Dear Berna Özdem ,

Acridine Orange dye is know to give green fluorescence when bind to DNA and Gives red color when permeate to lysosomes.

SO, when you open a bi-axis (FITC/PE) dot plot on gated cells, you will get FITC only cells (Which are basically Dead cells ), the PE only cells are live cells (membrane is intact) and orange signal give you the count of apoptotic cells.

Hi Berna

You need to process all the cell pool with same way for best analysis for your study. By this test, if you find more apoptotic cells as compared to nt and vehicle treated cell pool from your drug, your drug could be potential anticancer drug.

thank you so much to all.İ will also add acridine orange to nt and vehicle.

But, according to this result ll is live cells lr pro apoptotic. cells but in tihs result nt live cells also looks dead apoptotic cells? how can i fix that? Actually i want to learn how can i analyze and use this flow cytometer. Do if you have any articles or some books i will be appreciate. thank you so much for your interest ☺

First book to read is Howard Schapiro's book (it is been mentioned as bible of cytometry) so that you understand the basics. Michael Ormerod's book which is available through commercial company web sites is also a good reference book.

For cell proliferation and apoptosis assays, I suggest you look up to Flow Cytometry special issues of Methods in Molecular Biology. Current Protocols for Flow Cytometry published by Wiley is a good source for protocols. I also suggest to follo Cytometry A and B journals. ISAC web site has a lot of sources.