We are collecting blood from mice sacrificed by cervical dislocation by removing an eye and let blood drop by one eye.
This method provide us around 300 to 500 µl of blood per animal.
We let the whole blood for 1h at room temperature coagulating in a usual 1.5mL eppis and centrifuge it 10 minutes at 1000g. The resulting supernatent is unfortunately red, but time to time (20% of the time) clear.
I would like to ask if you have another method to get more blood from a dead animal and how you process the blood to avoid to have red serum.
I usually get the blood by decapitation, ideally on isofluran anaesthesia. Mostly I use rats (gilotine) but in mice (scissors) I guess you can get more than 0.5 ml by decapitating. My skilled Ecolleague is able to get more blood by cutting open the thorax and aspiring the blood by a needle and syringe directly from heart.
Regarding the centrifuging, I have good experience with leaving the blood to clogg any time from 15 min to 2 hrs and centrifuge at 3000-4000 RPM for 10 min. Haemolythic serum I get only when I centrifuge twice (when I clumsily remove the content).
Good luck!
Jana
Serum is the watery, pale yellow part of blood. If it turned red colour, we could be explain the hemolysis will occur when animal test. Maybe check some experimental conditions，for：buffer solution（PH、isotonic solution），asepsis， centrifugal temperature ect.
good luck!
Xuejun
We put the mice in co2 raising chamber for 6 minutes, then check for vital signs to prove it's dead then before dislocate the neck with fine syringe 45 degree angle just under the sternum cardiac puncture the extract slowly peacefully 600 ml blood within 1 minute.then you dislocate the neck.
We solved the problem using cervical dislocation and within 10 seconds cut the head and let blood leak in a microcentrifuge tube. We let the blood 2 hours coagulating and then 10 minutes centrifugation at 1000g.
Thank you for your help!
@Nicolas: I would like to know how much blood can be derived from a mice by cutting his head and let blood leak? I need to extract plasma thereafter. I am facing problems in separating plasma from blood as after deriving blood by cardiac puncture using needles leads to hemolysis and good quality plasma is not derived.
Normally, i keep blood at room temperature for around 3-4 hours. Then centrifuse 3000rpm for 10 minutes. Collect serum. Gradually, increase RPM and time to get better serum.
Best regards,
Babu
anaesthesize with avertin or ketamine+xylezene .... do surgical pneumothorax, cardiac puncture on right atrium and slowly draw the plunger of 1 ml syringe. ( prevent fast drawing the plunger because pressure at the needle can cause RBC lysis). This way you can collect about 1 ml of blood. Now cut the top part of syringe and transfer the blood to eppendroff tube, let it stand at room temp for 30 min and cfg at 1500 rpm/ 15 min
If you need less volume at different time interval without sacrificing the animal, collect blood by facial vein puncture or saphenous vein puncture
BR
Yadav
Serum is the liquid fraction of whole blood that is collected after the blood is allowed to clot. After collection of the whole blood, allow the blood to clot by leaving it undisturbed at room temperature. This usually takes 15–30 minutes. Remove the clot by centrifuging at 1,000–2,000 x g for 10 minutes in a refrigerated centrifuge.
Wish you good luck