3 mm diamond knife

Cancer cell cultures were acquired and immediately fixed in neutral buffered 2.5% glutaraldehyde at room temperature (25°C). The specimens were post-fixed in 1% osmium tetroxide (OsO₄) for 10 minutes, dehydrated using a graded ethanol series, enbloch stained with 2% uranyl acetate in 50% ethanol for 30 minutes, and embedded in Spurr's epoxy resin. Semi-thin (1 μm) and ultra-thin (< 90 nm) sections were cut using a Diatome 3 mm diamond knife on the Leica EM UC6 ultramicrotome. Semi-thin sections were stained with toluidine blue and examined using an Olympus BX60 light microscope with an attached Olympus DP71 digital camera. All ultra-thin sections were stained using lead citrate to be viewed under TEM on a Jeol 1400 EM at 80 kV. Images for publication were compiled into using Photoshop CS5 software.

Briefly, cells were fixed in 2.5% glutaraldehyde at room temperature. The specimens were postfixed in 1% osmium tetroxide (OsO₄) for 10 min, dehydrated using a graded ethanol series, en bloc stained with 2% uranyl acetate in 50% ethanol for 30 min, and embedded in Spurr's epoxy resin. Thereafter, ultrathin (< 90 nm) sections were cut using a Diatome 3‐mm diamond knife on the Leica EM UC6 ultramicrotome. The ultrathin sections were stained using lead citrate to be viewed under TEM on a Jeol 1400 EM at 80 kV.