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I devloped an assay to detect surviving fractions as a screen for potential
AML-specific natural products, using calceinAM and PI. Noted these intermediate
groups in most of the experiments; also noted thiss phenomenon when using PI
alone as an dead-cell sxclusion dye in immunofluorescence, especially when log
amped. Never actually pursued an explanation, but it's possible these were
apoptotic cells. Whether or not this process is important in your system you'd
have to say, but certainly these intermediates may occur do to physical changes
in the cell membrane which may affford some level of permeability and allow
passage of lower amounts off PI in cells with INTACT and viable (by retention
of the viability probe) characteristics.
Mark A. KuKuruga
Karmanos Cancer Institute
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