Tem h600

Mycelia were prepared similar to the SEM method, but the concentrations of aqueous Triton X‐100 were 0 and 40 g l⁻¹. The mycelia were fixed with 1 ml of fixative (4% glutaraldehyde and 3% paraformaldehyde) for 4 h. The mycelia were collected by centrifugation at 8000 rpm for 5 min and washed three times with 0.1 M PBS buffer to remove residual fixative. The mycelia were fixed with 0.1 ml of 1% osmic acid overnight, washed four times with 0.1 M PBS buffer and dehydrated successively with 30%, 50%, 70%, 85%, 95% and 100% (v/v) ethanol. The mycelia were embedded in resin and further polymerized at 65 °C for 2 days. The treated samples were sliced using an ultra‐microtome, stained with uranium acetate and lead citrate, and observed using TEM (H‐600; Hitachi, Tokyo, Japan).

For TEM analysis, mice were deeply anesthetized by intraperitoneal injection of sodium pentobarbital and transcardially perfused with ice-cold 0.9% saline and 4% paraformaldehyde (PFA) in 0.1M phosphate buffered saline (PBS, pH 7.4). The substantia nigra pars compacta (SNc) of mice were then cut into 1 mm³ squares and postfixed in Trump’s solution (4% formaldehyde + 0.1% glutaraldehyde in 0.1M phosphate buffer) for 1 h at room temperature. Subsequently, the samples were kept in the 2% glutaraldehyde in 0.1M Na-cacodylate buffer (pH 7.4) overnight. Tissue was fixed in 1% osmium tetroxide and 1% aqueous uranyl acetate, dehydrated in a graded series of ethanol, and embedded in Embed 812/Araldite (EMS, Hatfield, PA). Thin sections (0.1 μm) were collected on copper grids, poststained with lead citrate and viewed at 80 kV with a JEOL 1400 transmission electron microscope (JEOL USA, Peabody, MA) or TEM H-600 (HITACHI, Tokyo, Japan).