This is a protocol for collecting blood from a mouse via the saphenous vein.


Category
Protocol
Record History
Added on May 3, 2010 at 9:53 AM by Lindner, Jill
Modified on June 25, 2012 at 12:27 PM by Lindner, Jill
Shared with (contributions)
VCSCBi: Training / Protocols Master

Reagents, supplies and equipment:

  • 20 - 22 g needle
  • 50 cc conical tube with "air" holes
  • alcohol wipes (optional)
  • #21 Surgical blade
  • Microvette CB 300 tubes
  • scissors
  • dry ice
  • adjustable micro pipette with tips (such as  Pipetteman)
  • refrigerated microcentrifuge

Procedure: (if you are right handed the tube and tail will be held by the left hand)

  1. Encourage the mouse to enter the 50 cc conical tube, nose first, leaving the tail and foot on the outside of the tube.
  2. Left vein bleed: gently pull left foot into the fully extended position and pinch the skin fold between the tail and the leg.  (Right vein bleed: gently pull the right foot into the fully extended position and pinch the skin fold between the abdomen and the leg.)
  3. The saphenous vein runs laterally along the side of the leg and is easily visualized. Gently trim the hair with scissors; then shave the remaining hair using the surgical blade.
  4. While continuing to pinch the skin fold, nick the vein with a 20 - 22 g needle and collect blood into a Microvette tube. Allowable collection volumes are based on mouse weight and frequency of bleeds.  Guidelines are as follows: a) Never take more than 15 ml/kg body weight of blood under any condition where the animal is expected to live.  Replacement fluids should be considered at this level, b) 10 ml/kg may be taken if the bleedings are at one-month intervals, or if plasmapheresis is performed weekly, c) 5 ml/kg may be taken if bleedings are no more frequent than every other week, d) 2.5 ml/kg of blood may be taken if bleedings are performed weekly.  For more information regarding allowable volumes consult the IACUC web site: https://www4.vanderbilt.edu/oor/iacuc.
  5. Apply direct pressure to the vein until bleeding stops.
  6. Spin samples at 5000 rpm X 5 minutes in a refrigerated microcentrifuge.
  7. Remove plasma with a Pipetteman and snap freeze on dry ice.
  8. Store plasma at -80ÂșC.

Written by Jill Lindner - 9/23/05