I need to collect only one blood sample from rats for serum separation.
My questions are:
1) Is tail nick better than terminal decapitation under general anesthesia since with decapitation blood may be mixed with tissue fluid?
2) What is the type of tube that would work the best in that case? Is it possible to use normal eppendorf or specialized tubes like Vacutainers/microvettes are better?
Thanks a lot!
How much blood do you need, and do you need the rat to survive? If you need less than 1 ml, you can take from the lateral tail vein. There is a nice open access JOVE video that shows how to do it. If you need a lot of blood, I would do cardiac puncture (after anesthesis). In either case, you would collect with a syringe and then put the blood in a normal Eppendorf tube or 15 ml polypropylene tube.
http://www.jove.com/video/52766/sampling-blood-from-the-lateral-tail-vein-of-the-rat
I would also go with the tail vein. But keep in mind that if you haven't done it before it will be a bit of a challenge. It usually takes quite a few tries to get it, but keep in mind that the more you poke the vein the harder it is to get the blood so if you don't get it in 2 tries it probably wont happen.
Dear Shanshan, thanks a lot for the tips! The video is really helpful.
I won't keep my rats after the blood collection, so that's why I though terminal decapitation should also work. I would need less than 1 mL blood so the tail vein method would be fine.
For small volume Microvette from SARSTED is one of the best option. collection blood with its capillary tip is easy !
If you don't save the rats the best and the quicker option is to do cardiac stick blood collection - could be done on either deep anesthetized rat or it could be done immediately after CO2 euthanasia; I would use a 3.0 ml syringe with a 21G 1" long needle; if you need the plasma you would need ETDA tubes (or prefilled EDTA,or another anticoagulant - sodium citrate...); to avoid hemolysis I would remove the needle when the blood from the syringe is transferred to the tubes; if you're concern about hemolysis or blood clots you can use 21G winged blood collection sets and vacutainer tubes.
Another option is to do the retro-orbital sinus blood collection on anesthetized rat - it's fast but you would need some previous training. Tail blood collection works best if you pre-warm the animal (or only the tail) which requires some experience too - in this case you don't need anesthesia, just some sorts of rat restraining devices (cylinders or decapicone).
I would suggest even the eye puncture with capillary tubes. Its easy and safe.
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