A Mouse Tail Snip Procedure for Peripheral Blood Collection and Plasma Isolation
Published: November 30, 2023
Abstract
Source: Coats, J. S. et al., Expression of Exogenous Cytokine in Patient-derived Xenografts via Injection with a Cytokine-transduced Stromal Cell Line. J. Vis. Exp. (2017)
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.
Açıklamalar
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 | JAX Mice | # 005557 |
Bu Makaleden Alıntı Yapın
A Mouse Tail Snip Procedure for Peripheral Blood Collection and Plasma Isolation. J. Vis. Exp. (Pending Publication), e21836, doi: (2023).