2.14. Transmission electron microscopy

To measure autophagosomes, cells were seeded and grouped as described above. Cells were harvested by centrifugation at 300 g, and then sectioned and fixed in 4% glutaraldehyde containing 2% paraformaldehyde in 30 mmol/L phosphate buffer, pH 7.4. After dehydration with alcohol, the slides were placed in embedding moulds saturated in propylene oxide and incubated at 60°C for 48 hours. The 70‐nm‐thin sections were prepared and detected by TEM (FEI, Eindhoven, The Netherlands) at 120 kV.