Em208s

Cells were seeded onto 4-chambered coverglass (Lab-Tek Chambered Coverglass System) (Nalgene/Nunc, Rochester, NY, USA) at a density of 2 × 10⁴ cells/ml (14,000 cells/well). Images were acquired using the Olympus EM208S transmission electron microscope.

The lung specimens fixed in glutaraldehyde overnight were washed with cacodylate buffer (pH 7.2) three times and postfixed in 1% osmium tetraoxide. Then, samples were dehydrated in ascending concentrations of ethanol and embedded in epoxy resin. Sections of 1 μm thickness were prepared and stained with toluidine blue for observation under light microscopy. Areas suitable for examination under electron microscopy were determined. Ultrathin sections (60–70 nm) were prepared and stained with uranyl acetate and lead citrate. Imaging was conducted with a transmission electron microscope (EM208S; Olympus, Tokyo, Japan). The number of vacuoles and the ratio to the cross-sectional area occupied on the sections were quantified using MetaMorph 6.1 software.

We used TEM to observe mitochondrial ultrastructure of liver tissues in three experimental groups. Liver tissues were sliced less than 1 mm³ thickness, the sections were pre-fixed in 2.5% glutaraldehyde at 4°C for 2 h. After washing with sodium cacodylate buffer, tissues were postfixed with 1% osmium tetroxide solution at 4°C for 1 h. The slices were dehydrated in graded concentrations of ethanol, infiltrated with propylene oxide and embedded. The slices were sheered into 60 nm sections. Imaging was captured with transmission electron microscope (Olympus EM208S, Japan).