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      Three dimensional microelectrodes enable high signal and spatial resolution for neural seizure recordings in brain slices and freely behaving animals

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          Abstract

          Neural recordings made to date through various approaches—both in-vitro or in-vivo—lack high spatial resolution and a high signal-to-noise ratio (SNR) required for detailed understanding of brain function, synaptic plasticity, and dysfunction. These shortcomings in turn deter the ability to further design diagnostic, therapeutic strategies and the fabrication of neuro-modulatory devices with various feedback loop systems. We report here on the simulation and fabrication of fully configurable neural micro-electrodes that can be used for both in vitro and in vivo applications, with three-dimensional semi-insulated structures patterned onto custom, fine-pitch, high density arrays. These microelectrodes were interfaced with isolated brain slices as well as implanted in brains of freely behaving rats to demonstrate their ability to maintain a high SNR. Moreover, the electrodes enabled the detection of epileptiform events and high frequency oscillations in an epilepsy model thus offering a diagnostic potential for neurological disorders such as epilepsy. These microelectrodes provide unique opportunities to study brain activity under normal and various pathological conditions, both in-vivo and in in-vitro, thus furthering the ability to develop drug screening and neuromodulation systems that could accurately record and map the activity of large neural networks over an extended time period.

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          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.
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            Modification of seizure activity by electrical stimulation: II. Motor seizure

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              Acute brain slice methods for adult and aging animals: application of targeted patch clamp analysis and optogenetics.

              The development of the living acute brain slice preparation for analyzing synaptic function roughly a half century ago was a pivotal achievement that greatly influenced the landscape of modern neuroscience. Indeed, many neuroscientists regard brain slices as the gold-standard model system for detailed cellular, molecular, and circuitry level analysis and perturbation of neuronal function. A critical limitation of this model system is the difficulty in preparing slices from adult and aging animals, and over the past several decades few substantial methodological improvements have emerged to facilitate patch clamp analysis in the mature adult stage. In this chapter we describe a robust and practical protocol for preparing brain slices from mature adult mice that are suitable for patch clamp analysis. This method reduces swelling and damage in superficial layers of the slices and improves the success rate for targeted patch clamp recordings, including recordings from fluorescently labeled populations in slices derived from transgenic mice. This adult brain slice method is suitable for diverse experimental applications, including both monitoring and manipulating neuronal activity with genetically encoded calcium indicators and optogenetic actuators, respectively. We describe the application of this adult brain slice platform and associated methods for screening kinetic properties of Channelrhodopsin (ChR) variants expressed in genetically defined neuronal subtypes.
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                Author and article information

                Contributors
                nisyed@ucalgary.ca
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                9 November 2021
                9 November 2021
                2021
                : 11
                : 21952
                Affiliations
                [1 ]GRID grid.22072.35, ISNI 0000 0004 1936 7697, Faculty of Medicine, Hotchkiss Brain Institute, , University of Calgary, ; 2500 University Dr. NW, Calgary, AB T2N 1N4 Canada
                [2 ]GRID grid.22072.35, ISNI 0000 0004 1936 7697, Biomedical Engineering Graduate Program, , University of Calgary, ; 2500 University Dr. NW, Calgary, AB T2N 1N4 Canada
                [3 ]GRID grid.22072.35, ISNI 0000 0004 1936 7697, Alberta Children’s Hospital Research Institute, , University of Calgary, ; 2500 University Dr. NW, Calgary, AB T2N 1N4 Canada
                [4 ]GRID grid.22072.35, ISNI 0000 0004 1936 7697, Department of Cell Biology and Anatomy, , University of Calgary, ; 2500 University Dr. NW, Calgary, AB T2N 1N4 Canada
                [5 ]GRID grid.22072.35, ISNI 0000 0004 1936 7697, Department of Electrical and Computer Engineering, , University of Calgary, ; 2500 University Dr. NW, Calgary, AB T2N 1N4 Canada
                [6 ]GRID grid.266100.3, ISNI 0000 0001 2107 4242, Departments of Neurosciences and Pediatrics, , University of California San Diego, Rady Children’s Hospital, ; San Diego, CA USA
                [7 ]GRID grid.22072.35, ISNI 0000 0004 1936 7697, Cumming School of Medicine, , University of Calgary, ; 3330-Hospital Drive, NW, Calgary, AB T2N 4N1 Canada
                Article
                1528
                10.1038/s41598-021-01528-4
                8578611
                34754055
                a0433858-9766-4623-9d45-e123293d7234
                © The Author(s) 2021

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 1 March 2021
                : 22 October 2021
                Categories
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                © The Author(s) 2021

                Uncategorized
                neural circuits,neuronal physiology,nanobiotechnology,biomedical engineering
                Uncategorized
                neural circuits, neuronal physiology, nanobiotechnology, biomedical engineering

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