I do not know which area should I gate in the area vs height graph to distinguish between the doublets and polyploid cells using flowcytometry? I am working with a Beckman Coulter Machine. I have done PI staining
Doublets should have a larger side scatter area than polyploid cells. Ideally you'll be able to remove most of these using a forward scatter area / forward scatter height gate to select for single cells before doing your PI gating.
I suggest you to read Michael Ormerod's Flow Cytometry book for basic information on DNA Cycle analysis. It was free online on FCSExpress web page.
You may also read the related chapters written by Zbigniew Darzynkiewics in Current Protocols in Flow Cytometry.
Gating strategy will effect the quality of your results. Just to remind you that, PI can bind to both DNA and RNA, you should be using RNase to prevent unnecessary binding.
Adam Figarski - yes, I can use a microscope but I am anyway doing cell cycle analysis for my samples. So, I believe polyploidy experiment can be checked in the same experiment.
Gulderen Yanikkaya Demirel - Thank you for the study material suggestions.
Yes, I am using RNase.
Sourya Subhra Nasker - Thanks
Thank you everyone for the suggestions. Please share in this space if you happen to learn something more regarding this.