Epon 812 r

Adrenal glands were diced into small pieces at approximately 1 mm³ and fixed in 0.1 M phosphate buffer at pH 7.2–7.4 with 2% (vol/vol) glutaraldehyde. After washing in 0.2 M phosphate buffer (pH 7.2) overnight, these specimens were post-fixed in cold 1% OsO4 (pH 7.3) for 2 h at 4°C. All tissue slices were dehydrated in different gradients of ethyl alcohols (30–100%) and then embedded in Epon 812 R (Merck, Whitehouse Station, NJ). Finally, ultrathin sections (50 nm) were stained with uranyl acetate and lead citrate and examined by an H-7650 transmission electron microscope (Hitachi High-Technologies Co., Japan) at 80 kV with an Ultrascan CCD camera (11 (link)). Ultrastructure morphometric assessments of mitochondria were conducted by using NIH ImageJ software.

Blocks of turtle pancreas approximately 1 mm³ in size were fixed in 2.5% glutaraldehyde in 0.1 MBPS solution (pH 7.4) at 4 °C overnight. Post-fixation of the turtle pancreas was done in 1% osmium tetroxide (Polysciences Inc. Warrington, PA, USA) for 1 h. The specimen washing was done in distilled water and stained en bloc with 2% aqueous uranyl acetate, then washed again in distilled water (3 × 5 min). Dehydration was done in ethanol, and specimens were embedded in Epon 812 R (Merck, Whitehouse Station, NJ). Ultrathin 50 nm-thick sections were mounted on copper grids and contrasted with uranyl acetate and lead acetate. Specimens were observed under the Hitachi TEM system (H-7650, Tokyo, Japan).