Imaging Ca²⁺ Signaling in Trypanosoma cruzi with Genetically Encoded Ca²⁺ Indicators

Genetically encoded calcium indicators (GECIs) are proteins used to monitor calcium ion (Ca²⁺) changes in living cells. They can be targeted to the cytosol or to organelles for continuous measurements in vivo. In this chapter, we present an overview of the methods that our lab used to detect Ca²⁺ changes in Trypanosoma cruzi elicited by stimulation of Ca²⁺ channels in the plasma membrane or in intracellular organelles, including selection of adequate GECIs, their transfection and selection, and their use for microscopic and fluorometric analysis.