low FSC granulocyte

Hi Abdul,

We do a lot of flow on PMNs and we see a similar pattern when the cells are 
de-granulating and/or in early apoptosis.  I have no idea why you suddenly 
began seeing this phenomenon but I would check your processing to determine 
if any changes have occurred.  Things such as storage time, time from draw 
until processing, aggressive handling, bad reagents, etc.  can all make a 
difference.  Make sure you check your buffers.

George Babcock
University of Cincinnati &
Shriners Hospitals for Children



Date: Sat 28 Jun 17:19:12 EDT 2008
From: "Abdul Khan" <khan.abdulh@googlemail.com>
Subject: low FSC granulocyte
To: cyto-inbox


Dear Flowers,

I am a PhD student and doing flow cytometry on peripheral blood
from healthy women. So far I have done quite a few samples and all of them
are quite consistent in terms of results, for example CD3, CD4, CD8, CD14,
CD66b expression. The attached FSC/SSC plot (FSC-SSC -1 file) looks very
unusual to me as you can see most of the granulocytes are having low FSC and
I am wondering if anyone have come across with this earlier and would
appreciate any help/comments. First I thought that there might be some
problem with the machine, so I did an experiment on my own blood (FSC-SSC -2
file) just to make sure that FACS machine is working fine and indeed I found
the same reproducible result.

I sincerely hope that someone here will be able to help me out in
understanding what could be wrong here! Thank you all.


Best wishes

Abdul Khan
PhD Student
University of Glasgow