Mice serum - (Nov/16/2006 )

Hi,
Does anyone have experience wthdrawing blood of mice (and keep him alive) for serum determinations? How much serum can you get?
I tried cutting tail and got 100 ul... after which mice looked not very good, and besides, after centrifugation serum was not clear and yield poor. Any suggestions would be appreciate it
thanks

Hi,

We make a cut with a blade in the base (instead of the tip) of the tail... We collect as much as possible by capillarity into blood collection microtubes. The serum looks Ok, however, we don't get more than 100ul either...

i do a cut in the middle of the tail, in the veinous. Mice are OK . I do it with a warm lamp, and then I help stasis to perform by pressing the wound a few minutes (I agree I don't have dozens of mice to bleed !)
I get also around 100-200 µL of blood, and this is far enough to titrate the antibodies in the serum.

thanks fpr your replies.
just one more question. After the extraction, do you leave the blood to coagulate? and what speed do you centrifuge to collect the serum?
thanks

I don´t know for mice, in case it helps, I centrifugue human blood for 10-15 min at 3000 rpm.
If you collect blood in a tube without an anticoagulant (such as EDTA, heparin or citrate) you will obtain serum after centrifugation. In this case, it´s recommended to leave the blood coagulate during a few minutes before centrifugation. On the other side, using a tube with one of these substances, blood won´t coagulate so after centrifugation you will obtain plasma. The difference is that plasma contains coagulation proteins and serum doesn´t. All these information is for human serum (which I work with) but I think it can be useful also for mice.

I agree with Pumuki.

we use retroorbital eyebleed

it's easy once you get the knack, and you can get 100-200 ul from a mouse (dependent on % body weight, of course) without hampering their survival

We use retroorbital eyebleed too. It is easy and health of mouse is not compromised and you rae able to collect blood later too. I wonder if it is possible to collect blood again and again every week from the tail.

From retro-orbital method, we collect nearly 0.5-0.75ml of blood in eppendorf and we get around 200ul of serum after spinning at 15000rpm for 5 mins.

I normallly collect blood from mice by retroorbital...around 500ul...then leave the blood to coagulate at room temp for 45min before centrifuge for 20min at max speed at 4degree.
The serum is around 100ul-200ul.

Thanks a lot for your replies again,
Minnie and Nabin, Do your mice remain healthy after 500 ul blood extraction? seems to be a lot, considering that not more than 10% of the total blood should be withdrawn, but I suppose the quantity is reasonable to obtain that amount of serum. One more thing,
In the retroorbital bleeding, do you anesthesize your mice? some ppl do some other dont, i read that is ethical and necessary, but not always practiced.