Low resolution signal
I recently performed a cytof run that shows poor resolution between positive and negative populations mostly due to high intensity of the negative population than usual (Run 2 attachment). I'm attaching one good run (Run 1 attachment) done previously with the same panel and protocol. The only difference is this time I stained 170Er CTLA-4 intracellularly and previously was surface stain. I didn't think it would make a difference and hence the antibody was not retitrated for intracellular staining. During the run, I noticed high streaking in the Ir191 and 193 DNA channels but all other channels had a reasonably low background. I am not sure if that is contributing to the high intensity of the negative population.
It would great to get your thoughts and if there is a way to salvage the experiment since I still have leftover stained sample (in 1.6% pfa in pbs) that could be acquired again.
Thank you!
Mansi