Fluka epoxy embedding medium kit

To investigate the cellular ultrastructure, glioma cells were washed three times with ice-cold phosphate-buffered saline (PBS; Sigma-Aldrich Corporation). The cells were collected after centrifugation (1,500 rpm for 5 min) and prefixed with 2.5% glutaraldehyde, then postfixed in 1% osmium tetroxide, dehydrated in ethanol gradients, impregnated with epoxy embedding resin (Fluka Epoxy Embedding Medium Kit; Sigma-Aldrich) and cut with an ultramicrotome (Leica EM UC6). Thin sections were poststained with uranyl acetate and lead citrate, and evaluated by a JEM-1220 (Jeol) TEM at 80 keV, with a Morada 11 megapixels’ camera (Olympus Soft Imaging Solutions).

U87 and U118 glioma cells were seeded in six-well plates (1×10⁵ cells per well) and incubated for 24 hours. Cells cultured in medium without the addition of GO and rGO were used as the control. Graphene was introduced to the cells at increasing concentrations (5, 10, 20, 50, and 100 μg/mL). Cell morphology was recorded using an optical microscope 24 hours after exposure.
To investigate the cellular ultrastructure, the glioma cells were washed three times with ice-cold phosphate-buffered saline (Sigma-Aldrich). The cells were collected after centrifugation (1,200 rpm for 10 minutes) and prefixed with 2.5% glutaraldehyde, then post-fixed in 1% osmium tetroxide, dehydrated in ethanol gradients, impregnated with epoxy embedding resin (Fluka Epoxy embedding medium kit; Sigma-Aldrich), and cut with an ultramicrotome (EM UC6, Leica Microsystems GmbH, Wetzlar, Germany). Thin sections were post-stained with uranyl acetate and lead citrate and evaluated by TEM.

Brain tissues from the control group and the groups treated with 50 μg/L and 500 μg/L pG were fixed for TEM examination in a fixative consisting of 1% glutaraldehyde in phosphate-buffered saline at pH 7.2. After fixation, the samples were postfixed in 1% osmium tetroxide and dehydrated in a graded series of ethanol. The tissues were embedded in an epoxy embedding resin (Fluka Epoxy Embedding Medium Kit; Sigma-Aldrich Co., St Louis, MO, USA). Ultrathin sections (100 nm) were cut with an ultramicrotome (Leica EM UC6; Leica Microsystems Nussloch GmbH, Nussloch, Germany) and stained with uranyl acetate and lead citrate. The samples were viewed using the TEM at 80 keV (JEOL), and images were taken with a Morada 11 megapixel camera (Olympus Soft Imaging Solutions GmbH).