Name: construction of microdrive arrays for chronic neural recordings in awake behaving mice
Uploaded: 2013-07-05T13:00:00
Description: Watch this Scientific Journal Video about Construction of Microdrive Arrays for Chronic Neural Recordings in Awake Behaving Mice Video at JoVE.com

Vi takker Daniel Carpi for hans hjælp og tidlig bidrag til dette projekt. Vi takker også Lucrecia Novoa for hendes hjælp med illustrationer og billeder. Dette arbejde blev støttet af NIH / NIAID programmet tilskud 5P01AI073693-03.

1.. Tetrode Fabrication <ol><li> Start med at bruge isolerede 12,5 um (0,0005 &quot;) diameter core platin-iridium wire fra Californien Fin Wire. Længden af ​​wiren skal skæres i passende længde for målet struktur. F.eks tråden skæres til mindst 30 cm lang til målretning ryggens subiculum eller hippocampus. </li><li> Fold tråden over på midten, så at der er to parallelle tråde, der vil være 15 cm i længden. Drapere midtpunktet af denne ledning over en vandret arm til dannelse fire parallelle tråde p?...

<ol>
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L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+hippocampal-prefrontal+synchrony+in+a+genetic+mouse+model+of+schizophrenia.">Impaired hippocampal-prefrontal synchrony in a genetic mouse model of schizophrenia.</a> Nature. 464 (7289), 763-767 (2010).</li><li>Engel, A. K., Moll, C. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Invasive+recordings+from+the+human+brain:+clinical+insights+and+beyond.">Invasive recordings from the human brain: clinical insights and beyond.</a> Nat. Rev. Neurosci. 6 (1), 35-47 (2005).</li><li>Cash, S. S., Halgren, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+human+K-complex+represents+an+isolated+cortical+down-state.">The human K-complex represents an isolated cortical down-state.</a> Science. 324 (5930), 1084-1087 (2009).</li></ol>

Vi har beskrevet en række teknikker til konstruktion lette og kompakte Microdrives til optagelse af ekstracellulære enhed og marken potentielle aktivitet i mus. Ved at bygge brugerdefinerede Microdrives med baser gammeldags fra akrylglas (methylmethacrylat), kan det centrale system, der let tilpasses til flere drev og for målretning af en bred vifte af neurale områder. Vi har med succes ændret systemet til optagelse fra flere hjerne targets og med større arrays til optagelser i mus. Med yderligere modifikation kan...

Anvendelsen af mikroelektrode teknik til optagelse ekstracellulære neurale signaler in vivo har en lang og værdsat tradition i neurovidenskab 1, 2. Evnen til at optage elektriske aktivitet fra mange områder af hjernen i frit opfører dyr er dog en nyere teknologi, der bliver mere og mere almindeligt som de softwarepakker til erhvervelse, analyse og diskrimination af neurale signaler bliver mere sofistikerede og brugervenlige 3, 4.. De teknologiske fremskridt på software siden er også blevet ledsaget af reduktioner i vægt og hovedparten af ​​implantabelt udstyr, som er blevet skaleret ned tilstrækkeligt til optagelse i små pattedyr, såsom mus. Ved at bruge letvægts (hovedsagelig plast) komponenter, er forskere i stand til at konstruere Microdrives der giver mulighed for uafhængig positionering af elektroder eller tetrodes at målrette en bred vifte af hjernen regioner 5-7. Selv dybe hjernens strukturer, såsomamygdala 6 og striatum 5 kan rutinemæssigt målrettet med udvælgelsen af en passende lang drivskrue. Disse optagelse teknikker give forskere til at opnå high-fidelity neurale signaler, og er i register med den elektriske aktivitet i enkelte neuroner indspillet intracellulært 8, 9. Ved hjælp af disse typer af Microdrives, har vi med succes indspillet single-enheder fra mus i op til to måneder efter implantation 10. Desuden har den lette karakter af indretningerne (ca. 1,5-2,0 g) resulterede i adfærdsmæssige ydeevne, der er sammenlignelig med ikke-implanterede mus i mange adfærdsmæssige opgaver. Især har vi påvist, at implanterede mus udviser normale udførelse i den nye objekt anerkendelse opgave 10 og objektet sted opgave (upublicerede data). Brugen af ​​Microdrives koblet til flere tetrodes giver forskerne at overvåge og analysere neurale aktivitet på netværksniveausamtidig optagelse fra flere single-enheder i hjernen. Optagelse med disse tetrodes har flere store fordele for unit identifikation og gør det muligt høj nøjagtighed erhvervelse og diskrimination af multiple single-enheder 11. Vi beskriver, hvordan at fabrikere og guld-plade tetrode bundter og efterfølgende indlæse dem i køreklare elektrode luftfartsselskaber. En type drev luftfartsselskab, vi beskriver, er kommercielt tilgængelige, og den anden er en enkel, men let udvides, drev design, der kan rumme flere luftfartsselskaber og tetrode arrangementer uden en betydelig investering af ressourcer.

<table border="1">
		<tbody>
		 <tr>
			<td>Name of Reagent/Material</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		 </tr>
		 <tr>
			<td>0.0005" (12.5 &mu;M) diameter Platinum-Iridium wire </td>
			<td>California Fine Wire</td>
			<td>CFW#100-167</td>
			<td>HML VG insulated <a href="http://www.calfinewire.com" target="_blank">www.calfinewire.com</a></td>
		 </tr>
		 <tr>
			<td>0.002" (50 &mu;M) diameter Stableohm 675 wire</td>
			<td>California Fine Wire</td>
			<td>CFW# 100-188</td>
			<td>HML insulated Ni-Cr</td>
		 </tr>
		 <tr>
			<td>polyamide tubing</td>
			<td>Polymicro Technologies</td>
			<td>1068150020</td>
			<td>99 micron I.D., 166 micron O.D. <a href="http://www.polymicro.com" target="_blank">www.polymicro.com</a></td>
		 </tr>
		 <tr>
			<td>brass guides</td>
			<td>World Plastics Inc</td>
			<td></td>
			<td>3.3 x 6.6 mm</td>
		 </tr>
		 <tr>
			<td>Delrin blocks</td>
			<td>World Plastics Inc</td>
			<td></td>
			<td>3.13 x 2.5 mm</td>
		 </tr>
		 <tr>
			<td>Fillister head brass screws</td>
			<td>J.I. Morris Co.</td>
			<td>00-90 x 1/2</td>
			<td>drive screw <a href="http://www.jimorrisco.com" target="_blank">www.jimorrisco.com</a></td>
		 </tr>
		 <tr>
			<td>hex brass nuts</td>
			<td>J.I. Morris Co.</td>
			<td>00-90</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Fillister head brass screws</td>
			<td>J.I. Morris Co.</td>
			<td>000-120 x 3/32</td>
			<td>EIB mount and ground screw</td>
		 </tr>
		 <tr>
			<td>plexiglass acrylic</td>
			<td>Canal Street Plastics</td>
			<td></td>
			<td>5 mm thick, clear, <a href="http://www.cpcnyc.com" target="_blank"> www.cpcnyc.com</a></td>
		 </tr>
		 <tr>
			<td>cyanoacrylate</td>
			<td>Krazy Glue</td>
			<td></td>
			<td>2 g tube</td>
		 </tr>
		 <tr>
			<td>electronic interface board</td>
			<td>Neuralynx</td>
			<td>EIB-18</td>
			<td><a href="http://www.neuralynx.com" target="_blank">www.neuralynx.com</a></td>
		 </tr>
		 <tr>
			<td>non-cyanide gold solution</td>
			<td>SIFCO</td>
			<td>SIFCO 5355</td>
			<td><a href="http://www.sifcoasc.com" target="_blank">www.sifcoasc.com</a></td>
		 </tr>
		 <tr>
			<td>VersaDrive 4</td>
			<td>Neuralynx</td>
			<td></td>
			<td>four tetrode model</td>
		 </tr>
		 <tr>
			<td>tetrode assembly station</td>
			<td>Neuralynx</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>motorized tetrode spinner</td>
			<td>Neuralynx</td>
			<td>tetrode spinner 2.0</td>
			<td></td>
		 </tr>
		 <tr>
			<td>VersaDrive jig</td>
			<td>Neuralynx </td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>soldering iron</td>
			<td>Radio Shack </td>
			<td>64-2802B</td>
			<td><a href="http://www.radioshack.com" target="_blank">www.radioshack.com</a></td>
		 </tr>
		 <tr>
			<td>nanoZ</td>
			<td>Neuralynx</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>small bit drill/driver</td>
			<td>Ram Products </td>
			<td>Rampower 35</td>
			<td>with footpedal controller, <a href="http://www.ramprodinc.com" target="_blank">www.ramprodinc.com</a></td>
		 </tr>
		 <tr>
			<td>drill bits</td>
			<td>Small Parts, Inc.</td>
			<td></td>
			<td>3/32" bits, <a href="http://www.smallpartsinc.com" target="_blank">www.smallpartsinc.com</a></td>
		 </tr>
		 <tr>
			<td>dissecting microscope</td>
			<td>Olympus </td>
			<td>SZ-60</td>
			<td><a href="http://www.olympusamerica.com" target="_blank">www.olympusamerica.com</a></td>
		 </tr>
		 <tr>
			<td>heat gun</td>
			<td>Alphawire</td>
			<td>Fit gun 3</td>
			<td>use setting "1" only, <a href="http://www.alphawire.com" target="_blank">www.alphawire.com</a></td>
		 </tr>
		 <tr>
			<td>26 AWG copper wire</td>
			<td>Arcor Electronics</td>
			<td>F26</td>
			<td>for ground wires, <a href="http://www.arcorelectronics.com" target="_blank"> www.arcorelectronics.com</a></td>
		 </tr>
		 <tr>
			<td>soldering flux</td>
			<td>Eagle</td>
			<td>2 oz, #205</td>
			<td></td>
		 </tr>
		 <tr>
			<td>0.02" diameter solder</td>
			<td>Kester</td>
			<td>24-6337-0010</td>
			<td><a href="http://www.kester.com" target="_blank"> www.kester.com</a></td>
		 </tr>
		 <tr>
			<td>benchtop vise</td>
			<td>Vacu-Vise</td>
			<td>Model 300</td>
			<td></td>
		 </tr>
		 <tr>
			<td>fiber optic light</td>
			<td>Nikon </td>
			<td>MKII</td>
			<td>dual light arms, <a href="http://www.nikon.com" target="_blank"> www.nikon.com</a></td>
		 </tr>
		 <tr>
			<td>5-min epoxy</td>
			<td>Allied Electronics</td>
			<td></td>
			<td>25 ml, <a href="http://www.alliedelec.com" target="_blank"> www.alliedelec.com</a></td>
		 </tr>
		 <tr>
			<td>fine tweezers</td>
			<td>Roboz Surgical Instrument Co.</td>
			<td>RS-4907, RS-5010</td>
			<td>INOX material, <a href="http://www.roboz.com" target="_blank"> www.roboz.com</a></td>
		 </tr>
		 <tr>
			<td>micro dissecting scissors</td>
			<td>Roboz Surgical Instrument Co.</td>
			<td>RS-5880</td>
			<td></td>
		 </tr>
		</tbody>
 </table>
 Table 1. Materials and reagents used for constructing tetrodes and microdrives.

construction of microdrive arrays for chronic neural recordings in awake behaving mice

State-of-the-art elektrofysiologiske optagelser fra hjerner frit opfører dyr give forskere til samtidig at undersøge de lokale feltpotentialer (LFPs) fra populationer af neuroner og handling potentialer fra individuelle celler, som dyret engagerer sig i eksperimentelt relevante opgaver. Kronisk implanterede Microdrives tillader hjernen optagelser til strække sig over perioder på flere uger. Miniaturiserede drev og lette komponenter tillader disse langsigtede optagelser forekomme i små pattedyr, såsom mus. Ved hjælp tetrodes, der består af tæt flettede bundter af fire elektroder, hvor hver ledning har en diameter på 12,5 um, er det muligt at isolere fysiologisk aktive neuroner i overfladiske områder af hjernen, såsom hjernebarken, dorsal hippocampus, og subiculum, samt som dybere regioner som striatum og amygdala. Desuden er denne teknik sikrer stabile, high-fidelity neurale optagelser, da dyret udfordres med en variety adfærdsmæssige opgaver. Dette håndskrift beskriver flere teknikker, som er blevet optimeret til at optage fra musen hjernen. Først viser vi, hvordan at fabrikere tetrodes, indlæse dem i køreklare rør og guld-plade deres tips for at reducere deres impedans fra MOhm til kQ rækkevidde. For det andet, vi vise, hvordan man konstruere en brugerdefineret mikrodrev samling til transport og bevæge tetrodes lodret, med brug af billige materialer. For det tredje, vi viser trinene for at samle en kommercielt tilgængelig mikrodrev (Neuralynx VersaDrive), der er designet til at bære selvstændigt bevægelige tetrodes. Endelig præsenterer vi repræsentative resultater af lokale feltpotentialer og single-unit signaler opnået i den dorsale subiculum af mus. Disse teknikker kan let modificeres til at rumme forskellige typer af elektrode arrays og optagelse ordninger i musehjernen.

<p class="jove_content"&gt; Efter implantering af mikrodrevet og sænke elektroderne til de tilsigtede hjernen mål, en forstærket dataopsamlingssystem, såsom en Neuralynx Lynx-8, der er nødvendig til optagelse neurale signaler. Repræsentative neurale optagelser af lokale feltpotentialer (LFPs) samt single-unit action potentialer (ofte kaldet &quot;spikes&quot;) fra muse dorsale subiculum er vist i<strong&gt; Figur 2</strong&gt;. LFP signaler blev samplet ved 3 kHz og båndpasfiltrerede mellem 0,1-500 Hz (<strong&gt; 2A</strong&gt; Og<stron...

Watch this Scientific Journal Video about Construction of Microdrive Arrays for Chronic Neural Recordings in Awake Behaving Mice Video at JoVE.com

Construction of Microdrive Arrays for Chronic Neural Recordings in Awake Behaving Mice Video (Video) | JoVE

Udformningen og montage af Microdrives til in vivo elektrofysiologiske optagelser af hjernens signaler fra musen er beskrevet. Ved at binde mikroelektrode bundter til robuste køreklare luftfartsselskaber giver disse teknikker for langsigtede og stabile neurale optagelser. Det lette design giver mulighed for ubegrænset adfærdsmæssige resultater af dyret følgende drev implantation.

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Construction of Microdrive Arrays for Chronic Neural Recordings in Awake Behaving Mice

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A Novel Eksperimentel og analytisk tilgang til multimodal Neural dekodning hensigtserklæring Under Social Interaktion i frit-opfører Menneskelige Spædbørn

Understanding typical and atypical development remains one of the fundamental questions in developmental human neuroscience. Traditionally, experimental ...

A Pressure Injection System for Investigating the Neuropharmacology of Information Processing in Awake Behaving Macaque Monkey Cortex

A Pressure Injection System for Investigating the Neuropharmacology of Information Processing in Awake Behaving Macaque Monkey Cortex Video (Video) | JoVE

En Pressure Injection System til Undersøgelse af Neuropharmacology of Information Processing i Awake Behaving makak Monkey Cortex

The top-down modulation of feed-forward cortical information processing is functionally important for many cognitive processes, including the modulation ...

Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat

Using Laser Doppler Imaging and Monitoring to Analyze Spinal Cord Microcirculation in Rat Video (Video) | JoVE

Ved hjælp af Laser Doppler Imaging og overvågning til at analysere rygmarven mikrocirkulationen i Rat

Laser Doppler flowmetry (LDF) is a noninvasive method for blood flow (BF) measurement, which makes it preferable for measuring microcirculatory ...

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats

A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats Video (Video) | JoVE

En trådløs, tovejs grænseflade for In Vivo optagelse og Stimulation af neurale aktivitet i frit opfører rotter

In vivo electrophysiology is a powerful technique to investigate the relationship between brain activity and behavior at a millisecond and micrometer ...

A Behavioral Assay for Investigating the Role of Spatial Memory During Instinctive Defense in Mice

A Behavioral Assay for Investigating the Role of Spatial Memory During Instinctive Defense in Mice Video (Video) | JoVE

En adfærdsmæssige analyse for at undersøge rollen af rumlig hukommelse under instinktiv forsvar i mus

Evolution has selected a repertoire of defensive behaviors that are essential for survival across all animal species. These behaviors are often ...

The Unpredictable Chronic Mild Stress Protocol for Inducing Anhedonia in Mice

The Unpredictable Chronic Mild Stress Protocol for Inducing Anhedonia in Mice Video (Video) | JoVE

Den uforudsigelige kroniske Mild Stress protokol for at fremkalde Anhedonia i mus

Depression is a highly prevalent and debilitating condition, only partially addressed by current pharmacotherapies. The lack of response to treatment by ...

Novel Object Recognition and Object Location Behavioral Testing in Mice on a Budget

Novel Object Recognition and Object Location Behavioral Testing in Mice on a Budget Video (Video) | JoVE

Roman objekt anerkendelse og objekt placering adfærdsmæssige test i mus på en Budget

Ethologically relevant behavioral testing is a critical component of any study that uses mouse models to study the cognitive effects of various ...

Low-cost Protocol of Footprint Analysis and Hanging Box Test for Mice Applied the Chronic Restraint Stress

Low-cost Protocol of Footprint Analysis and Hanging Box Test for Mice Applied the Chronic Restraint Stress Video (Video) | JoVE

Low-cost protokol fodaftryk analyse og hængende boks Test for mus anvendes kronisk tilbageholdenhed Stress

Gait disturbance is frequently observed in patients with movement disorders. In mouse models used for movement disorders, gait analysis is important ...