How to Assess and Study Cell Death in Dictyostelium discoideum

In this chapter, we describe how to conveniently demonstrate, assess, and study cell death in Dictyostelium through simple cell culture, clonogenic tests, and photonic (with the help of staining techniques) and electronic microscopy. Cell death can be conveniently generated using minor modifications of the monolayer technique of Rob Kay et al., and either wild-type HMX44A Dictyostelium cells or the corresponding atg1 ⁻ autophagy gene mutant cells. Methods to follow cell death qualitatively and quantitatively facilitate detailed studies of vacuolar death in wild-type cells and of nonvacuolar, “condensed” death in atg1 ⁻ mutant cells.