In Situ Detection of Dormant Trypanosoma cruzi Amastigotes Using Bioluminescent-Fluorescent Reporters
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.
Key wordsClarification Dormancy Proliferation Bioluminescence Luciferase
We are grateful to Dr. Rick Tarleton, Wei Wang, Dylan Orr, as well as Julie Nelson from the CTEGD Flow Cytometry Core and Muthugapatti Kandasamy from the Biomedical Microscopy Core. This work was supported by US National Institutes of Health grants AI108265 and AI124692 to Rick Tarleton.
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