An Imaging Protocol to Discriminate Specialized Conduction Tissue During Congenital Heart Surgery.

We performed preclinical validation of intraoperative fiber-optic confocal microscopy (FCM) and assessed its safety and efficacy in an ovine model of the pediatric heart. Intraoperative imaging was performed using an FCM system (Cellvizio, Mauna Kea Technology, Paris, France) with specialized imaging miniprobe (GastroFlex UHD, Mauna Kea Technologies). Before imaging, we applied an extracellular fluorophore, sodium fluorescein, to fluorescently label extracellular space. We imaged arrested hearts of ovine (1-6 months) under cardiopulmonary bypass. Image sequences (1-10 seconds duration) were acquired from regions of the sinoatrial and atrioventricular node, as well as subepicardial and subendocardial working myocardium from atria and ventricle. The surgical process was evaluated for integration of the imaging protocol during the operative procedure. In addition, fluorescein cardiotoxicity studies (n = 3 animals) were conducted by comparing electrocardiogram (PR and QRS intervals) and ejection fraction at baseline and after topical application of fluorescein at 1:10, 1:100, and 1:1000 dilutions on a beating ovine heart. Our studies suggest that intraoperative FCM can be used to identify regions associated with specialized conducting tissue in ovine hearts in situ. The imaging protocol was integrated with conventional open heart surgical procedures with minimal changes to the operative process. Application of fluorescein in varying concentrations did not affect the normalized PR interval, QRS interval, and ejection fraction. These preclinical validation studies demonstrated both safety and efficacy of the proposed intraoperative imaging approach. The studies constitute an important step toward first-in-human clinical trials.