Well.. With 7AAD+AnnexinV you would get two populations in the UR window
in most situations. That is expected. One mid apoptotic phase and the
other late apoptotic phase. The midapoptotic phase would show slightly
weaker 7AAD staining and hence will appear lower than the late apoptotic
or dead cells. In a histogram, you would see it as a doublet peak which
is the typical 7AAD peak. Is this what you are seeing? If so, don't
worry. Your populations are correct.
If not properly compensated for spectral overlap, you will not see
this two populations in the UR.
Jayakumar R. Nair, Ph.D.,
Dept. of Immunology (Dr. Kelvin Lee's lab)
Cancer Cell Center, room 405.
Elm and Carlton St.
Roswell Park Cancer Institute,
NY 14263. USA
From: [Only registered users see links. ]
[mailto:[Only registered users see links. ].indiana.edu] On Behalf Of Koschutnig
Sent: Thursday, June 07, 2007 9:49 AM
To: [Only registered users see links. ]
Subject: [Cell-biology] annexin/7-AAD-staining
I have problems using the apoptosis kit ( BD Biosciences). Last week I
got quite strange pictures. It looks as if I have 2 populations in the
UR- window. I contacted our technican and she thought that the
compensation is wrong. But today I checked the whole configuration and
everything seems to be ok, when I use annexinv or 7 AAD alone. But
together it looks strange. I have attached you some pictures. (cells (
Hepg2) were treated with different oxysterols) If anybody could help me,
this wouls be wonderful. I really start feeling desperate...
best regards, Karin Koschutnig
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