Hello flowers I am posting this question for my lab, not having much expertise with this stain myself and while the local experts are frolicking at a conference in Monaco. Here is the question: We use Hoechst (for DNA) then Pyronin (for RNA) staining to define populations within the Go/G1 population by the amount of RNA. Additional info is that the Pyronin is used at 3.3 uM concentration, a non-toxic concentration. Lately we have seen a very odd phenomenon. When cord-blood samples were stained, the Hoechst single stain gives a normal cell-cycle pattern, but when the two stains are combined, most of the pyronin staining appears to be in sub-G0 peak on Hoechst. Additionally, when the same stain was run on peripheral blood cells, the stain was normal (i.e. that most of the pyronin stain fell on the G0/G1 peak and some on the S+G2/M). Are there any suggestions or ideas on what may be causing this sub-G0 peak? Any and all replies are greatly appreciated. Artur Plett IUPUI-Heme/Onc division
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