I have transfected cells with GFP, which emit in close range to FITC and used apoptosis kit that have annexin conjugated with FITC to measure apoptosis and PI to measure death. I am gating for GFP separately but feel it is probably false positive for apoptosis because the fluorescence spill over? Is this correct? need some thoughts? thank you
probably not a good idea to use a FITC (green) reagent in your GFP (also green) cells. I would choose another color of the annexin kit.
No. You need to choose another dye for your annexinV probe if you want to test GFP positive cells.
Fahad Alshehri Bartosz Wiernicki Aaron Dhanda Thank you for your answers much appreciated. What if I use the Cytek Aurora3 lasers? There are some claim that it can separate according to maximum expression band width with the right controls.
Asma El-Howati Yes, that is true. But why bother with unmixing GFP from FITC, if you can simply run the assay using AnnV-APC for instance. The cleaner the setup the better :)
Cheers.
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