2.4. Transmission Electron Microscope

An electron microscope was used to observe the autophagosomes in enterocyte cells to note the initiation of autophagy. To assess autophagic vacuoles in the jejunal cells, the jejunal tissue was cut into small pieces (approximate 1 mm³) and immediately fixed in 2.5% glutaraldehyde solution at 4 °C overnight. After being postfixed in 1% osmic acid and dehydrated with step-wise gradient ethanol (30, 50, 70, 80, 95, 100%), the samples were embedded in an epoxy resin. Samples then were cut on an LKB-NOVA ultramicrotome into 70 nm sections, which were examined under a HITACHI-600IV electron microscope (HITACHI, Tokyo, Japan)