Ht7800 120 kv transmission electron microscope

Experimental cells were pelleted by centrifuging (1000 rpm) before being fixed with 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer pH 7.4 for 30 min at 37 °C. The fixed cells stored in 4 °C in 0.1 M sodium cacodylate buffer were performed with a mixture of 1% OsO4, 1.5% K4Fe(CN)6 in 0.1 M sodium cacodylate pH 7.4 for 1 h at 4 °C and then overnight incubated in 0.25% uranyl acetate at 4 °C. After three water washes, samples were dehydrated in series of 15 min steps in 25%, 50%, 75%, 95% and 100% (v/v) ethanol and embedded in an epoxy resin (Sigma-Aldrich). The ultrathin sections (of 60–70 nm in thickness) of cells were prepared with a Leica EM UC7 ultramicrotome and counterstained with 1% uranyl acetate for 15 min and 1% lead citrate for 6 min. Thin sections were imaged using a HITACHI HT 7800 120kv transmission electron microscope operating at 80 kV.