I used to do TUNEL Staining on Chondrocytes and analyze using a Fluorescent Microscope in 2000-2001...it was our opinion that the TUNEL staining did not realistically represent true apoptotic cells (it may have been a time point issue) and we began to search for other methods. Though TUNEL protocols may have improved in the last seven years, I would trust your Annexin staining data. What protocol are you using for staining? The TUNEL is probably just giving you all dead cells. I'm sure someone with more expertise can chime in but I would forget the TUNEL data and use the Annexin V system. You could also try one of the Apoptosis kits Caspalux 8 or 9 from Oncoimmunin, I've heard they are quite good. Cheers, Eric Eric O'Connor Head of Flow Cytometry MRC Clinical Sciences Centre Imperial College Faculty of Medicine Hammersmith Hospital Campus DuCane Road, London W12 0NN Tel: +44(0) 208 383 8330 Mobile: +44(0) 781 575 7730 ________________________________ From: Sānia Alves dos Santos [mailto:saniasan@usp.br] Sent: 27 June 2008 12:52 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) 3061-7029Received on Mon Jun 30 14:18:00 2008
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