White light scanner

TTC staining was performed 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 infraction size according to previous guideline [4 (link)]. Similarly, TTC staining was also performed on the sham and I/R groups that were transfected with or without AAV9-CRT. After the mice were anesthetized with isoflurane vapor, the LAD was ligated again at the same level and 1% Evans blue was injected through the LV outflow tract. Each heart was immediately excised and cut evenly into 1-mm-thick slices from the apex to the bottom on frozen ice. The slices were incubated in 1% TTC solution at 37 ℃ for 10 min and fixed in 4% paraformaldehyde (PFA) overnight. The samples were then scanned using a white-light scanner (Canon, Tokyo, Japan). The area at risk (AAR) and infarct area of each slice were measured using the ImageJ software (Version 1.8, NIH). The relative infarct size was calculated as the ratio of the infarct area to the AAR.

TTC staining was conducted to measure cardiac infarction size [4 (link)]. After the mice were anesthetized with 2% isoflurane, the slipknot was tied again, and 1% Evans blue (w/v, Sigma, USA) was injected into the aortic root to perfuse the left ventricle. Then, the heart was rapidly excised, cut into 1 mm slices, and incubated with 1% TTC solution (Sigma, USA) at 37 °C for 10 min. Consecutive slices in each sample were scanned by a white light scanner (Canon, Japan). Area at risk (AAR) was defined as tissues not perfused by Evans blue. Infarcted myocardium was white, whereas viable myocardium was red. The infarction degree was calculated as the ratio of the infarction area to the AAR.