From: Jose A. Stoute (stoutej@net2000ke.com)
Date: Tue Dec 15 1998 - 07:49:17 EST
Dear all, I would like to get some advice on whether my
FACscan needs further fine tuning. I recently called the BD
engineer to come and recheck the alignment and sensitivity
of our machine. He says there is no problem with it but I am
not satisfied and would like to get a second opinion. Our
main application is flowcytometry of red blood cells and I
see two problems:
1. When we acquire using Log amplification for FSC and SSC
we see a lot of background and have to increase the
threshold on FSC to about 300. I understand that some
background using these settings is normal to some extent but
there was a time when my machine had no background at all
using the same settings. This background is not due to
impurities in our buffers because it is present even after
filtering through 0.22 um filters. Also, it became worse
after the engineer adjusted the machine.
2. The other problem is that the PMT gain that I have to use
to have unstained cells in the first decade of FL1 seems to
be too high, around 700. We found the same when we used the
FACscalibur across the hallway. The engineer said that he
had checked everything.
I would appreciate any suggestions as to how to deal with
the above problems.
--
Jose A. Stoute, MD
Unit 64109, Box 401
USAMRU-Kenya
APO AE 09831-4109
e-mail:stoutej@net2000ke.com
stoutej@workmail.com
Nairobi Tel 254-2-729303, Fax 254-2-714592
Kisumu Tel 254-35-22942/22903, Fax 254-35-22903
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