[Cytometry] 5 color compensation issue

Haviland, David L David.L.Haviland at uth.tmc.edu
Tue Feb 3 10:23:12 EST 2009


Yes.   If you compare the emission profile of PE-Cy5.5 to PE you are going to find overlapping emission spectra that can't be compensated.   A better choice for your user would have been PE-Cy5.   I had this once myself and kindly asked the user not to bring that combination in my flow room again.   It was all made clear to my user when I showed her the emmission profiles of the reagents in question. 

Hope this helps...

David



-----Original Message-----
From: cytometry-bounces at lists.purdue.edu on behalf of Zhugong Liu
Sent: Mon 2/2/2009 9:29 PM
To: cytometry at lists.purdue.edu
Subject: [Cytometry] 5 color compensation issue
 
Dear all,

We are running 5-color flow on FC500 and have some compensation issues.
The 5-color cocktail includes antibodies with the following
fluorechromes: FITC, PE, PE-Cy5.5, PE-Cy7 and APC. Do you think there
might be some reagent interactions in this panel combo, especially
PE-Cy5.5 and other dyes? Our single controls were well compensated using
the software-generated matrices, however the fully stained samples were
not. 

Thanks for any comment/suggestion.

Best,
Jason

-------------------------------------------------------
Jason Liu, PhD
Asst. Director, Flow Cytometry
Tolerance Assays and Data Analysis
UCSF/Immune Tolerance Network
3 Bethesda Metro Center, Suite 400
Bethesda, MD 20814
Phone: 240-235-6177
Fax: 240-235-6198
Email: zliu at immunetolerance.org

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