Wireless, battery-free optoelectronic systems as subdermal implants for local tissue oximetry

Monitoring regional tissue oxygenation in animal models and potentially in human subjects can yield insights into the underlying mechanisms of local O2-mediated physiological processes and provide diagnostic and therapeutic guidance for relevant disease states. Existing technologies for tissue oxygenation assessments involve some combination of disadvantages in requirements for physical tethers, anesthetics, and special apparatus, often with confounding effects on the natural behaviors of test subjects. This work introduces an entirely wireless and fully implantable platform incorporating (i) microscale optoelectronics for continuous sensing of local hemoglobin dynamics and (ii) advanced designs in continuous, wireless power delivery and data output for tether-free operation. These features support in vivo, highly localized tissue oximetry at sites of interest, including deep brain regions of mice, on untethered, awake animal models. The results create many opportunities for studying various O2-mediated processes in naturally behaving subjects, with implications in biomedical research and clinical practice.Center for Bio-Integrated Electronics at Northwestern University; Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF) [ECCS-1542205]; Materials Research Science and Engineering Center [DMR-1720139]; State of Illinois; Northwestern University; Developmental Therapeutics Core at Northwestern University; Robert H. Lurie Comprehensive Cancer Center [NCI CA060553]Open access journalThis item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at [email protected]

Developing Next-generation Brain Sensing Technologies - A Review.

Advances in sensing technology raise the possibility of creating neural interfaces that can more effectively restore or repair neural function and reveal fundamental properties of neural information processing. To realize the potential of these bioelectronic devices, it is necessary to understand the capabilities of emerging technologies and identify the best strategies to translate these technologies into products and therapies that will improve the lives of patients with neurological and other disorders. Here we discuss emerging technologies for sensing brain activity, anticipated challenges for translation, and perspectives for how to best transition these technologies from academic research labs to useful products for neuroscience researchers and human patients

Toward biomaterial-based implantable photonic devices

Optical technologies are essential for the rapid and efficient delivery of health care to patients. Efforts have begun to implement these technologies in miniature devices that are implantable in patients for continuous or chronic uses. In this review, we discuss guidelines for biomaterials suitable for use in vivo. Basic optical functions such as focusing, reflection, and diffraction have been realized with biopolymers. Biocompatible optical fibers can deliver sensing or therapeutic-inducing light into tissues and enable optical communications with implanted photonic devices. Wirelessly powered, light-emitting diodes (LEDs) and miniature lasers made of biocompatible materials may offer new approaches in optical sensing and therapy. Advances in biotechnologies, such as optogenetics, enable more sophisticated photonic devices with a high level of integration with neurological or physiological circuits. With further innovations and translational development, implantable photonic devices offer a pathway to improve health monitoring, diagnostics, and light-activated therapies. Keywords: biomaterials; biocompatible; biodegradable; optics; photonicsUnited States. Department of Defense (Award FA9550-13-1-0068)National Institutes of Health (U.S.) (Award P41-EB015903)National Institutes of Health (U.S.) (Award R01-CA192878)National Science Foundation (U.S.) (Award CBET-1264356)National Science Foundation (U.S.) (Award ECCS-1505569

Novel Class of Chronic and Implantable Wireless Optofluidic (Wof) Neural Devices for in Vivo Drug Delivery and Optogenetics in Freely Moving Animals

Chronic in-vivo studies through multimodal modulation of neurons in the deep brain can render pivotal insights not only in identifying their roles for a specific activity, but also to decipher their contribution in various neurodegenerative diseases. To prevent askew effects, probes were implanted to directly reach the target sites and the spatiotemporal resolution was further improved by targeting key neural circuits using optogenetics and pharmacology. Multimodal optofluidic probes allow integrated access to the target region without the need for multiple surgeries thus reducing tissue damage as well as simplifying combinatorial stimulations. Conventional tools have relied on rigid metal cannulas and silica optical fibers, but they cause adverse tissue inflammation due to their large probe sizes, rigid materials and associated tethers which severely limit their chronic integration. However, minimally invasive, mechanically compliant and untethered probes are shown to significantly limit tissue scarring in freely moving animals as shown in recent advances in soft optofluidic probes controlled by wireless head mounts, however, they didn&rsquo;t last long primarily due to exhaustion of the drugs and power. Moreover, they were limited to the brain due to their bulky head mounts with limited range, directionality, and selectivity. Here we introduce two wireless optofluidic (wOF) neural devices each solving unique challenges faced by previous state-of-the-art devices and commercial pumps. Firstly, is the Lego wOF device which enables: 1) uninterrupted drug supply using replaceable &ldquo;Lego&rdquo; drug cartridges and 2) an easy-to-use smartphone app that allows long wireless range (~100m), isotropic wireless access, no Line of Sight (LOS) handicap, Over The Air (OTA) updates, high target specificity and scalable closed loop systems within a large group as well as intuitive control through an easy to use app on any commercial handheld smartphone. Second is the fully implantable wOF device which enables: 1) both central and peripheral stimulation in any space critical location inside the body (92% smaller and 88% lighter than previous systems), and 2) battery-free operation using a soft, stretchable energy harvester that can conformally adhere to any curvilinear surface inside the body. Successful in-vivo studies in mice demonstrate their innate capability for programmable and chronic wireless pharmacology and optogenetics.</p

Dissection of Affective Catecholamine Circuits Using Traditional and Wireless Optogenetics

Parsing the complexity of the mammalian brain has challenged neuroscientists for thousands of years. In the early 21st century, advances in materials science and neuroscience have enabled unprecedented control of neural circuitry. In particular, cell-type selective manipulations, such as those with optogenetics and chemogenetics, routinely provide answers to previously intractable neurobiological questions in the intact, behaving animal. In this two-part dissertation, I first introduce new minimally invasive, wireless technology to perturb neural activity in the ventral tegmental area dopaminergic system of freely moving animals. I report a series of novel devices for studying and perturbing intact neural systems through optogenetics, microfluidic pharmacology, and electrophysiology. Unlike optogenetic approaches that rely on rigid, glass fiber optics coupled to external light sources, these novel devices utilize flexible substrates to carry microscale, inorganic light emitting diodes (μ-ILEDs), multimodal sensors, and/or microfluidic channels into the brain. Each class of device can be wirelessly controlled, enabling studies in freely behaving mice and achieving previously untenable control of catecholamine neural circuitry. In the second part of this dissertation, I apply existing cell-type selective approaches to dissect the role of the locus coeruleus noradrenergic (LC-NE) system in anxiety-like and aversive behaviors. The LC-NE system is one of the first systems engaged following a stressful event. While LC-NE neurons are known to be activated by many different stressors, the underlying neural circuitry and the role of this activity in generating stress-induced anxiety has not been elucidated until now. I demonstrate that increased tonic activity of LC-NE neurons is both necessary and sufficient for stress-induced anxiety; a behavior which is driven by LC projections to the basolateral amygdala. Furthermore, this activity and behavior is elicited by corticotropin releasing hormone-containing afferent inputs into the LC from the central amygdala. These studies position the LC-NE system as a critical mediator of acute stress-induced anxiety and offer a potential intervention for preventing stress-related affective disorders. Together these two objectives provide a rich technological toolbox for neuroscientists and yield important knowledge of how small catecholamine structures with widespread forebrain innervation can selectively mediate higher order behaviors

Tools for interfacing, extracting, and analyzing neural signals using wide-field fluorescence imaging and optogenetics in awake behaving mice

Imaging of multiple cells has rapidly multiplied the rate of data acquisition as well as our knowledge of the complex dynamics within the mammalian brain. The process of data acquisition has been dramatically enhanced with highly affordable, sensitive image sensors enable high-throughput detection of neural activity in intact animals. Genetically encoded calcium sensors deliver a substantial boost in signal strength and in combination with equally critical advances in the size, speed, and sensitivity of image sensors available in scientific cameras enables high-throughput detection of neural activity in behaving animals using traditional wide-field fluorescence microscopy. However, the tremendous increase in data flow presents challenges to processing, analysis, and storage of captured video, and prompts a reexamination of traditional routines used to process data in neuroscience and now demand improvements in both our hardware and software applications for processing, analyzing, and storing captured video. This project demonstrates the ease with which a dependable and affordable wide-field fluorescence imaging system can be assembled and integrated with behavior control and monitoring system such as found in a typical neuroscience laboratory. An Open-source MATLAB toolbox is employed to efficiently analyze and visualize large imaging data sets in a manner that is both interactive and fully automated. This software package provides a library of image pre-processing routines optimized for batch-processing of continuous functional fluorescence video, and additionally automates a fast unsupervised ROI detection and signal extraction routine. Further, an extension of this toolbox that uses GPU programming to process streaming video, enabling the identification, segmentation and extraction of neural activity signals on-line is described in which specific algorithms improve signal specificity and image quality at the single cell level in a behaving animal. This project describes the strategic ingredients for transforming a large bulk flow of raw continuous video into proportionally informative images and knowledge

Implantable Neural Probes for Electrical Recording and Optical Stimulation of Cellular Level Neural Circuitry in Behaving Animals.

In order to advance the understanding of brain function, it is critical to monitor how neural circuits work together and perform computational processing. For the past few decades, a wide variety of neural probes have been developed to study the electrophysiology of the brain. This work is focused on two important objectives to improve the brain-computer interface: 1) to enhance the reliability of recording electrodes by optimizing the shank structure; 2) to incorporate optical stimulation capability in addition to electrical recording for applications involving optogenetics. For the first objective, a flexible 64-channel parylene probe was designed with unique geometries for reduced tissue reactions. In order to provide the mechanical stiffness necessary to penetrate the brain, the miniaturized, flexible probes were coated with a lithographically patterned silk fibroin, which served as a biodegradable insertion shuttle. Because the penetration strength is independent from the properties of the probe itself, the material and geometry of the probe structure can be optimally designed without constraints. These probes were successfully implanted into the layer-V of motor cortex in 6 rats and recorded neural activities in vivo for 6 weeks. For the second objective, either optical waveguides or μLEDs were monolithically integrated on the probe shanks for optogenetic applications. Compared to existing methods, this work can offer high spatial-temporal resolution to record and stimulate from even subcellular neural structures. In the experiments using wild type animals, despite optimized recording of spontaneous neural activities, the cells never responded to illumination. In contrast, for the ChR2 expressed animals, light activation of neural activities was extremely robust and local, which phase-locked to the light waveform whenever the cell was close to the light source. In particular, the probes integrated with μLEDs were capable of driving different neural circuit behaviors using two adjacent μLEDs separated only by a 60-μm-pitch. With 3 μLEDs integrated at the tip of each of the 4 probe shanks, this novel optogenetic probe can provide more than 480 million (12!) different spiking sequences at the sub-cellular resolution, which is ideal to manipulate high density neural network with versatility and precision.PhDElectrical EngineeringUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttp://deepblue.lib.umich.edu/bitstream/2027.42/111604/1/wufan_1.pd

Optogenetic Brain Interfaces

The brain is a large network of interconnected neurons where each cell functions as a nonlinear processing element. Unraveling the mysteries of information processing in the complex networks of the brain requires versatile neurostimulation and imaging techniques. Optogenetics is a new stimulation method which allows the activity of neurons to be modulated by light. For this purpose, the cell-types of interest are genetically targeted to produce light-sensitive proteins. Once these proteins are expressed, neural activity can be controlled by exposing the cells to light of appropriate wavelengths. Optogenetics provides a unique combination of features, including multimodal control over neural function and genetic targeting of specific cell-types. Together, these versatile features combine to a powerful experimental approach, suitable for the study of the circuitry of psychiatric and neurological disorders. The advent of optogenetics was followed by extensive research aimed to produce new lines of light-sensitive proteins and to develop new technologies: for example, to control the distribution of light inside the brain tissue or to combine optogenetics with other modalities including electrophysiology, electrocorticography, nonlinear microscopy, and functional magnetic resonance imaging. In this paper, the authors review some of the recent advances in the field of optogenetics and related technologies and provide their vision for the future of the field.United States. Defense Advanced Research Projects Agency (Space and Naval Warfare Systems Center, Pacific Grant/Contract No. N66001-12-C-4025)University of Wisconsin--Madison (Research growth initiative; grant 101X254)University of Wisconsin--Madison (Research growth initiative; grant 101X172)University of Wisconsin--Madison (Research growth initiative; grant 101X213)National Science Foundation (U.S.) (MRSEC DMR-0819762)National Science Foundation (U.S.) (NSF CAREER CBET-1253890)National Institutes of Health (U.S.) (NIH/NIBIB R00 Award (4R00EB008738)National Institutes of Health (U.S.) (NIH Director’s New Innovator award (1-DP2-OD002989))Okawa Foundation (Research Grant Award)National Institutes of Health (U.S.) (NIH Director’s New Innovator Award (1DP2OD007265))National Science Foundation (U.S.) (NSF CAREER Award (1056008)Alfred P. Sloan Foundation (Fellowship)Human Frontier Science Program (Strasbourg, France) (Grant No. 1351/12)Israeli Centers of Research Excellence (I-CORE grant, program 51/11)MINERVA Foundation (Germany

DEVELOPEMENT OF WIDEFIELD MULTI-CONTRAST OPTICAL METHODS FOR IN VIVO MICROVASCULAR SCALE IMAGING

Traditional in vivo optical imaging methods rely on a single contrast mechanism, thereby limiting one’s ability to characterize more than one biological variable. However, most biological systems are complex and are comprised of multiple variables. Therefore, optical methods that employ multiple contrast mechanisms and are capable of visualizing multiple biological variables would permit a more comprehensive understanding of biological systems. Multi-contrast optical imaging, therefore, has great potential for both fundamental and applied biomedical research. The goal of this dissertation is to develop optical methods to enable multi-contrast imaging in vivo over a wide field of view while retaining a microvascular scale spatial resolution. We present the integration of three types of optical imaging contrast mechanisms: fluorescence (FL), intrinsic optical signals (IOS) and laser speckle contrast (LSC). Fluorescence enables tracking pre-labelled molecules and cells, IOS allow quantification of blood volume and/or intravascular oxygen saturation, and LSC permits assessment of tissue perfusion. Together, these contrast mechanisms can be harnessed to provide a more complete picture of the underlying physiology at the microvascular spatial scale. We developed two such microvascular resolution optical multi-contrast imaging methods, and demonstrated their utility in multiple biomedical applications. First, we developed a multi-contrast imaging system that can interrogate in vivo both neural activity and its corresponding microvascular scale hemodynamics in the brain of a freely moving rodent. To do this, we miniaturized an entire benchtop optical imaging system that would typically occupy 5 x 5 x 5 feet, into just 5 cm3. Our miniaturized microscope weighs only 9 g. The miniature size and light weight permitted us to mount our microscope on a rodent’s head and image brain activity in vivo with multiple contrast mechanisms. We used our microscope to study the functional activation of the mouse auditory cortex, and to investigate the alteration of brain function during arousal from deep anesthesia. Our miniaturized microscope is the world’s first rodent head-mountable imaging system capable of interrogating both neural and hemodynamic brain activity. We envision our microscope to usher an exciting new era in neuroscience research. Second, we developed an optical imaging system to extensively characterize microvascular scale hemodynamics in vivo in an orthotopic breast tumor model. We specifically designed it as a benchtop based system to allow ample space for surgical preparation and small animal manipulation. Using it, we continuously monitored in vivo microvascular scale changes in tissue perfusion, blood volume and intravascular oxygen saturation of an orthotopic breast tumor microenvironment for multiple hours over a field of view encompassing the entire tumor extent. This unique dataset enabled us for the first time to characterize the temporal relationship between different tumor hemodynamic variables at the scale of individual microvessels. We envision our work to inspire a whole new avenue of experimental cancer research where the role of a tumor’s hemodynamic microenvironment is extensively characterized at its native (i.e. microvascular) spatial scale. In summary, this dissertation describes the design, implementation and demonstration of two microvascular resolution, wide-field, multi-contrast optical imaging systems. We believe these methods to be a new tool for broadening our understanding of biology

MULTI-MODAL OPTICAL NEUROIMAGING AND APPLICATIONS

Optical imaging tools provide superior details than MRI, PET in monitoring the physiological and pathological state of brain in preclinical models. By combining different optical imaging modalities, a variety of physiological parameters (e.g. cerebral hemodynamics, metabolism and neuronal activity) could be detected simultaneously; such simultaneous imaging is expected to profoundly enhance our understanding of normal brain regulation and its disruption from neurovascular disorders. As a part of this thesis, I designed a multi-modal optical imaging system that could perform simultaneous laser speckle contrast imaging, wide-field fluorescence imaging and optical intrinsic signal imaging. The principles, processing methods and applications of these imaging modalities are presented in Chapter 1. The system was first applied to study a new atherothrombotic stroke model and to evaluate the recovery of stroke from different treatment protocols in mice (Chapter 2). Cerebral blood flow changes and thrombus formations were imaged by laser speckle contrast imaging and wide-field fluorescence imaging, respectively. We concluded that the combination treatment of tissue plasminogen activator and cathepsin G inhibitor improved the neurological outcomes of ischemic brain injury from induced atherothrombotic stroke. To investigate brain activity in high-resolution by optical imaging tools, cranial window preparation is an essential procedure to allow optical access to the brain. We also employed the optical imaging system to investigate the effects of cranial windows on monitoring neurovasculature by laser speckle contrast imaging (Chapter 3). Open-skull and thin-skull cranial window procedures were performed in separate experiments, and the neurovasculature underlying the cranial windows were monitored for fourteen days. The differences between two window types were systematically compared by parameters such as contrast-to-noise ratio and microvessel density. Finally, the last part of my thesis was to miniaturize the multi-modal bench-top imaging system to a head-mounted microscope, which allows imaging on awake freely moving animals. The natural physiological state of brain activities can be detected without the confounding effects of anesthetics. The current version of the microscope weighs less than 5 g and is able to perform laser speckle contrast imaging, wide-field fluorescence imaging and optical intrinsic signal imaging simultaneously. We are currently testing the miniaturized microscope to study a brain tumor murine model. Finally, I describe the current progress of miniaturized optical neuroimaging systems on awake moving animals in Chapter 4 of this thesis