Materials Needed:

Blood collection (saphenous):

• 50ml conical tube
• 25G needles (5/8 inch)
• capillary tube
• bulb to expel blood (Globe Scientific Inc. pipette bulb cat #51675)
• 2x2 gauze
• small mouse clippers
• 1.5 ml eppendorf tubes with 140microliters PBS
• rack to hold eppendorf tubes

Blood collection (retro-orbital):

• isoflurane anesthetic machine
• anesthetic chamber & mask
• capillary tube with heparin
• bulb to expel blood (Globe Scientific Inc. pipette bulb cat #51675)
• 2x2 gauze
• 1.5 ml eppendorf tubes with 140microliters PBS
• rack to hold eppendorf tubes

Blood collection technique - lateral saphenous vein

1. Make mouse holder by poking holes in bottom of 50ml conical tube.
2. Place mouse in conical tube with a rear leg extended outward. 
3. Shave leg to aid in visualization of lateral saphenous vein.
4. Use needle to prick lateral saphenous vein and allow blood to form droplet.
5. Hold capillary tube to droplet.
6. Once capillary tube is full, immediately add contents to Eppendorf tube with PBS using bulb to expel blood.
7. Use gauze to apply direct pressure over lateral saphenous vein to stop bleeding.  Once bleeding has stopped move mouse back to cage.
8. Discard empty capillary tube in sharps container and cap Eppendorf
9. Refrigerate Eppendorf tube overnight to allow complete clotting.
10. Centrifuge briefly if required, draw off 1:5 diluted serum* from clot.
11. Ship on -20C ice packs to RADIL.

Blood collection technique - retro-orbital sinus

1. Anesthetize mouse in isoflurane chamber.
2. Remove mouse from chamber, hold mouse such that eye slightly bulges, then gently rotate capillary tube in medial canthus until blood fills capillary tube.
3. Once capillary tube is full, immediately add contents to Eppendorf tube with PBS using bulb to expel blood, then discard capillary tube in sharps container and cap tube.
4. Use gauze to apply direct pressure over closed eye to stop bleeding. Once bleeding has stopped move mouse back to cage and allow to recover from anesthesia.
5. Refrigerate Eppendorf tube overnight to allow complete clotting.
6. Centrifuge briefly if required, draw off 1:5 diluted serum* from clot.
7. Ship on -20C ice packs to RADIL.

*Whole blood (~40% cellular by volume) diluted 1:3 yields serum diluted approximately 1:5