<meta charset="utf8"></head><body>Induzione e registrazione coerenti dell'attività convulsiva utilizzando CMOS-HD-MEA

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>2D Workbench</td>
			<td>Cloudray</td>
			<td>LM04CLLD26B</td>
			<td></td>
		</tr>
		<tr>
			<td>4-Aminopyridine</td>
			<td>Sigma-Aldrich</td>
			<td>275875</td>
			<td></td>
		</tr>
		<tr>
			<td>Accura Chip</td>
			<td>3Brain</td>
			<td>Accura HD-MEA</td>
			<td>CMOS-HD-MEA chip</td>
		</tr>
		<tr>
			<td>Agarose</td>
			<td>Thermo Fisher Scientific</td>
			<td>BP160-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Vibration isolation table</td>
			<td>Kinetic Systems</td>
			<td>91010124</td>
			<td></td>
		</tr>
		<tr>
			<td>Beaker for the slice holding chamber, 270 mL</td>
			<td>VWR</td>
			<td>10754-772</td>
			<td></td>
		</tr>
		<tr>
			<td>BioCam</td>
			<td>3Brain</td>
			<td>BioCAM DupleX</td>
			<td>CMOS-HD-MEA platform</td>
		</tr>
		<tr>
			<td>Brainwave Software</td>
			<td>3Brain</td>
			<td>Version 4</td>
			<td>CMOS-HD-MEA software</td>
		</tr>
		<tr>
			<td>Calcium Chloride</td>
			<td>Thermo Fisher Scientific</td>
			<td>BP510-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Carbogen</td>
			<td>Airgas</td>
			<td>X02OX95C2003102</td>
			<td></td>
		</tr>
		<tr>
			<td>Carbogen</td>
			<td>Airgas</td>
			<td>12005</td>
			<td></td>
		</tr>
		<tr>
			<td>Carbogen Stones</td>
			<td>Supelco</td>
			<td>59277</td>
			<td></td>
		</tr>
		<tr>
			<td>Compresstome</td>
			<td>Precissionary</td>
			<td>VF-300-0Z</td>
			<td></td>
		</tr>
		<tr>
			<td>Computer</td>
			<td>Dell</td>
			<td>Precission3650</td>
			<td></td>
		</tr>
		<tr>
			<td>Crocodile Clip Grounding Cables</td>
			<td>JWQIDI</td>
			<td>B06WGZG17W</td>
			<td></td>
		</tr>
		<tr>
			<td>Detergent</td>
			<td>Metrex</td>
			<td>10-4100-0000</td>
			<td></td>
		</tr>
		<tr>
			<td>D-Glucose</td>
			<td>Macron Fine Chemicals</td>
			<td>4912-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Dihydrogen Sodium Phosphate</td>
			<td>Thermo Fisher Scientific</td>
			<td>BP329-500</td>
			<td></td>
		</tr>
		<tr>
			<td>DinoCam</td>
			<td>Dino-Lite</td>
			<td>AM73915MZTL</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Thermo Fisher Scientific</td>
			<td>A407P-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td>Fine Science Tools</td>
			<td>11980-13</td>
			<td></td>
		</tr>
		<tr>
			<td>Hot plate</td>
			<td>Thermo Fisher Scientific</td>
			<td>SP88857200</td>
			<td></td>
		</tr>
		<tr>
			<td>Ice Machine</td>
			<td>Hoshizaki</td>
			<td>F801MWH</td>
			<td></td>
		</tr>
		<tr>
			<td>Inflow and outflow needles</td>
			<td>Jensen Global</td>
			<td>JG 18-3.0X</td>
			<td></td>
		</tr>
		<tr>
			<td>Inline Solution Heater</td>
			<td>Warner Instruments</td>
			<td>SH-27B</td>
			<td></td>
		</tr>
		<tr>
			<td>Isofluorine</td>
			<td>Dechra</td>
			<td>08PB-STE22002-0122</td>
			<td></td>
		</tr>
		<tr>
			<td>Kim Wipes</td>
			<td>Thermo Fisher Scientific</td>
			<td>06-666</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnesium Chloride</td>
			<td>Thermo Fisher Scientific</td>
			<td>FLM33500</td>
			<td></td>
		</tr>
		<tr>
			<td>Micropipets</td>
			<td>Gilson</td>
			<td>F144069</td>
			<td></td>
		</tr>
		<tr>
			<td>Mili-Q Water Filter</td>
			<td>Mili-Q</td>
			<td>ZR0Q008WW</td>
			<td></td>
		</tr>
		<tr>
			<td>Paintbrush</td>
			<td>Daler Rowney</td>
			<td>AF85 Round: 0</td>
			<td></td>
		</tr>
		<tr>
			<td>Paper Filter</td>
			<td>Whatman</td>
			<td>EW-06648-24</td>
			<td></td>
		</tr>
		<tr>
			<td>Parafilm</td>
			<td>American National Can</td>
			<td>PM996</td>
			<td></td>
		</tr>
		<tr>
			<td>Perfusion System</td>
			<td>Multi Channel System</td>
			<td>PPS2</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipetor</td>
			<td>Thermo Fisher Scientific</td>
			<td>FB14955202</td>
			<td></td>
		</tr>
		<tr>
			<td>Platinum Harp</td>
			<td>3Brain</td>
			<td>3Brain</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium Chloride</td>
			<td>Thermo Fisher Scientific</td>
			<td>P330-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Razor blade</td>
			<td>Personna</td>
			<td>BP9020</td>
			<td></td>
		</tr>
		<tr>
			<td>Scale</td>
			<td>Metter Toledo</td>
			<td>AB204</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>Solingen</td>
			<td>92008</td>
			<td></td>
		</tr>
		<tr>
			<td>Slice Holding Chamber</td>
			<td>Custom</td>
			<td>Custom</td>
			<td>Custom 3D Printer Design, available upon request</td>
		</tr>
		<tr>
			<td>Sodium Bicarbonate</td>
			<td>Macron Fine Chemicals</td>
			<td>7412-06</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Chloride</td>
			<td>Thermo Fisher Scientific</td>
			<td>S271-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Temperature Control Box</td>
			<td>Warner Instruments</td>
			<td>TC344B</td>
			<td></td>
		</tr>
		<tr>
			<td>Transfer Pipettes</td>
			<td>Genesee Scientific</td>
			<td>30-200</td>
			<td></td>
		</tr>
		<tr>
			<td>Tubing</td>
			<td>Tygon</td>
			<td>B-44-3 TPE</td>
			<td></td>
		</tr>
		<tr>
			<td>Vibratome VZ-300</td>
			<td>Precissionary</td>
			<td>VF-00-VM-NC</td>
			<td></td>
		</tr>
		<tr>
			<td>Weigh Boat</td>
			<td>Electron Microscopy Sciences</td>
			<td>70040</td>
			<td></td>
		</tr>
	</tbody>
</table>

high-quality seizure-like activity from acute brain slices using a complementary metal-oxide-semiconductor high-density microelectrode array system

Commercially available high-density microelectrode array (HD-MEA) systems, which include an MEA chip with thousands of recording points1,2 and an MEA platform to amplify and digitize the data, are an emerging tool for electrophysiological research. These HD-MEA systems use complementary metal-oxide-semiconductor (CMOS) technology to record electrophysiological data with high sensitivity from cell cultures and ex vivo brain slice preparations. These MEA systems afford unprecedented spatial and temporal resolution to neurophysiological research via high electrode density and quality signal-to-noise ratios3. This technology has mostly been used to study extracellular action potentials, but it can also capture high-quality local field potentials (LFPs) from various neuronal brain slice preparations4,5,6,7,8,9,10,11,12,13,14,15. Due to the above-mentioned high-resolution recording capability of CMOS-HD-MEA systems, users can track electrophysiological activity with great spatial accuracy16,17,18. This capability is particularly relevant to tracking propagation patterns of network LFPs5,12,15,19,20,21. Therefore, CMOS-HD-MEA systems can provide an unprecedented understanding of the propagation patterns of physiological and pathological activity from various cell culture and brain slice preparations. Of particular note, these capabilities of CMOS-HD-MEA systems can allow researchers to contrast seizure patterns of different brain regions simultaneously and assay how various anti-epileptic compounds affect these patterns. By doing so, it provides an innovative method for studying ictogenesis and ictal propagation and for understanding how pharmacology disrupts pathological network activity7,10,14. Therefore, these novel capacities of CMOS-HD-MEA systems can contribute significantly to the research of neurological disorders, as well as aid in drug discovery research5,7,11,22. We aim to provide details on using CMOS-HD-MEA systems to study seizure-like activity.
When using CMOS-HD-MEA systems to study LFPs, such as epileptiform activity in acute brain slices, users can face many challenges, including debilitating electrical noise, keeping the slice healthy during experimentation, and detecting a quality signal from a two-dimensional (2D) CMOS-MEA chip that records only from the surface of the brain slice. This protocol describes basic steps for properly grounding the MEA platform and other equipment used in experimentation, a crucial step that may require individual customization for each lab setup. In addition, we discuss steps to aid in keeping the brain slice healthy during long recordings in the submerged chambers used with CMOS-HD-MEA systems23,24,25. Additionally, in contrast to more common electrophysiological recording methods, which record from deep in the brain slice, most CMOS-HD-MEA systems use 2D chips that do not penetrate into the slice. Therefore, these systems require a healthy neuronal outer layer to produce the majority of the recorded LFP signals. Other challenges include visualizing the massive amount of data generated by thousands of electrodes. To overcome these challenges, we recommend a simple but effective protocol that increases the likelihood of achieving high-quality network epileptiform activity that propagates across the brain slice. We also include a brief description of a publicly available graphic user interface (GUI) we developed with associated resources to aid in data visualization10.
Previous publications have provided related protocols for the use of MEA recording systems26,27,28,29. However, this work aims to assist experimenters using CMOS-HD-MEA systems with 2D chips, specifically those seeking to study high-quality epileptiform activity from brain slices. In addition, we compare two of the most common solution manipulations for induction of seizure-like activity, namely the 0 Mg2+ and 4-AP paradigms, to help users identify the most appropriate convulsant media for their specific application. Although the protocol is focused primarily on the generation of seizure-like activity, it can be modified to explore other electrophysiological phenomena using brain slices.

<meta charset="utf8"></head><body>Autore in primo piano: Svelare le dinamiche delle crisi epilettiche e nuove terapie per lo stato epilettico utilizzando i sistemi di array di microelettrodi ad alt

Attività simile a una crisi epilettica di alta qualità da fette cerebrali acute utilizzando un sistema di array di microelettrodi ad alta densità a semiconduttore di ossido di metallo complementare

Complementary metal-oxide-semiconductor high-density microelectrode array (CMOS-HD-MEA) systems can record neurophysiological activity from cell cultures ...

high-quality-seizure-like-activity-from-acute-brain-slices-using

Without Section

Research

High-Quality Seizure-Like Activity from Acute Brain Slices Using a Complementary Metal-Oxide-Semiconductor High-Density Microelectrode Array System

JoVE Journal

Neuroscience

1.4K Views.  Brigham Young University. Nel laboratorio Parrish, siamo desiderosi di capire come iniziano, si propagano e terminano le convulsioni. Siamo particolarmente interessati a esplorare nuove terapie per lo stato epilettico, una condizione pericolosa per la vita in cui una crisi non termina automaticamente. Nella nostra ricerca utilizziamo sistemi di array di microelettrodi ad alta densità CMOS.Queste tecnologie avanzate ci consentono di registrare dati elettrofisiologici ad alta ri...

Video: Autore in primo piano: Svelare le dinamiche delle crisi epilettiche e nuove terapie per lo stato epilettico utilizzando i sistemi di array di microelettrodi ad alt

Complementary metal-oxide-semiconductor high-density microelectrode array (CMOS-HD-MEA) systems can record neurophysiological activity from cell cultures and ex vivo brain slices in unprecedented electrophysiological detail. CMOS-HD-MEAs were first optimized to record high-quality neuronal unit activity from cell cultures but have also been shown to produce quality data from acute retinal and cerebellar slices. Researchers have recently used CMOS-HD-MEAs to record local field potentials (LFPs) from acute, cortical rodent brain slices. One LFP of interest is seizure-like activity. While many users have produced brief, spontaneous epileptiform discharges using CMOS-HD-MEAs, few users reliably produce quality seizure-like activity. Many factors may contribute to this difficulty, including electrical noise, the inconsistent nature of producing seizure-like activity when using submerged recording chambers, and the limitation that 2D CMOS-MEA chips only record from the surface of the brain slice. The techniques detailed in this protocol should enable users to consistently induce and record high-quality seizure-like activity from acute brain slices with a CMOS-HD-MEA system. In addition, this protocol outlines the proper treatment of CMOS-HD-MEA chips, the management of solutions and brain slices during experimentation, and equipment maintenance.

Here, we outline a protocol for using complementary metal-oxide-semiconductor high-density microelectrode array systems (CMOS-HD-MEAs)&#160;to record seizure-like activity from ex vivo brain slices.

Ricerca

Neuroscienze

Equipment Preparation for CMOS-HD-MEA Experimentation

Preparazione dell'attrezzatura per la sperimentazione CMOS-HD-MEA

Chip Preparation and Placement of Brain Slice to Record High-Quality Seizure-like Activity Using CMOS-HD-MEA System

Preparazione e posizionamento del chip della fetta cerebrale per registrare un'attività simile a una crisi di alta qualità utilizzando il sistema CMOS-HD-MEA 