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Tissues collected for histology were fixed in Bouin's (Sigma St. Louis, MO) for 1 hour, embedded in paraffin and sectioned.
The samples were fixed in 4% formaldehyde and embedded in paraffin for histologic investigation, or fixed for 4 hours with 2% formaldehyde and 2.5% glutaraldehyde in 0.1 M phosphate buffer (pH 7.3), followed by 1% osmium tetroxide for 1 hour, embedded in Epon-Araldite resin and processed for TEM.
Femurs were fixed in 10% formalin for 24 hours, decalcified in EDTA/Formalin for additional 24 hours embedded into paraffin and sectioned.
The two middle parts were fixed in 4% neutral-buffered formalin for 48 hours, embedded in paraffin, and cut into 4-µm-thick sections for histology and immunostaining.
Immediately after MRI, tissues were fixed by perfusion with 4% paraformaldehyde, post fixed for 24 hours, embedded in paraffin and cut into 5 µm thick sections for histological analysis.
The scores for each individual parameter were then added together to give a total score between 0 and 12. Colonic specimens obtained as described were fixed in formalin for at least 24 hours, embedded into paraffin, and cut into 4 5 µm sections.
At the end of experimental procedures, hearts were fixed in 10% formalin for 24 hours, embedded in paraffin and cut into 5 µm sections that were mounted onto slides and stained with Picrossirius Red for measurement of collagen scar, left ventricular cavity perimeter (LV cavity) and thickness scar.
They were then fixed in formalin for 24 48 hours, embedded in paraffin and sectioned.
Autopsied tissues were fixed in 10% buffered formalin for 24 48 hours, embedded in paraffin, and sectioned 4 μm thick onto Vectabond treated slides.
All palpable tumors were fixed in 4% paraformaldehyde for 2 to 4 hours, embedded in paraffin and processed for staining with hematoxylin/eosin.
The surgical samples were fixed in 10% neutral buffered formalin for 12 24 hours, embedded in paraffin, cut and stained with Hematoxylin-Eosin.
