I want assess the rate of intracellular parasite killing in side the infected macrophages in in vitro experiment. what method is opt for assess live or dead parasites inside the macrophages? Please help me..
You may use a vital stain such as acridine orange to stain the parasites, and other stain for the phagocytic cell. Acridine orange stains degraded DNA in orange and intact DNA in green. It is also useful to stain engulfed apoptotic cells and organelles such as lysosomes. Hope it helps!
If you use a transgenic parasite model, it would be life easier. For instance use reporter molecule construct like GFP in your parasite and after transfection,the GFP signal can be detected using FACS from macrophages harboring parasites. Before FACS, you can also check for the release of nitric oxide.
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