In situ detection of dormant trypanosoma cruzi amastigotes using bioluminescent-fluorescent reporters

Abstract

Chagas disease agent, Trypanosoma cruzi, is capable to persist after prolonged drug treatment using effective drugs. The reason of treatment failure is not known, but recent development of highly sensible bioluminescence imaging coupled to tissue clarification techniques has made possible the detection of individual amastigotes within chronically infected murine tissues and the study of their replicative status. In this chapter, we provide a step-by-step explanation for these protocols that allowed the visualization of nonproliferating amastigotes in tissues of chronically infected mice for the first time.