How to Do Intracarotid Artery Injection in Mouse?
(Protocol for intracarotid artery injection in mouse)
Materials:
1. Stereomicroscope
2. Microscissors, microforceps
3. Surgical scissors, forceps, and scalpel
4. 70% ethanol wipes and Betadine swabs
5. An autoclip wound applier and stainless-steel autoclips
6. Q-tips
7. 27- or 30-gauge needles
8. A 1-cc syringe
9. 4.0 silk suture
10. Tumor cells
Anesthetics:
Nembutal (Pentobarbitol sodium)
Methods:
1. Anesthetize a mouse by intraperitoneal injection of pentobarbital sodium (50mg/kg body wt).
2. Clip the hairs on the ventral side of neck skin.
3. Restrain the mouse on the back on a plastic board with the head stabilized with a rubber band placed between the teeth of the upper jaw.
4. Disinfect the surgical site with Betadine and 70% ethanol.
5. Incise the skin along the midline for about 1.5 cm long.
6. Expose the trachea by blunt dissection.
7. Move the mouse under a dissecting microscope.
8. Bluntly separate the thin layer muscle in the carotid muscular triangle to expose the right common carotid artery.
9. Separate it from the vagal nerve and carotid vein.
10. Place a 4-0 silk suture under the proximal part of the common carotid artery and tie it tightly.
11. Place a second ligature under the common carotid artery distal to the expected injection site and tie it loosely.
12. Insert a small sterile cotton ball of 3-4 mm in diameter under the artery between the two sutures to elevate the common carotid artery to control the blood regurgitation from distal part.
13. Nick the common carotid artery proximal to the cotton ball with a pair of microscissors.
14. Insert the tip of a needle* connected to a 1-cc syringe.
15. Inject the tumor cells slowly.
16. Retract the needle.
17. Tighten the second ligature.
18. Remove the cotton ball.
19. Close the skin with wound autoclips.
20. Keep the mouse warm during recovery.
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