Materials Required

1. Isoflurane
2. 70% Alcohol
3. Animal Model : White Mice
4. Anesthesia Chamber
5. Cotton
6. Dropper
7. Microhematocrit Tube
8. Test tubes
9. Restrainer
10. Syringe
11. Needle

Procedure

Inhalant Anaesthesia Using Isoflurane for Mouse and Rat

(Same procedure can be performed using methoxyflurane)

1. 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.
2. Move the glass chamber and isoflurane to a fume hood/laminar flow.
3. 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.
4. Close container lid immediately. Allow the isoflurane to evaporate to vapour phase over 5-10 minutes.
5. 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.
6. 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.
7. Allow the animal to become recumbent and reach slow and regular breathing.
8. 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

1. Manually restrain and anesthetize the animal.
2. Introduce the end of the microhematocrit tube at the medial canthus of the orbit.
3. 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.
4. Remove the microhematocrit tube from orbit and dab excess blood from the site with a gauze sponge or swab moistened in saline or PBS.
5. Observe the animal for recovery from anaesthesia.

Blood Collection from Tail Vein of Mouse and Rat Using Microhematocrit Tube

1. Restrain the animal.
2. Warm the tail with a heat lamp or immerse in warm water to dilate the vessels.
3. Visualize a sampling site of the lateral tail vein at approximately midpoint on the length of the tail.
4. Extend the tail with one hand, and with the other hand insert needle 3 to 4 mm into the lateral tail vein.
5. Collect blood from the hub of the needle with the microhematocrit tube.
6. 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

1. Restrain the animal.
2. Warm tail with the heat lamp or immerse in warm water to dilate the blood vessels.
3. Visualize a sampling site of the lateral tail vein in the distal one-third of the tail.
4. While extending the tail, swiftly incise across the lateral tail vein with a new scalpel blade.
5. Collect blood drops into 12- to 50-ml centrifuge tube.
6. 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.