Reagents, supplies and
- 20 - 22 g needle
- 50 cc conical tube with "air" holes
- alcohol wipes (optional)
- #21 Surgical blade
- Microvette CB 300 tubes
- dry ice
- adjustable micro pipette with tips (such as
- refrigerated microcentrifuge
you are right handed the tube and tail will be held by the left
- Encourage the mouse to enter the 50 cc conical tube, nose
first, leaving the tail and foot on the outside of the tube.
- 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.)
- 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.
- 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
- Apply direct pressure to the vein until bleeding stops.
- Spin samples at 5000 rpm X 5 minutes in a refrigerated
- Remove plasma with a Pipetteman and snap freeze on dry
- Store plasma at -80ºC.
Written by Jill Lindner - 9/23/05