Jem 1010 transmission electron microscope

A375 cells were seeded at 1 × 10⁵ cells per well in 6-well plates and allowed to grow for three days to optimize membrane structures in the culture, and the medium was changed on the second day. Cells were washed twice in serum-free RPMI before being treated with LTX-315 dissolved in serum-free RPMI at 3.5 and 17 μM, with serum-free RPMI as a negative control. Cells were then washed with PBS before fixation for 24 hours at 4°C with 4% formal aldehyde and 1% gluteralaldehyde in a Hepes buffer at pH 7.8. Dehydration and post-fixation protocols included incubation in a 5% buffered tannic acid and incubation in 1% osmium-reduced ferrocyanide. Ultrathin sections were prepared, and uranyl acetate (5%) and Reynolds's lead citrate were used for staining and contrasting. Samples were examined on a JEOL JEM-1010 transmission electron microscope, and images were taken with an Olympus Morada side-mounted TEM CCD camera (Olympus soft imaging solutions, GmbH, Germany).

Cells were seeded at 1 × 10⁵ cells per well in 6-well plates and allowed to grow for three days to optimize membrane structures in the culture. Cells were then treated with peptides dissolved in serum-free RPMI at 25 μM. Cells were then washed with PBS before fixation for 24 h at 4 °C with 4% formal aldehyde and 1% glutaraldehyde in a HEPES buffer at pH 7.8. Dehydration and post-fixation protocols included incubation in a 5% buffered tannic acid and incubation in 1% osmium-reduced ferrocyanide. Ultrathin sections were prepared, and uranyl acetate (5%) and Reynolds’s lead citrate were used for staining and contrasting^{13 (link)}. Samples were examined on a JEOL JEM-1010 transmission electron microscope, and images were taken with an Olympus Morada side-mounted TEM CCD camera (Olympus soft imaging solutions, GmbH, Germany).