BKG staining
I am having issues with single tube sample acquisition. My runs have been going perfectly (no clogging) but when I check the data in flowjo, I am seeing some unusual bkg. I have discussed this with Fluidigm and they cannot seem to figure out where it is coming from. My staining protocol involves Cisplatin for LD, Surface markers, Fox p3 transcription buffer fix/perm, intracellular/intranuclear staining, and finally 2%PFA+Ir overnight. The bkg is not in every sample and there is nothing unusual that took place with the sample acquisition. From 18 samples/week I am seeing maybe 2-3 with signifcant Bkg. At one point we thought it was bead contamination. I ordered new beads and still see it. It was also suggested that residual acids from nitric or wash, so in many cases, I only used water between samples. I am attaching a ppt.
Thanks so much for your help
Diane
(Nadeau lab)