. . . unless, of course, if instead of dead cell elimination one is interested in live/dead discrimination and enumeration. Example: label cells with a PE conjugated antibody . . . now, of the dead cells, how many are PE positive, and how many are just dead? mak. -- Mark A. KuKuruga, Managing Director University of Michigan Flow Core 7416 CCGC 0946 (734) 647-3216, fax (734) 936-7376 kukuru@umich.edu >>> "DAVID M CODER" <d_coder@MSN.com> 12/17/01 09:33AM >>> Either PI or 7-AAD will work. Moreover, with either DNA-binding dye, compensation in only direction is needed: You must make sure that PE-positive cells do not appear to be 7-AAD or PI-positive. Compensation 7-AAD or PI into PE positive space is unnecessary as those cells are eliminated from further analysis. The attached images illustrate compensated vs. uncompensated PE. For details see web page that I developed when I was directing the lab at the University of Washington: http://nucleus.immunol.washington.edu/Research_facilities/Apps/7aad.html Dave ---------------- David M. Coder, Ph.D. Consulting in all aspects of cytometry. email: d_coder@msn.com tel./messages: 206-499-3446 ----- Original Message ----- From: Jáksó Pál To: Cytometry Mailing List Sent: Monday, November 26, 2001 11:21 PM Subject: RE: 7AAD PROTOCOLS Dear Rachel, 7AAD is may be better in some application because of the lower crosstalking to the FL2 channel. Accordingly you can use it together with PE stains. PI is very hard to compensate from the FL2. But if you use only FITC as second stain (and nothing else) you can use both 7AAD and PI for dead cell labeling. Pal Jakso University of Pecs Faculty of Medicine Dept. of Pathology 7643, Hungary 12. Szigeti str. While this subject has come up: it seems like both 7AAD and propidium iodide can be (and are) used for detecting dead or apoptotic cells - are they interchangeable ? If people have distinct preferences, what is the difference ? thanks, Rachel
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