Protocol Citation: Michaela Selby, Alicia Avelar 2022. Isoflurane Anesthesia Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.36wgqj7m5vk5/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: October 21, 2022
Last Modified: October 21, 2022
Protocol Integer ID: 71690
Keywords: anesthesia, isoflurane, rat
Abstract
protocol describes how to use anesthesia machine with isoflurane to anesthetize a rat.
Guidelines
Ensure the anesthesia box that contains the rat has a fully closed and locked lid. Ensure that the nose cone used to maintain anesthesia is tightly around the rats nose. Perform procedure in a hood for safety and wear lab coat and gloves. Turn off the gas flow as soon as you are done using it for anesthesia.
Materials
Paper towels
A glove
Tape
Isoflurane cart (Tape, extra isoflurane, oxygen tanks, black wrench to open oxygen tank)
Rat
Safety warnings
- Make sure at least one of the valves are open
- Always check oxygen levels and isoflurane levels
- Turn off isoflurane using the dial when you are not using it
Before start
- Handle rat beforehand to reduce stress
- Bring rat from vivarium (or wherever it is kept) to surgery room in a clean cage
Setup
Setup
Make sure you have a surgery room booked
Some sort of table or surgical workbench to transfer rat to when fully anesthetized
Grab isoflurane cart and/or supplies and bring it to the surgery room
Place paper towels in the plastic container
Make sure the tubing is tightly sealed together (if tape is needed add tape)
Make sure all of the tubing is in the correct place and connected
Make sure the valves are parallel with the tubing (the pink and orange tubing)
Use larger thicker tape to secure white tubing down onto the surgery table (where you want it on the workbench)
Snip open a little hole in a glove and place it over the tubing for the rat's mouth as a nose cone for anesthesia during surgical procedures
Check isoflurane level (make sure it is between the minimum and maximum fill lines)
If the isoflurane level is low then unscrew the cap and slowly add in the isoflurane
Set isoflurane regulator on top-adjust dial as appropriate
Use the black wrench to open the oxygen tank
Turn to the right to open
Look at the oxygen regulator and turn the green knob so the silver ball is in the middle of the 1 on the regulator.
Place the rat in the plastic box and close the lid
Adjust isoflurane dial as needed
Start at 4 (if female)
Start at 5 (if male), heavier animals usually need more isoflurane to become anesthetized
Keep rat under for a few minutes or until the rat’s breathing slows down
Take the rat out of the plastic container and place it onto the surgery table
Do a toe pinch using forceps to make sure the animal does not feel any pain
If the rat has a pain reflex or a jerk motion of the limb then place back into the plastic container and continue anesthesia
If the rat has no pain reflex or jerk then continue
Put the tubing that has the glove with the slit over the rat's mouth and nose to maintain anesthesia
Then when the rat is fully anesthetized you can proceed with your surgery
Turning off machine
Turning off machine
Turn off the isoflurane dial (back to 0)
Take the black wrench to close the oxygen tank
Turn to the left to close
Troubleshooting
Troubleshooting
Check to make sure there is no tubing loose/disconnected
In case there is a leak
Or if something is not working
Adjust dial
More or less isoflurane depending on how the rat is doing
Cleanup and Transport
Cleanup and Transport
Press the green oxygen flush button
This clears out oxygen from the tubing
Remove the paper towels and dispose of them
Clean out the plastic box with paper towels and water only
Return isoflurane machine and supplies to its storage place
Place cage where the dirty cages go to be cleaned