Intraoperative NIR imaging of prolonged ischemia in the mouse heart. Normally perfused heart assessed with IRDye78-CA (top panels), after ligation of a branch of the LAD artery with a suture (S) and reinjection of IRDye78-CA (second panels). Occluded vessels (arrowheads), a blood flow defect (white arrows), and vascular engorgement (V) are now visible in the NIR fluorescence and pseudo-colored merged images. After a total of 30 minutes of ligation, removal of the suture and reinjection of IRDye78-CA (third panels), hyperemia of the previously ligated vessels is seen (white arrowheads). After injection of IR-786 (bottom panels), a large perfusion defect persists (white arrows). Data are representative of four independent experiments. (Figure reproduced with permission from Nakayama et al.47)