specificity of our dual-crossed transducer system. CONCLUSIONS: Our results demonstrate the ability of the dual-crossed transducer system to target highly specific regions of mice brains using ultrasound stimulation. The proposed system is a valuable tool to study the complex neurological circuitry of the brain noninvasively.

Wafer-Scale Multilayer Fabrication for Silk Fibroin-Based Microelectronics.

Silk fibroin is an excellent candidate for biomedical implantable devices because of its biocompatibility, controllable biodegradability, solution processability, flexibility, and transparency. Thus, fibroin has been widely explored in biomedical applications as biodegradable films as well as functional microstructures. Although there exists a large number of patterning methods for fibroin thin films, multilayer micropatterning of fibroin films interleaved with metal layers still remains a challenge. Herein, we report a new wafer-scale multilayer microfabrication process named aluminum hard mask on silk fibroin (AMoS), which is capable of micropatterning multiple layers composed of both fibroin and inorganic materials (e.g., metal and dielectrics) with high-precision microscale alignment. To the best of our knowledge, our AMoS process is the first demonstration of wafer-scale multilayer processing of both silk fibroin and metal micropatterns. In the AMoS process, aluminum deposited on fibroin is first micropatterned using conventional ultraviolet (UV) photolithography, and the patterned aluminum layer is then used as a mask to pattern fibroin underneath. We demonstrate the versatility of our fabrication process by fabricating fibroin microstructures with different dimensions, passive electronic components composed of both fibroin and metal layers, and functional fibroin microstructures for drug delivery. Furthermore, because one of the crucial advantages of fibroin is biocompatibility, we assess the biocompatibility of our fabrication process through the culture of highly susceptible primary neurons. Because the AMoS process utilizes conventional UV photolithography, the principal advantages of our process are multilayer fabrication with high-precision alignment, high resolution, wafer-scale large area processing, no requirement for chemical modification of the protein, and high throughput and thus low cost, all of which have not been feasible with silk fibroin. Therefore, the proposed fabrication method is a promising candidate for batch fabrication of functional fibroin microelectronics (e.g., memristors and organic thin film transistors) for next-generation implantable biomedical applications.

Capacitive Micromachined Ultrasonic Transducer (CMUT)-based Biosensor for Detection of Low Concentration Neuropeptide.

Accurate detection of neuropeptides in cerebrospinal fluid (CSF) plays an important role in both indepth studies and early diagnosis of neurological diseases. Here, we report a biosensor based on Capacitive Micromachined Ultrasonic Transducer (CMUT) which is capable of detecting low concentrations (pg $\sim $ ng/ml) of a neuropeptide involved with the progression of Alzheimer's diseases, somatostatin (SST). A 10-MHz CMUT was fabricated and utilized as a physical resonant sensor which detects the change in the concentration of analyte through the mass-loading mechanism. The resonant plate was sequentially coated with protein G and antibodies to provide specificity to SST; Cysteine-tagged protein G layer enables controlled immobilization of antibodies in a welloriented manner. The change in the resonant frequency of the CMUT sensor was measured after incubating the sensor in various concentrations of SST. The significant shifts in the resonant frequency were observed for SST concentrations in the range of 10 pg/ml $\sim 1$ ng/ml. Compared to the previously reported biosensors developed for SST detection, our sensor shows discernable responses for SST that are $\sim 6$ orders of magnitude lower in concentration. Thus, this work demonstrates the potential of the CMUT resonant sensor as a promising biosensor platform for detection of neuropeptides involved with neurodegenerative diseases that often exist in low concentrations in CSF.

Polypyrrole/Agarose Hydrogel-Based Bladder Volume Sensor with a Resistor Ladder Structure.

Chronic monitoring of bladder activity and urine volume is essential for patients suffering from urinary dysfunctions. However, due to the anatomy and dynamics of the bladder, chronic and precise monitoring of bladder activity remains a challenge. Here, we propose a new sensing mechanism that measures the bladder volume using a resistive ladder network with contact switches. Instead of measuring the impedance between the electrode continuously, the proposed sensor provides a digitized output ('on' or 'off') when the bladder volume reaches a certain threshold value. We present simple proof-of-concept sensors which compare the discrete-mode operation to the continuous-mode operation. In addition, by using multiple pairs of this contact-mode switch in a resistor ladder structure, we demonstrate monitoring of the bladder volume in four discrete steps using an idealized balloon and an ex vivo pig's bladder. We implemented the resistive ladder network using a conductive polypyrrole/agarose hydrogel composite which exhibits a Young's modulus comparable to that of the bladder wall. Compared to the continuous-mode operation, the proposed sensing mechanism is less susceptible to drift due to material degradation and environmental factors.

Neural Probes for Chronic Applications.

Developed over approximately half a century, neural probe technology is now a mature technology in terms of its fabrication technology and serves as a practical alternative to the traditional microwires for extracellular recording. Through extensive exploration of fabrication methods, structural shapes, materials, and stimulation functionalities, neural probes are now denser, more functional and reliable. Thus, applications of neural probes are not limited to extracellular recording, brain-machine interface, and deep brain stimulation, but also include a wide range of new applications such as brain mapping, restoration of neuronal functions, and investigation of brain disorders. However, the biggest limitation of the current neural probe technology is chronic reliability; neural probes that record with high fidelity in acute settings often fail to function reliably in chronic settings. While chronic viability is imperative for both clinical uses and animal experiments, achieving one is a major technological challenge due to the chronic foreign body response to the implant. Thus, this review aims to outline the factors that potentially affect chronic recording in chronological order of implantation, summarize the methods proposed to minimize each factor, and provide a performance comparison of the neural probes developed for chronic applications.