In this video, we demonstrate a tail-snipping procedure in a mouse model to collect peripheral blood. The collected blood is subsequently subjected to centrifugation to separate plasma, which can be used for further experiments.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Serial Blood Collection and Plasma Monitoring for Exogenous Human Cytokine in Mice
Blood collection via tail snip
Disinfect the biosafety hood and assemble the mouse restrainer, scissors, and other tools/supplies for the procedure.
Place the mouse in a restrainer and secure the tube to minimize mouse movement.
Disinfect the tail and surgical scissors with isopropyl alcohol. Apply topical anesthetic cream on the tail.
Using very sharp surgical scissors snip off about 0.5 – 1 mm of the tip of the mouse's tail. Collect approximately 80 µL of peripheral blood using heparinized capillary tubes. NOTE: If blood will be collected with this method more than six times, tail removal should be conservative with respect to the amount of tail removed. As snips progress up the tail the diameter increases, it hemorrhages faster, and healing takes longer.
2. Plasma separation
Transfer the blood using a bulb syringe (to expel it from the capillary tubes) into a labeled K2EDTA microtainer tube; invert 20 times to prevent coagulation. NOTE: Typically, blood clotting from the tail nip is fast. However, if hemorrhage persists longer than ~ 2 min use a styptic pencil/powder to aid coagulation.
Centrifuge blood collection tubes according to the manufacturer's instruction and aliquot the plasma for storage (-20 °C) until ready for further analysis.
Disclosures
The authors have nothing to disclose.
Materials
Animal Implantable Nano Transponder with Canula
Trovan
ID-100B(1.25)
EMLA lidocaine anesthetic cream (obtain by presciption through animal care facility)
perscription
perscription
BD ½ cc LO-DOSE U-100 Insulin Syringe 28G½
BD
#329461
BD Microtainer Tubes with K2EDTA
BD
#365974
Centrifuge
Beckman Coulter
Alegra X-15R
FisherBrand Capillary Tubes (Heparinized)
Fisher Scientific
22-260-950
Hemocytometer
Fisher
02-671-6
MACSQuant Analyzer 10
Miltenyi Biotec
130-096-343
Mouse Pie Cage
Braintree Scientific, Inc.
MPC-1
Mouse Tail Illuminator Restrainer
Braintree Scientific, Inc.
MTI STD
Pistol Grip Implanter
Trovan
IM-300(1.25)
"NSG mice" NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ, male or female, age 6 – 8 weeks