Animation link: https://vlab.amrita.edu/repo/BIOTECH/IMM/Mouseanaesthesiabloodcollection/index.swf
Materials Required
- Isoflurane
- 70% Alcohol
- Animal Model : White Mice
- Anesthesia Chamber
- Cotton
- Dropper
- Microhematocrit Tube
- Test tubes
- Restrainer
- Syringe
- Needle
Procedure
Inhalant Anaesthesia Using Isoflurane for Mouse and Rat
(Same procedure can be performed using methoxyflurane)
- Determine the volume of the anaesthesia chamber in litres. Multiply the volume by 0.15 to 0.2 ml to obtain the volume, in ml of Isoflurane to be used.
- Move the glass chamber and isoflurane to a fume hood/laminar flow.
- Soak a cotton ball with the required amount of the anesthetic solution and put it in the glass chamber. Above the cotton ball place a raised wire gauze platform.
- Close container lid immediately. Allow the isoflurane to evaporate to vapour phase over 5-10 minutes.
- Transfer animal from its cage into anaesthesia chamber by lifting it, by holding at the tail and placing it into the chamber. Close lid tightly after this.
- Monitor animal closely. Within approximately one minute for mice and 2 minutes for rats, the animal will become anesthetized. Initially, respiratory rate will increase and then decrease.
- Allow the animal to become recumbent and reach slow and regular breathing.
- Allow the animal to remain at a deep anesthetic plane for ~10 more seconds before proceeding into the desired experimental procedures.
Blood Collection from Orbital Sinus or Plexus of Mouse and Rat
- Manually restrain and anesthetize the animal.
- Introduce the end of the microhematocrit tube at the medial canthus of the orbit.
- Slowly, and with axial rotation, advance the tip of the microhematocrit tube gently towards the rear of the socket until blood flows into the tube.
- Remove the microhematocrit tube from orbit and dab excess blood from the site with a gauze sponge or swab moistened in saline or PBS.
- Observe the animal for recovery from anaesthesia.
Blood Collection from Tail Vein of Mouse and Rat Using Microhematocrit Tube
- Restrain the animal.
- Warm the tail with a heat lamp or immerse in warm water to dilate the vessels.
- Visualize a sampling site of the lateral tail vein at approximately midpoint on the length of the tail.
- Extend the tail with one hand, and with the other hand insert needle 3 to 4 mm into the lateral tail vein.
- Collect blood from the hub of the needle with the microhematocrit tube.
- Remove needle from the tail vein and apply a gauze sponge with gentle pressure on the bleeding site to ensure hemostasis.
Blood Collection from Tail Vein of Mouse and Rat Using Centrifuge Tube
- Restrain the animal.
- Warm tail with the heat lamp or immerse in warm water to dilate the blood vessels.
- Visualize a sampling site of the lateral tail vein in the distal one-third of the tail.
- While extending the tail, swiftly incise across the lateral tail vein with a new scalpel blade.
- Collect blood drops into 12- to 50-ml centrifuge tube.
- Apply gauze sponge with gentle pressure to the wound to ensure hemostasis.
Result
Blood is collected from the anesthetized mouse from orbital sinsus or plexes and tail vein using a microhematocrit tube and stored for further research purposes.
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