Microfill mv 122

Vascularization was assessed at the two‐ and eight‐week end‐points with the use of micro‐CT angiography. Immediately after overdose with barbiturates (Pentobarb) and assessment of pedal withdrawal reflex, a lateral incision was made through the integument and abdominal wall, and the diaphragm was cut along the entire length of the rib cage to expose the pleural cavity. The rib cage was cut, and sternum lifted away to expose the heart, and a cardiac puncture was performed. Specifically, a 23G catheter was inserted into the left ventricle and an incision was made in the right atrium to create an outlet for blood (exsanguination) and the solutions perfused in the circulatory system. Heparinized PBS (20 U mL⁻¹), followed by 10% neutral buffered formalin, was perfused using a peristaltic pump at 5 mL min⁻¹ for 2 min. After fixation, 5 mL of radiopaque contrast agent (Microfill MV‐122, Flow Tech) was prepared and perfused as per manufactures instructions, and the compound was polymerized for 6 h at 4 °C. Following Microfill polymerization, grafts were explanted and post‐fixed in 10% neutral buffered formalin for 24 h.