Histological Assessment of Myocardial Fibrosis and Cardiomyocyte Size

Wheat germ agglutinin (Thermo Fisher, Waltham, Massachusetts, USA) staining was used to evaluate cardiomyocyte cross-sectional area in mid-LV cross sections (5μm). Digital images were taken from x40 magnification of 5 random fields of view in each heart sample. Area of cardiomyocytes were analyzed using ImageJ. For myocardial fibrosis, trichrome (Newcomer Supply, Middleton, Wisconsin, USA) stained slides (1 per animal) were examined using ZEN microscope software (Carl Zeiss Microscopy, Cambridge, United Kingdom) and quantified using MATLAB 7.10.