Hi all,
I am currently dealing with a knockout cells, which have already showed decreased growth rate while I am growing the cells. I used PI staining for cell cycle analysis, the KO cells showed increased S phase, compared to wildtype control. But when I use Cell trace Far red probe (Thermo Fisher) to do the proliferation assay, there is almost no change between wt and KO.
I should see the daughter cells' peak compared to wt ones, but no. Anyone has ever done proliferation assay using this probe? Any advice or suggestion on this probe? Thank you so much, any information will be really appreciated.
Best,
Lu Wei
Dear Lu,
I think, it should be the result of your flow cytometer setting. If the alignment of flow cytometer was not adjusted properly and C.V of your dye channel (FL3 in most devices) be high, peaks of daughter cells go inside each others.
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