[Cytometry] 5 color compensation issue

Björnsson Sven Sven.Bjornsson at skane.se
Wed Feb 4 03:37:10 EST 2009


Hi Jason,

There should be no fundamental problems with the suggested combo on your FC500. The easy way to investigate spillover and filter characteristics is to run singel fluorochrome labeled samples on your FC500 with all compensation values set to zero and all voltages set to 500V. Then you look at the MFI of each fluorochrome in each channel and deduct what is the origin of you problems.

Cheers

Sven

Sven Bjornsson
Cytometry lab
University Hospital UMAS
Malmo
Sweden

-----Ursprungligt meddelande-----
Från: cytometry-bounces at lists.purdue.edu [mailto:cytometry-bounces at lists.purdue.edu] För Zhugong Liu
Skickat: den 3 februari 2009 04:29
Till: cytometry at lists.purdue.edu
Ämne: [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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