I am infecting BMDMs with GFP-expressing Salmonella. I need to remove as many extracellular bacteria as possible. In a trial experiment, we rinsed the BMDMs after 30 minutes of infection and then incubated for an additional 2 hrs, but there were numerous extracellular bacteria remaining; to many for my assay. Are there recommendations for improving clearance? Thanks!
Michael Davis,
Adding an antibiotic like gentamicin after initial infection might get rid of the extracellular bacteria without affecting its intracellular viability. Hope this helps!
Thanks Deepika. I've looked into that a bit. While I think it will help, I don't think it will be enough. For reasons particular to my project, my read-out is microscopy based. I need to get rid of the extracellular bacteria in order to visualize a qualitative difference between the two.
Hi Michael,
You can follow a similar query on researchgate https://www.researchgate.net/post/Best_way_to_remove_surface-bound_bacteria_or_beads_phagocytosis_assays and I think it might be handy to your experiments staining intracellular and extracellular bacteria with differential dyes.
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