Name: construction of an improved multi-tetrode hyperdrive for large-scale neural recording in behaving rats
Uploaded: 2018-05-09T15:00:00
Description: Watch this Scientific Journal Video about Construction of an Improved Multi-Tetrode Hyperdrive for Large-Scale Neural Recording in Behaving Rats at JoVE.com

Nous remercions le laboratoire Moser au Kavli Institute for neurosciences des systèmes et Centre Neural Computation, Norwegian University of Science and Technology, pour la chronique neural recensant des procédures chez le rat. Ce travail a été soutenu par les NIH grant NS098146 R21 et humaine Frontier Science programme à long terme bourse LT000211/2016-L à L. Lu.

1. des modèles 3D, la stéréolithographie
<ol><li>Techniques de stéreolithographie permet d’imprimer les hyperdrive pièces et accessoires. Chaque hyperdrive est composée de dix-huit navettes, dix-huit navette boulons et un chacun de toutes les autres pièces en plastique (Figure 1). Remarque : Les accessoires ne font pas partie de l’hyperdrive mais sont nécessaires pour la construction de l’hyperdrive.</li>
</ol>2. préparation des accessoires (<strong...

<ol>
	<li>O'Keefe, J., Recce, 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=Phase+relationship+between+hippocampal+place+units+and+the+EEG+theta+rhythm.">Phase relationship between hippocampal place units and the EEG theta rhythm.</a> Hippocampus. 3 (3), 317-330 (1993).</li><li>Wilson, M. A., McNaughton, B. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamics+of+the+hippocampal+ensemble+code+for+space.">Dynamics of the hippocampal ensemble code for space.</a> Science. 261 (5124), 1055-1058 (1993).</li><li>Gothard, K. M., Skaggs, W. E., Moore, K. M., McNaughton, B. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Binding+of+hippocampal+CA1+neural+activity+to+multiple+reference+frames+in+a+landmark-based+navigation+task.">Binding of hippocampal CA1 neural activity to multiple reference frames in a landmark-based navigation task.</a> J Neurosci. 16 (2), 823-835 (1996).</li><li>Gray, C. M., Maldonado, P. E., Wilson, M., McNaughton, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tetrodes+markedly+improve+the+reliability+and+yield+of+multiple+single-unit+isolation+from+multi-unit+recordings+in+cat+striate+cortex.">Tetrodes markedly improve the reliability and yield of multiple single-unit isolation from multi-unit recordings in cat striate cortex.</a> J Neurosci Methods. 63 (1-2), 43-54 (1995).</li><li>Buzsaki, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large-scale+recording+of+neuronal+ensembles.">Large-scale recording of neuronal ensembles.</a> Nat Neurosci. 7 (5), 446-451 (2004).</li><li>Fyhn, M., Hafting, T., Witter, M. P., Moser, E. I., Moser, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Grid+cells+in+mice.">Grid cells in mice.</a> Hippocampus. 18 (12), 1230-1238 (2008).</li><li>Skaggs, W. 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=EEG+sharp+waves+and+sparse+ensemble+unit+activity+in+the+macaque+hippocampus.">EEG sharp waves and sparse ensemble unit activity in the macaque hippocampus.</a> J Neurophysiol. 98 (2), 898-910 (2007).</li><li>Guo, P., Pollack, A. J., Varga, A. G., Martin, J. P., Ritzmann, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Extracellular+wire+tetrode+recording+in+brain+of+freely+walking+insects.">Extracellular wire tetrode recording in brain of freely walking insects.</a> J Vis Exp. (86), (2014).</li><li>Knierim, J. J., McNaughton, B. L., Poe, G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Three-dimensional+spatial+selectivity+of+hippocampal+neurons+during+space+flight.">Three-dimensional spatial selectivity of hippocampal neurons during space flight.</a> Nat Neurosci. 3 (3), 209-210 (2000).</li><li>Leutgeb, S., 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=Independent+codes+for+spatial+and+episodic+memory+in+hippocampal+neuronal+ensembles.">Independent codes for spatial and episodic memory in hippocampal neuronal ensembles.</a> Science. 309 (5734), 619-623 (2005).</li><li>Lansink, C. S., 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=A+split+microdrive+for+simultaneous+multi-electrode+recordings+from+two+brain+areas+in+awake+small+animals.">A split microdrive for simultaneous multi-electrode recordings from two brain areas in awake small animals.</a> J Neurosci Methods. 162 (1-2), 129-138 (2007).</li><li>Kloosterman, F., 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=Micro-drive+array+for+chronic+in+vivo+recording:+drive+fabrication.">Micro-drive array for chronic in vivo recording: drive fabrication.</a> J Vis Exp. (26), (2009).</li><li>. Google Patents Available from: <a target="_blank" href="https://www.google.com/patents/US5928143">https://www.google.com/patents/US5928143</a> (1999)</li><li>Redish, A. D., 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=Independence+of+firing+correlates+of+anatomically+proximate+hippocampal+pyramidal+cells.">Independence of firing correlates of anatomically proximate hippocampal pyramidal cells.</a> J Neurosci. 21 (5), RC134 (2001).</li><li>Schmitzer-Torbert, N., Redish, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+activity+in+the+rodent+dorsal+striatum+in+sequential+navigation:+separation+of+spatial+and+reward+responses+on+the+multiple+T+task.">Neuronal activity in the rodent dorsal striatum in sequential navigation: separation of spatial and reward responses on the multiple T task.</a> J Neurophysiol. 91 (5), 2259-2272 (2004).</li><li>Nguyen, D. P., 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=Micro-drive+array+for+chronic+in+vivo+recording:+tetrode+assembly.">Micro-drive array for chronic in vivo recording: tetrode assembly.</a> J Vis Exp. (26), (2009).</li><li>Chang, E. H., Frattini, S. A., Robbiati, S., Huerta, P. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Construction+of+microdrive+arrays+for+chronic+neural+recordings+in+awake+behaving+mice.">Construction of microdrive arrays for chronic neural recordings in awake behaving mice.</a> J Vis Exp. (77), e50470 (2013).</li><li>Vandecasteele, M., 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=Large-scale+recording+of+neurons+by+movable+silicon+probes+in+behaving+rodents.">Large-scale recording of neurons by movable silicon probes in behaving rodents.</a> J Vis Exp. (61), e3568 (2012).</li><li>Siegle, J. H., 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=Chronically+implanted+hyperdrive+for+cortical+recording+and+optogenetic+control+in+behaving+mice.">Chronically implanted hyperdrive for cortical recording and optogenetic control in behaving mice.</a> Conf Proc IEEE Eng Med Biol Soc. 2011, 7529-7532 (2011).</li><li>Brunetti, P. M., 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=Design+and+fabrication+of+ultralight+weight,+adjustable+multi-electrode+probes+for+electrophysiological+recordings+in+mice.">Design and fabrication of ultralight weight, adjustable multi-electrode probes for electrophysiological recordings in mice.</a> J Vis Exp. (91), e51675 (2014).</li><li>. Google Patents Available from: <a target="_blank" href="https://www.google.com/patents/US4575330">https://www.google.com/patents/US4575330</a> (1986)</li><li>Ludvig, N., Potter, P. E., Fox, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simultaneous+single-cell+recording+and+microdialysis+within+the+same+brain+site+in+freely+behaving+rats:+a+novel+neurobiological+method.">Simultaneous single-cell recording and microdialysis within the same brain site in freely behaving rats: a novel neurobiological method.</a> J Neurosci Methods. 55 (1), 31-40 (1994).</li></ol>

Nous décrivons ici le processus de construction d’un hyperdrive nouvellement développé composé de dix-huit tétrodes indépendamment mobiles. Le lecteur peut être construit de pièces abordables achetées dans de nombreux magasins de matériel disponibles, combinées avec les composants créés par stéreolithographie impression. L’hyperdrive peut être chroniquement implanté sur le crâne d’un rat à l’aide de procédures chirurgicales standards et est capable d’enregistrer l’activité neuronale extrac...

Dans le domaine des neurosciences des systèmes, les scientifiques étudient les corrélats neurones sous-tendant les processus cognitifs tels que la navigation spatiale, la mémoire et prise de décision. Pour ces types d’études, il est essentiel de surveiller l’activité de neurones individuels nombreux pendant le comportement animal. Ces dernières décennies, deux progrès importants ont été réalisés pour répondre aux besoins expérimentaux pour enregistrement neuronal extracellulaire en petits animaux1,2,3. Tout d’abord le Θlaboration de la tétrode, un faisceau de quatre microfils utilisé pour enregistrer l’activité neurale des neurones simultanément1,2,4. Les amplitudes de signal différentiel d’activité à travers les quatre canaux d’une tétrode permet l’isolement de l’activité de neurones individuels de plusieurs cellules simultanément enregistré5. En outre, la souplesse de la microfils permet une plus grande stabilité de la tétrode réduisant au minimum le déplacement relatif entre la tétrode et la population de cellules cibles. Tétrodes sont maintenant largement utilisés au lieu d’une seule électrode pour de nombreuses études de cerveau chez diverses espèces, y compris les rongeurs1,2,6, primates7et insectes8. Deuxièmement l’élaboration d’un hyperdrive transportait des tétrodes indépendamment mobiles multiples, qui permet le contrôle simultané de l’activité neuronale de plus grandes populations de neurones provenant de plusieurs emplacements d’enregistrement3, 9,10,11,12.
La disponibilité d’un appareil d’enregistrement multi-tétrode fiable et abordable pour les petits animaux est limitée. L’hyperdrive classique, initialement développé par Bruce McNaughton13, a été utilisée avec succès pour des enregistrements neurones en comportement librement des rats dans nombreux laboratoires dans les dernières deux décennies9,10,14, 15. Toutefois, pour des raisons techniques, les composants originaux nécessaires pour construire le lecteur McNaughton sont désormais très difficiles à obtenir et ne sont pas compatibles avec les interfaces d’acquisition de données récemment améliorée. L’autre conception bien acceptée d’hyperdrive nécessite les microdrives d’être individuellement à la main, qui pourrait donner des résultats incohérents et consomment beaucoup de temps,12. Afin d’enregistrer l’activité neurale provenant de diverses régions du cerveau chez les rats se comporter, nous avons développé un nouvel hyperdrive utilisant la technologie stéreolithographie. Nous avons cherché à satisfaire aux exigences suivantes : (1) l’hyperdrive nouvelle doit permettre un déplacement précis des tétrodes dans le cerveau et fournir enregistrement stable de plusieurs régions cibles ; (2) l’hyperdrive nouvelle doit être compatible avec le système quickclip magnétiques récemment mis au point pour permettre une connexion facile ; et (3) le nouveau hyperdrive peut être fidèlement reproduit avec des matériaux facilement disponibles. Ici, nous fournissons une technique pour la construction de l’hyperdrive 3D-imprimable contenant dix-huit tétrodes indépendamment mobiles, basées sur la conception de McNaughton. Dans le protocole, nous décrivons les détails du processus de fabrication de l’hyperdrive nouvelle, que nous avons utilisée avec succès à des potentiels d’action simples-neurone Records et potentiels de champs locaux du cortex entorhinal postrhinal et médial au cours des semaines dans un librement se comporter de rat au cours des tâches de recherche de nourriture naturelles.

<table>
	<tbody>
		<tr>
			<td>Welding rod</td>
			<td>Blue Demon</td>
			<td>ER308L-035-01T</td>
			<td>Stainless steel, 0.035&#34; in diameter</td>
		</tr>
		<tr>
			<td>Screw</td>
			<td>McMaster</td>
			<td>91771A060</td>
			<td>Stainless steel, flat head, 0-80&#160;thread,&#160;5/8&#34;&#160;in length</td>
		</tr>
		<tr>
			<td>Screw</td>
			<td>McMaster</td>
			<td>91772A051</td>
			<td>Stainless steel, pan head, 0-80&#160;thread,&#160;5/32&#34;&#160;in length</td>
		</tr>
		<tr>
			<td>Screw</td>
			<td>McMaster</td>
			<td>92196A056</td>
			<td>Stainless steel, socket head, 0-80&#160;thread,&#160;5/16&#34;&#160;in length</td>
		</tr>
		<tr>
			<td>Screw</td>
			<td>McMaster</td>
			<td>92196A055</td>
			<td>Stainless steel, socket head, 0-80&#160;thread,&#160;1/4&#34;&#160;in length</td>
		</tr>
		<tr>
			<td>Screw</td>
			<td>McMaster</td>
			<td>95868A131</td>
			<td>Nylon,&#160; socket head, 2-56&#160;thread,&#160;3/16&#34;&#160;in length, black</td>
		</tr>
		<tr>
			<td>Screw nut</td>
			<td>McMaster</td>
			<td>90730A001</td>
			<td>Stainless steel, narrow hex,&#160; 0-80&#160;thread</td>
		</tr>
		<tr>
			<td>Shoulder screw</td>
			<td>McMaster</td>
			<td>90298A213</td>
			<td>Stainless steel, 8-32 thread, 3/16&#34; in diameter, 1/4&#34; in length</td>
		</tr>
		<tr>
			<td>Cup screw</td>
			<td>McMaster</td>
			<td>92313A105</td>
			<td>Stainless steel, 4-40 thread, 3/16&#34; in length</td>
		</tr>
		<tr>
			<td>Thumb screw</td>
			<td>McMaster</td>
			<td>94323A592</td>
			<td>Nylon, 8-32&#160;thread,&#160;3/8&#34;&#160;in length, black</td>
		</tr>
		<tr>
			<td>Magnet</td>
			<td>Apex</td>
			<td>M3X1MMDI</td>
			<td>Neodymium, 3 mm X 1 mm disc</td>
		</tr>
		<tr>
			<td>Metal tubing</td>
			<td>Small Parts</td>
			<td>B00137QHNS</td>
			<td>Stainless steel, 23 gauge, 0.0253&#34; OD, 0.013&#34; ID, 0.006&#34; wall</td>
		</tr>
		<tr>
			<td>Metal tubing</td>
			<td>New England Small Tube</td>
			<td>Custom-made</td>
			<td>Stainless steel, 30 gauge, 0.012/0.0125&#34; OD, 0.007/0.008&#34; ID, full hard</td>
		</tr>
		<tr>
			<td>Heat-shrink tubing</td>
			<td>McMaster</td>
			<td>7856K72</td>
			<td>0.09&#34; ID before shrinking, blue</td>
		</tr>
		<tr>
			<td>Silicone tubing</td>
			<td>A-M Systems</td>
			<td>807300</td>
			<td>0.040&#34; ID, 0.085&#34; OD</td>
		</tr>
		<tr>
			<td>Polyimide tubing</td>
			<td>A-M Systems</td>
			<td>823400</td>
			<td>0.0045&#34; ID, 0.0005&#34; wall</td>
		</tr>
		<tr>
			<td>Ground wire</td>
			<td>A-M Systems</td>
			<td>791500</td>
			<td>0.005&#34; bare, 0.008&#34; coated, half hard</td>
		</tr>
		<tr>
			<td>Tetrode wire</td>
			<td>California Fine Wire</td>
			<td>Custom-made</td>
			<td>0.0007&#34; in diameter, platinum-iridium (90%-10%), HML and VG coating</td>
		</tr>
		<tr>
			<td>EIB</td>
			<td>Neuralynx</td>
			<td></td>
			<td>EIB-72-QC-Large</td>
		</tr>
		<tr>
			<td>Gold pins</td>
			<td>Neuralynx</td>
			<td></td>
			<td>large EIB pins</td>
		</tr>
		<tr>
			<td>Tap</td>
			<td>Balax</td>
			<td>01302-000</td>
			<td>M1.2 thread size</td>
		</tr>
		<tr>
			<td>Tap</td>
			<td>McMaster</td>
			<td>2522A811</td>
			<td>0-80 thread size, bottoming</td>
		</tr>
		<tr>
			<td>Tap</td>
			<td>McMaster</td>
			<td>2522A771</td>
			<td>0-80 thread size, plug</td>
		</tr>
		<tr>
			<td>Tap</td>
			<td>McMaster</td>
			<td>26955A94</td>
			<td>3/8&#34;-24&#160;thread size, bottoming</td>
		</tr>
		<tr>
			<td>Tap</td>
			<td>McMaster</td>
			<td>2522A713</td>
			<td>2-56&#160;thread size</td>
		</tr>
		<tr>
			<td>Tap</td>
			<td>McMaster</td>
			<td>2522A715</td>
			<td>4-40&#160;thread size</td>
		</tr>
		<tr>
			<td>Tap</td>
			<td>McMaster</td>
			<td>2522A718</td>
			<td>8-32&#160;thread size</td>
		</tr>
		<tr>
			<td>Die</td>
			<td>McMaster</td>
			<td>2576A457</td>
			<td>3/8&#34;-24&#160;thread size, 1&#34; OD</td>
		</tr>
		<tr>
			<td>Drill bit</td>
			<td>McMaster</td>
			<td>30585A82</td>
			<td>Wire gauge&#160;65,&#160;0.035&#34; in diameter</td>
		</tr>
		<tr>
			<td>Drill bit</td>
			<td>McMaster</td>
			<td>30585A83</td>
			<td>Wire gauge&#160;66,&#160;0.033&#34; in diameter</td>
		</tr>
		<tr>
			<td>Drill bit</td>
			<td>McMaster</td>
			<td>30585A87</td>
			<td>Wire gauge&#160;70,&#160;0.028&#34; in diameter</td>
		</tr>
		<tr>
			<td>Drill bit</td>
			<td>McMaster</td>
			<td>30585A88</td>
			<td>Wire gauge&#160;71,&#160;0.026&#34; in diameter</td>
		</tr>
		<tr>
			<td>Drill bit</td>
			<td>McMaster</td>
			<td>30585A91</td>
			<td>Wire gauge&#160;73,&#160;0.024&#34; in diameter</td>
		</tr>
		<tr>
			<td>Drill bit</td>
			<td>McMaster</td>
			<td>8870A23</td>
			<td>3/16&#34;&#160;in diameter</td>
		</tr>
		<tr>
			<td>Dremel disc</td>
			<td>Wagner</td>
			<td>31M</td>
			<td>Diamond coated, 22 mm in diameter, 0.17 mm in thickness</td>
		</tr>
		<tr>
			<td>Steel wire</td>
			<td>Precision Brand</td>
			<td>21212</td>
			<td>0.012&#34; in diameter, full hard</td>
		</tr>
		<tr>
			<td>Steel wire</td>
			<td>Precision Brand</td>
			<td>21007</td>
			<td>0.007&#34; in diameter, full hard</td>
		</tr>
		<tr>
			<td>Steel wire</td>
			<td>A-M Systems</td>
			<td>792700</td>
			<td>0.003&#34; in diameter, half hard</td>
		</tr>
		<tr>
			<td>Super glue</td>
			<td>Loctite</td>
			<td>LT-40640</td>
			<td># 406</td>
		</tr>
		<tr>
			<td>Super glue</td>
			<td>Loctite</td>
			<td>LT-41550</td>
			<td># 415</td>
		</tr>
		<tr>
			<td>Dental acrylic powder&#160;</td>
			<td>Teets</td>
			<td>223-3773</td>
			<td>Coral</td>
		</tr>
		<tr>
			<td>Dental acrylic liquid</td>
			<td>Teets</td>
			<td>223-4003</td>
			<td></td>
		</tr>
	</tbody>
</table>

construction of an improved multi-tetrode hyperdrive for large-scale neural recording in behaving rats

Surveiller les patrons d’activité d’une importante population de neurones au cours de plusieurs jours chez l’animal éveillé est une technique dans le domaine des neurosciences des systèmes. Un élément clé de cette technique consiste le positionnement précis des électrodes multiples dans les régions du cerveau désiré et le maintien de leur stabilité. Nous décrivons ici un protocole pour la construction d’un hyperdrive 3D-imprimable, qui comprend dix-huit tétrodes réglables séparément et qui est spécialement conçue pour in vivo enregistrement neuronal extracellulaire en comportement librement des rats. Les tétrodes attachés aux microdrives peuvent soit être individuellement avancés dans plusieurs régions du cerveau le long de la piste, ou peuvent être utilisés pour placer un tableau des électrodes dans une zone plus petite. Les tétrodes multiples permettent l’examen simultané d’action potentials de quelques dizaines de neurones individuels, ainsi que les potentiels de champs locaux de populations de neurones dans le cerveau au cours de comportement actif. En outre, la conception fournit pour la 3D plus simple rédaction des logiciels qui peuvent facilement être modifiés pour différents besoins expérimentaux.

Nous avons utilisé un hyperdrive nouvellement construite afin d’obtenir des résultats de l’étude. Le lecteur était équipé de tétrodes construits à partir d’ø 17 µm (0,0007"), fil enduit de polyimide platine iridié (90 % - 10 %). Les conseils des tétrodes ont été cultivées en platine noir solution pour réduire les impédances d’électrode à entre 100 et 200 kΩ à 1 kHz. L’hyperdrive fut implanté à 4,6 mm à gauche de la ligne médiane et 0,5 mm antérieure du ...

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

Nous présentons la construction d’un hyperdrive 3D-imprimable avec dix-huit tétrodes réglables indépendamment. L’hyperdrive est conçu pour enregistrer l’activité cérébrale en comportement librement des rats sur une période de plusieurs semaines.

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 ...

The overall goal of this procedure is to construct a recording device with multiple independently adjustable electrodes for the examination of brain activity in freely behaving rats. So this method is very exciting because it will help answer several questions related to special navigation, decision making, and other cognitive processes. The main advantage of this hyperdrive is that it provides reliable neuro recording in the brain using materials that are easy to obtain.Although this method was designed to be used specifically with rats, it can be used with any species of similar or larger head sizes such as non-human primates, thereby allowing investigations of many species. Those new to this method might struggle with the nuance involved in the preparation of the steps cause there's a certain level of experience and accuracy that's required to develop these tricks that can make a successful drive. Begin by expanding the holes in the 3D printed core using different sized drill bits.Use a diameter of 0.61 millimeter for the 12 ground wire through holes. For the 18 tetrode through holes, first use a 0.66 millimeter bit and then widen with a 0.71 millimeter bit. A 0.84 millimeter diameter bit is used for the 18 guide rod blind holes.Next, thread the two through holes on top of the core and the remaining eight blind holes with a 0.80 tap. Use a bottoming tap for the blind holes. Create external threads at the base of the core using a 3/8-24 die.Adjust the die properly so the hyperdrive nut will fit over the new threads. Depending on the desired number of ground wires, insert multiple six millimeter long segments of 23 gauge metal tubing into the ground wire holes in the core gluing them if necessary. File the outer ends of the ground wire cannulas until they are flushed with the core and then clean the cannulas with a 0.30 millimeter diameter steel wire.Next, fully insert 18 08015.88 millimeter long flathead screws head down into the slots in the core. Do not bend the screws or damage the threads during this process. Using the rod positioning complex into the core station, position 18 17 millimeter segments of 0.89 millimeter diameter welding rod over the guide rod holes in the core and hammer them down about five millimeters until flushed with the screws.Correct the positions of the welding rods and screws if necessary then tighten the central shoulder screw and the surrounding six screws in the rod positioning complex to secure the outward directions of the rods in the core. Screw the nut onto the core with the rod positioning complex and fit the core into the hyperdrive holder to allow easier positioning under a stereoscope. Fill the slots with dilute dental cement to secure the screws to the core.Fill two to three slots at a time before the dental cement gets too thick. Scrape away any excess dental cement on the core to maintain a proper fit with the shield then allow to air dry for 15 minutes. Next, glue the screws and rods into the core with thin super glue and allow to dry for 15 minutes.Clean and expand the two outer holes in the 3D printed shuttle with drill bits. Use a 0.61 millimeter diameter bit for the smaller hole and a 0.89 millimeter bit for the larger hole. Insert the shuttle bolt into the bolt holder base ensuring the proper orientation.Then close the bolt holder lid. Hold the bolt holder tightly and thread slowly through the hole in the lid with a 0.80 tap. Tap two or three times until smooth.Then insert the shuttle bolt into the shuttle from the side with the smaller opening. Place the shuttle-shuttle bolt complex upside down in the microdrive assembly station base. Next, smooth both ends of a 15 millimeter segment of 23 gauge metal tubing, then position the tubing over the 0.61 millimeter diameter hole guided by the slot in the station lid.Hammer the cannula into the hole until the upper end is flushed with the station lid. Remove the outer half of the upper tip of the cannula with a sanding wheel. Clean the cannula with a 0.30 millimeter diameter metal wire.Then glue the cannula onto the shuttle using thin super glue making sure not to glue the shuttle bolts to the shuttle. Prepare at least 18 microdrives. Test the microdrives on the microdrive rack.Make sure that the shuttle bolt can rotate smoothly in the shuttle and that the entire microdrive moves freely along the length of the threaded rod. To insert guide cannulas into the hyperdrive core, begin by removing the heat shrink tubes and slide a four millimeter segment of silicone tubing along the bundle to the soldered-unsoldered border. Wedge the slit in the hyperdrive spacer to widen the central hole allowing the spacer to slip around the silicone tube.Remove the wedge when the spacer sits at the center of the silicone tube. Next, organize the positions of the guide cannulas in the bundle by placing 10 centimeter long segments of 0.18 millimeter diameter metal wire through each cannula into a specific tetrode hole in the hyperdrive core preventing any crossover of the wires or cannulas in the process. Bend the end of the wires to hold them in place.Push the cannulas through their respective holes in the core being careful to avoid bending or crossing between them until the free end of each cannula is at least two millimeters outside the upper end of the tetrode hole. Secure the spacer by screwing the nut onto the core being careful to prevent the spacer from rotating. Apply a drop of very dilute dental cement from the top of the core onto the junction of the cannulas to secure their relative positions.Cut the guide wires from the soldered end of the bundle and remove them from the cannulas by retracting from the free end. Load the microdrives slowly and carefully onto each threaded rod of the core. Confirm that the 23 gauge microdrive cannula goes into the tetrode hole smoothly, the 30 gauge guide cannula goes into the 23 gauge microdrive cannula smoothly, and the shuttle bolt turn smoothly along the threaded rod.Screw the microdrives down to one to 1.5 millimeters above the lower end of the threaded rods. Then cut 18 pieces of polyamide tubing into 43 millimeter segments which is the length of the guide cannula bundle plus seven millimeters. Clean each tube with a 0.8 millimeter diameter steel wire.Invert the core, insert the polyamide tubes carefully into the guide cannulas from the soldered end and push them all the way in. Then flip the core upright and glue the upper end of the polyamide tube onto the microdrive cannula with thick super glue. Place the core upside down and let the glue dry for 15 minutes.After drying, cut the extra polyamide tubing at the upper end leaving 0.5 to one millimeter outside the microdrive cannula. To assemble the ground wires, first cut lengths of 25 to 30 millimeters from coated steel wire. Then strip two millimeters of the plastic insulation from both tips of the wires and insert one end of each into both sides of a six to eight millimeter long 30 gauge cannula.Flatten this end of each cannula to secure the connection to their respective wires. Finally, insert the round end of the 30 gauge cannula into the upper end of the ground wire cannula in the core and press to make the insertion tight. Then complete assembly of the multi-tetrode hyperdrive according to the instructions in the written protocol.This scatter plot shows the relationship between peak to peak amplitudes of spikes recorded from two electrodes of a tetrode located in the post rhinal cortex. Each dot corresponds to one spike. Clusters of spikes are likely to originate from the same cell.Four clusters are color coded as seen here. The mean spike waveforms from all four tetrode channels of the color coded cells shown in the previous scatter plot are seen here. Traces of local field potential in the theta frequency range recorded simultaneously from four different tetrodes located in the medial entorhinal cortex when the rat was freely foraging are seen here.While attempting this procedure, it's important to work with precision throughout the entire procedure to ensure that all parts fit together properly. Also use care and wear proper PPE when working with the demo tool to avoid injury. After watching this video, you should have a good understanding of how to construct these improved hyperdrives carrying multiple tetrodes for chronic neuro recordings in behaving rats.

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