Pannoramic midi 2 scanner system

Sections were prepared as described above. After deparaffinization and rehydration, the sections were stained with iron hematoxylin for 15 min, rinsed with tap water for 1 min, rinsed with 1% hydrochloric acid in 80% ethanol for 5 s, rinsed with 1% ammonia solution for 5 s, and rinsed with tap water for 1 min. The sections were then stained with Masson’s fuchsin-Ponceau mixture for 25 min and rinsed with tap water for 1 min. Next, the slides were treated for 10 min with phosphotungstic acid orange G and Masson’s light green solution for 4 min. After rinsing with tap water for 1 min, 1% glacial acetic acid was added for 1 min, and the slides were washed with tap water for 1 min. Dehydration in an ethanol gradient and clearing with xylene were conducted, and the slides were mounted with neutral balsam. The stained sections were observed and photographed under a Pannoramic MIDI II scanner system (3DHISTECH, Budapest, Hungary). The images were captured with CaseViewer (version 2.1, 3DHISTECH, Budapest, Hungary).

The injured hearts collected at different time points were fixed overnight in 4% paraformaldehyde, dehydrated, cleared, and embedded in paraffin wax. Sections with a thickness of 5 μm were prepared for staining. The sections were deparaffinized in xylene (3 × 5 min) and rehydrated with successive 3 min washes in 100%, 100%, 100%, 90%, 80%, and 70% ethanol with one final wash with tap water. The sections were then stained with hematoxylin medium for 15 min, rinsed with tap water for 1 min, rinsed with 1% hydrochloric acid in 80% ethanol for 5 s, rinsed with a 1% ammonia solution for 5 s, and rinsed with tap water for 1 min. The sections were then stained with eosin medium for 3 min and rinsed with tap water for 1 min. After dehydration in an ethanol gradient and clearing with xylene, the slides were mounted with neutral balsam. The stained sections were observed and photographed under a Pannoramic MIDI II scanner system (3DHISTECH, Budapest, Hungary). The images were captured with CaseViewer (version 2.1, 3DHISTECH, Budapest, Hungary).