[Cytometry] 5 color compensation issue

Michelle Tourigny michellet at ichr.uwa.edu.au
Tue Feb 3 21:19:04 EST 2009


We have had many problems with PE-Cy7 becoming unstable and screwing up
the compensations.
michelle

Michelle R. Tourigny, Ph.D.
Research Assistant, Instrument Specialist
Telethon Institute for Child Health Research
100 Roberts Road, Subiaco
Western Australia, 6008 AUSTRALIA

Ph 61 8 9489 7878, Fax 61 8 9489 7700
email michellet at ichr.uwa.edu.au 


-----Original Message-----
From: cytometry-bounces at lists.purdue.edu
[mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Carol Oxford
Sent: Wednesday, 4 February 2009 12:57 AM
To: Zhugong Liu; cytometry at lists.purdue.edu
Subject: Re: [Cytometry] 5 color compensation issue

Hi Jason,

We run these together all the time, and all but one of our instruments 
are set up to run this combination- we even add PETR and PE-Cy5 to the 
mix.  I'm not familiar with the exact filters on the FC500, it's 
possible that changing the filter setup might improve things.  If you 
run the appropriate compensation controls and the FMO controls, you 
should be able to get this to work.  It takes a bit of panel 
optimization to make sure that your positive population is well 
separated and above the spread of the other dyes in the mix, but it 
routinely works well for us.

Carol

Zhugong Liu wrote:
> Dear all,
>
> We are running 5-color flow oFC500 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
>
> _______________________________________________
> Cytometry mailing list
> Cytometry at lists.purdue.edu
> https://lists.purdue.edu/mailman/listinfo/cytometry
>
>   

-- 


----------------------------------------------------
Carol Oxford                                           
Manager, UC Davis Optical Biology Lab       
Tupper Hall, rm 3425                             
University of California                          
Davis, California  95616                        
(530) 752-7205                                                     

Manager, Stem Cell Sorting Facility
CBST
(916) 703-9307

cloxford at ucdavis.edu
http://ccresources.ucdmc.ucdavis.edu            

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