Wireless Optofluidic Systems for Programmable In Vivo Pharmacology and Optogenetics.

In vivo pharmacology and optogenetics hold tremendous promise for dissection of neural circuits, cellular signaling, and manipulating neurophysiological systems in awake, behaving animals. Existing neural interface technologies, such as metal cannulas connected to external drug supplies for pharmacological infusions and tethered fiber optics for optogenetics, are not ideal for minimally invasive, untethered studies on freely behaving animals. Here, we introduce wireless optofluidic neural probes that combine ultrathin, soft microfluidic drug delivery with cellular-scale inorganic light-emitting diode (µ-ILED) arrays. These probes are orders of magnitude smaller than cannulas and allow wireless, programmed spatiotemporal control of fluid delivery and photostimulation. We demonstrate these devices in freely moving animals to modify gene expression, deliver peptide ligands, and provide concurrent photostimulation with antagonist drug delivery to manipulate mesoaccumbens reward-related behavior. The minimally invasive operation of these probes forecasts utility in other organ systems and species, with potential for broad application in biomedical science, engineering, and medicine.

Stretchable multichannel antennas in soft wireless optoelectronic implants for optogenetics.

Optogenetic methods to modulate cells and signaling pathways via targeted expression and activation of light-sensitive proteins have greatly accelerated the process of mapping complex neural circuits and defining their roles in physiological and pathological contexts. Recently demonstrated technologies based on injectable, microscale inorganic light-emitting diodes (µ-ILEDs) with wireless control and power delivery strategies offer important functionality in such experiments, by eliminating the external tethers associated with traditional fiber optic approaches. Existing wireless µ-ILED embodiments allow, however, illumination only at a single targeted region of the brain with a single optical wavelength and over spatial ranges of operation that are constrained by the radio frequency power transmission hardware. Here we report stretchable, multiresonance antennas and battery-free schemes for multichannel wireless operation of independently addressable, multicolor µ-ILEDs with fully implantable, miniaturized platforms. This advance, as demonstrated through in vitro and in vivo studies using thin, mechanically soft systems that separately control as many as three different µ-ILEDs, relies on specially designed stretchable antennas in which parallel capacitive coupling circuits yield several independent, well-separated operating frequencies, as verified through experimental and modeling results. When used in combination with active motion-tracking antenna arrays, these devices enable multichannel optogenetic research on complex behavioral responses in groups of animals over large areas at low levels of radio frequency power (<1 W). Studies of the regions of the brain that are involved in sleep arousal (locus coeruleus) and preference/aversion (nucleus accumbens) demonstrate the unique capabilities of these technologies.

Fabrication of stretchable single-walled carbon nanotube logic devices.

The fabrication of a stretchable single-walled carbon nanotube (SWCNT) complementary metal oxide semiconductor (CMOS) inverter array and ring oscillators is reported. The SWCNT CMOS inverter exhibits static voltage transfer characteristics with a maximum gain of 8.9 at a supply voltage of 5 V. The fabricated devices show stable electrical performance under the maximum strain of 30% via forming wavy configurations. In addition, the 3-stage ring oscillator demonstrates a stable oscillator frequency of ∼3.5 kHz at a supply voltage of 10 V and the oscillating waveforms are maintained without any distortion under cycles of pre-strain and release. The strains applied to the device upon deformation are also analyzed by using the classical lamination theory, estimating the local strain of less than 0.6% in the SWCNT channel and Pd electrode regions which is small enough to keep the device performance stable under the pre-strain up to 30%. This work demonstrates the potential application of stretchable SWCNT logic circuit devices in future wearable electronics.

High performance stretchable UV sensor arrays of SnO2 nanowires.

A high performance, stretchable UV sensor array was fabricated based on an active matrix (AM) device that combined field effect transistors of SWCNTs and SnO2 nanowires. The AM devices provided spatial UV sensing via the individual sensors in the array. SnO2 NW UV sensors showed an average photosensitivity of ∼10(5) and a photoconductive gain of ∼10(6) under very low UV (λ = 254 nm) power intensities of 0.02-0.04 mW cm(-2). The UV sensing performance was not deteriorated by a prestrain of up to 23% induced by radial deformation, consistent with the mechanical analysis.

Fabrication of a Flexible, Wireless Micro-Heater on Elastomer for Wearable Gas Sensor Applications.

Thin-film microdevices can be applied to various wearable devices due to their high flexibility compared to conventional bulk-type electronic devices. Among the various microdevice types, many IoT-based sensor devices have been developed recently. In the case of such sensor elements, it is important to control the surrounding environment to optimize the sensing characteristics. Among these environmental factors, temperature often has a great influence. There are cases where temperature significantly affects the sensor characteristics, as is the case for gas sensors. For this purpose, the development of thin-film-type micro-heaters is important. For this study, a wirelessly driven thin-film micro-heater was fabricated on the flexible and stretchable elastomer, a polydimethylsiloxane (PDMS); the antenna was optimized; and the heater was driven at the temperature up to 102 degrees Celsius. The effect of its use on gas-sensing characteristics was compared through the application of the proposed micro-heater to a gas sensor. The heated SnO2 nanowire gas sensor improved the performance of detecting carbon monoxide (CO) by more than 20%, and the recovery time was reduced to less than half. It is expected that thin-film-type micro-heaters that can be operated wirelessly are suitable for application in various wearable devices, including those for smart sensors and health monitoring.

Stretchable, Multi-Layered Stack Antenna for Smart/Wearable Electronic Applications.

The development of microelectronics has been achieved by improving its performance through miniaturization. This was possible through the development of silicon-based semiconductor process technology, but recently, the demand for wearable or flexible devices has increased. These devices are made using various functional elements based on materials that are difficult to utilize with semiconductor devices that contain existing hard silicon-based materials and are bent or flexibly stretched. In this study, wireless antennas suitable for wearable devices were implemented in a stretchable form. It was possible to stably receive a wireless signal, even with a strain of 20% or more, and power light-emitting diodes (LEDs), microheaters, etc. By devising a multi-layered stack antenna without the existing semiconductor process, it was possible to improve the antenna's reception performance. It is expected that this can be applied in various ways to smart wireless sensors and wearable biomedical devices using the near-field communication (NFC) of smartphones.

Micromechanics and advanced designs for curved photodetector arrays in hemispherical electronic-eye cameras.

The fabrication of a hemispherical electronic-eye camera with optimized designs based upon micromechanical analysis is reported. The photodetector arrays combine layouts with multidevice tiles and extended, non-coplanar interconnects to improve the fill factor and deformability, respectively. Quantitative comparison to micromechanics analysis reveals the key features of these designs. Color images collected with working cameras demonstrate the utility of these approaches.

Experimental and modeling studies of imaging with curvilinear electronic eye cameras.

Model calculations and the experimental measurements of the imaging properties of planar, hemispherical, and elliptic parabolic electronic eye cameras are compared. Numerical methods for comprehensive full field calculations of image formation are enabled by use computationally efficient modes. Quantitative agreement between these calculations and experimentally measured images of test patterns reveals advantages of curvilinear camera systems, and provides guidelines for future designs.

Soft, Wirelessly Powered Humidity Sensor Based on SnO2 Nanowires for Wireless/Wearable Sensor Application.

Humidity, along with temperature, is one of the most important environmental variables in people's lives. The control of humidity is an important matter that is related to material properties and stability in various industries, as well as basic living. In order to detect humidity, changes in the physical, chemical, and electrical properties of materials related to humidity are used, and studies using various methods are conducted. In this study, a field-effect transistor (FET) device was fabricated on a soft polymer substrate with SnO2 nanowires (NWs), whose electrical properties change in response to water molecules. The SnO2 NWs, synthesized by chemical vapor deposition (CVD), were transferred onto a polymer substrate, using a sliding transfer method. The NW FET device, which was connected to an aluminum (Al)-based radio frequency (RF) receiving antenna, was wirelessly operated as a humidity sensor, based on the change in electrical properties of SnO2 NWs according to the relative humidity (RH). It was configured with a wireless antenna and light emitting diode (LED) indicator to implement a soft wirelessly powered humidity sensor that senses high RH and is expected to be used as a wearable electronic/sensor in the future.

Fabrication of nanowire channels with unidirectional alignment and controlled length by a simple, gas-blowing-assisted, selective-transfer-printing technique.

A printing-based lithographic technique for the patterning of V(2)O(5) nanowire channels with unidirectional orientation and controlled length is introduced. The simple, directional blowing of a patterned polymer stamp with N(2) gas, inked with randomly distributed V(2)O(5) nanowires, induces alignment of the nanowires perpendicular to the long axis of the line patterns. Subsequent stamping on the amine-terminated surface results in the selective transfer of the aligned nanowires with a controlled length corresponding to the width of the relief region of the polymer stamp. By employing such a gas-blowing-assisted, selective-transfer-printing technique, two kinds of device structures consisting of nanowire channels and two metal electrodes with top contact, whereby the nanowires were aligned either parallel (parallel device) or perpendicular (serial device) to the current flow in the conduction channel, are fabricated. The electrical properties demonstrate a noticeable difference between the two devices, with a large hysteresis in the parallel device but none in the serial device. Systematic analysis of the hysteresis and the electrical stability account for the observed hysteresis in terms of the proton diffusion in the water layer of the V(2)O(5) nanowires, induced by the application of an external bias voltage higher than a certain threshold voltage.

Curvilinear electronics formed using silicon membrane circuits and elastomeric transfer elements.

Materials and methods to achieve electronics intimately integrated on the surfaces of substrates with complex, curvilinear shapes are described. The approach exploits silicon membranes in circuit mesh structures that can be deformed in controlled ways using thin, elastomeric films. Experimental and theoretical studies of the micromechanics of such curvilinear electronics demonstrate the underlying concepts. Electrical measurements illustrate the high yields that can be obtained. The results represent significant experimental and theoretical advances over recently reported concepts for creating hemispherical photodetectors in electronic eye cameras and for using printable silicon nanoribbons/membranes in flexible electronics. The results might provide practical routes to the integration of high performance electronics with biological tissues and other systems of interest for new applications.

Preparation and implementation of optofluidic neural probes for in vivo wireless pharmacology and optogenetics.

This Protocol Extension describes the fabrication and technical procedures for implementing ultrathin, flexible optofluidic neural probe systems that provide targeted, wireless delivery of fluids and light into the brains of awake, freely behaving animals. As a Protocol Extension article, this article describes an adaptation of an existing Protocol that offers additional applications. This protocol serves as an extension of an existing Nature Protocol describing optoelectronic devices for studying intact neural systems. Here, we describe additional features of fabricating self-contained platforms that involve flexible microfluidic probes, pumping systems, microscale inorganic LEDs, wireless-control electronics, and power supplies. These small, flexible probes minimize tissue damage and inflammation, making long-term implantation possible. The capabilities include wireless pharmacological and optical intervention for dissecting neural circuitry during behavior. The fabrication can be completed in 1-2 weeks, and the devices can be used for 1-2 weeks of in vivo rodent experiments. To successfully carry out the protocol, researchers should have basic skill sets in photolithography and soft lithography, as well as experience with stereotaxic surgery and behavioral neuroscience practices. These fabrication processes and implementation protocols will increase access to wireless optofluidic neural probes for advanced in vivo pharmacology and optogenetics in freely moving rodents.This protocol is an extension to: Nat. Protoc. 8, 2413-2428 (2013); doi:10.1038/nprot.2013.158; published online 07 November 2013.

Fully implantable, battery-free wireless optoelectronic devices for spinal optogenetics.

The advent of optogenetic tools has allowed unprecedented insights into the organization of neuronal networks. Although recently developed technologies have enabled implementation of optogenetics for studies of brain function in freely moving, untethered animals, wireless powering and device durability pose challenges in studies of spinal cord circuits where dynamic, multidimensional motions against hard and soft surrounding tissues can lead to device degradation. We demonstrate here a fully implantable optoelectronic device powered by near-field wireless communication technology, with a thin and flexible open architecture that provides excellent mechanical durability, robust sealing against biofluid penetration and fidelity in wireless activation, thereby allowing for long-term optical stimulation of the spinal cord without constraint on the natural behaviors of the animals. The system consists of a double-layer, rectangular-shaped magnetic coil antenna connected to a microscale inorganic light-emitting diode (µ-ILED) on a thin, flexible probe that can be implanted just above the dura of the mouse spinal cord for effective stimulation of light-sensitive proteins expressed in neurons in the dorsal horn. Wireless optogenetic activation of TRPV1-ChR2 afferents with spinal µ-ILEDs causes nocifensive behaviors and robust real-time place aversion with sustained operation in animals over periods of several weeks to months. The relatively low-cost electronics required for control of the systems, together with the biocompatibility and robust operation of these devices will allow broad application of optogenetics in future studies of spinal circuits, as well as various peripheral targets, in awake, freely moving and untethered animals, where existing approaches have limited utility.

Flexible Near-Field Wireless Optoelectronics as Subdermal Implants for Broad Applications in Optogenetics.

In vivo optogenetics provides unique, powerful capabilities in the dissection of neural circuits implicated in neuropsychiatric disorders. Conventional hardware for such studies, however, physically tethers the experimental animal to an external light source, limiting the range of possible experiments. Emerging wireless options offer important capabilities that avoid some of these limitations, but the current size, bulk, weight, and wireless area of coverage is often disadvantageous. Here, we present a simple but powerful setup based on wireless, near-field power transfer and miniaturized, thin, flexible optoelectronic implants, for complete optical control in a variety of behavioral paradigms. The devices combine subdermal magnetic coil antennas connected to microscale, injectable light-emitting diodes (LEDs), with the ability to operate at wavelengths ranging from UV to blue, green-yellow, and red. An external loop antenna allows robust, straightforward application in a multitude of behavioral apparatuses. The result is a readily mass-producible, user-friendly technology with broad potential for optogenetics applications.

Ultrathin Injectable Sensors of Temperature, Thermal Conductivity, and Heat Capacity for Cardiac Ablation Monitoring.

Knowledge of the distributions of temperature in cardiac tissue during and after ablation is important in advancing a basic understanding of this process, and for improving its efficacy in treating arrhythmias. Technologies that enable real-time temperature detection and thermal characterization in the transmural direction can help to predict the depths and sizes of lesion that form. Herein, materials and designs for an injectable device platform that supports precision sensors of temperature and thermal transport properties distributed along the length of an ultrathin and flexible needle-type polymer substrate are introduced. The resulting system can insert into the myocardial tissue, in a minimally invasive manner, to monitor both radiofrequency ablation and cryoablation, in a manner that has no measurable effects on the natural mechanical motions of the heart. The measurement results exhibit excellent agreement with thermal simulations, thereby providing improved insights into lesion transmurality.

Soft materials in neuroengineering for hard problems in neuroscience.

We describe recent advances in soft electronic interface technologies for neuroscience research. Here, low modulus materials and/or compliant mechanical structures enable modes of soft, conformal integration and minimally invasive operation that would be difficult or impossible to achieve using conventional approaches. We begin by summarizing progress in electrodes and associated electronics for signal amplification and multiplexed readout. Examples in large-area, surface conformal electrode arrays and flexible, multifunctional depth-penetrating probes illustrate the power of these concepts. A concluding section highlights areas of opportunity in the further development and application of these technologies.

Optodynamic simulation of ß-adrenergic receptor signalling.

Optogenetics has provided a revolutionary approach to dissecting biological phenomena. However, the generation and use of optically active GPCRs in these contexts is limited and it is unclear how well an opsin-chimera GPCR might mimic endogenous receptor activity. Here we show that a chimeric rhodopsin/ß2 adrenergic receptor (opto-ß2AR) is similar in dynamics to endogenous ß2AR in terms of: cAMP generation, MAP kinase activation and receptor internalization. In addition, we develop and characterize a novel toolset of optically active, functionally selective GPCRs that can bias intracellular signalling cascades towards either G-protein or arrestin-mediated cAMP and MAP kinase pathways. Finally, we show how photoactivation of opto-ß2AR in vivo modulates neuronal activity and induces anxiety-like behavioural states in both fiber-tethered and wireless, freely moving animals when expressed in brain regions known to contain ß2ARs. These new GPCR approaches enhance the utility of optogenetics and allow for discrete spatiotemporal control of GPCR signalling in vitro and in vivo.

Ultraminiaturized photovoltaic and radio frequency powered optoelectronic systems for wireless optogenetics.

OBJECTIVE: Wireless control and power harvesting systems that operate injectable, cellular-scale optoelectronic components provide important demonstrated capabilities in neuromodulatory techniques such as optogenetics. Here, we report a radio frequency (RF) control/harvesting device that offers dramatically reduced size, decreased weight and improved efficiency compared to previously reported technologies. Combined use of this platform with ultrathin, multijunction, high efficiency solar cells allows for hundred-fold reduction of transmitted RF power, which greatly enhances the wireless coverage. APPROACH: Fabrication involves separate construction of the harvester and the injectable µ-ILEDs. To test whether the presence of the implantable device alters behavior, we implanted one group of wild type mice and compared sociability behavior to unaltered controls. Social interaction experiments followed protocols defined by Silverman et al. with minor modifications. MAIN RESULTS: The results presented here demonstrate that miniaturized RF harvesters, and RF control strategies with photovoltaic harvesters can, when combined with injectable µ-ILEDs, offer versatile capabilities in optogenetics. Experimental and modeling studies establish a range of effective operating conditions for these two approaches. Optogenetics studies with social groups of mice demonstrate the utility of these systems. SIGNIFICANCE: The addition of miniaturized, high performance photovoltaic cells significantly expands the operating range and reduces the required RF power. The platform can offer capabilities to modulate signaling path in the brain region of freely-behaving animals. These suggest its potential for widespread use in neuroscience.

Distinct Subpopulations of Nucleus Accumbens Dynorphin Neurons Drive Aversion and Reward.

The nucleus accumbens (NAc) and the dynorphinergic system are widely implicated in motivated behaviors. Prior studies have shown that activation of the dynorphin-kappa opioid receptor (KOR) system leads to aversive, dysphoria-like behavior. However, the endogenous sources of dynorphin in these circuits remain unknown. We investigated whether dynorphinergic neuronal firing in the NAc is sufficient to induce aversive behaviors. We found that photostimulation of dynorphinergic cells in the ventral NAc shell elicits robust conditioned and real-time aversive behavior via KOR activation, and in contrast, photostimulation of dorsal NAc shell dynorphin cells induced a KOR-mediated place preference and was positively reinforcing. These results show previously unknown discrete subregions of dynorphin-containing cells in the NAc shell that selectively drive opposing behaviors. Understanding the discrete regional specificity by which NAc dynorphinerigic cells regulate preference and aversion provides insight into motivated behaviors that are dysregulated in stress, reward, and psychiatric disease.