Transmission Electron Microscopy

Samples of fresh spinal cord (1 mm × 1 mm × 1 mm) were carefully collected from the rats, then primarily fixed in 2.5% glutaraldehyde for 24 hours. Then, they were washed with Sorenson’s Phosphate Buffer (pH=7.4) and post-fixed in 1% osmium tetroxide for 1 hour. After post-fixation and dehydration, propylene oxide was used to wash the tissues, and then the tissues were embedded in epoxy resin embedding media. Following this procedure, the tissue blocks were sectioned using the ultramicrotome (LKB Nova, Sweden) to produce section around 60 nanometers in thickness. These sections were en bloc stained with uranyl acetate and lead citrate and examined using transmission electron microscope (HITACHI, Japan).