Coulter XL Fluorescence Compensation

Antony Bakke (bakkea@ccmail.ohsu.edu)
Wed, 15 Feb 95 11:40:47 PST

Brian, we have a new Coulter XL in a clinical lab and have been
using it for about 3 months. We previously had 8 years
experience with a FACScan. The XL operates well. We have only
seen differences with compensation when switching from unwashed
to washed samples. Coulter is making a big push to reduce the
labor for a sample and are therefore emphacizing the no-wash
method. Without washing there is considerable remaining labeled
antibody in the solution and this registers as increased
fluorescence even though it is not attached to a cell.
Depending on the antibody reagents used and their concentration,
this could drastically change the compensation. Therefore, the
no-wash method can only be used with well characterized
antibodies at a minimal concentration as in the Coulter CytoStat
reagents. When we wash the cells to remove excess antibody, the
results are very similar to those on the FACScan. The
compensation values are even similar.

On another note there are some problems with the XL. Although
Coulter has promised some things to make it more clinically
automated, these have not happened yet. There are three areas
that concern me: 1) the autogating does not work because it was
set up for only two standard deviations from the cluster mean.
Therefore you can never get a value above 95% for lymphocyte
recovery and often get 85-90%. 2) the XL only reads the Coulter
standard barcode and this is limiting it your institution does
not print this barcode. 3) the XL does not interface with
networks well. You cannot save data to a network server and
easily access it from other work stations. All of these
problems are supposed to be fixed in the new software which is
due out in April. Again Coulter tends to promise more than it
delivers.

I would also look at the new BD instrument. They are supposed
to have a dual laser FACSort with an autoloader. For clinical
use I think it would be useful to have the ability to quickly
sort a few cells for the pathologist to look at. However, BD is
behind in making the move into a clinical setting so they do not
have software to read barcodes and produce worklists. I do
think that their autogating software is good.

Hope this is helpful. Tony Bakke (bakkea@ohsu.edu)


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