Cardiac Blood Collection via Subclavian Puncture in Mice

Cardiac Blood Collection via Subclavian Puncture in Mice. Blood sampling in mice plays a significant role in scientific research. Cardiac puncture and large-vessel blood collection can obtain large blood volumes in terminal procedures.

However, cardiac puncture has received increasing attention due to ethical limitations and higher associated risks. In this study, we employed the subclavian puncture method for cardiac blood collection. Compared with the traditional abdominal puncture approach, this method effectively avoids liver damage, thereby improving sample quality and the reliability of experimental data.

When drawing blood, the syringe needle should be facing downward. Insert the needle from the left midclavicular line direction, aiming the needle towards the xiphoid process. Experiment preparation.

In intracardiac blood collection in mice, the one-milliliter syringe is prone to needle dislodgement due to its thin barrel and long plunger stroke. The two-milliliter syringe offers better handling than the five-milliliter syringe, making it the optimal choice for this procedure. Use heparin solution to rinse the syringe to prevent blood clotting.

Select healthy mice and weigh them. Administer inhalation anesthesia. Then, confirm that the mice have reached an adequate level of anesthesia.

Steps for cardiac blood collection in mice. After the mice were anesthetized, they were laid flat on the operating table. The chest area was sterilized with 75%ethanol.

Immobilize its forelimbs and wipe off excess alcohol. Hold the syringe with the needle facing downward. Locate the xiphoid process and the left clavicle.

Insert the two-milliliter syringe from the left clavicular midline with the needle directed toward the xiphoid process, ensuring the syringe remains parallel to the operating table. Control the insertion depth to 1 to 1.5 centimeters, ensuring that the syringe remains parallel to the operating table at all times. After confirming the correct positioning of the needle, slowly retract the syringe plunger to draw blood.

When the needle successfully enters the heart chamber, you will observe pulsatile blood flow into the syringe. During the blood-collection process, hand stability to prevent needle displacement. If difficulties in blood collection occur, adjust the syringe angle as needed to optimize blood flow and increase the success rate.

After blood collection, transfer the blood to heparin-coated microcentrifuge tubes. Centrifuge at four degrees Celsius and 2, 000 xg for 15 minutes. Carefully use a pipette to transfer the supernatant plasma to new microcentrifuge tubes and label appropriately.

In this study, we conducted repeated experiments on 14 C57BL/6 mice weighing between 21 and 29 grams. Each blood-collection volume ranged from 700 to 1, 200 microliter. All the experimental results are as follows, and no significant differences were observed between the genders.

Then, we measured the levels of ALT and AST in the plasma, and the results were all within the normal range. Cardiac puncture offers distinct advantages. Blood is directly drawn from the heart, which effectively avoids contamination from tissue, fluids, or other sources.

It requires only a single operation, reducing the stress response caused by repeated sampling in experimental animals. The subclavian puncture method features a well-defined needle trajectory and avoids penetrating the diaphragm or other complex anatomical layers. It also prevents liver damage, thus minimizing alterations in serum proteins, coagulation factors, and metabolic enzyme levels.