Surgical Implant Procedure and Wiring Configuration for Continuous Long-Term EEG/ECG Monitoring in Rabbits

Summary

The study outlines a protocol for the design, manufacturing, and placement of a novel continuous EEG/ECG recording system in rabbits. This hard-wired system allows for continuous EEG/ECG recordings from nine distinct electrodes on the head and chest, which facilitates comprehensive assessments of electrical activity in the brain and heart.

Abstract

A multi-system approach that includes the continuous collection of electroencephalogram (EEG) and electrocardiogram (ECG) recordings facilitates a comprehensive assessment of electrical activity in the brain and heart, particularly leading up to and surrounding episodic events. Building on our previous protocol that facilitates the collection of intermittent EEG/ECG recordings from conscious restrained rabbits, we developed a protocol for the collection of continuous EEG/ECG recordings from unrestrained rabbits. This new method enables high-quality 24/7 recordings in the housing cage, which captures the normal range of physiological states and enables the rabbits to move, eat, drink, and sleep freely. It enables comprehensive assessments of the prevalence, incidence, and susceptibility to EEG/ECG abnormalities, seizures, arrhythmias, and sudden death. This procedure involves the surgical placement of subdermal electrodes and the design and manufacturing of a robust wiring system that is impervious to damage by the rabbits. The surgical procedure includes two incisions and the sub-dermal tunneling of nine electrodes and wires that exit through a custom-made port. The external connector attaches to a wire harness that includes an electrical swivel, which allows for free range of rabbit motion throughout its cage. Additionally, the wires are enclosed in a metal sheath suspended by a retractor cable from the ceiling of the rabbit's cage. The wires are then connected to an amplifier/digitizer for the acquisition of high-quality continuous EEG/ECG recordings.

Introduction

We previously developed a protocol to collect acute (< 5 h) video/EEG/ECG recordings from rabbits. It involves the use of nine subdermal EEG and ECG pin electrodes, placement of the rabbit in a restrainer, and the investigator closely monitoring the rabbit for the entire duration of the recording¹. While this system provides high-quality video/EEG/ECG recordings, it has several limitations. Arrhythmias, epileptic seizures, and other EEG/ECG abnormalities may be triggered by various physiological and environmental substrates, so these events may be missed when conducting intermittent recordings. Thus, this recording platform does not facilitate comprehensive and accurate assessments of neuro-cardiac electrical function throughout the day, during various physiological states, or in the rabbits' normal housing environment.

Continuous video/EEG/ECG monitoring is important for evaluating cardiac and neuronal electrical function during various physiological states. It facilitates accurate assessment of the prevalence and incidence of epileptiform activity and arrhythmias, as well as the concordance and progression of multi-system changes surrounding episodic events. For example, circadian fluctuations in autonomic function, as well as stress and activity-mediated autonomic changes, greatly influence heart rate and ECG metrics and can provide a trigger for arrhythmias^2,3. In certain types of epilepsy (e.g., juvenile myoclonic epilepsy), seizures are more likely to occur during sleep or within 1-2 h of waking up⁴. The dynamic changes in electrical activity in the brain during sleep can influence the susceptibility, origin, and spread of seizures⁵.

Stress, activity, and motor seizures can cause muscle artifacts and dislodgement of the electrodes, which influences the quality and stability of the recordings. In particular, during pro-convulsant studies, rabbits experience myoclonic, clonic, tonic, and clonic-tonic seizures. The tonic phase involves the involuntary extension of the neck, which can dislodge the pin electrodes and thus interrupt the signal during a critical phase of the experiment. As such, the temporary system involving pin electrodes has critical limitations that must be addressed. Moreover, as the rabbits are in a restrainer, documenting the full extent of the motor manifestations is difficult.

Therefore, it is important to develop a procedure to implant electrodes for long-term continuous and stable multi-system recordings when the animal is in its normal unrestrained environment. This will facilitate detailed assessments during a wide range of physiological states and in response to environmental triggers. It will also be valuable for capturing rare episodic events. As the setup does not require the investigator to be present during the recording, it increases the efficiency and throughput of the research platform. It also minimizes the frequency of people disturbing the rabbits. Finally, as this procedure allows for the animals to stay in their natural housing environment throughout the recording, it improves the quality of life for the rabbits and improves overall animal welfare.

Protocol

The study outlines a protocol for the design, manufacturing, and placement of a novel continuous EEG/ECG recording system in rabbits. The protocol is approved by the Institutional Animal Care and Use Committee (IACUC) at SUNY Upstate Medical University. All IACUC requirements for animal welfare, surgical guidelines, and study endpoints are followed. The investigators and the faculty/staff of the Department of Laboratory Animal Research at SUNY Upstate perform regular welfare checks and general care. Any injuries, discomfort, infection, or adverse effects are dealt with immediately.

1. Construction of an implantable electrode assembly

1. Carry out the in-house fabrication of the implantable electrode assembly in a sanitary environment, which includes a clean lab bench, nitrile exam gloves, and a procedure mask.
2. If adapting an existing wire bundle with a quick connect plug and DIN connectors, remove the male and female ends of the plugs (Supplementary Figure 1A). Save the plugs and the long wire with the DIN connectors for the later stages of the wire assembly (Supplementary Figure 1B).
3. Add or remove wires from both plugs and the sheathed wire bundle with DIN connectors to ensure it has the number of electrodes needed (9 for this example setup; Supplementary Figure 1A).
4. Gather 5 EEG (colors red, orange, yellow, green, and blue) and 4 ECG (colors black, pink, white, and gray) electrodes. Using heat shrink tubing and a heat gun, bundle the wires at the correct lengths for the animal model from the head of each electrode (Supplementary Figure 2A).
   NOTE: For a New Zealand White rabbit (1.25 to 13 months of age and 0.8 to 4.17 kg): EEG wires are 25 cm from the head of the electrode, black and pink ECG wires are 40 cm (left arm and left leg), white ECG wire is 35 cm (right arm), and gray ECG wire is 30 cm (left leg). This provides ample length to tunnel the wires and electrodes to the desired location, as well as provides extra length for rabbit growth and movement.
5. Pass the wire bundle through the cable passthrough bushing (Supplementary Figure 2B).
   NOTE: Plasma sterilization is performed on the assembled implants prior to surgery.
6. Cut the side of the wires away from the electrode approximately 1-1.5 cm from the end of the cable passthrough bushing and strip a short ~5 mm segment of coating off the ends of each wire (Supplementary Figure 2B).
7. Attach one end of a butt connector to each wire exiting the passthrough bushing, crimping down for a secure attachment. Place a piece of heat shrink tubing on each plug wire. Insert the stripped end of a wire from the plug into the un-crimped side of the butt connector and attach it to its corresponding same-color wire (Supplementary Figure 2C).
   NOTE: Do not crimp over the heat shrink tubing, as it can tear and leave the conductive metal of the butt connectors exposed. Soldering can be done in place of butt connectors if the selected wire is compatible.
8. Cover and heat shrink each butt connector/connection (no metal exposed) with small heat shrink tubing to isolate each channel.
9. Slide a piece of large heat shrink tubing over the plug to the base of the cable passthrough bushing, covering all wires and leaving minimal wire exposed at the base of the plug (Supplementary Figure 2D). Use the heat gun and protective gloves to heat shrink the tubing.
   NOTE: Be sure not to shrink the tube over the necessary locking mechanism for the plug.
10. Generate a mold that serves to shape the acrylic base on the side of the port with the electrodes (Supplementary Figure 3).
    NOTE: Use a 50 mL conical tube cap, remove the threads with a sander, and make four slits in a group and then five more slits 110°-120° from the first group of slits (Supplementary Figure 3A). Drill a 0.5-inch hole in the base of the mold (tube cap). Apply masking tape to the inside of the mold. Cut the tape from the slits.
11. Lower the assembled port with passthrough bushing into the mold with the plug (red) passing through the hole in the bottom of the mold. The unit is in place when the cable passthrough bushing sits flush with the bottom of the mold (Supplementary Figure 3B).
12. Arrange the wires such that the five EEG electrodes lay within the five slits on the left side of the mold and the four ECG wires lay within the four slits on the right side of the mold. Cover the slits on the inside of the mold with masking tape.
13. In a well-ventilated area, mix the dental acrylic in a glass vial using two parts resin powder and one part resin liquid. Using a transfer pipette, mix the powder and liquid until the mixture is viscous but still flows through a transfer pipette.
14. Use a transfer pipette to distribute the acrylic mixture into the mold, ensuring that all wires are covered. Allow the acrylic to begin to harden, but remove the acrylic base from the tube cap before the acrylic fully sets.
15. File any rough edges from the acrylic port base. Supplementary Figure 3C illustrates the final product of the wires embedded in the acrylic, which go through the passthrough bushing, connect to the plug, and are all sealed with heat shrink tubing.

2. Wire harness assembly

1. Obtain the plug that is complementary to the plug on the port. Obtain the swivel and retain nine pre-connected wires on each side (Supplementary Figure 4).
2. On the small swivel side, trim the wires to 5 cm in length and strip 5 mm of the wire coating off the end of each (Supplementary Figure 5A).
3. Cut a 2 cm piece of small heat shrink tubing and place it over each wire of the complementary plug (Supplementary Figure 5A).
4. Solder the wires from the plug to the matching color wire on the small side of the swivel (Supplementary Figure 5A).
5. For each wire, slide the heat shrink tubing over the solder connections. Use heat gun to affix.
6. Pass a long piece of large heat shrink tubing over the plug and slide to cover the entire length of the bundle. The heat shrink tubing must be long enough to cover 2.5 mm of the plug and as much of the small side of the swivel as possible without impeding its ability to rotate. Use a heat gun to affix (Supplementary Figure 5B).
7. On the large side of the swivel, strip 5 mm of coating off the end of each wire. Slide a > 45 cm piece of metal sheathing over the wire bundle (this is easiest if started from the cut end, not the end with DIN connectors), and pass until the cut wires are exposed.
   NOTE: Cut 2 cm long pieces of small heat shrink tubing and place them over each of the wires on the large side of the swivel (Supplementary Figure 5B). Cut a larger piece of shrink wrap (5 cm in length) and slide it over the entire bundle of wires. In the following steps, keep all heat shrink tubing away from heat/soldering.
8. Attach the wires from the larger end of the swivel to the cut electrode wires (end with DIN connectors) through wire twisting and soldering (Supplementary Figure 5B).
9. Affix the small heat shrink tubing with a heat gun over each connection. To prevent pressure or pinching of the wires on the large side of the swivel, add an extra-large piece of adhesive heat shrink tubing directly over the large end of the swivel (covering 8 mm of the swivel) and on the wires (covering 2 cm of wire). Use a heat gun to affix (Supplementary Figure 5C).
10. Slide the metal sheathing over the newly connected wires and swivel (Supplementary Figure 6A). If the metal sheathing does not slide over the swivel, use pliers or a grinder to remove and smooth the swivel's collar so that it makes a tight fit in the metal sheathing. This should require the user to twist the sheathing over the swivel.
11. Continue twisting it until the sheathing reaches the base of the plug but does not inhibit the swivel from spinning. This is important so the sheathing stays affixed to the swivel (Supplementary Figure 6B).
12. Stretch the metal sheathing to the maximum length and note this location on the nylon sheathing/wire bundle.
13. Cut a 1 cm long piece of 15 mm (outer diameter, OD) rubber tubing and slide it over the nylon sheeting and bundle of wires (starting at the DIN connector side, Supplementary Figure 6C).
14. Place a 3 cm piece of large shrink tubing over the rubber tubing. Using a heat gun, affix the rubber and shrink tubing at this marked location. This will function as a stopper to prevent the wires from pulling through the metal sheathing.
15. Add two pieces of heat shrink tubing (1 cm in length) overtop the unstretched metal sheathing at 20 cm and 24 cm from the end of the plug. Use a heat gun to affix (Supplementary Figure 6C). These sites are later used to secure the retractor cable bracket. Supplementary Figure 7 illustrates the completed wire harness, which includes the plug, wires, swivel, metal sheathing, stopper, and retractor cable attachment points.

3. Surgical procedure

1. Administer oral antibiotics (e.g., doxycycline at 5 mg/kg) 24 h and 48 h prior to the surgical procedure, as well as daily for 5 days post-operation.
2. Calculate the dosage of ketamine (40 mg/kg) and xylazine (6 mg/kg) based on weight (no restrictions on weight or sex, but it is recommended that rabbits be at least weaned to cut down on the chance of the mother or littermates disturbing the electrode implant). Deliver these medications by intramuscular (IM) injection.
3. Once the rabbit is fully sedated (unresponsive to physical stimuli), apply protective moistening gel to the rabbit's eyes. Close the eyelids over the gel to distribute evenly.
4. Use an electric shaver (coarse, then fine attachment) to remove fur from between the rabbit's eyes to 5 cm caudal to the inferior angle of the scapulae. Remove fur around the rabbit's right flank. Continue to shave the rabbit's ventral fur from between the front limbs down to the inferior border of the ribs.
5. Initiate inhaled maintenance anesthesia (2%-3% isoflurane). Start at 1.5-2 L/min (LPM) with simultaneous oxygen administration at 1.5 LPM. Place a veterinary anesthesia mask that makes a tight fit over the rabbit's nose and mouth. Ensure that the rubber gasket seals appropriately around the animal's face without the rabbit's nose pushing up against the back of the mask.
6. Place a SpO₂ probe on the rabbit's paw and confirm physiological values and waveforms. Position the animal on its left side so the right flank is accessible. Perform the rest of the surgical procedure under aseptic technique, which includes the surgeons donning  a sterile gown, hair net, shoe covers, procedure mask, scrubbing in, and using sterile gloves. Sterilize all tools, solutions, and implants.
   NOTE: Investigator should check that the rabbit is staying warm while anesthetized. A heated procedure table or heating pads, along with surgical drapes, are recommended.
7. The non-sterile surgical assistant distributes betadine and isopropyl alcohol to sterile basins. Coat the area in betadine followed by isopropyl alcohol and repeat this 3x, starting at the intended surgical site and radiating outwards in a circular pattern. Lay sterile drapes over the rabbit such that the intended surgical site is still exposed.
8. Perform ECG lead placement as described below.
   1. Make 4 small marks using a sterile-tip marker at the intended locations for ECG electrode placement. The intended locations include the upper chest wall medial to the (1) right (RA) and (2) left axilla (LA), and at the (3) right (RL) and (4) left (LL) anterior aspect of the inferior border of the lowest ribs (along the midclavicular line; Supplementary Figure 8A). This will create Einthoven's Triangle lead configuration with a ground at the right inferior location.
   2. Make a 2.5-3.5 cm longitudinal incision on the animal's right flank, about 1 cm dorsal to the rabbit's right front limb (Supplementary Figure 8B).
   3. Using hemostats, bluntly dissect ventrally such that a subdermal pocket is made across the entire chest. Ensure that this pocket extends to the four marked locations for each of the four ECG electrodes.
   4. Using the non-needle end of the 3-0 monofilament suture, tie 3 sets of surgical knots around the base of the electrode (Supplementary Figure 9A). Supplementary Figure 9B illustrates the ideal way to load the needle into the hemostats: by grabbing it at the base of the suture so the needle is parallel to the length of the hemostats. This is to help pass the needle through the sub-dermal pocket without getting caught on nearby tissue.
   5. Pass the needle now attached to an electrode through the subdermal pocket and exit the skin at the marked location (Supplementary Figure 10A). Tenting the skin using forceps or a suture and gentle upward traction on the skin increases the space within the sub-dermal pocket and reduces the chance of the needle getting caught on connective tissue.
   6. Apply gentle traction until the electrode remains at the marked location (Supplementary Figure 10B). Release traction and palpate to verify that the electrode stays in place. If connective tissue is preventing the electrode from sitting at the marked location, further blunt dissect the path of the wire and electrode.
      NOTE: Applying excess force/traction on the suture to encourage the electrode to sit where desired will not work and could cause excess damage to the suture site or break the suture.
   7. To secure the electrode at the marked location, pass the 3-0 monofilament suture and needle through the rabbit's skin directly adjacent to the exit site of the suture (Supplementary Figure 11A).
   8. Pull the suture partially through the skin, such that a small loop remains (Supplementary Figure 11B).
   9. Tie a standard surgical knot using the singular string (with the needle attached) on one side and the loop (as if it were a singular string) on the other (Supplementary Figure 11C). The resulting tie should resemble Supplementary Figure 11D. Trim the remaining excess suture.
      NOTE: Avoid tying the external suture too tight or creating a dimple in the skin. If needed, place the tip of a small pair of sharp scissors within the knot as it is tightened. This will leave a loose attachment, which reduces the risk of the electrode later growing through the skin.
   10. Repeat this process for the remaining ECG electrodes. Apply surgical glue to each of the external knots.
9. Perform EEG lead placement as described below.
   1. Rotate the animal into the prone position and ensure the anesthesia face mask is secured and physiological monitoring devices are properly positioned. Coat the area in betadine followed by isopropyl alcohol and repeat this 3x, starting at the intended surgical site and radiating outwards in a circular pattern. Lay sterile drapes over the rabbit such that the intended surgical site is still exposed.
   2. Using a sterile tip marker, place five small dots at the intended location for EEG electrode placement at the right frontal (RF), left frontal (LF), right occipital (RO), left occipital (LO), and center (Cz) reference sites (Supplementary Figure 12). From the incision on the rabbit's flank, blunt dissect toward the inferior angle of the scapulae and then to the base of the head.
   3. Make a small transverse incision on the head, 1.5 cm in length, extending between the anterior aspect of the base of the ears (Supplementary Figure 12). Incise with caution to avoid damaging any underlying musculature.
   4. Blunt dissect caudally from this incision until a subdermal pocket is made that connects to the incision on the flank. Blunt dissect cranially until there is a subdermal pocket made on the head of the animal that extends to the locations for the placement of each of the EEG electrodes. Be careful not to apply excess force here, as the orbits and several muscles could be damaged.
   5. Pass rubberized hemostats through the incision on the head to the incision at the flank. Grab hold of the EEG electrodes (1-2 electrodes at a time), pull through the sub-dermal pocket, and exit at the incision on the head. Repeat the process of tying a suture around the base of each electrode, as previously described in step 3.8.4 and shown in Supplementary Figure 9A.
   6. Place and secure the EEG electrodes. Pass the needle through the subdermal pocket on the head and pierce through the skin at the marked locations. Pull the suture into place and secure it with the methods described in protocol steps 3.8.7-3.8.9 and illustrated in Supplementary Figure 11.
   7. Repeat for all five electrodes. It is recommended to first place the frontal electrodes, followed by Cz, followed by the occipital electrodes. Apply surgical glue to all external suture knots.
   8. Insert large forceps into the incision on the flank, spread the forceps open, and lift the skin at the site marked for the port, which is caudal to the inferior angle of the scapulae.
   9. Use a 6 mm punch biopsy to create a circular hole. Gently rotate the biopsy blade in either direction until the hole is complete. Remove the incised skin within the punch biopsy (Supplementary Figure 13A).
   10. Cover the wire plug with plastic/rubber to prevent blood/tissue from getting into the plug. Pass the port through the incision on the flank and push the plug side of the port through the hole made by the punch biopsy. Supplementary Figure 13B illustrates the port exiting the skin and the colors denote the wires that are tunneled under the skin and traveling to the head (EEG) and chest (ECG).
   11. Press the skin down to the base of the port so that everything is taut. Place a rubber gasket over the port and push down until it sits flush against the skin. Tuck excess wire into the subdermal pocket on the back and flank, taking care to blunt dissect a large enough pocket to accommodate and evenly distribute the excess wiring. Ensure that wires do not bunch up in any area or pass directly under the port, as this may cause irritation.
   12. Gently bring the skin together and close all incision sites using simple interrupted sutures. Apply surgical glue to all closing 3-0 monofilament sutures and along the length of the incisions. Allow time for the surgical glue to dry.
       NOTE: Begin to titrate down the inhaled anesthesia and later turn it off after the acrylic hardens (3.9.14). Keep the SpO2 probe in place for at least 15 min for postoperative monitoring.
   13. Mix a 2:1 ratio by volume of acrylic powder and liquid using a transfer pipette. Mix until it appears thin enough to pass through a transfer pipette but thick enough to stay at the port site.
   14. Apply acrylic mixture to the outside of the port, creating a small dome of acrylic that seals the port site to the skin. Allow time for the acrylic to solidify. Place sterile gauze over the incision site on the rabbit's flank and loosely wrap the rabbit with vet wrap (i.e., loose enough to easily fit 2-3 fingers underneath the wrap). The incision on the head may be left uncovered.
   15. Allow the rabbit to recover in a cage with padding, a blanket, and a heat source. Rabbit must be supervised throughout this process. When the rabbit begins to wake up (nose twitching, some movement), administer a subcutaneous dose of sustained release buprenorphine (0.1 mg/kg). Continue direct supervision until the rabbit has regained consciousness (standing and independently navigating the cage).
   16. Allow the rabbit time to heal (at least 72 h) before connecting the port on the rabbit to the wire harness for data collection.
       NOTE: Follow all proper post-operative care steps to ensure proper healing and comfort during this time, including, at minimum, daily observations of food and water intake, bowel movement, urination, incision site healing, energy levels, and appearance of pain or discomfort. Remove all sutures at the incision sites 10 days post-op.

4. EEG/ECG measurement

1. Pass the wire harness through the top center of the rabbit's cage (3/4-inch diameter; Supplementary Figure 14).
2. Plug the DIN connectors into an appropriate monitoring device. Affix a retractor cable to the top inside of the cage near the hole for the wire harness.
3. Extend the retractor cable wire with a clip on the end and attach it to the wire harness at the location between the 2 pieces of shrink tubing (Supplementary Figure 15).
4. Secure the wires to the top outside the cage so they do not slide into it (Supplementary Figure 14, e.g., tape).
5. Connect the plug on the wire harness to the plug on the rabbit's back (Supplementary Figure 16). Record signal (Figure 1).

Results

Using this surgical technique, there were not any cases of mortality during or after the surgery. There is a risk of infection and incomplete wound healing, particularly if the rabbit scratches or bites at the incision site. Pre-/post-operative antibiotics, covering the incision with a dry sterile piece of gauze, and fitting the rabbit with a vest help protect the surgical site and allow for better healing. All recordings are conducted in the rabbit's standard housing cage, which is what ...

Discussion

We present a detailed protocol for the generation of a custom wire harness, the in-house fabrication of an implant, and the procedure for the sterile implantation of the EEG/ECG electrodes, wires, and port. This setup facilitates the collection of continuous, high quality EEG/ECG recordings in an efficient, safe, and financially feasible way. This protocol generates a system in which both EEG and ECG data can be collected in rabbits without the need for restrainers or constant supervision. Continuous long-term EEG a...

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
3:1 heat shrink wrap with adhesive	N/A	Wirefly	Adhesive lined polyolefin tubing designed to shrink 1/3 diameter when heated. Isolate electrodes when connecting wires and plugs, connecting pieces of the wire harness together
Absorbent Pads	-	any	Absorbent pads for Fluid collection
BD E-Z Scrub 116 Surgical Scrub Brush/Sponge	371163	BD E-Z Scrub	3% PCMX Surgical Scrub Brush/Sponge To scrub prior to operating for aseptic technique
Biopsy Punches	1315	ARGENT, McKesson	6 mm circular cutting blade for circular incision for port
Buprenorphine	-	-	0.3 mg/mL Buprenorphine For post-operative pain control
Butt connectors	X003C7S38P	SVAAR	Tinned copper butt splice 26-22 AWG non-insulated butt connectors Connecting carbon fiber wires from the electrodes to the plug
Cautery Pen	https://jorvet.com/product/cautery-kit-accessories/	Jorgensen Laboratories, Inc.	Hand-held Bovie Cautery Pen For controlling bleeding
Coaxial cable through-wall bushing	COAX BUSH WH	Commercial Electric	Plastic bushing with 6.9mm inner diameter opening and 1cm+ post Wire port
Cohesiant Wrap	https://healthypets.com/vetone-cohesiant-wrap-blue-2x5yd/	AmerisourceBergen	4in x 5yd self-adherent wrap For post-operative wound protection
Disposable Scalpel with Handle	https://www.securos.com/	Securos Surgical AmerisourceBergen	Sterile disposable scalpels For making incisions
Electrical Slip Ring		Taidacent	High Speed Ball Conductive Slip Ring 12 way 2A Collector ring rotary joint rotary connector, Electrical slip ring to allow free movement of animal without binding up the wire harness
Eye lube pro	https://domespharma.us/eye-lube-pro-detailer-final-v6/	Sentrx	Lubricating Gel Prevents eye drying/damage during anesthesia
GLUture	503763	World Precision Instrument	Topical Tissue Adhesive For closure of incision sites
Isoflurane	ANADA # 200-129	Isospire	Isoflurane For anesthesia
Isopropyl alcohol	500205	VETone	70% isopropyl alcohol Topical antiseptic
Jet Liquid	1403	Lang Dental Mfg. Co., Inc.	Fast curing acrylic resin liquid Dental acrylic resin liquid for sealing the bushing and the wires for the wire exit port.  Also used to seal the port to the skin.
Jet Tooth Shade Powder	1420	Lang Dental Mfg. Co., Inc.	Fast curing provisional crown and bridge acrylic resin powder. Dental acrylic powder for sealing the bushing and the wires for the wire exit port.  Also used to seal the port to the skin.
Ketamine	-	-	100 mg/mL Ketamine For sedation
Metal shower sheathing	HOS-960M	Waterpik	Ultra-Flexible Replacement Metal Shower Hose, Extra Long for Handheld Shower Heads, 96-inch, Chrome Metal sheathing for wire harness
MR Conditional/CT Electrodes Array and Extension Set	MRQW32415	RhythmLink	Quick connect system with male-female plugs and 8 leads. Plugs used for port-wire harness connection.  Wires and DIN connectors used for the wire harness
Polypropylene Blue Monofilament Sutures	P8665	SECUROPRO	3-0, FS-2, 18” reverse cutting suture. Close any surgical incisions
Retractor cable	‎0487-013	West Coast Manufacturing Company	Industrial Retractable Reel with 48" Stainless Steel Cable, Steel Bracket Back, Chrome Front, 8 oz. Retraction, Plastic Hose Clamp. Pulls excess wire harness up and away from animal
Rubber gasket	E-3603 NSF-51	Saint Gobain	Clear rubber tubing cut to 1.5 cm width. Gasket for protection/maintaining a seal.
Small Webb Electrodes	SWEBB1215	RhythmLink	6mm, flat disc [WEBB*] electrode, silver-chloride, wire, Subdermal electrodes
Sterile Latex Surgical Gloves	10122	AmerisourceBergen	Sterile Latex Gloves, Powder-Free, Donned for sterile surgical technique
Sterile marker	DYNJSM01	Medline	Sterile Marker, Mark incision/electrode sites
Sterile surgical drapes with adhesive	-	any	Sterile surgical drapes with adhesive strip on one side, Create a fixed sterile field
Veterinary anesthesia masks	-	any	Plastic cones with rubber gaskets to seal over rabbit’s face. Various sizes. Delivery of inhaled anesthesia
Veterinary Betadine Surgical Scrub	NDC 67618-154-16	Betadine	Povidone-iodine, 7.5%, Antiseptic Sudsing Skin Cleanser
Xylazine	-	-	100 mg/mL Xylazine, For sedation