Langendorff Retrograde Heart Perfusion: An Ex Vivo Technique to Perfuse Isolated Mouse Heart

To begin, place an anesthetized mouse in the supine position on a dissection board. Make an incision to access the thoracic cavity and expose the heart. Locate the aorta and its branches - the left common carotid artery and brachiocephalic artery. Transect these arteries to isolate the heart.

Immediately, immerse the excised heart into a Petri dish containing a nutrient-rich oxygenated buffer solution. Concomitantly, insert a cannula into the heart such that its tip reaches the ascending aorta. Use a suture to ligate the cannula. Connect the cannula to a syringe containing oxygenated perfusion solution.

Inject the perfusion solution into the heart. This step allows the blood to leave the coronary arteries and effuse from the pulmonary veins. Next, connect the cannulated heart to the Langendorff apparatus. Use peristaltic pump to perform perfusion at a constant flow rate.

Under these conditions, the buffer perfuses into the heart. This creates cannulation pressure that closes the aortic valves, causing the buffer to move through the coronary arteries. Eventually, the buffer perfuses the heart muscles retrogradely and later effuses through the pulmonary veins. A successfully perfused heart is now ready for further downstream applications.