Blood collection through subclavian vein puncture in mice. Blood sample collection from mice is essential in most research laboratories. The conventional approaches for blood collection in mice is tail cutting, when less than 100 microliter sample is needed.
However, if more than 100 microliter of blood are required at a non-terminal time point, retro-orbital, submandibular bleeding, or submental blood collection, are the most commonly-considered techniques. In some locations the jugular vein incision or catheterization were adopted as alternative methods. Nevertheless, the above methods are hurtful to mice.
On the basis of our formal researches, here we introduce a new economical and gentle strategy for blood collection from subclavian vein in mice. The safety and feasibility, and obtained blood volume with this technique, are presented and discussed. After general anesthesia, lie the mouse in supine position.
Clean the infraclavicular space with 75%ethanol. Confirm the clavicle and suprasternal fossae before puncture. Identify the puncture site in the middle of clavicle.
Move the needles superior and upward toward the suprasternal fossae, just posterior to the clavicle. After moving three to four millimeters forward, keep negative pressure in the syringe, and slowly draw back the needle. Blood will drain into the syringe during the course of withdrawal.
After blood collection, press the puncture site softly for one to two minutes to stop bleeding. Prepare required materials, including 75%ethanol, adhesive tape, epilating agent, tubes, 1.0 milliliter syringe connected with needle, electronic scale, heparin, and saline. After satisfactory general anesthesia, place the mouse on the operating table in supine position, and two to four centimeters away from the edge of the table to facilitate vein puncture.
Fix the limbs in a comfortable position, as shown in Figure 2. No intubation or mechanical ventilation was needed in the whole procedure. Apply epilating agent around the infraclavicular space with cotton swab.
Three minutes later, wash the epilating agent with wet cotton swab, to remove the fur and any visible dirt. Sterilize the infraclavicular space with 75%ethanol, and then dry with clean gauze. Identify the location of clavicle bone and suprasternal fossae with one finger.
Locate the puncture site, close to the middle of the clavicle bone, and caught onto it. Put the left index finger lateral to the puncture site to fix the skin and subcutaneous tissue. Move the needle upward in a cranial lay toward the suprasternal fossae.
Once the needle enter into the subcutaneous tissue, the ring finger of right hand must move backward to form negative pressure. Move forward the syringe for three to four millimeters, but stop if there is no blood drain into the syringe. Then slowly draw back the syringe, and keep negative pressure in it.
In most occasions the blood will enter the syringe at the course of drawing back. Once the blood enter into the syringe, fix the syringe, and keep negative pressure, until required volume of blood collected in the syringe. After blood collection, withdraw the puncture needle, and press the puncture site with cotton swab slightly for one to two minutes to stop bleeding, then send mice back to cages.
Transfer the blood sample into our heparin tube. We repeated this procedure in 10 Quimi mice. Nine procedures succeeded in the right side.
One case failed within three attempts in the right side, and blood collection succeeded in the left side. The time course ranged between 35 to 126 seconds. Blood collection volume was set to around 200 microliters.
Blood collection was succeeded with one to four attempts. Blood sampling succeeded at the first attempt in two mice, and at four attempts in one mouse, at two to three attempts in other mice. All data were illustrated in Table 1.
All animals survived, and recovered within 30 minutes after the blood sampling. There was no significant difference between sex for all observed parameters. In conclusion, subclavian vein puncture is a safe and effective method for blood collection in mice.
Besides tail vein cutting and retro-orbital plexus blood collection, this method is feasible and suitable for observational researches at several time points in mice.
