RE: monos, monos, monos....

From: Newsom, Brian S. (BSNEWSOM@txccc.org)
Date: Tue Nov 16 1999 - 17:44:04 EST


It falls in the FITC channel.

Brian Newsom

-----Original Message-----
From: Maciej Simm [mailto:simmmmer@yahoo.com]
Sent: Monday, November 15, 1999 2:39 PM
To: cyto-inbox
Subject: monos, monos, monos....



Hello everyone. First, a newbie dye-chemistry question: 
is the emission for Rhodamine 123 similar to PE or FITC or PerCP? 
Assuming the basic FACScalibur laser configuration ( I don't remember
the wavelenghts). 

I tried to resolve monocytes using a perCP cd4 antibody. In
"unstimulated" (as in background level of activation) PBMC's CD4
can resolve between lymphocytes and monocytes b/c of level of 
expression. I know there are more efficient ways of doing this  
4ex. CD14 and CD33. But in this experiment (oxidative burst assay)
I was not able to detect the FL3 signal at all! It was suggested
thereafter that CD4 gets dropped during activation and that could
be my problem. 

Has anyone tried adding a PerCP marker to the burst test to enhance
gating? I would apprieciate any suggestions. 

Maciej

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