Fully integrated silicon probes for high-density recording of neural activity.

Sensory, motor and cognitive operations involve the coordinated action of large neuronal populations across multiple brain regions in both superficial and deep structures. Existing extracellular probes record neural activity with excellent spatial and temporal (sub-millisecond) resolution, but from only a few dozen neurons per shank. Optical Ca2+ imaging offers more coverage but lacks the temporal resolution needed to distinguish individual spikes reliably and does not measure local field potentials. Until now, no technology compatible with use in unrestrained animals has combined high spatiotemporal resolution with large volume coverage. Here we design, fabricate and test a new silicon probe known as Neuropixels to meet this need. Each probe has 384 recording channels that can programmably address 960 complementary metal-oxide-semiconductor (CMOS) processing-compatible low-impedance TiN sites that tile a single 10-mm long, 70 × 20-µm cross-section shank. The 6 × 9-mm probe base is fabricated with the shank on a single chip. Voltage signals are filtered, amplified, multiplexed and digitized on the base, allowing the direct transmission of noise-free digital data from the probe. The combination of dense recording sites and high channel count yielded well-isolated spiking activity from hundreds of neurons per probe implanted in mice and rats. Using two probes, more than 700 well-isolated single neurons were recorded simultaneously from five brain structures in an awake mouse. The fully integrated functionality and small size of Neuropixels probes allowed large populations of neurons from several brain structures to be recorded in freely moving animals. This combination of high-performance electrode technology and scalable chip fabrication methods opens a path towards recording of brain-wide neural activity during behaviour.

Multifield and inverse-contrast switching of magnetocaloric high contrast ratio MRI labels.

PURPOSE: Demonstrating multifield and inverse contrast switching of magnetocaloric high contrast ratio MRI labels that either have increasing or decreasing moment versus temperature slopes depending on the material at physiological temperatures and different MRI magnetic field strengths. METHODS: Two iron-rhodium samples of different purity (99% and 99.9%) and a lanthanum-iron-silicon sample were obtained from commercial vendors. Temperature and magnetic field-dependent magnetic moment measurements of the samples were performed on a vibrating sample magnetometer. Temperature-dependent MRI of different iron-rhodium and lanthanum-iron-silicon samples were performed on 3 different MRI scanners at 1 Tesla (T), 4.7T, and 7T. RESULTS: Sharp, first-order magnetic phase transition of each iron-rhodium sample at a physiologically relevant temperature (~37°C) but at different MRI magnetic fields (1T, 4.7T, and 7T, depending on the sample) showed clear image contrast changes in temperature-dependent MRI. Iron-rhodium and lanthanum-iron-silicon samples with sharp, first-order magnetic phase transitions at the same MRI field of 1T and physiological temperature of 37°C, but with positive and negative slope of magnetization versus temperature, respectively, showed clear inverse contrast image changes. Temperature-dependent MRI on individual microparticle samples of lanthanum-iron-silicon also showed sharp image contrast changes. CONCLUSION: Magnetocaloric materials of different purity and composition were demonstrated to act as diverse high contrast ratio switchable MRI contrast agents. Thus, we show that a range of magnetocaloric materials can be optimized for unique image contrast response under MRI-appropriate conditions at physiological temperatures and be controllably switched in situ.

Magnetocaloric materials as switchable high contrast ratio MRI labels.

PURPOSE: To develop switchable and tunable labels with high contrast ratio for MRI using magnetocaloric materials that have sharp first-order magnetic phase transitions at physiological temperatures and typical MRI magnetic field strengths. METHODS: A prototypical magnetocaloric material iron-rhodium (FeRh) was prepared by melt mixing, high-temperature annealing, and ice-water quenching. Temperature- and magnetic field-dependent magnetization measurements of wire-cut FeRh samples were performed on a vibrating sample magnetometer. Temperature-dependent MRI of FeRh samples was performed on a 4.7T MRI. RESULTS: Temperature-dependent MRI clearly demonstrated image contrast changes due to the sharp magnetic state transition of the FeRh samples in the MRI magnetic field (4.7T) and at a physiologically relevant temperature (~37°C). CONCLUSION: A magnetocaloric material, FeRh, was demonstrated to act as a high contrast ratio switchable MRI contrast agent due to its sharp first-order magnetic phase transition in the DC magnetic field of MRI and at physiologically relevant temperatures. A wide range of magnetocaloric materials are available that can be tuned by materials science techniques to optimize their response under MRI-appropriate conditions and be controllably switched in situ with temperature, magnetic field, or a combination of both.

Detachable glass microelectrodes for recording action potentials in active moving organs.

Here, we describe new detachable floating glass micropipette electrode devices that provide targeted action potential recordings in active moving organs without requiring constant mechanical constraint or pharmacological inhibition of tissue motion. The technology is based on the concept of a glass micropipette electrode that is held firmly during cell targeting and intracellular insertion, after which a 100-µg glass microelectrode, a "microdevice," is gently released to remain within the moving organ. The microdevices provide long-term recordings of action potentials, even during millimeter-scale movement of tissue in which the device is embedded. We demonstrate two different glass micropipette electrode holding and detachment designs appropriate for the heart (sharp glass microdevices for cardiac myocytes in rats, guinea pigs, and humans) and the brain (patch glass microdevices for neurons in rats). We explain how microdevices enable measurements of multiple cells within a moving organ that are typically difficult with other technologies. Using sharp microdevices, action potential duration was monitored continuously for 15 min in unconstrained perfused hearts during global ischemia-reperfusion, providing beat-to-beat measurements of changes in action potential duration. Action potentials from neurons in the hippocampus of anesthetized rats were measured with patch microdevices, which provided stable base potentials during long-term recordings. Our results demonstrate that detachable microdevices are an elegant and robust tool to record electrical activity with high temporal resolution and cellular level localization without disturbing the physiological working conditions of the organ.NEW & NOTEWORTHY Cellular action potential measurements within tissue using glass micropipette electrodes usually require tissue immobilization, potentially influencing the physiological relevance of the measurement. Here, we addressed this limitation with novel 100-µg detachable glass microelectrodes that can be precisely positioned to provide long-term measurements of action potential duration during unconstrained tissue movement.

Quantum dot-based multiphoton fluorescent pipettes for targeted neuronal electrophysiology.

Targeting visually identified neurons for electrophysiological recording is a fundamental neuroscience technique; however, its potential is hampered by poor visualization of pipette tips in deep brain tissue. We describe quantum dot-coated glass pipettes that provide strong two-photon contrast at deeper penetration depths than those achievable with current methods. We demonstrated the pipettes' utility in targeted patch-clamp recording experiments and single-cell electroporation of identified rat and mouse neurons in vitro and in vivo.

Sensitive enhancement of vessel wall imaging with an endoesophageal Wireless Amplified NMR Detector (WAND).

PURPOSE: To improve the imaging quality of vessel walls with an endoesophageal Wireless Amplified NMR Detector (WAND). METHODS: A cylindrically shaped double-frequency resonator has been constructed with a single metal wire that is self-connected by a pair of nonlinear capacitors. The double-frequency resonator can convert wirelessly provided pumping power into amplified MR signals. This compact design makes the detector easily insertable into a rodent esophagus. RESULTS: The detector has good longitudinal and axial symmetry. Compared to an external surface coil, the WAND can enhance detection sensitivity by at least 5 times, even when the distance separation between the region of interest and the detector's cylindrical surface is twice the detector's own radius. Such detection capability enables us to observe vessel walls near the aortic arch and carotid bifurcation with elevated sensitivity. CONCLUSION: A cylindrical MRI detector integrated with a wireless-powered amplifier has been developed as an endoesophageal detector to enhance detection sensitivity of vessel walls. This detector can greatly improve the imaging quality for vessel regions that are susceptible to atherosclerotic lesions. Magn Reson Med 78:2048-2054, 2017. © 2016 International Society for Magnetic Resonance in Medicine.

Robotic multi-probe single-actuator inchworm neural microdrive.

A wide range of techniques in neuroscience involve placing individual probes at precise locations in the brain. However, large-scale measurement and manipulation of the brain using such methods have been severely limited by the inability to miniaturize systems for probe positioning. Here, we present a fundamentally new, remote-controlled micropositioning approach composed of novel phase-change material-filled resistive heater micro-grippers arranged in an inchworm motor configuration. The microscopic dimensions, stability, gentle gripping action, individual electronic control, and high packing density of the grippers allow micrometer-precision independent positioning of many arbitrarily shaped probes using a single piezo actuator. This multi-probe single-actuator design significantly reduces the size and weight and allows for potential automation of microdrives. We demonstrate accurate placement of multiple electrodes into the rat hippocampus in vivo in acute and chronic preparations. Our robotic microdrive technology should therefore enable the scaling up of many types of multi-probe applications in neuroscience and other fields.

Multi-array silicon probes with integrated optical fibers: light-assisted perturbation and recording of local neural circuits in the behaving animal.

Recordings of large neuronal ensembles and neural stimulation of high spatial and temporal precision are important requisites for studying the real-time dynamics of neural networks. Multiple-shank silicon probes enable large-scale monitoring of individual neurons. Optical stimulation of genetically targeted neurons expressing light-sensitive channels or other fast (milliseconds) actuators offers the means for controlled perturbation of local circuits. Here we describe a method to equip the shanks of silicon probes with micron-scale light guides for allowing the simultaneous use of the two approaches. We then show illustrative examples of how these compact hybrid electrodes can be used in probing local circuits in behaving rats and mice. A key advantage of these devices is the enhanced spatial precision of stimulation that is achieved by delivering light close to the recording sites of the probe. When paired with the expression of light-sensitive actuators within genetically specified neuronal populations, these devices allow the relatively straightforward and interpretable manipulation of network activity.

Possible magneto-mechanical and magneto-thermal mechanisms of ion channel activation in magnetogenetics.

The palette of tools for perturbation of neural activity is continually expanding. On the forefront of this expansion is magnetogenetics, where ion channels are genetically engineered to be closely coupled to the iron-storage protein ferritin. Initial reports on magnetogenetics have sparked a vigorous debate on the plausibility of physical mechanisms of ion channel activation by means of external magnetic fields. The criticism leveled against magnetogenetics as being physically implausible is based on the specific assumptions about the magnetic spin configurations of iron in ferritin. I consider here a wider range of possible spin configurations of iron in ferritin and the consequences these might have in magnetogenetics. I propose several new magneto-mechanical and magneto-thermal mechanisms of ion channel activation that may clarify some of the mysteries that presently challenge our understanding of the reported biological experiments. Finally, I present some additional puzzles that will require further theoretical and experimental investigation.

Multimodal in vivo brain electrophysiology with integrated glass microelectrodes.

Electrophysiology is the most used approach for the collection of functional data in basic and translational neuroscience, but it is typically limited to either intracellular or extracellular recordings. The integration of multiple physiological modalities for the routine acquisition of multimodal data with microelectrodes could be useful for biomedical applications, yet this has been challenging owing to incompatibilities of fabrication methods. Here, we present a suite of glass pipettes with integrated microelectrodes for the simultaneous acquisition of multimodal intracellular and extracellular information in vivo, electrochemistry assessments, and optogenetic perturbations of neural activity. We used the integrated devices to acquire multimodal signals from the CA1 region of the hippocampus in mice and rats, and show that these data can serve as ground-truth validation for the performance of spike-sorting algorithms. The microdevices are applicable for basic and translational neurobiology, and for the development of next-generation brain-machine interfaces.

Stripe sensor tomography.

We introduce a general concept of tomographic imaging for the case of an imaging sensor that has a stripelike shape. We first show that there is no difference, in principle, between two-dimensional tomography using conventional electromagnetic or particle radiation and tomography where a stripe sensor is mechanically scanned over a sample at a sequence of different angles. For a single stripe detector imaging, linear motion and angular rotation are required. We experimentally demonstrate single stripe sensor imaging principle using an elongated inductive coil detector. By utilizing an array of parallel stripe sensors that can be individually addressed, two-dimensional imaging can be performed with rotation only, eliminating the requirement for linear motion, as we also experimentally demonstrate with parallel coil array. We conclude that imaging with a stripe-type sensor of particular width and thickness (where the width is much larger than the thickness) is resolution limited only by the thickness (smaller parameter) of the sensor. We give examples of multiple sensor families where this imaging technique may be beneficial such as magnetoresistive, inductive, superconducting quantum interference device, and Hall effect sensors, and, in particular, discuss the possibilities of the technique in the field of magnetic resonance imaging.

Evaluating the potential of using quantum dots for monitoring electrical signals in neurons.

Success in the projects aimed at providing an advanced understanding of the brain is directly predicated on making critical advances in nanotechnology. This Perspective addresses the unique interface of neuroscience and nanomaterials by considering the foundational problem of sensing neuron membrane voltage and offers a potential solution that may be facilitated by a prototypical nanomaterial. Despite substantial improvements, the visualization of instantaneous voltage changes within individual neurons, whether in cell culture or in vivo, at both the single-cell and network level at high speed remains complex and problematic. The unique properties of semiconductor quantum dots (QDs) have made them powerful fluorophores for bioimaging. What is not widely appreciated, however, is that QD photoluminescence is exquisitely sensitive to proximal electric fields. This property should be suitable for sensing voltage changes that occur in the active neuronal membrane. Here, we examine the potential role of QDs in addressing the important challenge of real-time optical voltage imaging.

Stray field magnetic resonance tomography using ferromagnetic spheres.

The methodology for obtaining two- and three-dimensional magnetic resonance images by using azimuthally symmetric dipolar magnetic fields from ferromagnetic spheres is described. We utilize the symmetric property of a geometric sphere in the presence of a large externally applied magnetic field to demonstrate that a complete two- or three-dimensional structured rendering of a sample can be obtained without the motion of the sample relative to the sphere. Sequential positioning of the integrated sample-sphere system in an external magnetic field at various angular orientations provides all the required imaging slices for successful computerized tomographic image reconstruction. The elimination of the requirement to scan the sample relative to the ferromagnetic tip in this imaging protocol is a potentially valuable simplification compared to previous scanning probe magnetic resonance imaging proposals.

Nanomagnetic planar magnetic resonance microscopy "lens".

The achievement of three-dimensional atomic resolution magnetic resonance microscopy remains one of the main challenges in the visualization of biological molecules. The prospects for single spin microscopy have come tantalizingly close due to the recent developments in sensitive instrumentation. Despite the single spin detection capability in systems of spatially well-isolated spins, the challenge that remains is the creation of conditions in space where only a single spin is resonant and detected in the presence of other spins in its natural dense spin environment. We present a nanomagnetic planar design where a localized Angstrom-scale point in three-dimensional space is created above the nanostructure with a nonzero minimum of the magnetic field magnitude. The design thereby represents a magnetic resonance microscopy "lens" where potentially only a single spin located in the "focus" spot of the structure is resonant. Despite the presence of other spins in the Angstrom-scale vicinity of the resonant spin, the high gradient magnetic field of the "lens" renders those spins inactive in the detection process.

Magnetic nanostructures as amplifiers of transverse fields in magnetic resonance.

We introduce the concept of amplifying the transverse magnetic fields produced and/or detected with inductive coils in magnetic resonance settings by using the reversible transverse susceptibility properties of magnetic nanostructures. First, we describe the theoretical formalism of magnetic flux amplification through the coil in the presence of a large perpendicular DC magnetic field (typical of magnetic resonance systems) achieved through the singularity in the reversible transverse susceptibility in anisotropic single domain magnetic nanoparticles. We experimentally demonstrate the concept of transverse magnetic flux amplification in an inductive coil system using oriented nanoparticles with uni-axial magnetic anisotropy. We also propose a composite ferromagnetic/anti-ferromagnetic core/shell nanostructure system with uni-directional magnetic anisotropy that, in principle, provides maximal transverse magnetic flux amplification.

Composite nanowire-based probes for magnetic resonance force microscopy.

We present a nanowire-based methodology for the fabrication of ultrahigh sensitivity and resolution probes for atomic resolution magnetic resonance force microscopy (MRFM). The fabrication technique combines electrochemical deposition of multifunctional metals into nanoporous polycarbonate membranes and chemically selective electroless deposition of optical nanoreflector onto the nanowire. The completed composite nanowire structure contains all the required elements for an ultrahigh sensitivity and resolution MRFM sensor with (a) a magnetic nanowire segment providing atomic resolution magnetic field imaging gradients as well as large force gradients for high sensitivity, (b) a noble metal enhanced nanowire segment providing efficient scattering cross-section from a sub-wavelength source for optical readout of nanowire vibration, and (c) a nonmagnetic/nonplasmonic nanowire segment providing the cantilever structure for mechanical detection of magnetic resonance.