FIG. 15.

In vivo transfection of rat cardiomyocytes by direct DNA injection. The animal is anesthetized with a mixture of N₂O (0.2 L/min), O₂ (0.2 L/min), and sevoflurane (2%–3%), and the heart is exposed through a right-lateral thoracotomy. The free wall of the right and/or the left ventricle is injected three to four times each, delivering a total of 20 μg of reporter plasmid(s)/ventricle in 100 μL of saline. In this example injection of the right ventricle is shown, with the 29-gauge needle bent at the tip at an almost right angle to allow easy injection of the thin right ventricle wall. The thorax is then closed and the animal is sacrificed 5 days later. Expression of luciferase reporter and normalization genes is determined in ventricle homogenates. Courtesy of Dr. Warner Simonides.