Source: Bosinski, C. et al., Multi-system Monitoring for Identification of Seizures, Arrhythmias and Apnea in Conscious Restrained Rabbits. J. Vis. Exp. (2021).
This video demonstrates the procedure to implant EEG-ECG electrodes and attach respiratory probes in a rabbit model for acquiring and analyzing records of its bio signals.
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Implanting EEG-ECG electrodes and attaching respiratory monitors
2. Recording of video-EEG-ECG
Figure 1: Picture of EEG and ECG electrodes. (A) Bent ECG electrodes and straight EEG electrodes. (B) How to hook the ECG electrode in the subcutaneous tissue of the rabbit, so that it is through and through. Abbreviations (LL: Left limb, RA: Right arm, RL: Right limb, LA: Left arm, RF: Right frontal, LF: Left frontal, Cz: Center, RO: Right occipital, LO: Left Occipital).
Figure 2: Rabbit connected to equipment. (A) Location of ECG electrodes, Left Arm is indicated by a yellow dot. Right Arm is indicated by a white dot. Left Leg is indicated by a red dot. Ground anterior to the right leg is indicated by a green dot. (B) Rabbit in restrainer with ECG and EEG electrodes attached. (C) Juvenile rabbit in a restrainer with appropriate modifications to accommodate a smaller rabbit, including a booster beneath the rabbit, neck foam and cut PVC pipe. (D) Rabbit in restrainer with location of EEG electrodes. Right Frontal is indicated by an orange dot. Left Frontal is indicated by a red dot. Right Occipital is indicated by a yellow dot. Left Occipital is indicated by a blue dot. The reference is indicated by a black dot. (E) Rabbit in restrainer with photic stimulator and mirror booth setup. Light source is indicated by a white dot. (F) Marginal ear vein after rabbit's ear has been shaven and wiped with alcohol. (G) Rabbit with angiocatheter securely taped in the left marginal ear vein. Site of injection plug is indicated with a blue dot. (H) Rabbit with facemask attached to the capnography tubing by a T-piece that contains a one-way valve. (I) Diagram of the facemask and T-piece connected to the capnography tubing. During inspiration, room air is able to enter the T-piece through a one-way valve (green arrow). During expiration, CO2 leaves the T-piece by entering the capnography tubing (yellow arrow.) Because of the small amount of dead space, very little CO2 is retained in the T-piece and is generally less than 5 mmHg.
Figure 3: Simultaneous Rabbit Video-EEG-ECG-Capnography-Oximetry. (A) Simultaneous video-EEG-ECG recording of 3 rabbits. (B) Zoomed in view of simultaneous video-EEG-ECG recording from Rabbit #2. (LL: Left limb, RA: Right arm, LA: Left arm) (C) Simultaneous recording of capnography (yellow) and plethysmography (blue). Measurements showing inspired CO2, end tidal CO2, respiratory rate, pulse rate and pulse oximetry are included in the figure.
The authors have nothing to disclose.
Computer | Dell | Optiplex 5040 | Acquisition computer |
ECG Electrode | RhythmLink | RLSND116-2.5 | 13mm 35-degree bent (0.4 mm diameter) subdermal pin electrodes |
EEG Electrode | RhythmLink | RLSP513 | 5-twist 13mm straight (0.4mm diameter) subdermal pin electrodes |
EEGLAB (2020) | Swartz Center for Computational Neuroscience | Open Access | Can perform spectral analysis of EEG |
Ethernet-to-ethernet adapter | Linksys | USB3G16 | Adapter for connecting the camera to the computer |
Foam padding | Generic | N/A | Reduces pressure applied to the neck of small rabbits by the restrainer in order to prevent the adverse cardiorespiratory effects of neck compression |
MATLAB (R2019b, Update 5) | MathWorks | N/A | Required to run EEGLAB |
Microphone | Sony Stereo | ECM-D570P | Recording of audible manifestions of seizures |
Micropore Medical Tape, Paper, White | 3M | 1530-1 | Used to secure wires and create ear splint |
Natus NeuroWorks | Natus | LC101-8 | Acquisition and review software |
Photic Stimulator | Grass | PS22 | Stimulator to control frequency, delay, duration, intensity of the light pulses |
Plastic wire organizer / bundler | 12Vwire.com | LM-12-100-BLK | Bundle wires to cut down on noise |
PS 22 Photic Stimulator | Grass Instruments | BZA641035 | Strobe light with adjustable flash frequency, delay, and intensity |
PVC pipe | Generic | N/A | Prevents small rabbits from kicking their hind legs and causing spinal injury |
Rabbit Restrainer | Plas-Labs | 501-TC | Various size rabbit restrainers are available. 6" x 18" x 6" in this study. |
Rubber pad (booster) | Generic | N/A | Raises small rabbits up in the restrainer to prevent neck compression |
SpO2 ear clip | NONIN | 61000 | PureSAT/SpO2 |
SpO2 sensor adapter | NONIN | 13931 | XPOD PureSAT/SpO2 |
Webcol Alcohol Prep, Sterile, Large, 2-ply | Covidien | 5110 | To prepare ear vein before catheterization |
LabChart Pro (2019, Version 8.1.16) | ADInstruments | N/A | ECG Analysis |
Quantum Amplifier | Natus | 13926 | Amplifier / digitizer |
Quantum HeadBox Amplifier | Natus | 22134 | 64-pin breakout box |
SRG-X120 1080p PTZ Camera with HDMI, IP & 3G-SDI Output | Sony | SRG-X120 | Impela Camera |