Hi Chris, we use the blood from cardiac puncture for mostly haematological assays so we collect the blood in EDTA-coated Eppendorfs which is more suitable for haematological downstream application. If you do non-haematological, heparin coated is fine also. We do not buy pre-coated sample tubes but just add 10% 0.5M sterile EDTA of the expected blood volume (so 100ul) into a 2ml Eppendorf tube. Just before the puncture we draw this up in a syringe with the needle we are using for the puncture, pull the plunger all the way back to coat the syringe and push the EDTA back into the tube. Whatever is left in your needle and syringe will be enough to prevent clotting. We perform this procedure under terminal anaesthesia. Once the mouse is fast asleep (test paw reflexes) there are two techniques: one is to rest the mouse on its back with its nose in a tube connected to the isoflurane device (method of choice), which is also more relaxing for you as you don't have to worry about the mouse waking up, the other is to hold the mouse in a scruff and get the puncture (and the cervical dislocation) done before the effect of the iso wears off. The quickest way is to use medium gauge seized needles, insert the needle under aspiration under the sternum pointed slightly towards the left side and NOT go too deep. Once you have the blood remove the needle before you add it to the tube to prevent rupture of blood cells by sheering forces. Good luck!

You could treat your seringues and needles by performing a couple of passages with your seringue in an heparin-containing solution. Then, you perform your cardiac puncture and collect the blood in an EDTA-coated tube (Starsted). The tubes must be in constant shaking/rotation. If you want to collect the plasmas, you can perform a centrifugation at 4oC @ 3000 RPM for 10 minutes in a micro centrifuge. Immediately after collecting the plasma, put your tubes on dry ice until you store it @ -20oC or -80oC for long term storage.

Hi chris,I think Peter has given you the best answer. However, I recommend use mild anesthesia to mice before cardiac puncture. It would be recommended if it must be done before sacrificing the animal. If you are looking for time dependent studies (or you have to survive the animal), go for tail vein puncture method to collect blood. Tail of mouse has three prominent veins that can easily be penetrated to collect blood.

Hi Chris, we use the blood from cardiac puncture for mostly haematological assays so we collect the blood in EDTA-coated Eppendorfs which is more suitable for haematological downstream application. If you do non-haematological, heparin coated is fine also. We do not buy pre-coated sample tubes but just add 10% 0.5M sterile EDTA of the expected blood volume (so 100ul) into a 2ml Eppendorf tube. Just before the puncture we draw this up in a syringe with the needle we are using for the puncture, pull the plunger all the way back to coat the syringe and push the EDTA back into the tube. Whatever is left in your needle and syringe will be enough to prevent clotting. We perform this procedure under terminal anaesthesia. Once the mouse is fast asleep (test paw reflexes) there are two techniques: one is to rest the mouse on its back with its nose in a tube connected to the isoflurane device (method of choice), which is also more relaxing for you as you don't have to worry about the mouse waking up, the other is to hold the mouse in a scruff and get the puncture (and the cervical dislocation) done before the effect of the iso wears off. The quickest way is to use medium gauge seized needles, insert the needle under aspiration under the sternum pointed slightly towards the left side and NOT go too deep. Once you have the blood remove the needle before you add it to the tube to prevent rupture of blood cells by sheering forces. Good luck!

hello Chirs, i recommend that, Tail vein sampling is suitable for all strain. it is quick and simple to perform for competent individuals. the lateral tail vein is usually used and 50ul and 0.2ml blood can be obtained per sample depending on size of animal and specific requirement.

We used femoral vein blood collection extensively. Our studies required weekly or biweekly bleeds for 25 weeks and tail clip was just not feasible. Unless plasma is needed, serum collected in regular vials is the easiest and cheapest way to go. Also, the smallest commercially available heparinized vials are 1ml which is way over what you need in order to collect supernatant after cfg. Collection in cold vials held on ice is recommended if you have more than a few animals (prevents sample degradation).

so heparinized/EDTA coated vials are not required??

It depends what you plan on analyzing from that sample. If really needed you can coat your own. Some preparation variability is unavoidable. I personally prefer serum.

a whole panel of tests, some of which are listed here http://hilltoplabs.com/public/blood.html

So you coat your tubes in serum? What kind?

I apologize for the confusion! I prefer serum to detect from, not to coat with :-)

Unless potassium is of great interest, uncoated vials are fine. Clotting will occur once you collect whole blood in an uncoated vial; upon centrifugation your sample will separate its own serum from the clot. You can collect this serum and use it for most chemistry tests. Potassium can be overestimated if using serum for analysis because clotting process leads to K release from platelets.

Collection volume plays a role as well. The bigger the volume, the more difficult it is to collect without anticoagulant. Recurrent bleeds tend to provide rather small sample volumes. As a few mentioned above, cardiac puncture is terminal. Depending of the animal size, it will indeed provide a bigger sample volume.

How to collect mouse blood samples from the tail ? (detailed protocol if possible).

This paper is very useful in terms of providing information about the various routes of blood collection in mice.

http://www.osa.sunysb.edu/dlar/newitems/mouseblood.pdf

Ian

We use these capillary tubes for collecting the blood.

http://wilburnmedicalusa.com/-c-0/cholestech-ldx-capillary-tubes-52193?gclid=Cj0KEQjww42tBRCO-sfEiO3DvYMBEiQAHeqMKOZBe72csuDvYHYLdKQZlXWs54n6cQwosa3Bf_sez4YaAm8P8P8HAQ

We then store the blood in heparin tubes.

In terms of storing whole blood samples, after extraction as described above. What are the optimum conditions please and maximum time before testing?. 

We've recently begun collecting blood via submandibular bleeding. It's incredibly easy, humane (no anesthesia) and reliably gives more blood (0.2-0.5 ml or more) than the tail veins. It's highly recommended for long-term studies that require repeated blood draws. Here's the reference:

Article A rapid, simple, and humane method for submandibular bleedin...

As for collection, we use EDTA-coated tubes that we purchase.

collection of blood by cardiac puncture is the most prevalent method.

in rat isolated heart study, how we can get blood sample (near 5ml)? cardiac puncture is not recommended in this method because it could affect cardiac contractility. and also we could not collect blood from the cavity after heart removal because during heart removal we add cold saline on heart due to decrease the risk of ischemic events. thank you