Commonly Used Blood Withdrawal Methods
Blood withdrawal is a critical procedure in
biomedical research, pharmacology, and laboratory animal science. It provides
essential samples for diagnostic testing, pharmacokinetic studies, toxicology,
and other experimental investigations. While accuracy and sample quality are
important, the welfare of the animals must also be considered. Understanding
proper techniques ensures reliable results while minimizing stress and
complications.
Here is a step-by-step overview of commonly
used blood withdrawal methods in laboratory animals, along with the
materials required and approximate blood volumes.
1. Retro-orbital
Sinus Method
Materials Needed: Capillary tube
or fine needle, anesthesia (if required), cotton swab.
Step-by-Step Procedure:
1.
Restrain the animal gently but securely.
2.
Anesthetize if required (commonly done in mice and
rats).
3.
Insert a fine glass capillary tube or needle into the
medial canthus of the eye.
4.
Rotate gently to puncture the retro-orbital sinus.
5.
Collect the blood as it flows into the tube.
6.
Apply gentle pressure after removal to minimize
bleeding.
Blood
Volume: 0.2–0.3 ml (mice), up to 3 ml (rats).
Notes:
Provides a large sample quickly but requires skill to avoid ocular damage.
2. Tail Vein
Blood Withdrawal
Materials Needed: Needle
(25–27G), syringe or capillary tube, alcohol swab, warming lamp.
Step-by-Step Procedure:
1.
Warm the tail to dilate blood vessels.
2.
Securely restrain the animal.
3.
Clean the tail with alcohol.
4.
Insert a needle into the lateral tail vein.
5.
Collect the desired blood volume.
6.
Apply pressure to the puncture site after collection.
Blood
Volume: 0.05–0.2 ml per puncture.
Notes:
Useful for repeated sampling of small to moderate blood volumes.
3. Tail Snip Method
Materials Needed: Sterile
scissors or blade, alcohol swab, microtube.
Step-by-Step Procedure:
1.
Warm the tail to promote blood flow.
2.
Restrain the animal gently.
3.
Disinfect the tip of the tail.
4.
Use sharp scissors or a sterile blade to cut a small
portion of the tail tip.
5.
Collect the blood droplets into a microtube or onto
test strips.
6.
Apply gentle pressure or styptic solution to stop
bleeding.
Blood Volume: 0.02–0.1 ml.
Notes: Suitable for small volume
collections, commonly in mice.
4. Cardiac
Puncture Blood Withdrawal
Materials Needed: Syringe with fine
needle (23–25G), anesthesia, alcohol swab.
Step-by-Step Procedure:
1.
Anesthetize the animal completely (this is usually a
terminal procedure).
2.
Place the animal in a supine position.
3.
Insert a needle into the chest cavity between the ribs.
4.
Advance carefully until the heart is reached.
5.
Aspirate slowly to collect the blood.
Blood Volume: 0.5–1 ml (mice),
3–10 ml (rats).
Notes: Provides large volumes
but is terminal in nature.
5. Posterior Vena
Cava Blood Withdrawal
Materials Needed: Syringe and
needle (21–25G), anesthesia, surgical tools (if needed).
Step-by-Step Procedure:
1.
Anesthetize the animal.
2.
Place in dorsal recumbency and make a small abdominal
incision (if surgical approach).
3.
Insert the needle into the posterior vena cava with
caution.
4.
Withdraw the required blood volume.
5.
Close the incision or euthanize depending on study
design.
Blood Volume: 0.5–1 ml (mice),
3–6 ml (rats).
Notes: Yields substantial blood
volume, requires advanced surgical skills.
6. Saphenous Vein
Blood Collection
Materials Needed: Needle
(25–27G) or lancet, restraining device, alcohol swab.
Step-by-Step Procedure:
1.
Restrain the animal without anesthesia.
2.
Shave or clean the hind limb.
3.
Position the animal to expose the lateral saphenous
vein.
4.
Puncture the vein with a needle or lancet.
5.
Collect the blood droplets.
6.
Apply pressure to stop bleeding.
Blood Volume: 0.05–0.2 ml per
collection.
Notes: Minimally invasive and
suitable for repeated samples.
7. Jugular Vein
Blood Collection
Materials Needed: Syringe with
fine needle (23–25G), alcohol swab, restraining device.
Step-by-Step Procedure:
1.
Restrain the animal in dorsal recumbency.
2.
Shave or disinfect the neck region.
3.
Insert the needle into the jugular vein at a shallow
angle.
4.
Aspirate slowly to collect blood.
5.
Remove the needle and apply pressure.
Blood Volume: 0.2–0.8 ml (mice),
2–5 ml (rats).
Notes: Provides large volumes
quickly but requires skill to prevent complications.
8. Submandibular
Vein Blood Collection
Materials Needed: Lancet or
needle, restraining device, alcohol swab.
Step-by-Step Procedure:
1.
Restrain the animal without anesthesia (commonly in
mice).
2.
Hold the animal securely by the scruff.
3.
Use a lancet or needle to puncture the vein located
below the mandible.
4.
Collect the blood droplets as they emerge.
5.
Apply pressure to stop bleeding.
Blood Volume: 0.1–0.2 ml (mice).
Notes: Provides moderate volumes
and is quick, but should be performed carefully.
9. Dorsal
Metatarsal Vein Blood Collection
Materials Needed: Needle
(25–27G), syringe, alcohol swab.
Step-by-Step Procedure:
1.
Restrain the animal gently.
2.
Shave or disinfect the dorsal side of the foot.
3.
Locate the dorsal metatarsal vein.
4.
Insert a needle at a shallow angle.
5.
Collect the desired blood volume.
6.
Apply pressure afterward.
Blood Volume: 0.05–0.2 ml.
Notes: Less common method, used
for small volume collections.
You may like to read these links:
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3. List of All SOPs and Documents for the In-vivo Laboratory.
4. List of All SOPs and Documents for Clinical Research
5. List of Instruments and Equipment Used In Laboratory Animal Facility
6. Animal Facility Design - Small Laboratory Animals (Rat, Mice, Rabbit, Guinea Pig)
7. List of Chemicals Needed for In-vivo Laboratory
8. List of Physiological Data for Small Laboratory Animals
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