Evaluation of Micronuclei in V79 cells

From: anne.h.illi-love@abbott.com
Date: Thu Sep 27 2001 - 14:08:12 EST


Hello everyone,

I need help in analyzing samples that require logarithmic pulse area vs.
width.  I have tried unsuccessfully to reproduce results from a method
described in a journal article to evaluate micronuclei in V79 cells (D. Roman,
et al, "Evaluation of a New Procedure for the Flow Cytometric Analysis of In
Vitro, Chemically Induced Micronuclei in V79 Cells", Environ. & Mol. Mutagen.
32:387-396, 1998).  The method uses a two-step technique to destroy the
cellular membrane and cytoplasm, and release and stain the nuclei and
micronuclei with ethidium bromide.  It requires analysis of ethidium bromide
width versus area using a logarithmic scale to separate debris from
micronuclei.  The analysis was done using a BD FACSVantage flow cytometer.
However, we have a BD FACSCalibur which can only analyze width vs. area using
a linear scale.  Using the linear scale does not sufficiently separate the
populations.    Has anyone successfully accomplished this procedure without
using the logarithmic scale?  Any and all comments and suggestions are
appreciated.  Many thanks.

Anne Illi-Love
Department of Microscopy and Microanalysis
DR45M, Bldg. AP31
Abbott Laboratories, Abbott Park, IL  60064
(847) 937-0991
anne.h.illi-love@abbott.com



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