Ugo Consoli (uconsoli@mbox.vol.it)
Sun, 20 Oct 1996 23:16:10 +0200

We used Tunel in combination with MoAb and DNA content (Hoechst with a
second laser) with good results!
Tunel assay is very bright and can create few problems with PE
compensation, but is sufficient to increase green PMT to bring green
florescence in the second log and adjust for compensation!
We have observed a loss of orange fluorescence in apoptotic cells that we
thought was related to ethanol fixation. However, we observed the same loss
of fluorescence in apoptotic cells studied using changes in forward and
right scatter which require no fixation at all.
Is loss of fluorescence related to the apoptotic phenomenon itself?
Probably it is!
With Tunel/MoAb we were able to succesfully detect apoptosis in a selected
population of cell that represented 2% of the entire sample.
Annexin is also very good (and easy enough!) and can be combined with MoAb!
but you need to run your sample almost immediately. With Tunel we stored
samples for several weeks before analysis.

There is good and bad in everything!!!

good luck!

Ugo Consoli, M.D.
Institute of Hematology
University of Catania - Italy
Fax - (095) 731-1510

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