I checked many of the PI supplier companies, they all recommend not to wash after PI staining. But the reason is unstated. I am really curious to know what is the consequence of washing. By right, PI would get into the nucleus and intercalate with DNA strands very firmly. Shouldn't be a problem if being washed. Did anyone get an idea? Many thanks!
it seems that there exist protocols where (at least adherent cells) are washed in PBS after PI-staining, followed by FLUOR-MICR.: e.g. cf.:
http://www.cyto.purdue.edu/archive/flowcyt/research/cytotech/apopto/data/chap3.htm
(admitting that I do not know, why PI-suppliers like Thermofisher -this company mentioned only as one example - recommend NOT WASHING cells AFTER PI-stain....(e.g. cf.: https://tools.thermofisher.com/content/sfs/manuals/viability-staining.pdf ) Could it be that there is a different treatment regarding "Flow cytometry measurement" vs. "regular Fluorescence Microscopy"? It would be interesting what "experts" in this field have to say.... Best of luck ! NB: perhaps you should read (already on RG) also: https://www.researchgate.net/post/Is_there_a_simple_way_to_fluorescently_stain_surviving_cells_at_the_end_of_an_experiment....
Hello,
at least when dealing with sperm, PI will remain after washing, so you will not have any effect. Regards,
Bea
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