Thioflavin-S Staining for No-Reflow Assessment

Thioflavin-S staining was conducted 3 days after surgery on the sham group, I/R injury group, and I/R + pinacidil groups (0.1 and 0.5 mg/kg/day) to measure the benefits of pinacidil on no-reflow area according to previous study [29 (link)]. Similarly, Thioflavin-S staining was also performed on the sham and I/R groups that were transfected with or without AAV9-CRT. 2% thioflavin-S (MCE, USA) was dissolved in normal saline and injected into a different set of mice via the tail vein. After ten cardiac cycles, mouse hearts were immediately harvested, quickly washed with cold PBS, and fixed in 4% PFA overnight. The heart was cut into 1 mm slices, exposed to ultraviolet light, and photographed by a stereomicroscope (Leica, Germany). The no-reflow degree was calculated by the ratio of the dark area to the LV area.