Have you tried mixing the control cells with you triploid cells. This will hopefully correct for changes in PI staining since both the control cells and the triploid are exposed to the same PI concentration. Tim -----Original Message----- From: Cogswell, Andrew [mailto:CogswellA@mar.dfo-mpo.gc.ca] Sent: Thursday, February 05, 2004 7:43 AM To: cyto-inbox Subject: Ploidy Analysis Dear Flowers, I am currently assessing triploidy induction rates using Propidium iodide as a DNA specific flourecsent dye. I find that when I run controls containing diploid nuclei, the peak on my histograms tends to shift significantly to the right or left of where I set it on the FL2 (x) axis from one run to another. Therefore, when I run triploid samples I cannot be sure (without some kind of internal standard) where the actual peak lies, and whether it is in fact diploid. Any ideas for what I might do to standardize samples so that there is no question of my samples ploidy level? I am a new user, so please feel free to elaborate. Andrew T. Cogswell Science Branch, Maritimes Region Invertebrate Fisheries Division Fisheries and Oceans Canada 1 Challenger Drive Halifax, NS B2Y 4A2 Phone: 902-426-4353Received on Mon Feb 9 14:38:00 2004
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