Panoramic scan scanner

Hearts were harvested after the indicated timelines and fixed in 4 % paraformaldehyde overnight. Following fixation, hearts were rinsed in 50 % ethanol, dehydrated and embedded in paraffin. 2.5-µm-thick sections were cut from the cardiac samples. Masson’s trichrome with aniline blue stainings were prepared using a staining kit from Bio Optica (#04-010802, Milan, Italy). Hematoxylin and Eosin (H&E)-stained sections were made following standard protocols. A “TdT-mediated dUTP nick-end labeling” (TUNEL) staining kit from Promega (#G3250, Madison, Wisconsin, USA) was used to visualize cell death in cardiac sections 24 h after LAD ligation. Samples were counterstained with DAPI (4′,6-diamidino-2-phenylindole) and mounted in DAKO fluorescent mounting medium (S3023, DAKO; Denmark). All sections, including the fluorescently labeled TUNEL stainings, were scanned using a Panoramic SCAN scanner from 3D HISTECH (Budapest, Hungary).

The eyeballs were fixed in FAS eyeball fixator for 24 h. Paraffin-embedded eyeballs were cut into 3-μm-thick slices and stained with HE. A panoramic SCAN scanner (3D histech, Budapest, Hungary) was used to scan images and CaseViewer software was used to analyze them. We take 6-8 points from different locations of the retina to measure the thickness of each layer and count the number of outer nuclear layer (ONL) and inner nuclear layer (INL).