Questo studio è stato sostenuto dal National Institute on Drug Abuse Borse di DA 006.886 e 032.270 DA.

<ol><li> Cablare i motori passo-passo bipolari avvitando i fili nei connettori forniti con la scheda driver. Colori dei fili su motori passo passo bipolari sono standardizzate (Figura 1). Nota: I motori passo-passo descritte hanno una risoluzione di 1,8 ° / step, volto a 0,346 ° / step. Un stereotax standard ha 3 mm/360 ° di viaggio. La risoluzione finale è 2.88 micron / step. I motori sono anche in grado di intervenire frazionata. <ol><li> Collegare il filo verde ad A +, collegare il filo nero ad A-, collegare il filo rosso al B + e collegare il filo blu a B-. </li></ol></li><li> Far scorrere gli accoppiatori sui motori passo-passo, facendo attenzione ad allineare i fori di montaggio e fissarli utilizzando 12x Viti M3 Socket testa (20 mm) (Figura 2). <ol><li> Assicurarsi che i manicotti sono saldamente attaccati ai motori. Nota: I modelli 3D non comprendono i thread. Le parti sono etichettati con dimensioni filettatura, ma devono essere sfruttato dopo che sonofabbricati. </li></ol></li><li> Rimuovere le viti di fermo dalle impugnature pollice su tutti e 3 gli assi dello strumento stereotassico con una piccola chiave esagonale. Gli incavi per i pollici sono filettati, in modo da girare in senso antiorario per la rimozione. Conservare le rondelle in PTFE posto sul braccio (Figura 3). </li><li> Avvitare l&#39;estremità filettata dei collari sulle aste filettate braccia dello strumento stereotassico (Figura 3). <ol><li> Assicurarsi che non c&#39;è divario tra i collari e PTFE O-ring. Ciò garantisce che le coordinate vengono mantenuti quando il robot cambia direzione. </li><li> Fissare i collari sui fili delle braccia stereotassica con 3x NF10-32 (1/4 di pollice) Viti punto tazza set. </li><li> Far scorrere ogni motore e accoppiatore sopra i collari e braccia stereotassica. Assicurarsi che i motori si siedono a filo con le armi, e i fori delle viti insieme sui collari allineati con la parte piatta degli alberi motore (Figura 4). </li><li> Fissare il couplers alla stereotax utilizzando i fori di montaggio e 6x NF10-32 (1/2 pollice) Viti punto cup set (Figura 4). </li><li> Fissare i collari per gli alberi motore con 3x NF10-32 (1/4 di pollice) viti di fermo (Figura 4). </li></ol></li><li> Preparare la scheda driver CNC impostando ciascuno dei perni del controller a mezzo passo. Nota: Questo driver motore passo-passo si presenta come un circuito esposto. Un caso può essere costruito, anche se non è necessario. Inoltre, può essere utilizzato un numero di differenti dispositivi di comando motore passo-passo bipolare. Se è così, in modo che le istruzioni sono seguite per la scheda specifica acquistato. <ol><li> Allineare tutte le 6 pin per motore passo-passo nello stesso modo. Half-passo consente il doppio della risoluzione passo in Gradi / step (Figura 5). </li><li> Capovolgere il pin 1 in posizione ON, pin 2 in posizione off, pin 3 alla posizione inzione, pin 4 nella posizione off, pin 5 in posizione ON, e pin 6 in posizione off (Figura 5). </li></ol></li><li> Collegare i motori (X - Y - Z) nel driver del motore passo-passo, con l&#39;alimentazione a 12 V. Il posizionamento corretto è segnato sul driver. Inoltre, collegare il driver passo passo alla porta seriale di un computer utilizzando un DB25cable (Figura 6). </li><li> Installare il software di fresatura CNC su un personal computer (questo dovrà essere situato in una zona chirurgica) seguendo le istruzioni predefinite. Una volta installato, aprire il software per iniziare la configurazione. <ol><li> Configurare il software per comunicare con i motori passo-passo. Nota: Le seguenti istruzioni devono essere utilizzati solo con il driver del motore passo-passo TB6560. </li><li> Fare clic attraverso i menu del software come segue. Aperto → Config → Porte e Pins→ Segnali di uscita. Compila la richiesta di abbinare la figura 7 e premi applica. </li><li> Fare clic attraverso i menu del software come segue. Aprire Config → → Porte e pin → segnali di ingresso. Compila la richiesta di abbinare la figura 8 e premi applica. </li><li> Fare clic attraverso i menu del software come segue. Aprire Config → → Porte e Spille → uscite motore. Compila la richiesta di abbinare Figura 9 e premi applica. </li><li> Fare clic attraverso i menu del software come segue. Aprire → sintonia Config → Motor. Compila la richiesta di abbinare Figura 10 e fare clic su Salva impostazioni dell&#39;Asse. Ripetere il passaggio precedente per tutti e 3 gli assi utilizzando gli stessi valori. </li></ol></li><li> Calibrare il stereotax alla scala del software CNC. Nota: Il software è progettato per le macchine di fresatura standard, quindi la sua unità di misura non sarà proporzionale alla corsa di uno strumento stereotassico.<ol><li> Impostare la velocità dei motori &#39;di 1 pollice per minuto, e &quot;jog&quot; Z-axis dello strumento stereotassico con PgUp / PgDn al millimetro. Nota: La velocità di jogging consigliata massima è 3.500 micron / sec e la velocità di taglio massimo consigliato è di 500 micron / sec. </li><li> Zero l&#39;asse Z, e il Jog stereotassico strumento 1 mm. La distanza percorsa sulla Z in Mach3 è il &quot;Scaling costante&quot;. Le coordinate della macchina sono determinate moltiplicando le coordinate del cranio (mm) dal &quot;Scaling costante&quot;. </li><li> Eseguire i test casuali di tutti e 3 gli assi programmando loro di percorrere alcune distanze note, e garantire i movimenti sono accurati. Se la corsa stereotax è troppo lontano o corto, modificare la costante ridimensionamento di conseguenza. Nota: Una volta che il ridimensionamento è completo, possono essere utilizzati i script personalizzati inclusi per generare g-code per gli ambulatori. Tuttavia, si consiglia vivamente agli utenti di familiarizzare con g-code prima di tentare di generare automaticamenteinterventi chirurgici. Questo è indispensabile per la risoluzione dei problemi e la modifica ambulatori automatizzati. </li></ol></li><li> Fissare il micro trapano del motore allo strumento stereotassico utilizzando il supporto extra large sonda. Nota: La velocità minima consigliata punta è di 40.000 rpm. </li><li> Generazione automatica di codice G per una craniotomia spigolo vivo con 3 fori cranio. <ol><li> Mettere tutti gli script personalizzati dalla tabella software in un&#39;unica cartella sul PC. </li><li> Aprire lo script &quot;SharpEdgeCraniotomy.m&quot; ed eseguire il codice. </li><li> Selezionare Sia al prompt &quot;Che tipo di chirurgia sarai Performing?&quot; (Figura 11). </li><li> Selezionare Personalizzata per definire gli angoli della finestra cranio. Compilare in ogni prompt per abbinare Figura 12. </li><li> Definire le posizioni X e Y degli angoli craniotomia. Ogni coordinata deve essere indicato in modo corretto, secondo l&#39;esempio in Figura 13 </strong&gt;. </li><li> Inserire 3 nel prompt di produrre 3 fori cranio (Figura 14). </li><li> Selezionare Definisci utilizzando le coordinate, inserisci le coordinate di ogni foro dal modello in Figura 15. Nota: Se le coordinate precise non sono importanti, c&#39;è la possibilità di puntare e fare clic collocare i buchi su un immagine di un teschio ratto. Le posizioni saranno generati automaticamente. </li><li> Definire i parametri di perforazione. Per il primo intervento chirurgico prove, accettare i valori predefiniti. Nota: Questi valori dipendono i motori passo-passo, e cranio dell&#39;animale. Ogni ratto razza e di destinazione posizione ha uno spessore cranio leggermente diverso. Per i pochi interventi chirurgici iniziali utilizzo di questo dispositivo, essere pronti a testare la profondità di foratura e rimuovere manualmente eventuali pezzi di cranio rimanenti. I valori possono essere modificate per interventi futuri (Figura 16). </li><li> Nominare il g-code, ma verrà generato e sa automaticamenteved nella directory di lavoro. </li></ol></li><li> Caricare il g-code nel software fresatura CNC e un teschio di prova negli strumenti stereotassica barre orecchio. <ol><li> Jogging manualmente la punta di Bregma utilizzando i tasti freccia. Utilizzare una velocità di corsa lenta (~ 5 pollici / m) per garantire l&#39;accuratezza. </li><li> Avviare la punta rotante a più di 38.000 giri al minuto. </li><li> Premere CycleStart, il stereotax eseguirà molti passaggi dello stesso taglio, a diverse profondità. Tra ogni passaggio, il stereotax metterà in pausa, in modo che il chirurgo può continuare o interrompere il taglio. Premere Continua Ciclo (Alt-R) per continuare a tagliare passaggi. </li></ol></li></ol>

<ol>
	<li>Yin, H. H., Knowlton, B. J., Balleine, B. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lesions+of+dorsolateral+striatum+preserve+outcome+expectancy+but+disrupt+habit+formation+in+instrumental+learning.">Lesions of dorsolateral striatum preserve outcome expectancy but disrupt habit formation in instrumental learning.</a> Eur. J. Neurosci. 19 (1), 181-189 (2004).</li><li>West, M. O., Woodward, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+technique+for+microiontophoretic+study+of+single+neurons+in+the+freely+moving+rat.">A technique for microiontophoretic study of single neurons in the freely moving rat.</a> J. Neurosci. Methods. 11 (3), 179-186 (1984).</li><li>Peoples, L. L., West, M. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phasic+firing+of+single+neurons+in+the+rat+nucleus+accumbens+correlated+with+the+timing+of+intravenous+cocaine+self-administration.">Phasic firing of single neurons in the rat nucleus accumbens correlated with the timing of intravenous cocaine self-administration.</a> J. Neurosci. 16 (10), 3459-3473 (1996).</li><li>Wolske, M., Rompre, P. P., Wise, R. A., West, M. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activation+of+single+neurons+in+the+rat+nucleus+accumbens+during+self-stimulation+of+the+ventral+tegmental+area.">Activation of single neurons in the rat nucleus accumbens during self-stimulation of the ventral tegmental area.</a> J. Neurosci. 13 (1), 1-12 (1993).</li><li>Bozza, T., McGann, J. P., Mombaerts, P., &amp;Wachowiak, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+imaging+of+neuronal+activity+by+targeted+expression+of+a+genetically+encoded+probe+in+the+mouse.">In vivo imaging of neuronal activity by targeted expression of a genetically encoded probe in the mouse.</a> Neuron. 42 (1), 9-21 (2004).</li><li>Pitts, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Office+of+Laboratory+Animal+Welfare.">Office of Laboratory Animal Welfare.</a> Institutional animal care and use committee guidebook. , (2002).</li><li>Yoon, T., Otto, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+contributions+of+dorsal+vs.+ventral+hippocampus+to+auditory+trace+fear+conditioning.">Differential contributions of dorsal vs. ventral hippocampus to auditory trace fear conditioning.</a> Neurobiol. Learn. Mem. 87 (4), 464-475 (2007).</li><li>Root, D. 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=Differential+roles+of+ventral+pallidum+subregions+during+cocaine+self-administration+behaviors.">Differential roles of ventral pallidum subregions during cocaine self-administration behaviors.</a> J. Comp. Neurol. 521 (3), 558-588 (2012).</li><li>Yang, G., Pan, F., Parkhurst, C. N., Grutzendler, J., Gan, W. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thinned-skull+cranial+window+technique+for+long-term+imaging+of+the+cortex+in+live+mice.">Thinned-skull cranial window technique for long-term imaging of the cortex in live mice.</a> Nat. Protoc. 5 (2), 201-208 (2010).</li><li>Feng, L., Sametsky, E. A., Gusev, A. G., Uteshev, V. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Responsiveness+to+nicotine+of+neurons+of+the+caudal+nucleus+of+the+solitary+tract+correlates+with+the+neuronal+projection+target.">Responsiveness to nicotine of neurons of the caudal nucleus of the solitary tract correlates with the neuronal projection target.</a> J. Neurophysiol. 108 (7), 1884-1894 (2012).</li><li>Babaei, P., Soltani Tehrani, ., B, A., Alizadeh, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transplanted+Bone+Marrow+Mesenchymal+Stem+Cells+Improve+Memory+in+Rat+Models+of+Alzheimer's+Disease.">Transplanted Bone Marrow Mesenchymal Stem Cells Improve Memory in Rat Models of Alzheimer's Disease.</a> Stem Cells Int. 2012, 369417 (2012).</li></ol>

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L&#39;uso di attrezzature di chirurgia automatizzato aiuta a eliminare alcuni dei problemi più comuni nella ricerca neuroscientifica. In primo luogo, i percorsi utensile sono al 100% riproducibili. Ogni taglio è garantito essere nella stessa posizione rispetto al bregma. In secondo luogo, occorre ridurre l&#39;errore sperimentatore. Anche se molti ricercatori sono chirurghi altamente qualificati, ci vuole una quantità eccezionale di pratica per diventare anche un chirurgo competente. Questo dispositivo permetterà ai...

Chirurgia stereotassica roditore è utilizzato in un&#39;ampia varietà di applicazioni Neuroscienze lesione 1, 2 ionoforesi, microconduttori impianto 3, 4 stimolazione, e sottile cranio immagini 5. Tuttavia, ci sono i principali ostacoli che devono affrontare coloro che desiderano applicare queste tecniche, tra cui la ripida curva di apprendimento per eseguire la chirurgia stereotassica preciso e la elevata probabilità di errore umano. Gli errori umani includono la misurazione e fallimenti di calcolo, così come la scarsa precisione e la riproducibilità di movimenti umani. Nel tentativo di ridurre questi errori confondenti, i chirurghi stereotassica trarrebbero beneficio da un sistema che assicura che tutte le procedure chirurgiche sono eseguite in modo identico tra i soggetti. La riduzione degli errori è anche un metodo attraverso il quale gli investigatori possono ridurre al minimo l&#39;uso di soggetti animali, un obiettivo primario dei National Institutes of Health per la sperimentazione animale 6. In un mondo ideale, tutti sambulatori tereotactic sarebbe perfettamente replicabile all&#39;interno di esperimenti, e tra i laboratori. Per risolvere questo problema, le aziende hanno sviluppato nuovi stereotaxics ultra-precisi e display digitali per la lettura misurazioni. Per rimuovere gli errori di movimento umano, motorizzati micro manipolatori e stereotaxics sono stati prodotti in commercio, ma il loro alto costo può essere proibitivo per un laboratorio con un budget limitato. Inoltre, il software è completamente proprietaria, e non può essere modificato dal ricercatore per accogliere un nuovo tipo di intervento chirurgico. Una soluzione economica al problema errore umano è quello di costruire un robot da stereotax modello esistente di un laboratorio, utilizzando attrezzature per l&#39;industria CNC standard. A causa di una fiorente comunità di hobbisti CNC, i materiali sono molto meno costosi di attrezzature scientifiche. Questo permette di costruire un accurato strumento stereotassico CNC, che è anche altamente flessibile ed economica. Con una conoscenza di base di lavorazione CNC e G-Code, indivials possono programmare qualsiasi intervento chirurgico stereotassica che si immagina, senza le limitazioni del software proprietario. E, al fine di accelerare la produzione di g-code per interventi chirurgici semplici, questo protocollo include un software che permette all&#39;utente di progettare interventi chirurgici (spigolo craniotomia, sottile windowing cranio, foratura e abbassando impianto) entro punta e clicca menù. Questi programmi di emettere un g-code completato che può essere eseguito direttamente dal software CNC. In tutto, un aggiornamento stereotassico motorizzata è ideale per coloro che hanno un interesse ad aumentare la precisione e la riproducibilità di interventi chirurgici, pur mantenendo la flessibilità e il basso costo di una piattaforma open source.

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			<td height="44" style="height:44px;width:141px;">1x Standard U Frame Stereotax</td>
			<td style="width:137px;">Kopf</td>
			<td style="width:117px;"><a href="http://www.kopfinstruments.com/Stereotaxic/900.html">Kopf</a></td>
			<td style="width:355px;">This protocol should work with most existing stereotaxic devices.</td>
		</tr>
		<tr height="65">
			<td height="65" style="height:65px;width:141px;">3x 12 V, 1.6 A, 233 oz-inch Geared Bipolar Stepper Motor</td>
			<td style="width:137px;">Phidgets</td>
			<td style="width:117px;"><a href="http://www.robotshop.com/geared-bipolar-stepper-motor-3v-233-oz-in.html">Robot Shop</a></td>
			<td style="width:355px;">Any high torque geared stepper motor should do.&#160;</td>
		</tr>
		<tr height="73">
			<td height="73" style="height:73px;width:141px;">1x 3 Axis CNC Stepper Motor Driver Board Controller</td>
			<td style="width:137px;">Toshiba</td>
			<td style="width:117px;"><a href="http://www.ebay.com/itm/CNC-Router-3-Axis-TB6560-3-5A-Stepper-Motor-Driver-Board-For-Engraving-Machine-/300981888819">Ebay</a></td>
			<td style="width:355px;">Any 3 Axis CNC driver should do. Linked Item includes Mach3 CNC software.&#160;</td>
		</tr>
		<tr height="94">
			<td height="94" style="height:94px;width:141px;">2x Arm Couplers: medial-lateral (ML) &#38; dorsal-ventral (DV)</td>
			<td style="width:137px;">custom machined</td>
			<td style="width:117px;"><a href="http://eden.rutgers.edu/~mrcoffey/images/3D%20Models%20-%20CNC%20Stereotax%20Parts.zip">Part Drawings</a></td>
			<td style="width:355px;">These must be machined by your local machine shop. (costs will vary)</td>
		</tr>
		<tr height="76">
			<td height="76" style="height:76px;width:141px;">1x anterior-posterior (AP) Coupler</td>
			<td style="width:137px;">custom machined</td>
			<td style="width:117px;"><a href="http://eden.rutgers.edu/~mrcoffey/images/3D%20Models%20-%20CNC%20Stereotax%20Parts.zip">Part Drawings</a></td>
			<td style="width:355px;">These must be machined by your local machine shop. (costs will vary)</td>
		</tr>
		<tr height="127">
			<td height="172" rowspan="3" style="height:172px;width:141px;">3x Motor to Stereotax Collar</td>
			<td rowspan="3" style="width:137px;">custom machined</td>
			<td style="width:117px;"><a href="http://eden.rutgers.edu/~mrcoffey/images/3D%20Models%20-%20CNC%20Stereotax%20Parts.zip">Part Drawings</a></td>
			<td rowspan="3" style="width:355px;">These must be machined by your local machine shop. (costs will vary)</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:117px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:117px;"><a href="http://eden.rutgers.edu/~mrcoffey/images/Collar.html">View in Browser</a></td>
		</tr>
		<tr height="43">
			<td height="65" rowspan="2" style="height:65px;width:141px;">12x NF10-32 Cup Point Set Screws</td>
			<td rowspan="2" style="width:137px;">McMaster Carr</td>
			<td style="width:117px;"><a href="http://www.mcmaster.com/#92313A829">&#189;&#8221; Length</a></td>
			<td rowspan="2" style="width:355px;">You will need 6 of each.</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:117px;"><a href="http://www.mcmaster.com/#92313A825">&#188;&#8221; Length</a></td>
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			<td height="46" style="height:46px;width:141px;">12x M3 Socket Head Screws (20 mm)</td>
			<td style="width:137px;">McMaster Carr</td>
			<td style="width:117px;"><a href="http://www.mcmaster.com/#92855A319">20mm Length</a></td>
			<td style="width:355px;">You will need 4 for each motor</td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;width:141px;">1x Micro-Motor Drill&#160;</td>
			<td style="width:137px;">Buffalo Dental</td>
			<td style="width:117px;"><a href="http://www.buffalodental.com/pdfnew/x50.pdf">X50</a></td>
			<td style="width:355px;">Any Micromotor drill will work.&#160; At least 38,000 rpm recommended</td>
		</tr>
		<tr height="53">
			<td height="53" style="height:53px;width:141px;">1x 12 V DC Power Supply</td>
			<td style="width:137px;">12 Volt Adapters</td>
			<td style="width:117px;"><a href="http://www.12vadapters.com/adapter/power-supply/12v/7-amp-7a.html">12v DC &#8211; 7 Amp</a></td>
			<td style="width:355px;">Any 12 V DC PSU should work (ensure amperage rating is higher than the sum of the motors&#8217; amperage).</td>
		</tr>
		<tr height="44">
			<td height="44" style="height:44px;width:141px;">1x Extra Large Probe Holder</td>
			<td style="width:137px;">Stoelting</td>
			<td><a href="http://www.stoeltingco.com/neuroscience/accessories/stereotaxic-probe-holders/extra-large-probe-holder-551.html">Stoelting</a></td>
			<td style="width:355px;"></td>
		</tr>
		<tr height="44">
			<td height="44" style="height:44px;width:141px;">1x Grade B Rat Skull</td>
			<td style="width:137px;">Skulls Unlimited</td>
			<td><a href="http://www.skullsunlimited.com/record_variant.php?id=4009">Skulls Unlimited</a></td>
			<td style="width:355px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:141px;">Mach 3 Mill</td>
			<td style="width:137px;">ArtSoft USA</td>
			<td style="width:117px;"><a href="http://www.machsupport.com/ccount/click.php?id=1">Trial Download</a></td>
			<td style="width:355px;">Any Standard CNC controlling software should work.</td>
		</tr>
		<tr height="44">
			<td height="44" style="height:44px;width:141px;">Surgery Designer</td>
			<td style="width:137px;">Kevin Coffey &#38; David Barker</td>
			<td style="width:117px;"><a href="http://www.mathworks.com/matlabcentral/fileexchange/40282-robot-assisted-stereotactic-surgery">MATLAB File Exchange</a></td>
			<td style="width:355px;">These codes are available to modify. We accept no responsibility for your use or modification of code.</td>
		</tr>
	</tbody>
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building an open-source robotic stereotaxic instrument

Questo protocollo include i disegni e il software necessari per aggiornare uno strumento stereotassico esistente a un robot (CNC) strumento stereotassico per circa 1.000 dollari (escluso il trapano), utilizzando l&#39;industria motori passo-passo standard e software di controllo CNC. Ogni asse ha il controllo di velocità variabile e possono essere azionati simultaneamente o indipendentemente. La flessibilità del robot e il sistema di codifica aperta (g-code), lo rendono in grado di svolgere attività personalizzate che non sono supportati da sistemi commerciali. Le sue applicazioni includono, ma non sono limitati a, fori, craniotomie bordi taglienti, cranio diradamento, ed elettrodi abbassamento o cannula. Al fine di accelerare la scrittura di g-codifica per interventi chirurgici semplici, abbiamo sviluppato script personalizzati che permettono agli individui di progettare un intervento chirurgico senza alcuna conoscenza di programmazione. Tuttavia, per gli utenti di ottenere il massimo del stereotax motorizzato, sarebbe utile per essere informati in programmazione matematica e G-Coding (semplice progsperonamento per la lavorazione CNC). La velocità di fresatura consigliata è superiore a 40.000 rpm. La risoluzione del motore passo-passo è 1,8 ° / Step, orientata a 0,346 ° / Step. Un stereotax standard ha una risoluzione di 2,88 micron / step. La velocità di taglio massima raccomandata è di 500 micron / sec. La velocità di jogging consigliata massima è 3.500 micron / sec. La dimensione massima consigliata punta da trapano è HP 2.

Il risultato finale della chirurgia creazione nei metodi sarà un teschio ratto con una craniotomia spigolo vivo, e 3 fori cranio (Figura 17). Si noti che il cranio utilizzato per dimostrare la chirurgia era molto più ampia del ratto cranio prototipo. Il bordo tagliente craniotomia può essere utilizzato per inserire una matrice microconduttori nel cervello, ad alta densità registrazioni neurali. Il stereotax CNC può essere usato per ridurre l&#39;array con grande precisione. Software è incluso in q...

Watch this Scientific Journal Video about Building An Open-source Robotic Stereotaxic Instrument at JoVE.com

Building An Open-source Robotic Stereotaxic Instrument

Questo protocollo include i disegni e il software necessari per aggiornare uno strumento stereotassico esistente a un robot (controllo numerico computerizzato); CNC strumento stereotassico per circa 1.000 dollari (escluso un trapano).

Costruire un Open-source Robotic Stereotassica Instrument

This protocol includes the designs and software necessary to upgrade an existing stereotaxic instrument to a robotic (CNC) stereotaxic instrument for ...

building-an-open-source-robotic-stereotaxic-instrument

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

14.8K Views.  Rutgers, The State University of New Jersey.  Questo protocollo include i disegni e il software necessari per aggiornare uno strumento stereotassico esistente a un robot (controllo numerico computerizzato); CNC strumento stereotassico per circa 1.000 dollari (escluso un trapano). 

Video: Building An Open-source Robotic Stereotaxic Instrument

Rodent Stereotaxic Surgery and Animal Welfare Outcome Improvements for Behavioral Neuroscience

Stereotaxic surgery for the implantation of cannulae into specific brain regions has for many decades been a very successful experimental technique to investigate the effects of locally manipulated neurotransmitter and signaling pathways in awake, behaving animals. Moreover, the stereotaxic implantation of electrodes for electrophysiological stimulation and recording studies has been instrumental to our current understanding of neuroplasticity and brain networks in behaving animals. Ever-increasing knowledge about optimizing surgical techniques in rodents1-4, public awareness concerning animal welfare issues and stringent legislation (e.g., the 2010 European Union Directive on the use of laboratory animals5) prompted us to refine these surgical procedures, particularly with respect to implementing new procedures for oxygen supplementation and the continuous monitoring of blood oxygenation and heart rate levels during the surgery as well as introducing a standardized protocol for post-surgical care. Our observations indicate that these modifications resulted in an increased survival rate and an improvement in the general condition of the animals after surgery (e.g. less weight loss and a more active animal). This video presentation will show the general procedures involved in this type of stereotaxic surgery with special attention to our several modifications. We will illustrate these surgical procedures in rats, but it is also possible to perform this type of surgery in mice or other small laboratory animals by using special adaptors for the stereotaxic apparatus6.

Stereotaxic surgery on rodents allows for targeted administration of drugs or electrical stimulation and recordings in awake, behaving animals. In this video presentation we will demonstrate recent procedural refinements to this long-standing procedure that successfully improved survival rate and reduced post-surgical weight loss.

Chirurgia Stereotassica roditori e animali Miglioramenti Esito di assistenza sociale per Behavioral Neuroscience

Stereotaxic surgery for the implantation of cannulae into specific brain regions has for many decades been a very successful experimental technique to ...

Ricerca

Neuroscienze

Stereotaxic Surgery for Excitotoxic Lesion of Specific Brain Areas in the Adult Rat

Many behavioral functions in mammals, including rodents and humans, are mediated principally by discrete brain regions. A common method for discerning the function of various brain regions for behavior or other experimental outcomes is to implement a localized ablation of function. In humans, patient populations with localized brain lesions are often studied for deficits, in hopes of revealing the underlying function of the damaged area. In rodents, one can experimentally induce lesions of specific brain regions.
Lesion can be accomplished in several ways. Electrolytic lesions can cause localized damage but will damage a variety of cell types as well as traversing fibers from other brain regions that happen to be near the lesion site. Inducible genetic techniques using cell-type specific promoters may also enable site-specific targeting. These techniques are complex and not always practical depending on the target brain area. Excitotoxic lesion using stereotaxic surgery, by contrast, is one of the most reliable and practical methods of lesioning excitatory neurons without damaging local glial cells or traversing fibers.
Here, we present a protocol for stereotaxic infusion of the excitotoxin, N-methyl-D-aspartate (NMDA), into the basolateral amygdala complex. Using anatomical indications, we apply stereotaxic coordinates to determine the location of our target brain region and lower an injection needle in place just above the target. We then infuse our excitotoxin into the brain, resulting in excitotoxic death of nearby neurons. While our experimental subject of choice is a rat, the same methods can be applied to other mammals, with the appropriate adjustments in equipment and coordinates.
This method can be used on a variety of brain regions, including the basolateral amygdala1-6, other amygdala nuclei6, 7, hippocampus8, entorhinal cortex9 and prefrontal cortex10. It can also be used to infuse biological compounds such as viral vectors1, 11. The basic stereotaxic technique could also be adapted for implantation of more permanent osmotic pumps, allowing more prolonged exposure to a compound of interest.

Targeted ablation of specific brain region(s) by infusion of an excitotoxin using stereotaxic coordinates is described. This technique could also be adapted for infusion of other chemicals into the rat brain.

La chirurgia stereotassica della lesione eccitotossico di specifiche aree cerebrali nel ratto adulto

Many behavioral functions in mammals, including rodents and humans, are mediated principally by discrete brain regions. A common method for discerning the ...

Stereotaxic Infusion of Oligomeric Amyloid-beta into the Mouse Hippocampus

Alzheimer&#8217;s disease is a neurodegenerative disease affecting the aging population. A key neuropathological feature of the disease is the over-production of amyloid-beta and the deposition of amyloid-beta plaques in brain regions of the afflicted individuals. Throughout the years scientists have generated numerous Alzheimer&#8217;s disease mouse models that attempt to replicate the amyloid-beta pathology. Unfortunately, the mouse models only selectively mimic the disease features. Neuronal death, a prominent effect in the brains of Alzheimer&#8217;s disease patients, is noticeably lacking in these mice. Hence, we and others have employed a method of directly infusing soluble oligomeric species of amyloid-beta - forms of amyloid-beta that have been proven to be most toxic to neurons - stereotaxically into the brain. In this report we utilize male C57BL/6J mice to document this surgical technique of increasing amyloid-beta levels in a select brain region. The infusion target is the dentate gyrus of the hippocampus because this brain structure, along with the basal forebrain that is connected by the cholinergic circuit, represents one of the areas of degeneration in the disease. The results of elevating amyloid-beta in the dentate gyrus via stereotaxic infusion reveal increases in neuron loss in the dentate gyrus within 1 week, while there is a concomitant increase in cell death and cholinergic neuron loss in the vertical limb of the diagonal band of Broca of the basal forebrain. These effects are observed up to 2 weeks. Our data suggests that the current amyloid-beta infusion model provides an alternative mouse model to address region specific neuron death in a short-term basis. The advantage of this model is that amyloid-beta can be elevated in a spatial and temporal manner.

Here, we present a protocol for direct stereotaxic brain infusion of amyloid-beta. This methodology provides an alternative in vivo mouse model to address the short-term effects of amyloid-beta on brain neurons.

Infuso stereotassico di Oligomeric amiloide-beta nelle Hippocampus mouse

Alzheimer&#8217;s disease is a neurodegenerative disease affecting the aging population. A key neuropathological feature of the disease is the ...

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

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method described here enables the investigator to measure long-lasting (from minutes to hours) high-quality intracellular responses from single Drosophila R1-R6 photoreceptors and Large Monopolar Cells (LMCs) to light stimuli. Because the recording system has low noise, it can be used to study variability among individual cells in the fly eye, and how their outputs reflect the physical properties of the visual environment. We outline all key steps in performing this technique. The basic steps in constructing an appropriate electrophysiology set-up for recording, such as design and selection of the experimental equipment are described. We also explain how to prepare for recording by making appropriate (sharp) recording and (blunt) reference electrodes. Details are given on how to fix an intact fly in a bespoke fly-holder, prepare a small window in its eye and insert a recording electrode through this hole with minimal damage. We explain how to localize the center of a cell's receptive field, dark- or light-adapt the studied cell, and to record its voltage responses to dynamic light stimuli. Finally, we describe the criteria for stable normal recordings, show characteristic high-quality voltage responses of individual cells to different light stimuli, and briefly define how to quantify their signaling performance. Many aspects of the method are technically challenging and require practice and patience to master. But once learned and optimized for the investigator's experimental objectives, it grants outstanding in vivo neurophysiological data.

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

Sharp microelectrodes enable accurate electrophysiological characterization of photoreceptor and visual interneuron output in living Drosophila. Here we show how to use this method to record high-quality voltage responses of individual cells to controlled light stimulation. This method is ideal for studying neural information processing in insect compound eyes.

Metodo elettrofisiologiche per la registrazione intracellulare di tensione Risposte di<em&gt; Drosophila</em&gt; Fotorecettori e Interneuroni a luce stimoli<em&gt; In Vivo</em

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

Non-restraining EEG Radiotelemetry: Epidural and Deep Intracerebral Stereotaxic EEG Electrode Placement

Implantable EEG radiotelemetry is of central relevance in the neurological characterization of transgenic mouse models of neuropsychiatric and neurodegenerative diseases as well as epilepsies. This powerful technique does not only provide valuable insights into the underlying pathophysiological mechanisms, i.e., the etiopathogenesis of CNS related diseases, it also facilitates the development of new translational, i.e., therapeutic approaches. Whereas competing techniques that make use of recorder systems used in jackets or tethered systems suffer from their unphysiological restraining to semi-restraining character, radiotelemetric EEG recordings overcome these disadvantages. Technically, implantable EEG radiotelemetry allows for precise and highly sensitive measurement of epidural and deep, intracerebral EEGs under various physiological and pathophysiological conditions. First, we present a detailed protocol of a straight forward, successful, quick and efficient technique for epidural (surface) EEG recordings resulting in high-quality electrocorticograms. Second, we demonstrate how to implant deep, intracerebral EEG electrodes, e.g., in the hippocampus (electrohippocampogram). For both approaches, a computerized 3D stereotaxic electrode implantation system is used. The radiofrequency transmitter itself is implanted into a subcutaneous pouch in both mice and rats. Special attention also has to be paid to pre-, peri- and postoperative treatment of the experimental animals. Preoperative preparation of mice and rats, suitable anesthesia as well as postoperative treatment and pain management are described in detail.

Non-restraining EEG radiotelemetry is a valuable methodological approach to record in vivo long-term electroencephalograms from freely moving rodents. This detailed protocol describes stereotaxic epidural and deep intracerebral electrode placement in different brain regions in order to obtain reliable recordings of CNS rhythmicity and CNS related behavioral stages.

Non restrittivo EEG Radiotelemetry: epidurale e Deep intracerebrale Stereotassica EEG Posizionamento degli elettrodi

Implantable EEG radiotelemetry is of central relevance in the neurological characterization of transgenic mouse models of neuropsychiatric and ...

Automated Analysis of C. elegans Swim Behavior Using CeleST Software

Dissecting the neuronal and neuromuscular circuits that regulate behavior remains a major challenge in biology. The nematode Caenorhabditis elegans has proven to be an invaluable model organism in helping to tackle this challenge, from inspiring technological approaches,&#160;building the human brain connectome, to actually shedding light on the specific molecular drivers of basic functional patterns. The bulk of the behavioral studies in C. elegans have been performed on solid substrates. In liquid, animals exhibit behavioral patterns that include movement at a range of speeds in 3D, as well as partial body movements, such as a posterior curl without anterior shape change, which introduce new challenges for quantitation. The steps of a simple procedure, and use of a software that enables high-resolution analysis of C. elegans swim behavior, are presented here. The software, named CeleST, uses a specialized computer program that tracks multiple animals simultaneously and provides novel measures of C. elegans locomotion in liquid (swimming). The measures are mostly grounded in animal posture and based on mathematics used in computer vision and pattern recognition, without computational requirements for threshold cut-offs. The software tool can be used to both assess overall swimming prowess in hundreds of animals from combined small batch trials and to reveal novel phenotypes even in well-characterized genetic mutants. The preparation of specimens for analysis with CeleST is simple and low-tech, enabling wide adaptation by the scientific community. Use of the computational approach described here should therefore contribute to the greater understanding of behavior and behavioral circuits in the C. elegans model.

Automated Analysis of C. elegans Swim Behavior Using CeleST Software

An efficient and simple methodology for computer-based analysis of nematode swimming behavior in liquid is described. The method requires little to no investment for C. elegans laboratories. The hardware used is standard, and the computer software for the behavioral analysis (CeleST) is an open source one.

Analisi automatizzata di<em&gt; C. elegans</em&gt; Comportamento Swim Utilizzando Celest Software

Dissecting the neuronal and neuromuscular circuits that regulate behavior remains a major challenge in biology. The nematode Caenorhabditis elegans has ...

High-density Electroencephalographic Acquisition in a Rodent Model Using Low-cost and Open-source Resources

Advanced electroencephalographic analysis techniques requiring high spatial resolution, including electrical source imaging and measures of network connectivity, are applicable to an expanding variety of questions in neuroscience. Performing these kinds of analyses in a rodent model requires higher electrode density than traditional screw electrodes can accomplish. While higher-density electroencephalographic montages for rodents exist, they are of limited availability to most researchers, are not robust enough for repeated experiments over an extended period of time, or are limited to use in anesthetized rodents.1-3 A proposed low-cost method for constructing a durable, high-count, transcranial electrode array, consisting of bilaterally implantable headpieces is investigated as a means to perform advanced electroencephalogram analyses in mice or rats.

Procedures for headpiece fabrication and surgical implantation necessary to produce high signal to noise, low-impedance electroencephalographic and electromyographic signals are presented. While the methodology is useful in both rats and mice, this manuscript focuses on the more challenging implementation for the smaller mouse skull. Freely moving mice are only tethered to cables via a common adapter during recording. One version of this electrode system that includes 26 electroencephalographic channels and 4 electromyographic channels is described below.

Instructions for the low-cost construction and surgical implantation of a chronic transcranial high-density electroencephalographic montage into mice are provided. Signal recording, extraction, and processing techniques are also described.

Ad alta densità elettroencefalografico Acquisizione in un modello di roditore Utilizzando risorse a basso costo e open-source

Advanced electroencephalographic analysis techniques requiring high spatial resolution, including electrical source imaging and measures of network ...

Stereotaxic Surgery for Genetic Manipulation in Striatal Cells of Neonatal Mouse Brains

Many genes are expressed in embryonic brains, and some of them are continuously expressed in the brain after birth. For such persistently expressed genes, they may function to regulate the developmental process and/or physiological function in neonatal brains. To investigate neurobiological functions of specific genes in the brain, it is essential to inactivate genes in the brain. Here, we describe a simple stereotaxic method to inactivate gene expression in the striatum of transgenic mice at neonatal time windows. AAV-eGFP-Cre viruses were microinjected into the striatum of Ai14 reporter gene mice at postnatal day (P) 2 by stereotaxic brain surgery. The tdTomato reporter gene expression was detected in P14 striatum, suggesting a successful Cre-loxP mediated DNA recombination in AAV-transduced striatal cells. We further validated this technique by microinjecting AAV-eGFP-Cre viruses into P2Foxp2fl/fl mice. Double labeling of GFP and Foxp2 showed that GFP-positive cells lacked Foxp2 immunoreactivity in P9 striatum, suggesting the loss of Foxp2 protein in AAV-eGFP-Cre transduced striatal cells. Taken together, these results demonstrate an effective genetic deletion by stereotaxically microinjected AAV-eGFP-Cre viruses in specific neuronal populations in the neonatal brains of floxed transgenic mice. In conclusion, our stereotaxic technique provides an easy and simple platform for genetic manipulation in neonatal mouse brains. The technique can not only be used to delete genes in specific regions of neonatal brains, but it also can be used to inject pharmacological drugs, neuronal tracers, genetically modified optogenetics and chemogenetics proteins, neuronal activity indicators and other reagents into the striatum of neonatal mouse brains.

We describe a protocol of stereotaxic surgery with a homemade head-fixed device for microinjecting reagents into the striatum of neonatal mouse brains. This technique allows genetic manipulation in neuronal cells of specific regions of neonatal mouse brains.

Chirurgia stereotassica per la manipolazione genetica in cellule striatali di cervelli di topo neonatale

Many genes are expressed in embryonic brains, and some of them are continuously expressed in the brain after birth. For such persistently expressed genes, ...

An Alternative Approach to Study Primary Events in Neurodegeneration Using Ex Vivo Rat Brain Slices

Despite&#160;numerous studies that attempt to develop reliable animal models&#160;which reflecting the primary processes underlying neurodegeneration, very few have been widely accepted. Here, we propose&#160;a new procedure&#160;adapted from the well-known ex vivo brain slice technique, which offers a closer in vivo-like scenario than in vitro preparations, for investigating the early events triggering cell degeneration, as observed in Alzheimer&#39;s disease (AD). This variation consists of simple and easily reproducible steps, which enable preservation of the anatomical cytoarchitecture of the selected brain region and its local functionality in a physiological milieu. Different anatomical areas can be obtained from the same brain, providing the opportunity to perform multiple experiments with the treatments in question in a site-, dose-, and time-dependent manner. Potential limitations&#160;which could affect the outcomes related to this methodology are related to the conservation of the tissue, i.e., the maintenance of its anatomical integrity during the slicing and incubation steps and the section thickness, which can influence the biochemical and immunohistochemical analysis. This approach can be employed for different purposes, such as exploring molecular mechanisms involved in physiological or pathological conditions, drug screening, or dose-response assays. Finally, this protocol could also reduce the number of animals employed in behavioral studies. The application reported here has been recently described and tested for the first time on ex vivo rat brain slices containing the basal forebrain (BF), which is one of the cerebral regions primarily affected in AD. Specifically, it has been demonstrated that the administration of a toxic peptide derived from the C-terminus of acetylcholinesterase (AChE) could prompt an AD-like profile, triggering, along the antero-posterior axis of the BF, a differential expression of proteins altered in AD, such as the alpha7 nicotinic receptor (&#945;7-nAChR), phosphorylated Tau (p-Tau), and amyloid beta (A&#946;).

An Alternative Approach to Study Primary Events in Neurodegeneration Using Ex Vivo Rat Brain Slices

We present a method which can&#160;provide further insights into the early events underlying neurodegeneration and based on the established ex-vivo brain technique, combining the advantages of in vivo and in vitro experiments. Moreover, it represents a unique opportunity for direct comparison of treated and untreated group in the same anatomical plane.

Un approccio alternativo per studiare eventi primari nella neurodegenerazione utilizzando fettine di cervello di ratto Ex Vivo

Despite&#160;numerous studies that attempt to develop reliable animal models&#160;which reflecting the primary processes underlying neurodegeneration, ...

Stereotaxic Surgery for Implantation of Microelectrode Arrays in the Common Marmoset (Callithrix jacchus)

Marmosets (Callithrix jacchus) are small non-human primates that are gaining popularity in biomedical and preclinical research, including the neurosciences. Phylogenetically, these animals are much closer to humans than rodents. They also display complex behaviors, including a wide range of vocalizations and social interactions. Here, an effective stereotaxic neurosurgical procedure for implantation of recording electrode arrays in the common marmoset is described. This protocol also details the pre- and postoperative steps of animal care that are required to successfully perform such a surgery. Finally, this protocol shows an example of local field potential and spike activity recordings in a freely behaving marmoset 1 week after the surgical procedure. Overall, this method provides an opportunity to study the brain function in awake and freely behaving marmosets. The same protocol can be readily used by researchers working with other small primates. In addition, it can be easily modified to allow other studies requiring implants, such as stimulating electrodes, microinjections, implantation of optrodes or guide cannulas, or ablation of discrete tissue regions.

Stereotaxic Surgery for Implantation of Microelectrode Arrays in the Common Marmoset Callithrix jacchus

This work presents a protocol to perform a stereotaxic, neurosurgical implantation of&#160;microelectrode arrays in the common marmoset. This method specifically enables electrophysiological recordings in freely behaving animals but can be easily adapted to any other similar neurosurgical intervention in this species (e.g., cannula for drug administration or electrodes for brain stimulation).

Costruire un Open-source Robotic Stereotassica Instrument

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