Microelectrodes:
Electrodes with an extremely small tip, used in a voltage clamp or other apparatus to stimulate or record bioelectric potentials of single cells intracellularly or extracellularly. (Dorland, 28th ed)
JoVE Bioengineering
Planar and Three-Dimensional Printing of Conductive Inks
1Department of Materials Science and Engineering, University of Illinois at Urbana-Champaign, 2Center for Micro- and Nanotechnology, Lawrence Livermore National Laboratory, 3Presently at the Interdisciplinary Center for Wide Band-gap Semiconductors, University Of California Santa Barbara
Planar and three-dimensional printing of conductive metallic inks is described. Our approach provides new avenues for fabricating printed electronic, optoelectronic, and biomedical devices in unusual layouts at the microscale.
JoVE General
Antifouling Self-assembled Monolayers on Microelectrodes for Patterning Biomolecules
1Department of Physics, Texas A&M University (TAMU), 2Department of Biomedical Engineering, Texas A&M University (TAMU)
We present a procedure for forming a poly(ethylene glycol) self-assembled monolayer (PEG-SAM) on a silicon substrate with gold microelectrodes. The PEG-SAM is formed in a single step and prevents biofouling on silicon and gold surfaces. Electrophoresis is then used for patterning biomolecules down to the nanoscale.
JoVE Neuroscience
Presynaptic Dopamine Dynamics in Striatal Brain Slices with Fast-scan Cyclic Voltammetry
1Department of Chemistry, Wayne State University,
Using fast-scan cyclic voltammetry to measure electrically evoked presynaptic dopamine dynamics in striatal brain slices.
JoVE General
Fabrication of Amperometric Electrodes
1Department of Pharmacological and Physiological Science, Saint Louis University School of Medicine, 2Yale University School of Medicine
This protocol describes how to generate carbon fiber electrodes. The electrodes are subsequently used to detect catecholamine release from vesicles with carbon fiber amperometry.
JoVE Neuroscience
Optical Imaging of Neurons in the Crab Stomatogastric Ganglion with Voltage-sensitive Dyes
1Institute of Neurobiology, Ulm University, 2School of Computing Science & Institute of Neuroscience, Newcastle University
Here we present the methodology for fast and high resolution fluorescent voltage-sensitive dye imaging of detailed activity of neurons in the crab stomatogastric ganglion.
JoVE Neuroscience
Electrophysiological Recordings from the Giant Fiber Pathway of D. melanogaster
1Institute of Healthy Ageing, and GEE, University College London - UCL, 2School of Biosciences, University of Kent
The Giant Fiber System is a simple neuronal circuit of adult Drosophila melanogaster containing the largest neurons in the fly. We describe the protocol for monitoring synaptic transmission through this pathway by recording post synaptic potentials in dorsal longitudinal (DLM) and tergotrochanteral (TTM) muscles following direct stimulation of the Giant Fiber interneurons.
JoVE Bioengineering
High efficiency, Site-specific Transfection of Adherent Cells with siRNA Using Microelectrode Arrays (MEA)
School of Biological and Health Systems Engineering, Arizona State University
The article details the protocol for site-specific transfection of scrambled sequence of siRNA in an adherent mammalian cell culture using a microelectrode array (MEA).
JoVE Neuroscience
Intact Histological Characterization of Brain-implanted Microdevices and Surrounding Tissue
1Weldon School of Biomedical Engineering, Purdue University, 2Department of Biological Sciences, Purdue University
Here we present a histological method for capturing, labeling, optically clearing, and imaging the intact brain tissue interface around chronically implanted microdevices in rodent brain tissue. Results from the techniques comprising this method are useful for understanding the impact of various penetrating brain-implants on their surrounding tissue.
JoVE Clinical and Translational Medicine
Intra-Operative Behavioral Tasks in Awake Humans Undergoing Deep Brain Stimulation Surgery
1Nayef Al-Rodhan Laboratories for Cellular Neurosurgery and Neurosurgical Technology, Harvard Medical School, 2Department of Neurosurgery , Massachusetts General Hospital
Deep brain stimulation surgery offers a unique opportunity to examine information encoding in the awake human brain. This article will describe intra-operative methods used to perform cognitive and behavioral tasks while simultaneously acquiring physiological data such as EMG, single-unit neuronal activity and/or local field potentials.
JoVE Neuroscience
Live-imaging of PKC Translocation in Sf9 Cells and in Aplysia Sensory Neurons
Neurology and Neurosurgery, McGill University
In this video, we demonstrate visualization of PKC translocation in living cells using fluorescently tagged PKCs.
JoVE General
Patterning Cells on Optically Transparent Indium Tin Oxide Electrodes
Department of Biomedical Engineering, University of California, Davis
Non-fouling PEG silane monolayer was desorbed from individually addressable ITO electrodes on glass by application of a reductive potential. Electrochemical stripping of PEG-silane layer from ITO microelectrodes allowed for cell adhesion to take place in a spatially defined fashion, with cellular patterns corresponding closely to electrode patterns.
JoVE Neuroscience
Simultaneous fMRI and Electrophysiology in the Rodent Brain
1Biomedical Engineering, Emory University, 2Biomedical Engineering, Georgia Institute of Technology, 3Biology, Emory University
We have developed a method for simultaneous functional magnetic resonance imaging and electrophysiological recording in the rodent brain, providing a platform for the investigation of the relationship between neural activity and the blood oxygenation level dependent (BOLD) MRI signal.
JoVE Neuroscience
Multi-electrode Array Recordings of Neuronal Avalanches in Organotypic Cultures
Section on Critical Brain Dynamics, National Institute of Mental Health
A robust way to study neuronal avalanches, i.e. scale-invariant spatio-temporal activity bursts, indicative of critical state dynamics in cortex. Avalanches emerge spontaneously in developing superficial layers of cultured cortex which allows for long-term measurements of the activity with planar integrated multi-electrode arrays (MEA) under precisely controlled conditions.
JoVE Neuroscience
Recording Electrical Activity from Identified Neurons in the Intact Brain of Transgenic Fish
Department of Physiology, University of California, Los Angeles
In this video, we will demonstrate how to record electrical activity from identified single neurons in a whole brain preparation, which preserves complex neural circuits. We use transgenic fish in which gonadotropin-releasing hormone (GnRH) neurons are genetically tagged with a fluorescent protein for identification in the intact brain preparation.
JoVE Neuroscience
Electrophysiological Measurements from a Moth Olfactory System
Department of Entomology, University of California, Davis
Insect olfactory systems provide unique opportunities for recording odorant-induced responses in the forms of electroantennograms (EAG) and single sensillum recordings (SSR), which are summed responses from all odorant receptor neurons (ORNs) located on the antenna and from those housed in individual sensilla, respectively.
JoVE Neuroscience
Voltage Biasing, Cyclic Voltammetry, & Electrical Impedance Spectroscopy for Neural Interfaces
1Weldon School of Biomedical Engineering, Purdue University, 2Biomedical Engineering, University of Wisconsin-Madison, 3Biomedical Engineering, University of Michigan, 4Department of Biological Sciences, Purdue University
The electrode-tissue interface of neural recording electrodes can be characterized with electrical impedance spectroscopy (EIS) and cyclic voltammetry (CV). Application of voltage biasing changes the electrochemical properties of the electrode-tissue interface and can improve recording capability. Voltage biasing, EIS, CV, and neural recordings are complementary.
JoVE General
Making Patch-pipettes and Sharp Electrodes with a Programmable Puller
1Department of Molecular and Cellular Physiology, Stanford University, 2Department of Molecular and Cellular Physiology, Stanford University School of Medicine
This video shows how to use a programmable puller to make patch pipettes and sharp electrodes for electrophysiology. The same procedure can be used to make a variety of glass tools, including injection needles.
JoVE Bioengineering
Micropipette Aspiration of Substrate-attached Cells to Estimate Cell Stiffness
1Section of Respiratory, Critical Care and Sleep Medicine, Department of Medicine, University of Illinois, 2Institute for Medicine and Engineering, University of Pennsylvania
Here we describe a quick and simple method to measure cell stiffness. The general principle of this approach is to measure membrane deformation in response to well-defined negative pressure applied through a micropipette to the cell surface. This method provides a powerful tool to study biomechanical properties of substrate-attached cells.
JoVE Neuroscience
Design and Assembly of an Ultra-light Motorized Microdrive for Chronic Neural Recordings in Small Animals
1Center for Brain Science, Harvard University, 2Program in Neuroscience, Harvard University, 3Department of Organismic and Evolutionary Biology, Harvard University
The design, fabrication and assembly of an ultra-light motorized microdrive is described. The device provides a cost-effective and easy-to-use solution for chronic recordings of single units in small behaving animals.
JoVE General
Electrophysiological Methods for Recording Synaptic Potentials from the NMJ of Drosophila Larvae
Here we describe electrophysiological methods for measuring synaptic transmission at the neuromuscular junction of Drosophila larva. Evoked release is initiated artificially by stimulating the motor neuron axons, and transmission through the NMJ can be measured by the postsynaptic response evoked in the muscle.
JoVE General
Measuring Cation Transport by Na,K- and H,K-ATPase in Xenopus Oocytes by Atomic Absorption Spectrophotometry: An Alternative to Radioisotope Assays
1Institute of Chemistry, Technical University of Berlin, 2The Vollum Institute, Oregon Health & Science University
We describe a method to quantify the activity of K+-countertransporting P-type ATPases by heterologous expression of the enzymes in Xenopus oocytes and measuring Rb+ or Li+ uptake into individual cells by atomic absorption spectrophotometry. The method is a sensitive and safe alternative to radioisotope flux experiments facilitating complex kinetic studies.
JoVE Bioengineering
Plasma Lithography Surface Patterning for Creation of Cell Networks
1Aerospace and Mechanical Engineering, University of Arizona, 2Biomedical Engineering IDP and BIO5 Institute, University of Arizona
A versatile plasma lithography technique has been developed to generate stable surface patterns for guiding cellular attachment. This technique can be applied to create cell networks including those that mimic natural tissues and has been used for studying several, distinct cell types.
JoVE Neuroscience
Manufacturing and Using Piggy-back Multibarrel Electrodes for In vivo Pharmacological Manipulations of Neural Responses
Department of Physiology & Biophysics, University of Colorado Medical Campus
Iontophoresis of neural agonists and antagonists during extracellular in vivo recordings is a powerful way to manipulate a neuron’s microenvironment. These manipulations can most easily be done via piggy-back multibarrel electrodes. Here we describe how to manufacture them and use them during auditory recordings.
JoVE Neuroscience
Physiological, Morphological and Neurochemical Characterization of Neurons Modulated by Movement
Department of Neural and Pain Sciences, University of Maryland
A technique is described to quantify the in vivo physiological response of mammalian neurons during movement and correlate the physiology of the neuron with neuronal morphology, neurochemical phenotype and synaptic microcircuitry.
JoVE Neuroscience
Flash Photolysis of Caged Compounds in the Cilia of Olfactory Sensory Neurons
1SISSA, International School for Advanced Studies, 2Istituto di Biofisica, Consiglio Nazionale delle Ricerche, 3SISSA Unit, Italian Institute of Technology
Photolysis of caged compounds allows the production of rapid and localized increases in the concentration of various physiologically active compounds. Here, we show how to obtain patch-clamp recordings combined with photolysis of caged cAMP or caged Ca for the study of olfactory transduction in dissociated mouse olfactory sensory neurons.
JoVE Neuroscience
Behavioral Determination of Stimulus Pair Discrimination of Auditory Acoustic and Electrical Stimuli Using a Classical Conditioning and Heart-rate Approach
School of Psychological Science, La Trobe University
The application of a classical fear conditioning behavioral paradigm for auditory prosthetic research in rats is described. This paradigm provides a mechanism for identifying both detection of, and discrimination between, distinct acoustic and electrical stimuli using heart-rate as an outcome measure.
JoVE Bioengineering
Synthesis, Assembly, and Characterization of Monolayer Protected Gold Nanoparticle Films for Protein Monolayer Electrochemistry
1Department of Chemistry, Gottwald Center for the Sciences, University of Richmond, 2Department of Biochemistry and Molecular Biology, Gottwald Center for the Sciences, University of Richmond
Alkanethiolate stabilized gold colloids known as monolayer protected clusters (MPCs) are synthesized, characterized, and assembled into thin films as an adsorption interface for protein monolayer electrochemistry of simple redox protein like Pseudomonas aeruginosa azurin (AZ) and cytochrome c (cyt c).
JoVE Bioengineering
Hollow Microneedle-based Sensor for Multiplexed Transdermal Electrochemical Sensing
1Joint Department of Biomedical Engineering, University of North Carolina and North Carolina State University, 2Department of Biosensors and Nanomaterials, Sandia National Laboratories
This article details the construction of a multiplexed microneedle-based sensor. The device is being developed for in situ sampling and electrochemical analysis of multiple analytes in a rapid and selective manner. We envision clinical medicine and biomedical research uses for these microneedle-based sensors.
JoVE General
DNA Transfection of Mammalian Skeletal Muscles using In Vivo Electroporation
Department of Physiology, David Geffen School of Medicine, University of California, Los Angeles
We describe detailed procedures for the efficient transfection of plasmid DNA into the fibers of foot muscles of live mice using electroporation and the subsequent visualization of protein expression using fluorescence microscopy.
JoVE General
Xenotransplantation of Human Stem Cells into the Chicken Embryo
1Department of Physiology, University of Oslo, 2Norwegian Center for Stem Cell Research, University of Oslo
In this paper we present a method for transplanting human stem cells into various regions of the central nervous system of the chicken embryo. This provides an in vivo model for assessing the proliferation and differentiation of various types of human stem cells in embryonic tissue environments.
JoVE Bioengineering
Measurement of Bioelectric Current with a Vibrating Probe
Dermatology, University of California, Davis
The manufacture, calibration and use of non-invasive vibrating probes to measure bioelectric current in various biological systems is described.
JoVE Neuroscience
Simultaneous Electroencephalography, Real-time Measurement of Lactate Concentration and Optogenetic Manipulation of Neuronal Activity in the Rodent Cerebral Cortex
A procedure is described for manipulating the activity of cerebral cortical pyramidal neurons optogenetically while the electroencephalogram, electromyogram, and cerebral lactate concentration are monitored. Experimental recordings are performed on cable-tethered mice while they undergo spontaneous sleep/wake cycles. Optogenetic equipment is assembled in our laboratory; recording equipment is commercially available.
JoVE General
Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling
Department of Chemistry and Biochemistry, Worcester Polytechnic Institute
We will describe a method which measures the kinetics of ion transport of membrane proteins alongside site-specific analysis of conformational changes using fluorescence on single cells. This technique is adaptable to ion channels, transporters and ion pumps and can be utilized to determine distance constraints between protein subunits.
JoVE Neuroscience
Time-lapse Imaging of Neuroblast Migration in Acute Slices of the Adult Mouse Forebrain
The Cellular Neurobiology Unit, Centre de Recherche Université Laval Robert-Giffard
We describe a protocol for real-time videoimaging of neuronal migration in the mouse forebrain. The migration of virally-labeled or grafted neuronal precursors was recorded in acute live slices using wide-field fluorescent imaging with a relatively rapid acquisition interval to study the different phases of cell migration, including the durations of the stationary and migration phases and the speed of migration.
JoVE Editorial
December 2011: This Month in JoVE
Here are some highlights from the December 2011 Issue of Journal of Visualized Experiments (JoVE).
JoVE General
How to Culture, Record and Stimulate Neuronal Networks on Micro-electrode Arrays (MEAs)
1Department of Neurology, Emory University School of Medicine, 2Coulter Department of Biomedical Engineering, Laboratory for Neuroengineering, Georgia Institute of Technology and Emory, University School of Medicine, 3Emory University School of Medicine
This protocol provides the necessary information for setting up, caring for, recording from and electrically stimulating cultures on MEAs. In vitro networks provide a means for asking physiologically relevant questions at the network and cellular levels leading to a better understanding of brain function and dysfunction.
JoVE Neuroscience
Modeling Neural Immune Signaling of Episodic and Chronic Migraine Using Spreading Depression In Vitro
1Department of Neurology and Committee on Neurobiology, The University of Chicago Medical Center, 2Department of Neurology, The University of Chicago Medical Center
Migraine and its transformation to chronic migraine are immense healthcare burdens in need of improved treatment options. We seek to define how neural immune signaling modulates the susceptibility to migraine, modeled in vitro using spreading depression in hippocampal slice cultures, as a means to develop novel therapeutic targets.
JoVE Neuroscience
Membrane Potentials, Synaptic Responses, Neuronal Circuitry, Neuromodulation and Muscle Histology Using the Crayfish: Student Laboratory Exercises
1Department of Biology, University of Kentucky, 2Department of Physiology, University of Toronto
The experiments demonstrate an easy approach for students to gain experience in examining muscle structure, synaptic responses, the effects of ion gradients and permeability on membrane potentials. Also, a sensory-CNS-motor-muscle circuit is presented to show a means to test effects of compounds on a neuronal circuit.
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