Assessment of autophagosome formation by transmission electron microscopy.

Autophagy is a complex degradative process by which cytosolic material, including organelles, is randomly sequestered within double-membrane bound vesicles termed autophagosomes and targeted for degradation. Initially described as a nutrient stress adaptation response, the process of autophagy is now recognized as a central mechanism involved in many developmental processes. In this chapter, we provide guidelines to assess the initial steps of autophagy by monitoring autophagic body vacuolar accumulation. We employed a standard electron microscopy approach to observe the vacuoles of nutrient stressed fungal cells.