ABSTRACT Sparse sequences of neuronal activity are fundamental features of neural circuit computation; however, the underlying homeostatic mechanisms remain poorly understood. To approach these questions, we have developed a method for cellular-resolution imaging in organotypic cultures of the adult zebra finch brain, including portions of the intact song circuit. These in vitro networks can survive for weeks, and display mature neuron morphologies. Neurons within the organotypic slices exhibit a diversity of spontaneous and pharmacologically induced activity that can be easily monitored using the genetically encoded calcium indicator GCaMP6. In this study, we primarily focus on the classic song sequence generator HVC and the surrounding areas. We describe proof of concept experiments including physiological, optical, and pharmacological manipulation of these exposed networks. This method may allow the cellular rules underlying sparse, stereotyped neural sequencing to be examined with new degrees of experimental control. Highlights Organotypic brain slices from adult zebra finch (Taeniopygia guttata), expressing the calcium indicator GCaMP6, can be cultured and maintained for at least several weeks and display spontaneous and evoked calcium transients.
Applying an accurate parametric prediction model to identify abnormal or false pressurizer water levels (PWLs) is critical to the safe operation of marine pressurized water reactors (PWRs). Recently, deep-learning-based models have proved to be a powerful feature extractor to perform high-accuracy prediction. However, the effectiveness of models still suffers from two issues in PWL prediction: the correlations shifting over time between PWL and other feature parameters, and the example imbalance between fluctuation examples (minority) and stable examples (majority). To address these problems, we propose a cost-sensitive mechanism to facilitate the model to learn the feature representation of later examples and fluctuation examples. By weighting the standard mean square error loss with a cost-sensitive factor, we develop a Cost-Sensitive Long Short-Term Memory (CSLSTM) model to predict the PWL of PWRs. The overall performance of the CSLSTM is assessed by a variety of evaluation metrics with the experimental data collected from a marine PWR simulator. The comparisons with the Long Short-Term Memory (LSTM) model and the Support Vector Regression (SVR) model demonstrate the effectiveness of the CSLSTM.
Objective. The vision of bioelectronic medicine is to treat disease by modulating the signaling of visceral nerves near various end organs. In small animal models, the nerves of interest can have small diameters and limited surgical access. New high-resolution methods for building nerve interfaces are desirable. In this study, we present a novel nerve interface and demonstrate its use for stimulation and recording in small nerves. Approach. We design and fabricate micro-scale electrode-laden nanoclips capable of interfacing with nerves as small as 50 µm in diameter. The nanoclips are fabricated using a direct laser writing technique with a resolution of 200 nm. The resolution of the printing process allows for incorporation of a number of innovations such as trapdoors to secure the device to the nerve, and quick-release mounts that facilitate keyhole surgery, obviating the need for forceps. The nanoclip can be built around various electrode materials; here we use carbon nanotube fibers for minimally invasive tethering. Main results. We present data from stimulation-evoked responses of the tracheal syringeal (hypoglossal) nerve of the zebra finch, as well as quantification of nerve functionality at various time points post implant, demonstrating that the nanoclip is compatible with healthy nerve activity over sub-chronic timescales. Significance. Our nerve interface addresses key challenges in interfacing with small nerves in the peripheral nervous system. Its small size, ability to remain on the nerve over sub-chronic timescales, and ease of implantation, make it a promising tool for future use in the treatment of disease.
Objective. Most preparations for making neural recordings degrade over time and eventually fail due to insertion trauma and reactive tissue response. The magnitudes of these responses are thought to be related to the electrode size (specifically, the cross-sectional area), the relative stiffness of the electrode, and the degree of tissue tolerance for the material. Flexible carbon fiber ultra-microelectrodes have a much smaller cross-section than traditional electrodes and low tissue reactivity, and thus may enable improved longevity of neural recordings in the central and peripheral nervous systems. Only two carbon fiber array designs have been described previously, each with limited channel densities due to limitations of the fabrication processes or interconnect strategies. Here, we describe a method for assembling carbon fiber electrodes on a flexible polyimide substrate that is expected to facilitate the construction of high-density recording and stimulating arrays. Approach. Individual carbon fibers were aligned using an alignment tool that was 3D-printed with sub-micron resolution using direct laser writing. Indium deposition on the carbon fibers, followed by low-temperature microsoldering, provided a robust and reliable method of electrical connection to the polyimide interconnect. Main results. Spontaneous multiunit activity and stimulation-evoked compound responses with SNR >10 and >120, respectively, were recorded from a small (125 µm) peripheral nerve. We also improved the typically poor charge injection capacity of small diameter carbon fibers by electrodepositing 100 nm-thick iridium oxide films, making the carbon fiber arrays usable for electrical stimulation as well as recording. Significance. Our innovations in fabrication technique pave the way for further miniaturization of carbon fiber ultra-microelectrode arrays. We believe these advances to be key steps to enable a shift from labor intensive, manual assembly to a more automated manufacturing process.
Abstract Most preparations for making neural recordings degrade over time and eventually fail due to insertion trauma and reactive tissue response. The magnitudes of these responses are thought to be related to the electrode size (specifically, the cross-sectional area) and the relative stiffness of the electrode material. Carbon fiber ultramicroelectrodes have a much smaller cross-section than traditional electrodes and thus may enable improved longevity of neural recordings in the central and peripheral nervous systems. Only two carbon fiber array designs have been described previously, each with limited channel densities due to limitations of the fabrication processes or interconnect strategies. Here, we describe a method for assembling carbon fiber electrodes on a flexible polyimide substrate that will facilitate the construction of high-density recording and stimulating arrays for acute use in peripheral nerves. Fibers were aligned using an alignment tool that was 3D-printed with sub-micron resolution using direct laser writing. Indium deposition on the carbon fibers provided a robust and reliable method of electrical connection to the polyimide traces. Spontaneous action potentials and stimulation-evoked compound responses with SNR > 10 and > 120, respectively, were recorded from a small (125 μm) peripheral nerve. We also improved the typically poor charge injection capacity of small diameter carbon fibers can be improved by electrodepositing 100 nm thick iridium oxide films, making the carbon fiber arrays suitable for electrical stimulation as well as recording.
Abstract Coordinated skills such as speech or dance involve sequences of actions that follow syntactic rules in which transitions between elements depend on past actions. Canary songs are comprised of repeated syllables, called phrases, and the ordering of these phrases follows long-range rules, where the choice of what to sing depends on song structure many seconds prior. The neural substrates that support these long-range correlations are unknown. Using miniature head-mounted microscopes and cell-type-specific genetic tools, we observed neural activity in the premotor nucleus HVC as canaries explore various phrase sequences in their repertoire. We find neurons that encode past transitions, extending over 4 phrases and spanning up to 3 seconds and 40 syllables. These neurons preferentially encode past actions rather than future actions, can reflect more than a single song history, and occur mostly during the rare phrases that involve history-dependent transitions in song. These findings demonstrate that network dynamics in HVC reflect preceding behavior context relevant to flexible transitions.
Optical techniques for recording and manipulating neural activity have traditionally been constrained to superficial brain regions due to light scattering. New techniques are needed to extend optical access to large 3D volumes in deep brain areas, while retaining local connectivity.We have developed a method to implant bundles of hundreds or thousands of optical microfibers, each with a diameter of 8 μm. During insertion, each fiber moves independently, following a path of least resistance. The fibers achieve near total internal reflection, enabling optically interfacing with the tissue near each fiber aperture.At a depth of 3 mm, histology shows fibers consistently splay over 1 mm in diameter throughout the target region. Immunohistochemical staining after chronic implants reveals neurons in close proximity to the fiber tips. Models of photon fluence indicate that fibers can be used as a stimulation light source to precisely activate distinct patterns of neurons by illuminating a subset of fibers in the bundle. By recording fluorescent beads diffusing in water, we demonstrate the recording capability of the fibers.Our histology, modeling and fluorescent bead recordings suggest that the optical microfibers may provide a minimally invasive, stable, bidirectional interface for recording or stimulating genetic probes in deep brain regions-a hyper-localized form of fiber photometry.
'/%3e%3cuse xlink:href='%23a' transform='rotate(23.036 .093 25.536)'/%3e%3cuse xlink:href='%23a' transform='rotate(45.87 1.273 16.18)'/%3e%3cuse xlink:href='%23a' transform='rotate(69.945 .996 12.078)'/%3e%3cuse xlink:href='%23a' transform='rotate(20.66 -19.689 31.932)'/%3e%3c/svg%3e)