Hi Sania, In the later stages of apoptosis, membranes become permeable to PI. So it could be that the PI+ cells you are seeing are apoptotic , in which case your total apoptosis measured with both methods would be similar. I would recommend looking at ethanol-fixed PI stained cells under the fluorescent scope in order to identify apoptotic cells, which typically undergo characteristic nuclear changes. Good luck, Tom Thomas W. Mc Closkey, Ph. D. Associate Director, Cellular Immunology Research and Development ICON Central Laboratory 123 Smith Street Farmingdale, NY 11735 Tel: + (1) 631-306-9789 Fax: + (1) 631-694-2524 Email: mccloskeyt@iconlabinc.com <mailto:mccloskeyt@iconlabinc.com> Web: www.icolabs.com ________________________________ From: Sānia Alves dos Santos [mailto:saniasan@usp.br] Sent: Friday, June 27, 2008 8:52 AM To: cyto-inbox Subject: Annexin x Tunel Hi Flowers I'm working with hepatocyte culture, studying the frequency of apoptotic death in cells invaded by bacteria. Using the Tunel staining I've obtaining higher values in comparison to negative control cells (30%). Using Annexin staining the values are very low, about 1-5% of apoptotic cells. Otherwise the PI staining indicate necrosis in about 20% of cells. How can I interprete these results? Thanks in advance, Sania Alves dos Santos Laboratory of Bacteriology Faculty of Medicine University of Sao Paulo tel: (5511) 3061-7029 ********************************************************************** This e-mail transmission may contain confidential or legally privileged information from ICON Central Laboratories, Inc. intended only for the use of the individual(s) or entity named on the transmission sheet. If you are not the intended recipient, you are here by notified that any disclosure, copying, distribution, use or taking of action in reliance upon the contents of this e-mail is strictly prohibited. If you have received this e-mail transmission in error, please notify us immediately by telephone or e-mail so that ICON Central Laboratories, Inc. can arrange for the return of the documents to us at no cost to you, and then please delete the message. Thank You, ICON Legal Department, ICON Central Laboratories, Inc. (631) 777-8833 **********************************************************************Received on Mon Jun 30 14:58:00 2008
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