[Cytometry] 5 color compensation issue

Pál Jáksó pal.jakso at aok.pte.hu
Tue Feb 3 03:11:42 EST 2009


Dear Jason,

I've never used PE-Cy5.5 but according to its spectrum, it has a  
significant crosstalk to the PE channel.
We use FITC, PE, PerCP, PE-Cy7 and APC 5-color combo and it works fine  
on our Partec CyFlow space. Depending on your filter setup in your  
FC500 you may consider PerCP instead of PE-Cy5.5.

Bests,

Pal Jakso

University of Pecs
Faculty of Medicine
Dept. of Pathology
12. Szigeti str.
H-7643 Pecs, HUNGARY

On Feb 3, 2009, at 4:29 AM, Zhugong Liu wrote:

> 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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