PI and sample tubing

From: ray hester (rhester@jaguar1.usouthal.edu)
Date: Mon Aug 07 2000 - 11:03:43 EST


Hi,

I don't remember the following having been discussed (but then my memory is
not that great!):

For years I've noticed when we run propidium iodide-stained samples for
ploidy analysis that the PI fluorescence (488 Ex, 585/42 EM) gradually
increases for the first 10 or 15 secs and then finally stabilizes (samples
are in the staining buffer with PI; sheath fluid is PBS).

Several years ago I was at a _major_ manufacturer's instrument training
session and they were touting the fact that analyses on their instruments
were not susceptible to this gradual increase because of their (proprietary)
sample tubing and sure enough we ran PI stained samples (I can't remember
exactly what they were) and plotted fluorescence vs time, there was no
variation in fluorescence intensity.

One can wait until the fluorescence seems to stabilize so that CVs are not
affected but it would be nice to know on the front end that this phenomenon
is not a potential factor.

Any suggestions about tubing and/or sheath fluid to prevent the above would
be appreciated.

Ray

Raymond B. Hester, Ph.D.
Research Cytometry Laboratory
CSAB 357, Biotechnical Services
University of South Alabama
Mobile, AL 36688
Voice:	  (334) 460-6029
FAX:	  (334) 460-6073
E-mail:   rhester@jaguar1.usouthal.edu





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