What is a Transcardial perfusion?

Transcardial perfusion, which utilizes the vasculature to achieve systemic delivery of fixative in the living animal, is a commonly used standard for tissue fixation. In this technique, the animal is anesthetized, the chest wall opened and the beating heart exposed.

How long does it take to perfuse a rat?

The entire procedure takes approximately 3 minutes from chest opening to the opening of the right atrium. (Solutions& Anesthesia used for the perfusion should be included in the description.) the heart, and then a needle (attached to a gravity perfusion system) will be inserted into in the left ventricle.

How do you perform a perfusion fixation?

Perfusion Surgery Make a 5-6 cm lateral incision through the integument and abdominal wall just beneath the rib cage. Carefully separate the liver from the diaphragm. Make a small incision in the diaphragm using the curved, blunt scissors.

Why do we perfuse mice?

The advantage of directly perfusing fixative through the circulatory system is that the chemical can quickly reach every corner of the organism using the natural vascular network. In order to utilize the circulatory system most effectively, care must be taken to match physiological pressures 3.

How to euthanize a rat after a perfusion?

Protocol. Rat should be fully anesthetized (e.g., unresponsive to toe pinch). Euthanize the rat by exsanguination. The easiest way to do this is by cutting the abdominal aorta. Avoid cardiac puncture, as the puncture site will leak during the perfusion procedure. Open the chest widely, taking care not to puncture the lungs.

How is the perfusion of a rat lung done?

Protocol. Retract one side of the chest wall laterally, bluntly dissect the pleura to free up the lung, gently push the lung away from the chest wall with a Q-tip, and cut the chest wall vertically along the spinal processes. Repeat for the other half of the chest wall. Be careful not to puncture the lung with the sharp edges of the cut ribs.

How is a fixative solution Perfused in a rat?

However, a more rapid and uniform fixation is usually obtained if the fixative solution is perfused via vascular system, either through the heart or through the abdominal aorta. The following procedures provide fixation of most rat organs with 4% paraformaldehyde. Materials

How much fix to flow in a perfusion for a mouse?

For rats, allow 200 to 250 cc of fix to flow. For mice, allow 50-100 cc of fix to flow. Consider the perfusion a dialysis process, exchanging formaldehyde and water, rather than a ‘flushing’ process. Close both clamps and remove perfusion instruments.