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Hmm, is this true for unwashed cells as well? I've been using a one-step
hypotonic lysing buffer that contains PI (50 ug/ml), but I've been varying
the time (up to several hours) before reading on the FACS scanner. As
such, would the PI saturate all nuclei, giving a reduced amount of
hypodiploid fluoresence and sending everything up to the G0/G1 peak (thus
giving abnormally low levels of apoptosis)?
Ryan.
_/ \__/ \__/ \__/ \__/ \__/ \__/ \__/rhung@vcn.bc.ca__/ \__/ \__/
\_Apoptosis=programmed cell death/ \__/ \rwhung@netinfo.ubc.ca \__/ \__
_/ --you can't live without it!/ \__/ \http://www.vcn.bc.ca/people/rhung
\__/ \__/ \__/ \__/ \__/ \__/ \__/ \My words Copyright (C) 1997 \__
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web