Nos gustaría agradecer Riceberg Justin para la asistencia técnica en las pruebas de la headstage. Este trabajo fue apoyado en parte por los Institutos Nacionales de Salud subvenciones MH65658, MH073689, y F30 AG034003-01A1, y el Mount Sinai School of Medicine.

1. Headstage Diseño y Fabricación general Un cabezal completado pre-amplificador del sistema se compone de los siguientes componentes: <ol><li> Una computadora diseñada placa de circuito integrado con cuatro canales de grabación y dos canales de estimulación. </li><li> Electrodos que transmiten las grabaciones y canales de estimulación para los conectores adecuados en la etapa de la cabeza los animales. </li><li> Un cable de amarre y una interfaz que transmite la grabación y canales de estimulación a un colector de anillos. </li><li> Dos formas de impermeabilización en la etapa de la cabeza y en el circuito integrado. </li></ol> 2. Diseño y maquetación de la Headstage Hemos diseñado una placa de circuito impreso integrado con cuatro seguidores de origen y pasan a través de dos canales de estimulación con un producto comercial de software CAD paquete (www.expresspcb.com). Una headstage está diseñado, y el patrón de diseño es de baldosas a través de una hoja más grande de la placa de circuito para proporcionar headstages muchos a un menor costo. El archivo CAD se transmite a la placa de circuito impreso (PCB) en el servicio de fabricación de doble cara, dos tablas de capa se imprimen con capas de cobre superior e inferior con todos los orificios pasantes recubiertos. Las placas usan estaño / soldadura de plomo chapado en rastros y las pastillas que corresponden al estándar de la industria FR-4 laminado. El PCB se pueden pedir y se envían el siguiente día hábil. Headstages individuales se cortan de la hoja grande de azulejos PCB y componentes electrónicos están conectados con soldadura. Un ejemplo de disposición de headstage utilizados en este protocolo se muestra (Figura 1). Por favor, consulte el sitio web del fabricante para obtener instrucciones detalladas y consejos sobre el diseño de un PCB. 3. Preparación y soldadura de componentes electrónicos Todos los componentes electrónicos utilizados en este protocolo son de la superficie del dispositivo de montaje (SMD) de tipo. La huella de cada tipo de dispositivo (resistencia, condensador, etc) fue elegido para minimizar el espacio desperdiciado en el headstage, pero más grandes componentes SMD se pueden utilizar si es necesario. <ol><li> Organizar los componentes en la placa de circuito y los preparan para la soldadura. </li><li> Coloque una pequeña cantidad de estaño en la plataforma de estaño / plomo para ser soldadas y acerque la punta de la estrecha cautín para derretir la soldadura y se adhieren a la plataforma. </li><li> Soldar los condensadores primera resistencias, a continuación, y luego los amplificadores operacionales. </li><li> Los amplificadores operacionales pueden ser destruidos por sobrecalentamiento. Se utilizó una pistola de calor variable de soldadura y el menor calor necesario para fundir la soldadura. Minimizar el tiempo de calentamiento usado para hacer una buena soldadura. Nos pareció que era conveniente crear un pequeño depósito de soldadura que desemboca en la plataforma de la huella mediante la aplicación de soldadura a lo largo de las señales de comunicación o el poder. </li><li> Prueba de que la resistencia a través de sus detractores se reúne el valor esperado con un óhmetro. </li><li> Inspeccione cada pad de cerca para asegurarse de que no es la soldadura de conexión o puente almohadillas adyacentes. Si hay un puente de soldadura, el calor y fundir la soldadura y el uso de una pinza para romper el puente. </li></ol> 4. Montaje y fabricación de interfaces implante y la cuerda <ol><li> Insertar un tubo de 23 Ga hipodérmica en uno de los puertos no utilizados de la toma de 9 pines ABS para el soporte. </li><li> Corte seis piezas de 1 cm de longitud de cable con aislamiento magnético para la conexión de los pines amphenol mujeres a la toma de beneficios. </li><li> Soldar los pines hembra para el cable magnético usando una pequeña cantidad de estaño en cada pin, y calentando el pin de la punta. </li><li> Suelde el otro extremo de cada cable magnético en las tomas de beneficios. </li><li> La posición del hombre - pines hembra millMAX sobre las almohadillas en la parte de sujeción de la headstage, y apretar las clavijas en el PCB para asegurar un ajuste de presión. Entonces, la soldadura de cada pin a cada pad en orden. </li><li> La posición de la tuerca en el conector de implante neural y empujar todos los bolos femeninos en las ranuras que desee en la base de plástico. </li><li> Aplique el pegamento acrílico (krazy pegamento) para la parte superior del conector hembra ABS y seguro a la base de la headstage. Pegue el tubo 23Ga al lado de la correa headstage de apoyo. </li></ol> 5. La impermeabilización de los Headstage <ol><li> Para impermeabilizar la headstage, encierran todo el circuito integrado y las superficies metálicas expuestas con 5 minutos de epoxy. 5-10 minutos para el secado inicial, y 60 minutos para el secado permanente. </li><li> Masilla cartel Tack se debe aplicar a la superficie de los 9 pines ABS toma alrededor de los clavos que sobresalen de las mujeres al final de los animales. </li><li> Para la impermeabilización de máxima, la toma de ABS y ABS plug deberán acoplarse y la tuerca debe apretarse ser. Una pequeña franja de parafina debe ser envuelto alrededor de la interfaz para evitar que el agua entre en contacto con los pines. </li></ol> Secretos para el Éxito <ol><li> Al soldar el amplificador operacional, no recalentar el envase de plástico. Si hayse está quemando o visible de fusión, retire el amplificador y reemplazarlo. La aplicación de calor excesivo puede dañar el amplificador operacional sin derretirse visible. Prueba de cada canal con un generador de funciones después de la soldadura. </li><li> Asegúrese de que no hay conexiones perdidas o puentes de soldadura en la headstage. Mecha de soldadura se puede utilizar para eliminar los componentes o callejeros soldadura. </li></ol> 6. Resultados representante Una buena grabación seguirá una señal de fuente de entrada sin cortar y salir durante el movimiento. Para probar la headstage, el usuario debe utilizar un generador de funciones como una entrada a cada pin de registro y evaluación de las formas de onda en el extremo Mill Max de salida de cabecera. El movimiento físico de los headstage no debe influir en la forma de formas de onda registradas. <img src="/files/ftp_upload/2155/2155fig1.jpg" alt="Figura 1" /> Figura 1. (A) Representación esquemática de la capa superior de la placa de circuito impreso (PCB). Las líneas rojas indican rastros de cobre. Las líneas amarillas indican la capa de serigrafía opcional para el posicionamiento de las piezas electrónicas. Se muestran la ubicación del amplificador operacional (central cuadrado amarillo), resistencias (pequeños cuadrados amarillos) y los condensadores (el más pequeño cuadrados amarillos). (B) Representación esquemática de la capa inferior de la PCB. Líneas verdes indican rastros de cobre. Gran zona verde llena representa placa de tierra. Un archivo de modelo de este esquema se proporciona para su descarga.

<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>

Agua laberinto tareas basadas en el comportamiento proporcionar herramientas eficaces para probar las habilidades de los animales y las representaciones cognitivas en tiempo real 1. El uso de preamplificadores headstage ligera y rentable es especialmente crítica en las aplicaciones de la neurofisiología del comportamiento implican el laberinto de agua. Debido a escapar de la agua fría proporciona la motivación intrínseca de los animales en estas tareas, su comportamiento puede ser probado en ensayos repe...

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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>
</tr>		</tbody>
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design and construction of a cost effective headstage for simultaneous neural stimulation and recording in the water maze

Preamplificadores Headstage y seguidores de origen son utilizados para estudiar la actividad neuronal en el comportamiento experimentos neurofisiología. Disponible en los productos comerciales suelen ser caros, no se puede personalizar fácilmente, y no sumergibles. Aquí se describe un método para diseñar y construir un cabezal a medida, circuitos integrados de traducción simultánea de 4 canales de grabación neural y la simulación de 2 canales en animales despiertos, de comportarse. El headstage está diseñado con un país libre, disponible en el mercado CAD-tipo de paquete de diseño, y puede ser modificado fácilmente para adaptarse a diferentes escalas (por ejemplo, para añadir los canales). Una placa de circuito impreso personalizado está construido con resistencias de montaje superficial, los condensadores y amplificadores operacionales para la construcción de la ganancia de la unidad fuente circuito seguidor. El headstage se hace a prueba de agua con una combinación de epoxi, parafina y una masilla de caucho sintético. Hemos utilizado con éxito este dispositivo para registrar los potenciales de campo locales y estimular las diferentes regiones del cerebro al mismo tiempo a través de canales independientes en ratas nadando en un laberinto de agua. El costo total es &lt;$ 30/unit y puede ser fabricado fácilmente.

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

Se presenta un método de bajo costo para diseñar y construir un cabezal de luz pre-amplificador del sistema de grabación simultánea neural y la capacidad de estimulación. Este dispositivo puede ser impermeabilizadas para su uso en animales que nadan.

Diseño y construcción de un cabezal rentable para la estimulación simultánea neural y de grabación en el laberinto de agua

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 vistas.  Mount Sinai School of Medicine. Se presenta un método de bajo costo para diseñar y construir un cabezal de luz pre-amplificador del sistema de grabación simultánea neural y la capacidad de estimulación. Este dispositivo puede ser impermeabilizadas para su uso en animales que nadan.

Video: 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.

Agua Morris laberinto de prueba para los déficits de aprendizaje y memoria en ratones de Alzheimer modelo de enfermedad

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

Investigación

Neurociencias

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.

Grabación óptica de la actividad neural suprathreshold con la resolución de una sola célula y único punta-

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 Cell Patch Clamp para investigar los mecanismos de la Estimulación Neural infrarrojos

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.

Fabricación de alta densidad, Contacto-Interface plana Electrodos nerviosas para aplicaciones de registro y estimulación

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.

Incremento dependiente del tiempo en la respuesta de la red a la estimulación de cultivos de células neuronales en matrices de microelectrodos

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.

Grabaciones simultáneas de potenciales de campo Local Cortical, Electrocardiograma, electromiograma y respirar a ritmo de una rata libremente móvil

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.

Construcción de un hiperimpulsor mejorado del tetrodo múltiples de grabación Neural a gran escala en comportarse ratas

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.

Construcción e implementación de matrices de microelectrodos de fibra de carbono para grabaciones in vivo crónicas y agudas

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 para modulación optogenética simultánea y grabación neuronal eléctrica

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