Mitochondrial Ultrastructure Analysis

Mitochondrial ultrastructure was assessed in several pieces obtained from different regions of the left ventricle that were pre-fixed in 2.5% glutaraldehyde in 0.1M sodium cacodylate buffer, post-fixed in 2% osmium tetroxide (OsO4) in 0.1M sodium cacodylate buffer, dehydrated in an ethyl alcohol series, embedded with epoxy resin, and thermally polymerized as previously described (Katragadda et al., 2009 (link)). Ultra thin-sections (60 nm) were cut by an ultramicrotome (Leica UC7, Leica Microsystems Inc., ON, Canada) and then stained with 4% uranyl acetate and Reinold's lead citrate. The contrasted sections were imaged under a Hitachi H-7650 transmission electron microscope at 80 kV equipped with a 16 megapixel EMCCD camera (XR111, Advanced Microscopy Technique, MA, USA) was used for viewing the sections (Cho et al., 2007 (link); Katragadda et al., 2009 (link)).