Whole hearts isolated from 32-week-old mice (NAGLU-/-, n = 5 and WT, n = 5) were fixed overnight in a bath of 4% aqueous buffered formalin, and processed for paraffin embedding. Coronal sections (10 μm thick) containing both right and left ventricles were processed as previously described [17 (link)–18 (link)]. Sequential sections from each heart were stained with haematoxylin and eosin (H&E) (Sigma-Aldrich), Masson's trichrome dye (Sigma-Aldrich) and periodic acid-Schiff Alcian blue-PAS (Dako). All sections were examined on a light microscope (Leitz, DIAPLAN), and images were acquired with a digital camera (Digital JVC, TK-C1380).
Plastic embedded sections were obtained from heart tissue specimens of 32-week-old mice fixed in 2.5% glutaraldehyde and postfixed in 1% osmium tetroxide in cacodylate buffer (0.2 M, pH 7.4). Tissues were washed and dehydrated in a graded series of ethanol solutions, cleared in propylene oxide, and embedded in Epon—Araldite resin. Semithin (0.5 μm) sections were stained with toluidine blue and examined under a light microscope.
Plastic embedded sections were obtained from heart tissue specimens of 32-week-old mice fixed in 2.5% glutaraldehyde and postfixed in 1% osmium tetroxide in cacodylate buffer (0.2 M, pH 7.4). Tissues were washed and dehydrated in a graded series of ethanol solutions, cleared in propylene oxide, and embedded in Epon—Araldite resin. Semithin (0.5 μm) sections were stained with toluidine blue and examined under a light microscope.
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