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Mouse Anesthesia and Blood Collection


Flash EOL

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




 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.



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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