Nous tenons à remercier Justin Riceberg d&#39;assistance technique dans le test du headstage. Ce travail a été soutenu en partie par les Instituts nationaux de Santé des subventions MH65658, MH073689, et F30-01A1 AG034003, et la Mount Sinai School of Medicine.

1. Conception Headstage et aperçu de fabrication Un headstage complété pré-amplificateur du système comprend les composants suivants: <ol><li> Un ordinateur conçu circuit intégré à quatre canaux d&#39;enregistrement et de deux canaux de stimulation. </li><li> Les électrodes qui transmettent les enregistrements et les canaux de stimulation pour les connecteurs appropriés sur la scène la tête des animaux. </li><li> Un câble d&#39;attache et d&#39;interface qui relaie l&#39;enregistrement et la stimulation des canaux d&#39;un collecteur à bagues. </li><li> Deux formes d&#39;étanchéité sur la scène la tête et sur le circuit intégré. </li></ol> 2. Conception et Mise en forme du Headstage Nous avons conçu un circuit imprimé intégré avec 4 suiveurs source et 2 passent par les canaux de stimulation en utilisant un logiciel de CAO disponibles dans le commerce paquet (www.expresspcb.com). Un headstage est conçu, et le modèle de conception est carrelé sur une grande feuille de circuits pour fournir de nombreux headstages à un coût inférieur. Le fichier CAO est transmis à la carte de circuit imprimé (PCB) de services de fabrication où recto-verso, deux planches sont imprimées en utilisant la couche couches de cuivre en haut et en bas avec tous les trous métallisés. Les conseils utilisent étain / plomb traces plaqués et plaquettes qui correspondent aux standards de l&#39;industrie FR-4 stratifié. Le CCP peut être commandé et livré le jour ouvrable suivant. Headstages individuels sont coupés de la feuille de PCB de grandes tuiles et composants électroniques sont reliés avec de la soudure. Un exemple de schéma de l&#39;headstage utilisé dans ce protocole est illustré (Figure 1). S&#39;il vous plaît voir le site Web du fabricant pour des instructions détaillées et des conseils sur la conception d&#39;un PCB. 3. Préparation et souder des composants électroniques Tous les composants électroniques utilisés dans ce protocole sont de la montagne de périphérique de surface (SMD) de type. L&#39;empreinte de chaque type de périphérique (résistance, condensateur, etc) a été choisie pour minimiser l&#39;espace perdu sur la headstage, mais plus de composants CMS peuvent être utilisées si nécessaire. <ol><li> Organiser vos composants sur le circuit et les préparer pour la soudure. </li><li> Placer une petite quantité de soudure sur le pavé étain / plomb à souder et mettre la pointe de la clôture de fer à souder pour faire fondre et adhérer la soudure au pad. </li><li> Souder les premiers condensateurs, résistances suivant, puis les amplificateurs opérationnels. </li><li> Les amplis op-peut être détruit par un chauffage plus. Nous avons utilisé un pistolet thermique variable de la soudure et le plus bas de la chaleur nécessaire pour faire fondre la soudure. Minimisez le temps de chauffage utilisé pour faire une bonne soudure. Nous avons trouvé utile de créer un petit réservoir de soudure qui se jette dans le pavé empreinte en appliquant la soudure le long de la trace de communication ou d&#39;alimentation. </li><li> Test de la résistance que les résistances au sein de votre rencontre à la valeur attendue aide d&#39;un ohmmètre. </li><li> Inspectez chaque pad en étroite collaboration afin de s&#39;assurer qu&#39;aucune soudure est de connecter ou de combler bloc adjacent. S&#39;il ya un pont de soudure, la chaleur et faire fondre la soudure et d&#39;utiliser une pince à rompre le pont. </li></ol> 4. Fabrication et le montage des interfaces d&#39;implant et Tether <ol><li> Insérez un tube 23 hypodermiques Ga dans l&#39;un des ports inutilisés du socket 9pin ABS pour le support mécanique. </li><li> Coupez six morceaux de 1 cm de longueur de fil magnétique isolé pour connecter les broches femelles Amphenol à la prise de l&#39;ABS. </li><li> Soudez les broches femelles au fil magnétique en utilisant une petite quantité de soudure dans chaque goupille, et en chauffant l&#39;épingle de la pointe. </li><li> Soudez l&#39;autre extrémité de chaque fil magnétique dans les sockets ABS. </li><li> Position du mâle - femelle sur broches MillMax les patins sur le côté d&#39;attache de l&#39;headstage, et presser les broches sur le PCB pour assurer un ajustement de pression. Puis, souder chaque broche à chaque pad dans l&#39;ordre. </li><li> Position de la bague sur le connecteur d&#39;implants neuronaux et pousser toutes les broches femelles dans les fentes désiré sur la base en plastique. </li><li> Appliquez de la colle acrylique (Krazy Glue) pour le haut de la prise du connecteur ABS et le fixer à la base de la headstage. Collez le tube 23Ga au côté d&#39;attache de l&#39;headstage de soutien. </li></ol> 5. Imperméabilisation du Headstage <ol><li> Pour imperméabiliser le headstage, envelopper l&#39;ensemble du circuit intégré et des surfaces métalliques exposées avec 5 époxy minute. Permettez 5-10 minutes pour le séchage initial, et 60 minutes pour le séchage des permanents. </li><li> Mastic Tack affiche devrait être appliqué à la surface de la prise 9 broches ABS autour des broches femelles en saillie de la fin des animaux. </li><li> Pour l&#39;imperméabilisation maximale, la prise et la fiche ABS ABS devrait être accouplés et l&#39;écrou doit être serré. Une petite bande de parafilm doit être enroulé autour de l&#39;interface pour empêcher l&#39;eau de contacter l&#39;connecteurs à broches. </li></ol> Les secrets du succès <ol><li> Lors de la soudure de l&#39;amplificateur opérationnel, ne pas surchauffer l&#39;emballage plastique. S&#39;il yabrûle visible ou de fusion, retirez le ampli-op et de le remplacer. L&#39;application trop de chaleur peut endommager l&#39;ampli op sans fondre visibles. Test de chaque canal avec un générateur de fonction après la soudure. </li><li> Assurez-vous qu&#39;il n&#39;ya pas de connexions errants ou des ponts de soudure sur le headstage. Soudez la mèche peut être utilisée pour supprimer des composants ou errants soudure. </li></ol> 6. Les résultats représentatifs Un bon enregistrement suivra une source de signal d&#39;entrée sans couper dans et hors pendant le mouvement. Pour tester la headstage, l&#39;utilisateur doit utiliser un générateur de fonctionner comme une entrée pour chaque broche d&#39;enregistrement et d&#39;évaluer les formes d&#39;onde à l&#39;extrémité de sortie max Moulin-tête. Physiquement déplaçant le headstage ne devrait pas influencer la forme des courbes enregistrées. <img src="/files/ftp_upload/2155/2155fig1.jpg" alt="Figure 1" /> Figure 1. Vue schématique (A) de la couche supérieure du circuit imprimé (PCB). Les lignes rouges indiquent des traces de cuivre. Les lignes jaunes indiquent option couche de sérigraphie pour le positionnement des pièces électroniques. On voit ici le placement amplificateur opérationnel (central carré jaune), des résistances (petits carrés jaunes) et les condensateurs (petits carrés jaunes). (B) Vue schématique de la couche de fond de PCB. Lignes vertes indiquent des traces de cuivre. Grand espace vert remplie représente la plaque au sol. Un fichier de modèle de ce schéma est fourni en téléchargement.

<ol>
	<li>Morris, R. G., Garrud, P., Rawlins, J. N., O'Keefe, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Place+navigation+impaired+in+rats+with+hippocampal+lesions.">Place navigation impaired in rats with hippocampal lesions.</a> Nature. 297, 681-683 (1982).</li><li>Rolston, J. D., Gross, R. E., Potter, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+low-cost+multielectrode+system+for+data+acquisition+enabling+real-time+closed-loop+processing+with+rapid+recovery+from+stimulation+artifacts.+Front.">A low-cost multielectrode system for data acquisition enabling real-time closed-loop processing with rapid recovery from stimulation artifacts. Front.</a> Neuroengineering. 2, 12-12 (2009).</li><li>Fee, M. S., Leonardo, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Miniature+motorized+microdrive+and+commutator+system+for+chronic+neural+recording+in+small+animals.">Miniature motorized microdrive and commutator system for chronic neural recording in small animals.</a> J. Neurosci. Methods. 112, 83-94 (2001).</li><li>Shirvalkar, P. R., Rapp, P. R., Shapiro, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bidirectional+changes+to+hippocampal+theta+gamma+oscillations+predict+memory+for+recent+spatial+episodes.">Bidirectional changes to hippocampal theta gamma oscillations predict memory for recent spatial episodes.</a> Proc Natl Acad Sci U S A. 107 (15), 7054-709 (2010).</li></ol>

Eau labyrinthe base des tâches comportementales fournir des outils efficaces pour sonder les capacités des animaux et des représentations cognitives en temps réel 1. L&#39;utilisation de matériaux légers et le coût des préamplificateurs headstage efficace est particulièrement critique dans les applications de neurophysiologie comportementale impliquant le labyrinthe d&#39;eau. Parce s&#39;échapper l&#39;eau froide fournit une motivation intrinsèque pour les animaux dans ces tâches, leur comporteme...

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		<div>
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		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Custom designed Integrated Circuit Board</td>
<td>&nbsp;</td>
<td>ExpressPCB.com</td>
<td>Custom</td>
<td>2 layer double sided PCB</td>
</tr>
<tr>
<td>Surface mount Resistors</td>
<td>&nbsp;</td>
<td>Mouser Electronics</td>
<td>RK73B1JTTDD156J</td>
<td>1/10 W, 15 M Ohm 0603, Thick film</td>
</tr>
<tr>
<td>SMD Capacitors </td>
<td>&nbsp;</td>
<td>Newark Electronics</td>
<td>0402YD104KAT2A</td>
<td>0.1 uF (0402)</td>
</tr>
<tr>
<td>SMD Operational Amplifier (quad FET)</td>
<td>&nbsp;</td>
<td>Mouser Electronics</td>
<td>595-TLC2274AIPW</td>
<td>Quad FET - low noise</td>
</tr>
<tr>
<td>9 pin ABS socket</td>
<td>&nbsp;</td>
<td>Ginder Scientific</td>
<td>GS09SKT-220</td>
<td>For animal connection</td>
</tr>
<tr>
<td>9-pin ABS Plug</td>
<td>&nbsp;</td>
<td>Ginder Scientific</td>
<td>GS09PLG-220</td>
<td>To be implanted in animal</td>
</tr>
<tr>
<td>Black Delrin Ring Nut</td>
<td>&nbsp;</td>
<td>Ginder Scientific</td>
<td>GS09BDN-220</td>
<td>&nbsp;</td>
<tr>
<td>Black Delrin Cap Nut</td>
<td>&nbsp;</td>
<td>Ginder Scientific</td>
<td>GS09BDC-220</td>
<td>For protecting animal's implant</td>
</tr>
<tr>
<td>Amphenol pins (male)</td>
<td>&nbsp;</td>
<td>Ginder Scientific</td>
<td>220-P02</td>
<td>&nbsp;</td>
<tr>
<td>Amphenol pins (female)</td>
<td>&nbsp;</td>
<td>Ginder Scientific</td>
<td>220-S02</td>
<td>&nbsp;</td>
<tr>
<td>5-minute epoxy</td>
<td>&nbsp;</td>
<td>Devcon Inc.</td>
<td>S-208 / 20845</td>
<td>&nbsp;</td>
<tr>
<td>Poster Tack Putty</td>
<td>&nbsp;</td>
<td>Elmer's</td>
<td>026000015318</td>
<td>&nbsp;</td>
<tr>
<td>Rosin Core Solder</td>
<td>&nbsp;</td>
<td>Radio Shack</td>
<td>64-009</td>
<td>&nbsp;</td>
<tr>
<td>Soldering Iron </td>
<td>&nbsp;</td>
<td>Weller</td>
<td>Pyropen WSTA6</td>
<td>(adjustable heat)</td>
</tr>
<tr>
<td>Metal tweezers</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>Size 5</td>
<td>&nbsp;</td>
<tr>
<td>Parafilm sheets</td>
<td>&nbsp;</td>
<td>American National Can</td>
<td>Parafilm 4" x 250'</td>
<td>&nbsp;</td>
<tr>
<td>Magnet wire</td>
<td>&nbsp;</td>
<td>Belden</td>
<td>8052</td>
<td>Enamel Coated</td>
</tr>
<tr>
<td>Ammeter/ Multimeter</td>
<td>&nbsp;</td>
<td>Fluke Mfg Co.</td>
<td>77 Multimeter</td>
<td>&nbsp;</td>
<tr>
<td>Millmax Double Row Headers</td>
<td>&nbsp;</td>
<td>Millmax / Mouser</td>
<td>575 - 002101</td>
<td>0.05"</td>
</tr>
<tr>
<td>Solder Wick</td>
<td>&nbsp;</td>
<td>Chemtronics</td>
<td>7-10L</td>
<td>0.75 in.</td>
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design and construction of a cost effective headstage for simultaneous neural stimulation and recording in the water maze

Préamplificateurs Headstage et les adeptes de source sont couramment utilisées pour étudier l&#39;activité neuronale dans des expériences de neurophysiologie comportementale. Disponible produits commerciaux sont souvent coûteux, pas facile à personnaliser, et non submersible. Nous décrivons ici une méthode pour concevoir et construire un sur mesure, headstage circuit intégré pour simultanée de 4 canaux d&#39;enregistrement neuronal et 2 canaux de simulation en éveil, les animaux se comportent. Le headstage est conçu en utilisant un libre, forfait disponible dans le commerce de conception CAD-type, et peuvent être facilement modifiés pour s&#39;adapter à différentes échelles (par exemple pour ajouter des chaînes). Une carte de circuit imprimé personnalisé est construit en utilisant les résistances à montage en surface, des condensateurs et des amplificateurs opérationnels pour construire le gain d&#39;unité source suiveuse circuit. Le headstage est imperméable à l&#39;eau avec une combinaison de résine époxyde, parafilm et un mastic caoutchouc synthétique. Nous avons utilisé avec succès ce dispositif pour enregistrer les potentiels de champ local et stimuler différentes régions du cerveau simultanément via des canaux indépendants chez les rats de nager dans un labyrinthe d&#39;eau. Le coût total est &lt;30/unit $ et peut être fabriqué facilement.

Watch this Scientific Journal Video about Design and Construction of a Cost Effective Headstage for Simultaneous Neural Stimulation and Recording in the Water Maze at JoVE.com

Design and Construction of a Cost Effective Headstage for Simultaneous Neural Stimulation and Recording in the Water Maze

Nous présentons une méthode à faible coût pour concevoir et construire un headstage la lumière pré-amplificateur du système avec enregistrement simultané de neurones et de la capacité de stimulation. Ce dispositif peut être imperméabilisé pour une utilisation chez les animaux de natation.

Conception et construction d&#39;un Headstage rentable pour la stimulation neuronale et enregistrement simultanés dans le labyrinthe de l&#39;eau

Headstage preamplifiers and source followers are commonly used to study neural activity in behavioral neurophysiology experiments. Available commercial ...

design-construction-cost-effective-headstage-for-simultaneous-neural

Research

JoVE Journal

Neuroscience

10.6K vues.  Mount Sinai School of Medicine. Nous présentons une méthode à faible coût pour concevoir et construire un headstage la lumière pré-amplificateur du système avec enregistrement simultané de neurones et de la capacité de stimulation. Ce dispositif peut être imperméabilisé pour une utilisation chez les animaux de natation.

Vidéo: Design and Construction of a Cost Effective Headstage for Simultaneous Neural Stimulation and Recording in the Water Maze

Morris Water Maze Test for Learning and Memory Deficits in Alzheimer&#39;s Disease Model Mice

The Morris Water Maze (MWM) was first established by neuroscientist Richard G. Morris in 1981 in order to test hippocampal-dependent learning, including acquisition of spatial memoryand long-term spatial memory 1. The MWM is a relatively simple procedure typically consisting of six day trials, the main advantage being the differentiation between the spatial (hidden-platform) and non-spatial (visible platform) conditions 2-4. In addition, the MWM testing environment reduces odor trail interference 5. This has led the task to be used extensively in the study of the neurobiology and neuropharmacology of spatial learning and memory. The MWM plays an important role in the validation of rodent models for neurocognitive disorders such as Alzheimer&#x2019;s Disease 6, 7. In this protocol we discussed the typical procedure of MWM for testing learning and memory and data analysis commonly used in Alzheimer&#x2019;s disease transgenic model mice.

Morris Water Maze Test for Learning and Memory Deficits in Alzheimer&#039;s Disease Model Mice

The Morris Water Maze is a behavioral task to test hippocampal-dependent learning and memory. It has been widely used in the study of neurobiology, neuropharmacology and neurocognitive disorders in rodent models.

Test Morris Water Maze pour les déficits d&#39;apprentissage et de mémoire chez la souris modèle de la maladie d&#39;Alzheimer

The Morris Water Maze (MWM) was first established by neuroscientist Richard G. Morris in 1981 in order to test hippocampal-dependent learning, including ...

Recherche

Neurosciences

Optical Recording of Suprathreshold Neural Activity with Single-cell and Single-spike Resolution

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also propagation of suprathreshold spiking activity involves populations of neurons. Empirical studies addressing cortical function directly thus require recordings from populations of neurons with high resolution. Here we describe an optical method and a deconvolution algorithm to record neural activity from up to 100 neurons with single-cell and single-spike resolution. This method relies on detection of the transient increases in intracellular somatic calcium concentration associated with suprathreshold electrical spikes (action potentials) in cortical neurons. High temporal resolution of the optical recordings is achieved by a fast random-access scanning technique using acousto-optical deflectors (AODs)1. Two-photon excitation of the calcium-sensitive dye results in high spatial resolution in opaque brain tissue2. Reconstruction of spikes from the fluorescence calcium recordings is achieved by a maximum-likelihood method. Simultaneous electrophysiological and optical recordings indicate that our method reliably detects spikes (&gt;97% spike detection efficiency), has a low rate of false positive spike detection (&lt; 0.003 spikes/sec), and a high temporal precision (about 3 msec) 3. This optical method of spike detection can be used to record neural activity in vitro and in anesthetized animals in vivo3,4.

Understanding the function of the vertebrate central nervous system requires recordings from many neurons because cortical function arises on the level of populations of neurons. Here we describe an optical method to record suprathreshold neural activity with single-cell and single-spike resolution, dithered random-access scanning. This method records somatic fluorescence calcium signals from up to 100 neurons with high temporal resolution. A maximum-likelihood algorithm deconvolves the underlying suprathreshold neural activity from the somatic fluorescence calcium signals. This method reliably detects spikes with high detection efficiency and a low rate of false positives and can be used to study neural populations in vitro and in vivo.

D&#39;enregistrement optique de l&#39;activité neuronale supraliminaire avec une résolution à cellule unique et un seul pic

Signaling of information in the vertebrate central nervous system is often carried by populations of neurons rather than individual neurons. Also ...

Whole Cell Patch Clamp for Investigating the Mechanisms of Infrared Neural Stimulation

It has been demonstrated in recent years that pulsed, infrared laser light can be used to elicit electrical responses in neural tissue, independent of any further modification of the target tissue. Infrared neural stimulation has been reported in a variety of peripheral and sensory neural tissue in vivo, with particular interest shown in stimulation of neurons in the auditory nerve. However, while INS has been shown to work in these settings, the mechanism (or mechanisms) by which infrared light causes neural excitation is currently not well understood. The protocol presented here describes a whole cell patch clamp method designed to facilitate the investigation of infrared neural stimulation in cultured primary auditory neurons. By thoroughly characterizing the response of these cells to infrared laser illumination in vitro under controlled conditions, it may be possible to gain an improved understanding of the fundamental physical and biochemical processes underlying infrared neural stimulation.

Infrared nerve stimulation has been proposed as an alternative to electrical stimulation in a range of nerve types, including those associated with the auditory system. This protocol describes a patch clamp method for studying the mechanism of infrared nerve stimulation in a culture of primary auditory neurons.

Whole Patch Clamp cellulaire pour étudier les mécanismes de la stimulation neurale infrarouge

It has been  demonstrated in recent years that pulsed, infrared laser light can be used to  elicit electrical responses in neural tissue, independent of ...

Simultaneous Electrophysiological Recording and Calcium Imaging of Suprachiasmatic Nucleus Neurons

Simultaneous electrophysiological and fluorescent imaging recording methods were used to study the role of changes of membrane potential or current in regulating the intracellular calcium concentration. Changing environmental conditions, such as the light-dark cycle, can modify neuronal and neural network activity and the expression of a family of circadian clock genes within the suprachiasmatic nucleus (SCN), the location of the master circadian clock in the mammalian brain. Excitatory synaptic transmission leads to an increase in the postsynaptic Ca2+ concentration that is believed to activate the signaling pathways that shifts the rhythmic expression of circadian clock genes. Hypothalamic slices containing the SCN were patch clamped using microelectrodes filled with an internal solution containing the calcium indicator bis-fura-2. After a seal was formed between the microelectrode and the SCN neuronal membrane, the membrane was ruptured using gentle suction and the calcium probe diffused into the neuron filling both the soma and dendrites. Quantitative ratiometric measurements of the intracellular calcium concentration were recorded simultaneously with membrane potential or current. Using these methods it is possible to study the role of changes of the intracellular calcium concentration produced by synaptic activity and action potential firing of individual neurons. In this presentation we demonstrate the methods to simultaneously record electrophysiological activity along with intracellular calcium from individual SCN neurons maintained in brain slices.

Procedures are described to perform simultaneous recordings of membrane potential or current and changes of intracellular calcium concentration. Suprachiasmatic nucleus neurons are filled with the calcium indicator bis-fura-2 using a patch clamp electrode in the whole cell patch clamp configuration.

Simultaneous electrophysiological and fluorescent imaging recording methods were used to study the role of changes of membrane potential or current in ...

Fabrication of High Contact-Density, Flat-Interface Nerve Electrodes for Recording and Stimulation Applications

Many attempts have been made to manufacture multi-contact nerve cuff electrodes that are safe, robust and reliable for long term neuroprosthetic applications. This protocol describes a fabrication technique of a modified cylindrical nerve cuff electrode to meet these criteria. Minimum computer-aided design and manufacturing (CAD and CAM) skills are necessary to consistently produce cuffs with high precision (contact placement 0.51 &#177; 0.04 mm) and various cuff sizes. The precision in spatially distributing the contacts and the ability to retain a predefined geometry accomplished with this design are two criteria essential to optimize the cuff&#39;s interface for selective recording and stimulation. The presented design also maximizes the flexibility in the longitudinal direction while maintaining sufficient rigidity in the transverse direction to reshape the nerve by using materials with different elasticities. The expansion of the cuff&#39;s cross sectional area as a result of increasing the pressure inside the cuff was observed to be 25% at 67 mm Hg. This test demonstrates the flexibility of the cuff and its response to nerve swelling post-implant. The stability of the contacts&#39; interface and recording quality were also examined with contacts&#39; impedance and signal-to-noise ratio metrics from a chronically implanted cuff (7.5 months), and observed to be 2.55 &#177; 0.25 k&#8486; and 5.10 &#177; 0.81 dB respectively.

This article provides a detailed description on the fabrication process of a high contact-density flat interface nerve electrode (FINE). This electrode is optimized for recording and stimulating neural activity selectively within peripheral nerves.

Fabrication de haute Contactez-Density, Flat-Interface Electrodes nerveuses pour l&#39;enregistrement et de stimulation Applications

Many attempts have been made to manufacture multi-contact nerve cuff electrodes that are safe, robust and reliable for long term neuroprosthetic ...

Time-dependent Increase in the Network Response to the Stimulation of Neuronal Cell Cultures on Micro-electrode Arrays

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network dynamics in neuronal cultures. In contrast with more traditional methods, such as patch-clamping, which can only record activity from a single cell, MEAs can record simultaneously from multiple sites in a network, without requiring the arduous task of placing each electrode individually. Moreover, numerous control and stimulation configurations can be easily applied within the same experimental setup, allowing for a broad range of dynamics to be explored. One of the key dynamics of interest in these in vitro studies has been the extent to which cultured networks display properties indicative of learning. Mouse neuronal cells cultured on MEAs display an increase in response following training induced by electrical stimulation. This protocol demonstrates how to culture neuronal cells on MEAs; successfully record from over 95% of the plated dishes; establish a protocol to train the networks to respond to patterns of stimulation; and sort, plot, and interpret the results from such experiments. The use of a proprietary system for stimulating and recording neuronal cultures is demonstrated. Software packages are also used to sort neuronal units. A custom-designed graphical user interface is used to visualize post-stimulus time histograms, inter-burst intervals, and burst duration, as well as to compare the cellular response to stimulation before and after a training protocol. Finally, representative results and future directions of this research effort are discussed.

Mouse neuronal cells cultured on multi-electrode arrays display an increase in response following electrical stimulation. This protocol demonstrates how to culture neurons, how to record activity, and how to establish a protocol to train the networks to respond to patterns of stimulation.

Augmentation de la réponse du réseau à la stimulation des cultures cellulaires neuronales sur les tableaux de microélectrodes

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network ...

Simultaneous Recordings of Cortical Local Field Potentials, Electrocardiogram, Electromyogram, and Breathing Rhythm from a Freely Moving Rat

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls body functions and internal organ rhythms when animals are exposed to emotional challenges and changes in their living environments. In general experiments, signals from different organs, such as the brain and the heart, are recorded by independent recording systems that require multiple recording devices and different procedures for processing the data files. This study describes a new method that can simultaneously monitor electrical biosignals, including tens of local field potentials in multiple brain regions, electrocardiograms that represent the cardiac rhythm, electromyograms that represent awake/sleep-related muscle contraction, and breathing signals, in a freely moving rat. The recording configuration of this method is based on a conventional micro-drive array for cortical local field potential recordings in which tens of electrodes are accommodated, and the signals obtained from these electrodes are integrated into a single electrical board mounted on the animal's head. Here, this recording system was improved so that signals from the peripheral organs are also transferred to an electrical interface board. In a single surgery, electrodes are first separately implanted into the appropriate body parts and the target brain areas. The open ends of all of these electrodes are then soldered to individual channels of the electrical board above the animal's head so that all of the signals can be integrated into the single electrical board. Connecting this board to a recording device allows for the collection of all of the signals into a single device, which reduces experimental costs and simplifies data processing, because all data can be handled in the same data file. This technique will aid the understanding of the neurophysiological correlates of the associations between central and peripheral organs.

This study introduces a method for the simultaneous recording of local field potentials in the brain, electrocardiograms, electromyograms, and breathing signals of a freely moving rat. This technique, which reduces experimental costs and simplifies data analysis, will contribute to the understanding of the interactions between the brain and peripheral organs.

Enregistrements simultanés des potentiels corticaux champ Local, électrocardiogramme, électromyogramme et respiration rythmique d’un Rat librement mobile

Monitoring the physiological dynamics of the brain and peripheral tissues is necessary for addressing a number of questions about how the brain controls ...

Construction of an Improved Multi-Tetrode Hyperdrive for Large-Scale Neural Recording in Behaving Rats

Monitoring the activity patterns of a large population of neurons over many days in awake animals is a valuable technique in the field of systems neuroscience. One key component of this technique consists of the precise placement of multiple electrodes into desired brain regions and the maintenance of their stability. Here, we describe a protocol for the construction of a 3D-printable hyperdrive, which includes eighteen independently adjustable tetrodes, and is specifically designed for in vivo extracellular neural recording in freely behaving rats. The tetrodes attached to the microdrives can either be individually advanced into multiple brain regions along the track, or can be used to place an array of electrodes into a smaller area. The multiple tetrodes allow for simultaneous examination of action potentials from dozens of individual neurons, as well as local field potentials from populations of neurons in the brain during active behavior. In addition, the design provides for simpler 3D drafting software that can easily be modified for differing experimental needs.

We present the construction of a 3D-printable hyperdrive with eighteen independently adjustable tetrodes. The hyperdrive is designed to record brain activity in freely behaving rats over a period of several weeks.

Construction d’un Hyperdrive tétrode multi amélioré pour un enregistrement neuronaux à grande échelle en comportement des Rats

Monitoring the activity patterns of a large population of neurons over many days in awake animals is a valuable technique in the field of systems ...

Construction and Implementation of Carbon Fiber Microelectrode Arrays for Chronic and Acute In Vivo Recordings

Multichannel electrode arrays offer insight into the working brain and serve to elucidate neural processes at the single-cell and circuit levels. Development of these tools is crucial for understanding complex behaviors and cognition and for advancing clinical applications. However, it remains a challenge to densely record from cell populations stably and continuously over long time periods. Many popular electrodes, such as tetrodes and silicon arrays, feature large cross-diameters that produce damage upon insertion and elicit chronic reactive tissue responses associated with neuronal death, hindering the recording of stable, continuous neural activity. In addition, most wire bundles exhibit broad spacing between channels, precluding simultaneous recording from a large number of cells clustered in a small area. The carbon fiber microelectrode arrays described in this protocol offer an accessible solution to these concerns. The study provides a detailed method for fabricating carbon fiber microelectrode arrays that can be used for both acute and chronic recordings in vivo. The physical properties of these electrodes make them ideal for stable and continuous long-term recordings at high cell densities, enabling the researcher to make robust, unambiguous recordings from single units across months.

Construction and Implementation of Carbon Fiber Microelectrode Arrays for Chronic and Acute In Vivo Recordings

This protocol describes a procedure for constructing carbon fiber microelectrode arrays for chronic and acute in vivo electrophysiological recordings in mouse (Mus musculus) and ferret (Mustela putorius furo) from multiple brain regions. Each step, following the purchase of raw carbon fibers to microelectrode array implantation, is described in detail, with emphasis on microelectrode array construction.

Construction et mise en œuvre de réseaux de microélectrodes en fibre de carbone pour les enregistrements in vivo chroniques et aigus

Multichannel electrode arrays offer insight into the working brain and serve to elucidate neural processes at the single-cell and circuit levels. ...

Optrode Array for Simultaneous Optogenetic Modulation and Electrical Neural Recording

During the last decade, optogenetics has become an essential tool for the investigation of neural signaling due to its unique capability of selective neural modulation or monitoring. As specific types of neuronal cells can be genetically modified to express opsin proteins, optogenetics enables optical stimulation or inhibition of the selected neurons. There have been several technological advances in the optical system for optogenetics. Recently, it was proposed to combine the optical waveguide for light delivery with electrophysiological recording to simultaneously monitor the neural responses to optogenetic stimulation or inhibition. In this study, an implantable optrode array (2x2 optical fibers) was developed with embedded multichannel electrodes. 
A light-emitting diode (LED) was employed as a light source, and a microfabricated microlens array was integrated to provide sufficient light power at the tip of the optical fibers. The optrode array system comprises the disposable part and the reusable part. The disposable part has optical fibers and electrodes, while the reusable part has the LED and electronic circuitry for light control and neural signal processing. The novel design of the implantable optrode array system is introduced in the accompanying video in addition to the procedure of the optrode implantation surgery, optogenetic light stimulation, and the electrophysiological neural recording. The results of in vivo experiments successfully showed time-locked neural spikes evoked by the light stimuli from hippocampal excitatory neurons of mice.

Here, we present the fabrication method of an optrode system with optical fibers for light delivery and an electrode array for neural recording. In vivo experiments with transgenic mice expressing channelrhodopsin-2 show the feasibility of the system for simultaneous optogenetic stimulation and electrophysiological recording.

Optrode Array pour la modulation optogénétique simultanée et l’enregistrement neuronal électrique

During the last decade, optogenetics has become an essential tool for the investigation of neural signaling due to its unique capability of selective ...