cell cycle problems

From: Dr M Salmon (salmonm@rheuma.bham.ac.uk)
Date: Wed Aug 18 1993 - 10:11:29 EST

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We have been using 7-actinomycin D to analyse cell cy[C[C[C[Cstatus of lymphocytes.
The big advantage of this dye over P.I. is the low compensation requirement
which allows you to study two other cell markers concurrently with FITC and PE.

Anyway, the technique seems to work fine giving a nice sharp G0/G1 spike, until
we run a large number of events.  This is essential for trying to characterise
the activated lymphocytes in our clinical samples; but the problem is, there
is a shift to the right of the G0/G1 peak after about 18,000 events.  We are
using a Coulter ELITE, the 7AAD is excited with an Argon laser, and detected
in PMT4 with a 640LP filter.

Has anyone encountered this before? If not, any bright ideas would be welcome.


Best Wishes & Have Fun


Mike Salmon
Email to salmonm@rheuma.bham.ac.uk           Voice: 021-414-6781

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