Name: Building An Open-source Robotic Stereotaxic Instrument (Video) | JoVE Journal
Uploaded: 2013-10-29T19:00:00.000+00:00
Duration: 11 min 40 s
Description: Learn to build an open-source robotic stereotaxic instrument for rodent neurosurgery using CNC machining and G-code generation. This cost-effective method enhances surgical precision and reproducibility for researchers.

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5 minutes

This study was supported by the National Institute on Drug Abuse Grants DA 006886, and DA 032270.

<ol>
	<li>Wire the bipolar stepper motors by screwing the wires into the connectors supplied with the driver board. Wire colors on bipolar stepper motors are standardized (Figure 1). 
	Note: The described stepper motors have a resolution of 1.8&#176;/step, geared to 0.346&#176;/step. A standard stereotax has 3 mm/360&#176; of travel. The final resolution is 2.88 &#956;m/step. The motors are also capable of fractional stepping.
	<ol>
		<li>Connect the green wire to A+, connect the black wire to A-, connect the red wire to B+ and connect the blue wire to B-.</li>
	</ol>
	</li>
	<li>Slide the couplers over the stepper motors, being careful to align the mounting holes, and secure them using 12x M3 Socket Head Screws (20 mm) (Figure 2).
	<ol>
		<li>Ensure that the couplers are firmly attached to the motors. 
		Note: The 3D models do not include threads. The parts are labeled with thread size, but they must be tapped after they are manufactured.</li>
	</ol>
	</li>
	<li>Remove the set screws from the thumb grips on all 3 axes of the stereotaxic instrument using a small hex key. The thumb grips are threaded, so turn them counter-clockwise for removal. Keep the PTFE washers in place on the arm (Figure 3).</li>
	<li>Screw the threaded end of the collars onto the threaded rods of the stereotaxic instrument&#39;s arms (Figure 3).
	<ol>
		<li>Ensure there is no gap between the collars and PTFE O-rings. This guarantees that coordinates are maintained when the robot changes directions.</li>
		<li>Secure the collars onto the threads of the stereotaxic arms using 3x NF10-32 (1/4 inch) cup point set screws.</li>
		<li>Slide each motor and coupler over the collars &#38; stereotaxic arms. Ensure that the motors sit flush with the arms, and the set screw holes on the collars line up with the flat portion of the motor shafts (Figure 4).</li>
		<li>Secure the couplers to the stereotax using the mounting holes and 6x NF10-32 (1/2 inch) cup point set screws (Figure 4).</li>
		<li>Secure the collars to the motor shafts using 3x NF10-32 (1/4 inch) set screws (Figure 4).</li>
	</ol>
	</li>
	<li>Prepare the CNC driver board by setting each of the controller pins to half stepping. 
	Note: This stepper motor driver comes as an exposed circuit board. A case may be built, although it is not necessary. Also, a number of different bipolar stepper motor drivers may be used. If so, ensure that the setup instructions are followed for the specific board purchased.
	<ol>
		<li>Align all 6 pins per stepper motor in the same way. Half-stepping allows for double the step resolution in Degrees/step (Figure 5).</li>
		<li>Flip pin 1 to the on position, pin 2 to the off position, pin 3 to the on position, pin 4 to the off position, pin 5 to the on position, and pin 6 to the off&#160;position (Figure 5).</li>
	</ol>
	</li>
	<li>Plug the motors (X - Y - Z) into the stepper motor driver, along with the 12 V power supply. The correct placement is marked on the driver. Also, attach the stepper driver to a computer&#39;s serial port using a DB25cable (Figure 6).</li>
	<li>Install CNC milling software onto a personal computer (this will need to be located in a surgical area) following the default instructions. Once installed, open the software to begin configuration.
	<ol>
		<li>Configure the software to communicate with the stepper motors. 
		Note: The following instructions are intended for use only with the TB6560 stepper motor driver.</li>
		<li>Click through the software&#39;s menus as follows. Open &#8594;Config&#8594; Ports and Pins&#8594;Output Signals. Fill in the prompt to match Figure 7 and hit apply.</li>
		<li>Click through the software&#39;s menus as follows. Open&#8594;Config&#8594;Ports and Pins&#8594;Input Signals. Fill in the prompt to match Figure 8 and hit apply.</li>
		<li>Click through the software&#39;s menus as follows. Open&#8594;Config&#8594;Ports and Pins&#8594;Motor Outputs. Fill in the prompt to match Figure 9 and hit apply.</li>
		<li>Click through the software&#39;s menus as follows. Open&#8594;Config&#8594;Motor tuning. Fill in the prompt to match&#160;Figure 10 and click&#160;Save Axis Settings. Repeat the previous step for all 3 axes using the same values.</li>
	</ol>
	</li>
	<li>Calibrate the stereotax to the scale of the CNC software. 
	Note: The software is designed for standard milling machines, so its unit of measure will not be proportional to the travel of a stereotaxic instrument.
	<ol>
		<li>Set the motors&#39; velocity to 1 inch per minute, and &#34;jog&#34; the stereotaxic instrument&#39;s Z-axis with PgUp/PgDn to the nearest millimeter. 
		Note: The maximum recommended jogging speed is 3,500 &#181;m/sec and the maximum recommended cutting speed is 500 &#181;m/sec.</li>
		<li>Zero the Z-axis, and Jog the stereotaxic instrument 1 mm. The distance traveled on the Z-axis in Mach3 is the &#34;Scaling Constant&#34;. Machine coordinates are determined by multiplying skull coordinates (mm) by the &#34;Scaling Constant&#34;.</li>
		<li>Perform random tests of all 3 axes by programming them to travel some known distances, and ensure the movements are accurate. If the stereotax travel is too far or short, modify the scaling constant accordingly. 
		Note: Once scaling is complete, the included custom scripts may be used to generate g-code for surgeries. However, it is highly recommended that users become familiar with g-code before attempting to auto-generate surgeries. This is imperative for troubleshooting and modifying automated surgeries.</li>
	</ol>
	</li>
	<li>Attach the micro motor drill to the stereotaxic instrument using the extra large probe holder. Note: The minimum recommended drill bit speed is 40,000 rpm.</li>
	<li>Auto-generate G-Code for a sharp edge craniotomy with 3 skull screw holes.
	<ol>
		<li>Place all of the custom scripts from the software table into a single folder on a PC.</li>
		<li>Open the script &#34;SharpEdgeCraniotomy.m&#34;and run the code.</li>
		<li>Select Both&#160;to the prompt &#34;What Type of Surgery Will You be Performing?&#34; (Figure 11).</li>
		<li>Select&#160;Custom, to define the corners of the skull window. Fill in each prompt to match Figure 12.</li>
		<li>Define the X and Y positions of the craniotomy corners. Each coordinate must be entered in correct order, according the example in Figure 13.</li>
		<li>Enter 3 in the prompt to produce 3 skull holes (Figure 14).</li>
		<li>Select Define Using Coordinates, and enter the coordinates of each hole from the template in Figure 15. 
		Note: If precise coordinates are not important, there is an option to point and click the holes&#39; positions onto an image of a rat skull. Positions will be automatically generated.</li>
		<li>Define the drilling parameters. For the first tests surgery, accept the default values. 
		Note: These values are dependent on the stepper motors, and the animal&#39;s skull. Every rat breed and target location has a slightly different skull thickness. For the initial few surgeries using this device, be prepared to test the drilling depths and remove any remaining skull pieces manually. The values can then be modified for future surgeries (Figure 16).</li>
		<li>Name the g-code; it will be automatically generated and saved to the working directory.</li>
	</ol>
	</li>
	<li>Load the g-code into the CNC milling software and a test skull into the stereotaxic instruments ear bars.
	<ol>
		<li>Manually jog the drill bit to Bregma using the arrow keys. Use a slow jog speed (~5 inch/m) to ensure accuracy.</li>
		<li>Start the drill bit rotating at greater than 38,000 rpm.</li>
		<li>Press&#160;CycleStart; the stereotax will perform many passes of the same cut, at different depths. Between each pass, the stereotax will pause, so the surgeon may continue or abort cutting. Press Continue Cycle (Alt-R) to continue cutting passes.</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/pubmed?term=((Lesions+of+dorsolateral+striatum+preserve+outcome+expectancy+but+disrupt+habit+formation+in+instrumental+learning%5BTitle%5D)+AND+%22Eur.+J.+Neurosci%22%5BJournal%5D)">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/pubmed?term=((A+technique+for+microiontophoretic+study+of+single+neurons+in+the+freely+moving+rat%5BTitle%5D)+AND+%22J.+Neurosci.+Methods%22%5BJournal%5D)">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/pubmed?term=((Phasic+firing+of+single+neurons+in+the+rat+nucleus+accumbens+correlated+with+the+timing+of+intravenous+cocaine+self-administration%5BTitle%5D)+AND+%22J.+Neurosci%22%5BJournal%5D)">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/pubmed?term=((Activation+of+single+neurons+in+the+rat+nucleus+accumbens+during+self-stimulation+of+the+ventral+tegmental+area%5BTitle%5D)+AND+%22J.+Neurosci%22%5BJournal%5D)">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., &Wachowiak, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((In+vivo+imaging+of+neuronal+activity+by+targeted+expression+of+a+genetically+encoded+probe+in+the+mouse%5BTitle%5D)+AND+%22Neuron%22%5BJournal%5D)">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. Office of Laboratory Animal Welfare. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aPitts%2C+M+%22Institutional+animal+care+and+use+committee+guidebook%22">Institutional animal care and use committee guidebook</a>. , (2002).</li>
<li>Yoon, T., Otto, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Differential+contributions+of+dorsal+vs.+ventral+hippocampus+to+auditory+trace+fear+conditioning%5BTitle%5D)+AND+%22Neurobiol.+Learn.+Mem%22%5BJournal%5D)">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/pubmed?term=((Differential+roles+of+ventral+pallidum+subregions+during+cocaine+self-administration+behaviors%5BTitle%5D)+AND+%22J.+Comp.+Neurol%22%5BJournal%5D)">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/pubmed?term=((Thinned-skull+cranial+window+technique+for+long-term+imaging+of+the+cortex+in+live+mice%5BTitle%5D)+AND+%22Nat.+Protoc%22%5BJournal%5D)">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/pubmed?term=((Responsiveness+to+nicotine+of+neurons+of+the+caudal+nucleus+of+the+solitary+tract+correlates+with+the+neuronal+projection+target%5BTitle%5D)+AND+%22J.+Neurophysiol%22%5BJournal%5D)">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,, Alizadeh, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Transplanted+Bone+Marrow+Mesenchymal+Stem+Cells+Improve+Memory+in+Rat+Models+of+Alzheimer%27s+Disease%5BTitle%5D)+AND+%22Stem+Cells+Int%22%5BJournal%5D)">Transplanted Bone Marrow Mesenchymal Stem Cells Improve Memory in Rat Models of Alzheimer's Disease.</a> Stem Cells Int. 2012, 369417(2012).</li>
</ol>

The authors have no competing financial interests to disclose.

The use of automated surgery equipment helps to eliminate some of the most common problems in neuroscience research. First, the tool paths are 100% reproducible. Every cut is guaranteed to be in the same location relative to Bregma. Second, it should reduce experimenter error. Although many researchers are highly skilled surgeons, it takes an exceptional amount of practice to become even a competent surgeon. This device will allow new students to quickly and easily perform highly accurate surgeries. Third, motorized surg...

Stereotactic rodent surgery is used in a wide variety of neuroscience applications, including lesioning1, iontophoresis2, microwire implantation3, stimulation4, and thin skull imaging5. However, there are major hurdles facing those who wish to apply these techniques, including the steep learning curve for performing accurate stereotactic surgery and the high probability of human error. Human errors include measurement and calculation failures, as well as the low accuracy and replicability of human movements. In an effort to reduce these confounding errors, stereotactic surgeons would benefit from a system that ensures that all surgical procedures are performed identically across subjects. The reduction of errors is also one method by which investigators can minimize the use of animal subjects, a primary goal of the National Institutes of Health for animal experimentation6. In an ideal world, all stereotactic surgeries would be perfectly replicable within experiments, and between labs. To address this issue, companies have developed new ultra-precise stereotaxics, and digital displays for reading measurements. To remove human movement errors, motorized micro manipulators and stereotaxics were produced commercially, but their high cost can be prohibitive to a laboratory with a limited budget. Also, their software is fully proprietary, and cannot be modified by the researcher to accommodate a new type of surgery.
An affordable solution to the human error problem is to build a robotic stereotax from a lab&#39;s existing model, using industry standard CNC equipment. Because of a burgeoning CNC hobbyist community, the materials are significantly less expensive than scientific equipment. This allows one to build an accurate CNC stereotaxic instrument, which is also highly flexible and inexpensive. With a basic knowledge of CNC machining and G-Code, individuals can program any stereotactic surgery that they imagine, without the limitations of proprietary software. And, in order to expedite the production of g-code for simple surgeries, this protocol includes software that allows the user to design surgeries (sharp edge craniotomy, thin skull windowing, hole drilling &#38; implant lowering) within point and click menus. These programs output a completed g-code that may be run directly from CNC software.
In all, a motorized stereotaxic upgrade is ideal for those who have an interest in increasing the accuracy and replicability of surgeries, while retaining the flexibility and low cost of an open source platform.

<table><tbody><tr><td>1x Standard U Frame Stereotax</td><td>Kopf</td><td>&lt;a href=&quot;http://www.kopfinstruments.com/Stereotaxic/900.html&quot;&gt;Kopf&lt;/a&gt;</td><td>This protocol should work with most existing stereotaxic devices.</td></tr><tr><td>3x 12 V, 1.6 A, 233 oz-inch Geared Bipolar Stepper Motor</td><td>Phidgets</td><td>&lt;a href=&quot;http://www.robotshop.com/geared-bipolar-stepper-motor-3v-233-oz-in.html&quot;&gt;Robot Shop&lt;/a&gt;</td><td>Any high torque geared stepper motor should do.&amp;nbsp;</td></tr><tr><td>1x 3 Axis CNC Stepper Motor Driver Board Controller</td><td>Toshiba</td><td>&lt;a href=&quot;http://www.ebay.com/itm/CNC-Router-3-Axis-TB6560-3-5A-Stepper-Motor-Driver-Board-For-Engraving-Machine-/300981888819&quot;&gt;Ebay&lt;/a&gt;</td><td>Any 3 Axis CNC driver should do. Linked Item includes Mach3 CNC software.&amp;nbsp;</td></tr><tr><td>2x Arm Couplers: medial-lateral (ML) &amp;amp; dorsal-ventral (DV)</td><td>custom machined</td><td>&lt;a href=&quot;http://eden.rutgers.edu/~mrcoffey/images/3D%20Models%20-%20CNC%20Stereotax%20Parts.zip&quot;&gt;Part Drawings&lt;/a&gt;</td><td>These must be machined by your local machine shop. (costs will vary)</td></tr><tr><td>1x anterior-posterior (AP) Coupler</td><td>custom machined</td><td>&lt;a href=&quot;http://eden.rutgers.edu/~mrcoffey/images/3D%20Models%20-%20CNC%20Stereotax%20Parts.zip&quot;&gt;Part Drawings&lt;/a&gt;</td><td>These must be machined by your local machine shop. (costs will vary)</td></tr><tr><td>3x Motor to Stereotax Collar</td><td>custom machined</td><td>&lt;a href=&quot;http://eden.rutgers.edu/~mrcoffey/images/3D%20Models%20-%20CNC%20Stereotax%20Parts.zip&quot;&gt;Part Drawings&lt;/a&gt;</td><td>These must be machined by your local machine shop. (costs will vary)</td></tr><tr><td>&lt;a href=&quot;http://eden.rutgers.edu/~mrcoffey/images/Collar.html&quot;&gt;View in Browser&lt;/a&gt;</td><td></td><td></td><td></td></tr><tr><td>12x NF10-32 Cup Point Set Screws</td><td>McMaster Carr</td><td>&lt;a href=&quot;http://www.mcmaster.com/#92313A829&quot;&gt;&amp;frac12;&amp;rdquo; Length&lt;/a&gt;</td><td>You will need 6 of each.</td></tr><tr><td>&lt;a href=&quot;http://www.mcmaster.com/#92313A825&quot;&gt;&amp;frac14;&amp;rdquo; Length&lt;/a&gt;</td><td></td><td></td><td></td></tr><tr><td>12x M3 Socket Head Screws (20 mm)</td><td>McMaster Carr</td><td>&lt;a href=&quot;http://www.mcmaster.com/#92855A319&quot;&gt;20mm Length&lt;/a&gt;</td><td>You will need 4 for each motor</td></tr><tr><td>1x Micro-Motor Drill&amp;nbsp;</td><td>Buffalo Dental</td><td>&lt;a href=&quot;http://www.buffalodental.com/pdfnew/x50.pdf&quot;&gt;X50&lt;/a&gt;</td><td>Any Micromotor drill will work.&amp;nbsp; At least 38,000 rpm recommended</td></tr><tr><td>1x 12 V DC Power Supply</td><td>12 Volt Adapters</td><td>&lt;a href=&quot;http://www.12vadapters.com/adapter/power-supply/12v/7-amp-7a.html&quot;&gt;12v DC &amp;ndash; 7 Amp&lt;/a&gt;</td><td>Any 12 V DC PSU should work (ensure amperage rating is higher than the sum of the motors&amp;rsquo; amperage).</td></tr><tr><td>1x Extra Large Probe Holder</td><td>Stoelting</td><td>&lt;a href=&quot;http://www.stoeltingco.com/neuroscience/accessories/stereotaxic-probe-holders/extra-large-probe-holder-551.html&quot;&gt;Stoelting&lt;/a&gt;</td><td></td></tr><tr><td>1x Grade B Rat Skull</td><td>Skulls Unlimited</td><td>&lt;a href=&quot;http://www.skullsunlimited.com/record_variant.php?id=4009&quot;&gt;Skulls Unlimited&lt;/a&gt;</td><td></td></tr><tr><td>Mach 3 Mill</td><td>ArtSoft USA</td><td>&lt;a href=&quot;http://www.machsupport.com/ccount/click.php?id=1&quot;&gt;Trial Download&lt;/a&gt;</td><td>Any Standard CNC controlling software should work.</td></tr><tr><td>Surgery Designer</td><td>Kevin Coffey David Barker</td><td>&lt;a href=&quot;http://www.mathworks.com/matlabcentral/fileexchange/40282-robot-assisted-stereotactic-surgery&quot;&gt;MATLAB File Exchange&lt;/a&gt;</td><td>These codes are available to modify. We accept no responsibility for your use or modification of code.</td></tr></tbody></table>

building an open-source robotic stereotaxic instrument

Building An Open-source Robotic Stereotaxic Instrument

This protocol includes the designs and software necessary to upgrade an existing stereotaxic instrument to a robotic (CNC) stereotaxic instrument for around $1,000 (excluding a drill), using industry standard stepper motors and CNC controlling software. Each axis has variable speed control and may be operated simultaneously or independently. The robot&#39;s flexibility and open coding system (g-code) make it capable of performing custom tasks that are not supported by commercial systems. Its applications include, but are not limited to, drilling holes, sharp edge craniotomies, skull thinning, and lowering electrodes or cannula. In order to expedite the writing of g-coding for simple surgeries, we have developed custom scripts that allow individuals to design a surgery with no knowledge of programming. However, for users to get the most out of the motorized stereotax, it would be beneficial to be knowledgeable in mathematical programming and G-Coding (simple programming for CNC machining).
The recommended drill speed is greater than 40,000 rpm. The stepper motor resolution is 1.8&#176;/Step, geared to 0.346&#176;/Step. A standard stereotax has a resolution of 2.88 &#956;m/step. The maximum recommended cutting speed is 500 &#956;m/sec. The maximum recommended jogging speed is 3,500 &#956;m/sec. The maximum recommended drill bit size is HP 2.

The end result of the surgery designed in the methods will be a rat skull with a sharp edge craniotomy, and 3 skull holes (Figure 17). Note that the skull used to demonstrate the surgery was much wider than the prototypical rat skull. The sharp edge craniotomy may be used to insert a microwire array into the brain, for high density neural recordings. The CNC stereotax may also be used to lower the array with great precision. Software is included in this protocol that allows the surgeon to define the para...

This protocol includes the designs and software necessary to upgrade an existing stereotaxic instrument to a robotic (computer numeric controlled; CNC) stereotaxic instrument for around $1,000 (excluding a drill).

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

The overall goal of this procedure is to expand the accessibility and flexibility of robotic rodent neurosurgery by providing researchers with plans to build a flexible, inexpensive, and open source robotic stereotaxic instrument, as well as the software necessary to design and share surgeries. This is accomplished by first machining and assembling a computer numeric controlled stereotaxic instrument. The second step is to determine the scaling ratio of the stereo attacks.By measuring the stereotaxic distance traveled per motor distance, a toolpath will be automatically generated from skull coordinates and will be exported to G code. The final step is to test the surgeries using skulls of the animal species. It is designed for surgery should be modified according to the results of the test.Ultimately, the stereotaxic instrument can be used to quickly perform and share live rodent neurosurgery. Current applications include but are not limited to craniotomies, thin skull, windows, and electrode or cannula. Lowering Robotic surgical instruments can increase the accuracy and replicability of rodent neurosurgery.However, commercial models can be prohibitively expensive. The following protocol provides plans to upgrade an existing stereo attacks to a motorized stereo attacks. G code used to control the motors is open source and may be modified to meet surgeons highly specific needs.The included MATLAB scripts will help early adopters to automatically generate surgeries using simple point and click menus. While only a few generated surgeries are currently supported, the robot may perform the vast majority of neurosurgery techniques used in animal models. Robotic surgeries have many advantages such as eliminating variables at certain times.In the past 30 years, my lab has suffered through periods of low survival rates in our survival surgeries. These are frustrating periods because of the number of variables that have to be eliminated one at a time. Robotic surgery would eliminate some of those variables and simplify our troubleshooting process.In general, it promotes more consistent surgeries and faster recovery times and fewer animal deaths. Begin this procedure by wiring the bipolar stepper motors to the connectors supplied with the driver board. Connect the green wire to a plus the black wire to A minus the red wire to B plus and the blue wire to B minus.Next slide. The couplers over the stepper motors being careful to align the mounting holes. Then secure them using the 1220 millimeter M three socket head screws and ensure that the couplers are firmly attached to the motors.Note that the 3D models for the couplers and collar do not include threads. The holes that are labeled with a thread size must be tapped after they're manufactured. After that, remove the set screws from the thumb grips on all three axes of the stereotaxic instrument using a small hex key.The thumb grips are threaded, so turn them counterclockwise for removal while keeping the PTFE washers in place on the arm. Then screw the threaded end of the collars onto the threaded rods of the stereotaxic instruments arms ensure that there is no gap between the colors and the P-T-F-E-O rings. This guarantees that the coordinates are maintained when the robot changes directions.Subsequently, secure the collars onto the threads of the stereotaxic arms using three one quarter inch nf 10 32 cup point set screws. Slide each motor and coupler over the collars and stereotaxic arms. Ensure that the motors sit flush with the arms and the set screw holes on the collars line up with the flat portion of the motor shafts.Then secure the couplers to the stereotypes using the mounting holes and six half inch NF 10 32 cut point set screws Afterward, secure the collars to the motor shafts using three quarter inch NF 10 32 set screws. Now prepare the CNC driver board by setting each of the three step controllers to half stepping, which allows for double the step resolution for each of the three step controllers. Flip pin one to the on position pin two to the off position pin three to the on position pin four to the off position, pin five to the on position and pin six to the off position.Plug the x, y and Z motors into the stepper motor driver along with the 12 volt power supply. Then attach the stepper driver to a computer serial port using a DB 25 cable. Now open the CNC milling software to begin configuration for the stepper motors communication.Please note that the following instructions are intended for use only with a TB 6 5 6 0 stepper motor driver. Click config, then ports and and output signals. Fill in the prompt and hit apply.Then click config ports and pins and input signals. Fill in the prompt and hit apply. Next, click config.Then ports and pins and motor outputs. Fill in the prompt and hit apply. After that, click config and motor tuning.Fill in the prompt and click save axis settings and repeat these steps for all three AEs using the same values. Next, calibrate the stereotypes to the scale of the CNC software by setting the motor's velocity to one inch per minute and jog the stereotaxic instruments Z axis with page up page down to the nearest millimeter, zero the Z axis and jog the stereotaxic instrument one millimeter. The distance traveled on the Z axis in Mach three is the scaling constant.After performing random tests of all three axes by programming them to travel some known distances to ensure the movements are accurate, attach the micro motor drill to the stereotaxic instrument using the extra large probe holder. First place all of the custom scripts from the software table into a single folder on a pc. Then open the script sharp edge craniotomy M and run the code.Select both at the prompt. What type of surgery will you be performing? Subsequently, select custom to define the corners of the skull window, define the x and Y positions of the craniotomy corners.Each coordinate must be entered in correct order After completing the form, hit okay. Next, enter three in the prompt to produce three skull holes. Then select define using coordinates and enter the coordinates of each hole from the template.After that, define the drilling parameters for the first test surgery. Accept the default values. Name the G code.It will be automatically generated and saved to the working directory. Load the G code into the CNC milling software and a test skull into the stereotaxic instruments. Ear bars manually jug the drill bit torema using the keyboard's arrow keys, use a slow jog speed to ensure accuracy.Now start the micro motor drill and set the speed dial on the drill controller to 50, 000 RPM. Press cycle start. The stereo attacks will perform many passes of the same cut at different depths between each pass.The stereo attacks will pause so the surgeon may continue or abort.Cutting. Press continue cycle to continue cutting passes. Here is a skull showing the end result of running the previously designed surgery, and this skull shows a skull screw inserted into a hole.Produced the drill bits size will determine the hole's, diameter, and consequentially the screw size. This figure shows the skull containing a thin skull window indicated by the arrows. Note that in the right panel, light seems to penetrate the thin skull window.Uniformly Robotic surgical devices can help solve some of the most common problems in neuroscience today. First, tool paths are 100%reproducible. Second, it can reduce experiment or error.Third, motorized surgical devices can reduce the number of animals needed to perform an experiment, and finally, they're more precise and accurate than the human hand. We have included custom software that allows researchers to automatically generate simple surgeries. It's important to remember that the motorized stereo attacks is not constrained by this software.Researchers may code complex tool paths to best suit their unique neurosurgeries. Still, the accuracy of the robot depends on quality machining, proper setup, and proper understanding of how A CNC machine operates. As long as researchers are willing to take the time to understand the functioning of this motorized stereotactic instrument, they may perform surgeries more accurately with better replicability and less training.Overall, automated surgery provides many benefits. Motorized stereotactics exist. They're commercially available, but they're extremely expensive.The system that Kevin and David have put together here is of the same high quality, but for a very low cost, and they're making the software and the system freely available to the whole research community.

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Research

JoVE Journal

Neuroscience

Source: Lee, H. et. al., Successful In vivo Calcium Imaging with a Head-Mount Miniaturized Microscope in the Amygdala of Freely Behaving Mouse. J. Vis. Exp. (2020)This video demonstrates in vivo calcium imaging in a mouse using a gradient refractive index, or GRIN lens, and a miniaturized microscope. The setup combines head fixation within a frictionless mobile cage system, stimulating neurons and triggering calcium ion influx. The GRIN lens captures calcium signaling in the brain, enabling...

Real-Time Microendoscopic Calcium Imaging in a Freely Moving Mouse

Viral vectors allow for targeted gene manipulation. We demonstrate a method for conditional gene expression or ablation in the mouse spinal cord, using stereotaxic injection of a viral vector into the dorsal horn, a prominent site of synaptic contact between primary somatosensory afferents and neurons of the central nervous system.

Stereotaxic Injection of a Viral Vector for Conditional Gene Manipulation in the Mouse Spinal Cord

The SEEG methodology is simplified and made faster with a stereotactic robot. Careful attention must be paid to the registration of the preoperative volumetric MRI to the patient prior to use of the robot in the OR. The robot streamlines the procedure, leading to decreased operative times and accurate implantations.

Operative Technique and Nuances for the Stereoelectroencephalographic (SEEG) Methodology Utilizing a Robotic Stereotactic Guidance System

Described here is a stereotactic procedure that can target challenging and difficult-to-reach brain regions (due to spatial limitations) using an angled coronal approach. This protocol is adaptable to both mouse and rat models and can be applied to diverse neuroscientific applications, including cannula implantation and microinjections of viral constructs.

Adaptable Angled Stereotactic Approach for Versatile Neuroscience Techniques

Focused ultrasound with microbubble agents can open the blood brain barrier focally and transiently. This technique has been used to deliver a wide range of agents across the blood brain barrier. This article provides a detailed protocol for the localized delivery to the rodent brain with or without MRI guidance.

A Benchtop Approach to the Location Specific Blood Brain Barrier Opening using Focused Ultrasound in a Rat Model

The blood-brain barrier (BBB) can be temporarily disrupted with microbubble-mediated focused ultrasound (FUS). Here, we describe a step-by-step protocol for high-throughput BBB opening in vivo using a modular FUS system accessible for non-ultrasound experts.

A High-Throughput Image-Guided Stereotactic Neuronavigation and Focused Ultrasound System for Blood-Brain Barrier Opening in Rodents

This protocol demonstrates the use of a robotic platform for microinjection into single neural stem cells and neurons in brain slices. This technique is versatile and offers a method of tracking cells in tissue with high spatial resolution.

Manipulation of Single Neural Stem Cells and Neurons in Brain Slices using Robotic Microinjection

The human depth perception of 3D stereo videos depends on the camera separation, point of convergence, distance to, and familiarity of the object. This paper presents a robotized method for rapid and reliable test data collection during live open-heart surgery to determine the ideal camera configuration.

Robotized Testing of Camera Positions to Determine Ideal Configuration for Stereo 3D Visualization of Open-Heart Surgery

Here, we present a unique, 3D-printable implant for rats, named TD Drive, capable of symmetric, bilateral wire electrode recordings, currently in up to ten distributed brain areas simultaneously.

The TD Drive: A Parametric, Open-Source Implant for Multi&#45;Area Electrophysiological Recordings in Behaving and Sleeping Rats

This protocol presents robot-assisted stereo electroencephalography (sEEG), a method for the stereotactic implantation of intracerebral electrodes used for invasive seizure monitoring in patients with refractory epilepsy. Techniques incorporating stereotactic robotic guidance for sEEG are safe and precise and may improve surgical efficiency, especially when implanting greater numbers of electrodes.

Robotic-Guided Stereoelectroencephalography for Invasive Epilepsy Monitoring

Learn to build an open-source robotic stereotaxic instrument for rodent neurosurgery using CNC machining and G-code generation. This cost-effective method enhances surgical precision and reproducibility for researchers.

Building An Open-source Robotic Stereotaxic Instrument

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