Annexin V help

From: Andrew Oberyszyn (oberyszyn.2@osu.edu)
Date: Mon Mar 27 2000 - 13:48:30 EST 

Hi everybody in Flow land,
I hope someone can steer me on this one.
I have a user who is looking at Annexin V- FITC/PI staining of Caco-2
cells.  The problem that I am encountering is that the untreated, Annexin
V- FITC only stained cells are a log higher than untreated/unstained cells
(see attached Powerpoint slide).  I looked under a flourescent microscope
and saw considerable surface staining in the Annexin only samples.  The
user is using the Pharmingen kit (5 ul of Annexin V).
I have not seen this happen (Annexin V so much higher than unstained) in
other cell lines that I've analyzed.  Would it be "legal" to adjust the
volts to bring the Annexin V Only stained population into the negative
region or is it more correct to just set the gate as I've indicated on the
attached slide (marked with an *)?
Am I missing something obvious?  Please help!

Thanks in advance!
Andy
(:)-(:)-(:)-(:)-(:)-(:)-(:)-(:)-(:)-(:)-(:)-(:)-(:)-(:) Andy Oberyszyn, M.S. The Ohio State University Analytical Cytometry Laboratory 416 Comprehensive Cancer Center 410 West 12th Avenue Columbus, Ohio 43210 Tel: 614/292-FLOW(3569) Fax: 614/292-7335 E-Mail: cytometry@osu.edu Web Page: <http://cytometry.med.ohio-state.edu>http://cytometry.med.ohio-state.edu (-):(-):(-):(-):(-):(-):(-):(-):(-):(-):(-):(-):(-):(-) "What if the Hokey Pokey is really what it's all about?!?"

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