Chagas disease, caused by the protozoan parasite Trypanosoma cruzi, is a life-threatening neglected illness endemic to Latin America. Despite efforts, it has recently become a global concern due to human migration from endemic to non-endemic areas. Exploring parasite pathways to interrupt the T. cruzi life cycle may lead to new treatments. In particular, autophagy plays a relevant role throughout its life cycle. In this context, the modulation of autophagy could provide multiple targets for new trypanocidal drugs. In this chapter, we describe various techniques developed to study autophagy in T. cruzi, such as fluorescence and electron microscopy to visualize autophagic structures in living or fixed parasites, and Western blotting to analyze autophagosome formation by tagging the LC3-like protein TcAtg8.1. The interpretation of these techniques allows for the distinction between changes in autophagic flux and specific differences in the steps of this pathway. Understanding the dynamics of autophagy in T. cruzi could assist researchers in exploring its potential role as a therapeutic target for Chagas disease.