fixation, how long

From: Mary-Lou Zuzarte (ZUZARTE@IMT.Uni-Marburg.DE)
Date: Tue Oct 09 2001 - 16:49:33 EST


Dear flow users,

I would like your opinion to a situation concerning fixation of cells for
DNA analysis. We are working with a number of human cancer cell lines in
the study of cell cycle and apoptosis. We have found that a 1 hour fixation
with cold EtOH is not enough to obtain an adequate DNA profile. We have, to
be sure that the protocol is consistantly followed, fixed the cells
overnight. I think That 1 hour is not enough because we see massive
variations in the instrument settings of the Calibur. The cells are stained
with PI.
I would really appreciate some feedback to this opinion.

Thank you in advance,

Marylou Zuzarte
FACS & Confocal Facility
 Prof. Rolf Muller,
Inst. f. Molekularbiologie & Tumorforschung
 Emil-Mannkopf Str 2
 35037 Marburg, Germany

Tel: 49-(0)6421-2865925

Zuzarte@IMT.uni-marburg.de



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