Name: construction of an improved multi-tetrode hyperdrive for large-scale neural recording in behaving rats
Uploaded: 2018-05-09T15:00:00
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हम सिस्टम तंत्रिका विज्ञान और केंद्र के लिए Kavli संस्थान में मोजर लैब धंयवाद तंत्रिका गणना, नार्वे विज्ञान और प्रौद्योगिकी के विश्वविद्यालय, चूहों में जीर्ण तंत्रिका रिकॉर्डिंग प्रक्रियाओं के लिए । यह काम NIH ग्रांट R21 NS098146, और मानव सीमांत विज्ञान कार्यक्रम लंबी अवधि के फैलोशिप LT000211/2016-एल के एल लू के द्वारा समर्थित किया गया था ।

1.3d मॉडल का Stereolithography
<ol><li>hyperdrive भागों और सहायक उपकरण मुद्रित करने के लिए stereolithographic तकनीकों का उपयोग करें । प्रत्येक hyperdrive अठारह शटल, अठारह शटल बोल्ट के शामिल है, और सभी अंय प्लास्टिक के टुकड़ों में से एक (1 चि...

<ol>
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(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. 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यहां, हम अठारह स्वतंत्र रूप से जंगम tetrodes के शामिल एक नव विकसित hyperdrive के निर्माण की प्रक्रिया का वर्णन । ड्राइव सस्ती कई उपलब्ध हार्डवेयर दुकानों पर खरीदा भागों से निर्माण किया जा सकता है, stereolithographic मुद्रण के ?...

सिस्टम तंत्रिका विज्ञान के क्षेत्र में, वैज्ञानिकों का अध्ययन तंत्रिका ऐसे स्थानिक नेविगेशन, स्मृति, और निर्णय लेने के रूप में अंतर्निहित संज्ञानात्मक प्रक्रियाओं को संबद्ध करता है । अध्ययन के इन प्रकार के लिए, यह पशु व्यवहार के दौरान कई व्यक्ति ंयूरॉंस की गतिविधि पर नजर रखने के लिए महत्वपूर्ण है । पिछले दशकों में, दो महत्वपूर्ण अग्रिमों छोटे जानवरों1,2,3में extracellular तंत्रिका रिकॉर्डिंग के लिए प्रयोगात्मक जरूरतों को पूरा किया गया है । पहले tetrode के विकास, चार microwires के एक बंडल के लिए एक साथ1,2,4ंयूरॉंस की तंत्रिका गतिविधि रिकॉर्ड करते थे । एक tetrode के चार चैनलों भर में गतिविधि के अंतर संकेत आयाम कई एक साथ दर्ज की कोशिकाओं से व्यक्तिगत ंयूरॉन गतिविधि के अलगाव के लिए अनुमति देता है5। इसके अलावा, microwires के लचीले प्रकृति tetrode और लक्ष्य सेल जनसंख्या के बीच रिश्तेदार विस्थापन को कम करने tetrode की अधिक से अधिक स्थिरता की अनुमति देता है । Tetrodes अब व्यापक रूप से उपयोग किया जाता है के बजाय विभिन्न प्रजातियों में कई मस्तिष्क अध्ययन के लिए एक एकल इलेक्ट्रोड, कुतर1,2,6, रहनुमाओं7, और कीड़े8सहित. दूसरा एक hyperdrive के विकास के कई स्वतंत्र रूप से चल tetrodes है, जो कई रिकॉर्डिंग स्थानों से ंयूरॉंस की बड़ी आबादी से तंत्रिका गतिविधि के एक साथ निगरानी के लिए अनुमति देता है3, 9,10,11,12.
छोटे जानवरों के लिए एक विश्वसनीय और सस्ती बहु-tetrode रिकॉर्डिंग डिवाइस की उपलब्धता सीमित है । क्लासिक hyperdrive, शुरू में ब्रूस McNaughton13द्वारा विकसित, सफलतापूर्वक पिछले दो दशकों में कई प्रयोगशालाओं में चूहों व्यवहार में स्वतंत्र रूप से तंत्रिका रिकॉर्डिंग के लिए इस्तेमाल किया गया है9,10,14, 15. हालांकि, तकनीकी कारणों के लिए, मूल McNaughton ड्राइव बनाने के लिए आवश्यक घटक अब प्राप्त करने के लिए बहुत मुश्किल है और हाल ही में सुधार डेटा अधिग्रहण इंटरफेस के साथ संगत नहीं हैं । hyperdrive के अंय अच्छी तरह से स्वीकार किए जाते है डिजाइन microdrives व्यक्तिगत दस्तकारी, जो असंगत परिणाम उपज सकता है और पर्याप्त समय12उपभोग की आवश्यकता है । आदेश में चूहों व्यवहार में विभिंन मस्तिष्क क्षेत्रों से तंत्रिका गतिविधि रिकॉर्ड करने के लिए, हम stereolithographic प्रौद्योगिकी का उपयोग कर एक नया hyperdrive विकसित की है । हम निंनलिखित आवश्यकताओं को पूरा करने की मांग की: (1) नई hyperdrive मस्तिष्क में tetrodes के सटीक विस्थापन की अनुमति और कई लक्ष्य क्षेत्रों से स्थिर रिकॉर्डिंग प्रदान करना चाहिए; (2) नई hyperdrive हाल ही में आसान कनेक्शन की अनुमति देने के लिए विकसित चुंबकीय quickclip प्रणाली के साथ संगत होना चाहिए; और (3) नई hyperdrive सही सामग्री आसानी से उपलब्ध के साथ reproduced किया जा सकता है । यहां, हम 3 डी प्रिन्ट hyperdrive युक्त अठारह स्वतंत्र रूप से जंगम tetrodes, McNaughton डिजाइन पर आधारित के निर्माण के लिए एक तकनीक प्रदान करते हैं । प्रोटोकॉल में, हम नए hyperdrive, जो हम सफलतापूर्वक इस्तेमाल किया है के निर्माण की प्रक्रिया के विवरण का वर्णन एकल ंयूरॉन कार्रवाई क्षमता और स्थानीय क्षेत्र postrhinal और औसत दर्जे का entorhinal cortices से एक स्वतंत्र रूप से हफ्तों में क्षमता प्राकृतिक चारा कार्यों के दौरान चूहे बर्ताव ।

<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

कई दिनों से अधिक न्यूरॉन्स की एक बड़ी आबादी के गतिविधि पैटर्न की निगरानी में जाग जानवरों प्रणालियों तंत्रिका विज्ञान के क्षेत्र में एक मूल्यवान तकनीक है. इस तकनीक का एक प्रमुख घटक वांछित मस्तिष्क क्षेत्रों और उनके स्थिरता के रखरखाव में कई इलेक्ट्रोड के सटीक स्थान के होते हैं । यहां, हम एक 3 डी मुद्रण योग्य hyperdrive, जो अठारह स्वतंत्र रूप से समायोज्य tetrodes भी शामिल है के निर्माण के लिए एक प्रोटोकॉल का वर्णन है, और विशेष रूप से vivo में extracellular तंत्रिका रिकॉर्डिंग स्वतंत्र रूप से व्यवहार चूहों में के लिए बनाया गया है । microdrives से जुड़ी tetrodes या तो व्यक्तिगत रूप से ट्रैक के साथ कई मस्तिष्क क्षेत्रों में उन्नत किया जा सकता है, या एक छोटे क्षेत्र में इलेक्ट्रोड की एक सरणी जगह के लिए इस्तेमाल किया जा सकता है. एकाधिक tetrodes कार्रवाई क्षमता के व्यक्तिगत ंयूरॉंस के दर्जनों से एक साथ परीक्षा के लिए अनुमति देते हैं, के रूप में के रूप में अच्छी तरह से स्थानीय क्षेत्र सक्रिय व्यवहार के दौरान मस्तिष्क में ंयूरॉंस की आबादी से क्षमता । इसके अलावा, डिजाइन सरल 3 डी मसौदा सॉफ्टवेयर है कि आसानी से प्रयोगात्मक जरूरतों को अलग करने के लिए संशोधित किया जा सकता है के लिए प्रदान करता है ।

हम परीक्षण के परिणाम प्राप्त करने के लिए एक नव निर्मित hyperdrive का इस्तेमाल किया । ड्राइव ø 17 µm (०.०००७ "), polyimide लेपित प्लेटिनम-इरिडियम (90%-10%) तार से निर्माण tetrodes से सुसज्जित किया गया था । tetrodes के सुझावों क?...

Watch this Scientific Journal Video about Construction of an Improved Multi-Tetrode Hyperdrive for Large-Scale Neural Recording in Behaving Rats at JoVE.com

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

हम अठारह स्वतंत्र रूप से समायोज्य tetrodes के साथ एक 3 डी मुद्रण योग्य hyperdrive के निर्माण वर्तमान । hyperdrive को स्वतंत्र रूप से कई हफ्तों की अवधि में चूहों व्यवहार में मस्तिष्क गतिविधि रिकॉर्ड बनाया गया है ।

चूहों का बर्ताव करने में बड़े पैमाने पर तंत्रिका रिकॉर्डिंग के लिए एक बेहतर बहु-Tetrode Hyperdrive का निर्माण

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.

Research

JoVE Journal

Neuroscience

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

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It is well known that anesthesia alters neural response properties in various regions of the brain.13.  In the auditory system, fundamental response ...

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डिजाइन और छोटे जानवरों में एक अल्ट्रा प्रकाश चिरकारी तंत्रिका रिकॉर्डिंग के लिए Motorized Microdrive की विधानसभा

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Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

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बहु इकाई रिकॉर्डिंग Locust में तंत्रिका गतिविधि विशेषताएँ तरीके (<em&gt; Schistocerca Americana</em&gt;) घ्राण सर्किट

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Neural Circuit Recording from an Intact Cockroach Nervous System

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The cockroach ventral nerve cord preparation is a tractable system for neuroethology experiments, neural network modeling, and testing the physiological ...

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

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Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method ...

Recording Spatially Restricted Oscillations in the Hippocampus of Behaving Mice

चूहों व्यवहार के हिप्पोकैम्पस में स्थानिक प्रतिबंधित दोलनों रिकॉर्डिंग

The local field potential (LFP) emerges from ion movements across neural membranes. Since the voltage recorded by LFP electrodes reflects the summed ...