Standard Operating Procedure RevisedNovember 2017

Saphenous Blood Collectionin the Mouse Page 1 - 2

Purpose:to describe the procedure to perform blood collection using the saphenous vein in the mouse. The saphenous vein blood collection is used for repeat blood sampling from the mouse. Individual samples should not exceed (0.007 ml/gm taken bi-monthly). See attachment for blood collection.

Policy: followed by CCAC guidelines and regulations.

Responsibility: this procedure lies with the trained technologist(s) and researchers assigned to the study.

MATERIALS

Needle (e.g. 23G)

Scalpel blade (e.g. #20)

Gauze

50 ml conical tube (holes are drilled into the conical end to facilitate air flow).

Collection tubes (e.g. capillary tubes, small Microtainer tubes, Starstedt
Microvette tubes).

PROCEDURES

Set up work surface with the above outlined materials.

Gently place the animal into the conical tube.

Note:Anesthesia is not required.

Extend the left hind leg and hold it in position by placing your thumb and indexfinger over the fold of skin between the tail and thigh.

By applying firm pressureto this area, you can keep the leg in the proper position while occluding thesaphenous vein.

If you need to collect blood from the right saphenous vein, holdthe fold of skin between the abdomen and cranial thigh surface of the right hindleg.

Using a small scalpel blade, shave a small area of dryhair (approximately 1cm) off the lateral surface of the left hind leg proximal to the hind foot.

Thesaphenous vein should now be visible on the surface of the thigh.

You can use a damp gauze to smooth away any hair that is covering the bleeding site.

Use a needle to puncture the saphenous vein.

A drop a blood will appear and can then be collected into a collection tube.

If theflow of blood stops, flex the foot and/or briskly rub the puncture site with a dry gauze swab.

Avoid re-puncture of the site where ever possible. If it becomesnecessary the same puncture site should be penetrated. You can collectmultiple samples by this method; just remember that a maximum volume of 100 microlitres only may be collected once every two weeks.

When the blood collection is completed, flex the foot towards the knee in order
to reduce the flow of blood back to the puncture site.

By using a gauze sponge, apply firm pressure to the puncture site until
hemostasis is achieved.

Once hemostasis is achieved, remove the mouse from the conical tube andreturn it to its cage.

If several small blood samples are required throughout the day, just remove thescab off the puncture site to initiate blood flow. If the flow doesn't start, flex thefoot and/or briskly rub the puncture site with a dry gauze swab.

Label the collection tube .as per facility standards.

Discard all garbage into the appropriate containers and clean the work area.

Footnote: There is a tendency for the formation of large subcutaneous hematomas with this technique. Therefore, this technique is not recommended if there is a more suitable alternate available.

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