IACUC / LARC STANDARD PROCEDURES

Sub-mandibular Blood Collection in Mice

Description of procedure:

Blood collection from the submandibular vein is an alternative to retro-orbital blood collection. It allows for the maximum allowable sample volume with minimal trauma to the animal. Multiple samples can be taken daily by alternating sides.

This technique can be performed on conscious animals if appropriate justification for why anesthesia can not be used is included in the approved IACUC protocol.

In general, 1% of the animal's body weight can be safely collected every 14 days. If blood must be drawn more frequently, 0.05% of the body weight may be removed each week. This volume may be divided into several draws.

Only trained personnel may perform this method.

Supplies:

• Isoflurane anesthesia system
• 22 gauge needles
• Gauze sponges
• Collection tube

1. Anesthetize mouse with isoflurane
2. Remove the mouse from the induction chamber and restrain the mouse with the non-dominant hand by grasping the loose skin over the shoulders and behind the ears; the skin should be taut over the mandible
3. Puncture the vein with a 22 gauge needle slightly behind the mandible, but in front of the ear canal (animals <25 grams use a 25 gauge needle). A swift lancing motion is used to puncture the vessel. Only the tip of the needle should enter the vessel. Blood will flow immediately.
4. Collect sample with a pipette or any type of collection tube.
5. When the sample has been collected apply, immediately pressure with a gauze sponge until bleeding has stopped.
6. Once the animal has recovered fully from the anesthesia, it is returned to the home cage.

Literature search words required:

Literature search was performed for refinement of this Standard Procedure on 5-22-07

Key Words	Search Site	Years Covered
Blood collection in rodents	Pubmed	2000-2007
Submandibular blood collection

Agents:

This procedure requires gas anesthesia

All agents administered to animals should be listed in the "Agents" section of RIO.

Adverse Effects:

Procedure, Agent or Phenotype	Potential Adverse Effects	Management
Inserting the needle too deeply.	Hematoma	euthanasia

Monitoring Parameters:

Monitoring Parameters	Frequency	PI/Lab will Document
General appearance and signs of active bleeding or hematoma at collection site	Several times in the first 30-60 minutes after blood is collected, then 1 day after each collection	No

Describe the conditions, complications, and criteria (e.g. uncontrolled infection, loss of more than 15% body weight, etc.) that would lead to removal of an animal from the study, and describe how this will be accomplished (e.g. stopping treatment, euthanasia).

Hematoma at collection site or uncontrolled bleeding - animal euthanized

For all investigators housing animals with tumor formation, skin lesions, neurological deficits, or that are in Category E, list the expected characteristics/clinical presentations and endpoints of the animal model and the criteria for euthanasia. Note: The IACUC also requires such lists to be posted in the respective animal rooms and monitored by the IACUC compliance staff and LARC, to assure PI adherence to the endpoints listed.